EIF4E inhibitors and uses thereof

The present invention provides compounds inhibiting eIF4E activity, and compositions and methods of using thereof. Compounds of the present invention, and pharmaceutically acceptable compositions thereof, are useful for treating a variety of diseases, disorders or conditions, associated with eIF4E. Such diseases, disorders, or conditions include cellular proliferative disorders (e.g., cancer) such as those described herein.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to compounds and methods useful for inhibition of Eukaryotic initiation factor 4E (eIF4E). The invention also provides pharmaceutically acceptable compositions comprising compounds of the present invention and methods of using said compositions in the treatment of various disorders.

BACKGROUND OF THE INVENTION

Eukaryotic initiation factor 4E (eIF4E) is a 24 kDa protein that plays a key role in the initiation of translation of mRNA. At the initiation of mRNA translation, eIF4E binds to the 7-methylguanosine cap at the 5′ end of mRNAs, and forms a complex (called eIF4F) with the scaffolding protein eIF4G and the helicase eIF4A. The formation of this complex is required for the initiation of cap-dependent translation and therefore the binding of eIF4E to eIF4G is a critical event in this process.

A number of studies have suggested that eIF4E is involved in tumorigenesis and is a potential target in the field of oncology.

SUMMARY OF THE INVENTION

It has now been found that compounds of the present invention, and pharmaceutically acceptable compositions thereof, are effective as eIF4E inhibitors. In one aspect, the present invention provides a compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein each variable is as defined and described herein.

Compounds of the present invention, and pharmaceutically acceptable compositions thereof, are useful for treating a variety of diseases, disorders or conditions, associated with eIF4E. Such diseases, disorders, or conditions include cellular proliferative disorders (e.g., cancer) such as those described herein.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

1. General Description of Certain Embodiments of the Invention

Compounds of the present invention, and pharmaceutical compositions thereof, are useful as inhibitors of eIF4E. Without wishing to be bound by any particular theory, it is believed that compounds of the present invention, and pharmaceutical compositions thereof, may inhibit the activity of eIF4E and thus treat certain diseases, such as cancer.

It has now been found that compounds of this invention, and pharmaceutically acceptable compositions thereof, are effective as eIF4E inhibitors. In one aspect, the present invention provides a compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein:L1is a bond, O, or S;R1is an optionally substituted ring selected from phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring, a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or an 8-10 membered bicyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur;L2is a C1-8bivalent straight or branched hydrocarbon chain, wherein 1, 2, or 3 methylene units of the hydrocarbon chain are optionally and independently replaced with —N(R)— or —Cy—;each R is independently selected from hydrogen or optionally substituted —C1-6aliphatic;—Cy— is an optionally substituted bivalent ring selected from 3-6 membered monocyclic, saturated or partially unsaturated, carbocyclylene, or 3-6 membered monocyclic, saturated or partially unsaturated, heterocyclylene having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur;R2is —COOR; andR3is R, halogen, or an optionally substituted ring selected from phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring, a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or an 8-10 membered bicyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

2. Compounds and Definitions

As used herein, the term “bicyclic ring” or “bicyclic ring system” refers to any bicyclic ring system, i.e. carbocyclic or heterocyclic, saturated or having one or more units ofunsaturation, having one or more atoms in common between the two rings of the ring system. Thus, the term includes any permissible ring fusion, such as ortho-fused or spirocyclic. As used herein, the term “heterobicyclic” is a subset of “bicyclic” that requires that one or more heteroatoms are present in one or both rings of the bicycle. Such heteroatoms may be present at ring junctions and are optionally substituted, and may be selected from nitrogen (including N-oxides), oxygen, sulfur (including oxidized forms such as sulfones and sulfonates), phosphorus (including oxidized forms such as phosphates), boron, etc. In some embodiments, a bicyclic group has 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. As used herein, the term “bridged bicyclic” refers to any bicyclic ring system, i.e. carbocyclic or heterocyclic, saturated or partially unsaturated, having at least one bridge. As defined by IUPAC, a “bridge” is an unbranched chain of atoms or an atom or a valence bond connecting two bridgeheads, where a “bridgehead” is any skeletal atom of the ring system which is bonded to three or more skeletal atoms (excluding hydrogen). In some embodiments, a bridged bicyclic group has 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Such bridged bicyclic groups are well known in the art and include those groups set forth below where each group is attached to the rest of the molecule at any substitutable carbon or nitrogen atom. Unless otherwise specified, a bridged bicyclic group is optionally substituted with one or more substituents as set forth for aliphatic groups. Additionally or alternatively, any substitutable nitrogen of a bridged bicyclic group is optionally substituted. Exemplary bicyclic rings include:

As used herein, the term “bivalent C1-8(or C1-6) saturated or unsaturated, straight or branched, hydrocarbon chain”, refers to bivalent alkylene, alkenylene, and alkynylene chains that are straight or branched as defined herein.

As used herein, the term “cyclopropylenyl” refers to a bivalent cyclopropyl group of the following structure:

Each R●is independently selected from C1-4aliphatic, —CH2Ph, —O(CH2)0-1Ph, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and wherein each R●is unsubstituted or where preceded by halo is substituted only with one or more halogens; or wherein an optional substituent on a saturated carbon is a divalent substituent independently selected from ═O, ═S, ═NNR*2, ═NNHC(O)R*, ═NNHC(O)OR*, ═NNHS(O)2R*, ═NR*, ═NOR*, —O(C(R*2))2-3O—, or —S(C(R*2))2-3S—, or a divalent substituent bound to vicinal substitutable carbons of an “optionally substituted” group is —O(CR*2)2-3O—, wherein each independent occurrence of R* is selected from hydrogen, C1-6aliphatic or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

When R* is C1-6aliphatic, R* is optionally substituted with halogen, —R●, -(haloR●), —OH, —OR●, —O(haloR●), —CN, —C(O)OH, —C(O)OR●, —NH2, —NHR●, —NR●2, or —NO2, wherein each R●is independently selected from C1-4aliphatic, —CH2Ph, —O(CH2)0-1Ph, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and wherein each R●is unsubstituted or where preceded by halo is substituted only with one or more halogens.

As used herein, the term “inhibitor” is defined as a compound that binds to and/or inhibits eIF4E with measurable affinity. In certain embodiments, an inhibitor has an IC50and/or binding constant of less than about 100 □M, less than about 50 □M, less than about 22.5 uM, less than about 15 uM, or less than about 7.5 uM.

The terms “measurable affinity” and “measurably inhibit,” as used herein, means a measurable change in eIF4E activity between a sample comprising a compound of the present invention, or composition thereof, and eIF4E, and an equivalent sample comprising eIF4E, in the absence of said compound, or composition thereof.

3. Description of Exemplary Embodiments

or a pharmaceutically acceptable salt thereof, wherein:L1is a bond, O, or S;R1is an optionally substituted ring selected from phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring, a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or an 8-10 membered bicyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur;L2is a C1-8bivalent straight or branched hydrocarbon chain, wherein 1, 2, or 3 methylene units of the hydrocarbon chain are optionally and independently replaced with —N(R)— or —Cy—;each R is independently selected from hydrogen or optionally substituted —C1-6aliphatic;—Cy— is an optionally substituted bivalent ring selected from 3-6 membered monocyclic, saturated or partially unsaturated, carbocyclylene, or 3-6 membered monocyclic, saturated or partially unsaturated, heterocyclylene having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur;R2is —COOR; andR3is R, halogen, or an optionally substituted ring selected from phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring, a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or an 8-10 membered bicyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

As defined generally above, L1is a bond, O, or S.

In some embodiments, L1is selected from those depicted in Table 1, below.

As defined generally above, R1is an optionally substituted ring selected from phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring, a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or an 8-10 membered bicyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

In some embodiments, R1is an optionally substituted phenyl.

In some embodiments, R1is phenyl substituted with 1-2 halogen. In some embodiments, R1is

In some embodiments, R1is

In some embodiments, R1is selected from those depicted in Table 1, below.

As defined generally above, L2is a C1-8bivalent straight or branched hydrocarbon chain, wherein 1, 2, or 3 methylene units of the hydrocarbon chain are optionally and independently replaced with —N(R)— or —Cy—.

In some embodiments, L2is a C1-8bivalent straight or branched hydrocarbon chain, wherein 1, 2, or 3 methylene units of the hydrocarbon chain are optionally and independently replaced with —N(R)—.

In some embodiments, L2is a C1-8bivalent straight or branched hydrocarbon chain, wherein 1, 2, or 3 methylene units of the hydrocarbon chain are optionally and independently replaced with —Cy—.

In some embodiments, L2is C1-8bivalent straight or branched hydrocarbon chain, wherein 1 methylene unit of the hydrocarbon chain is optionally replaced with —N(R)— or —Cy—.

In some embodiments, L2is

In some embodiments, L2is

In some embodiments, L2is

In some embodiments, L2is

In some embodiments, L2is selected from those depicted in Table 1, below.

As defined generally above, each R is independently selected from hydrogen or optionally substituted —C1-6aliphatic.

In some embodiments, R is hydrogen.

In some embodiments, R is optionally substituted —C1-6aliphatic. In some embodiments, R is optionally substituted —C1-6alkyl. In some embodiments, R is optionally substituted —C1-3alkyl.

In some embodiments, R is methyl. In some embodiments, R is ethyl. In some embodiments, R is propyl or isopropyl.

In some embodiments, R is selected from those depicted in Table 1, below.

As defined generally above, each —Cy— is independently an optionally substituted bivalent ring selected from a 3-6 membered monocyclic, saturated or partially unsaturated, carbocyclic ring, or a 3-6 membered monocyclic, saturated or partially unsaturated, heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

In some embodiments, —Cy— is an optionally substituted 3-6 membered monocyclic, saturated or partially unsaturated, bivalent carbocyclic ring. In some embodiments, —Cy— is an optionally substituted 5 or 6 membered monocyclic, saturated or partially unsaturated, bivalent carbocyclic ring.

In some embodiments, —Cy— is an optionally substituted 3-6 membered monocyclic, saturated or partially unsaturated, bivalent heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, —Cy— is an optionally substituted 5 or 6 membered monocyclic, saturated or partially unsaturated, bivalent heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, —Cy— is an optionally substituted 5 or 6 membered monocyclic, saturated or partially unsaturated, bivalent heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen and oxygen.

In some embodiments, —Cy— is a 3-6 membered monocyclic saturated bivalent carbocyclic ring optionally substituted 1-4 times with unsubstituted —C1-6alkyl.

In some embodiments, —Cy— is a 3-6 membered monocyclic saturated bivalent heterocyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, optionally substituted 1-4 times with unsubstituted —C1-6alkyl. In some embodiments, —Cy— is a 3-6 membered monocyclic saturated bivalent heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen and oxygen, optionally substituted 1-4 times with unsubstituted —C1-6alkyl.

In some embodiments, —Cy— is selected from those depicted in Table 1, below.

In some embodiments, R2is selected from those depicted in Table 1, below.

As defined generally above, R3is R, halogen, or an optionally substituted ring selected from phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring, a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or an 8-10 membered bicyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

In some embodiments, R3is

In some embodiments, R3is

In some embodiments, R3is selected from those depicted in Table 1, below.

Exemplary compounds of the invention are set forth in Table 1, below.

In some embodiments, the present invention provides a compound of Table 1, or a pharmaceutically acceptable salt thereof.

4. Formulation and Administration

According to another embodiment, the invention provides a composition comprising a compound of this invention, or a pharmaceutically acceptable derivative thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of compound in compositions of this invention is such that is effective to measurably inhibit eIF4E, or a mutant thereof, in a biological sample or in a patient. In certain embodiments, the amount of compound in compositions of this invention is such that is effective to measurably inhibit eIF4E, or a mutant thereof, in a biological sample or in a patient. In certain embodiments, a composition of this invention is formulated for administration to a patient in need of such composition. In some embodiments, a composition of this invention is formulated for oral administration to a patient.

In some embodiments, the invention provides a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

In some embodiments, the invention provides a pharmaceutical composition comprising a compound of Table 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

In some embodiments, a compound of the invention, or a pharmaceutically acceptable derivative or composition thereof, is administered in a single composition as a single dosage form.

A “pharmaceutically acceptable derivative” means any non-toxic salt, ester, salt of an ester or other derivative of a compound of this invention that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this invention or an inhibitorily active metabolite or residue thereof.

As used herein, the term “inhibitorily active metabolite or residue thereof” means that a metabolite or residue thereof is also an inhibitor of eIF4E, or a mutant thereof.

Most preferably, pharmaceutically acceptable compositions of this invention are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions of this invention are administered without food. In other embodiments, pharmaceutically acceptable compositions of this invention are administered with food.

4.2. Co-Administration with One or More Other Therapeutic Agent

Depending upon the particular condition, or disease, to be treated, additional therapeutic agents that are normally administered to treat that condition, may also be present in the compositions of this invention. As used herein, additional therapeutic agents that are normally administered to treat a particular disease, or condition, are known as “appropriate for the disease, or condition, being treated.”

In some embodiments, the present invention provides a method of treating a disclosed disease or condition comprising administering to a patient in need thereof an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof and co-administering simultaneously or sequentially an effective amount of one or more additional therapeutic agents, such as those described herein. In some embodiments, the method includes co-administering one additional therapeutic agent. In some embodiments, the method includes co-administering two additional therapeutic agents. In some embodiments, the combination of the disclosed compound and the additional therapeutic agent or agents acts synergistically.

A compound of the current invention may also be used in combination with known therapeutic processes, for example, the administration of hormones or radiation. In certain embodiments, a provided compound is used as a radiosensitizer, especially for the treatment of tumors which exhibit poor sensitivity to radiotherapy.

A compound of the current invention can be administered alone or in combination with one or more other therapeutic compounds, possible combination therapy taking the form of fixed combinations or the administration of a compound of the invention and one or more other therapeutic compounds being staggered or given independently of one another, or the combined administration of fixed combinations and one or more other therapeutic compounds. A compound of the current invention can besides or in addition be administered especially for tumor therapy in combination with chemotherapy, radiotherapy, immunotherapy, phototherapy, surgical intervention, or a combination of these. Long-term therapy is equally possible as is adjuvant therapy in the context of other treatment strategies, as described above. Other possible treatments are therapy to maintain the patient's status after tumor regression, or even chemopreventive therapy, for example in patients at risk.

One or more other therapeutic agent may be administered separately from a compound or composition of the invention, as part of a multiple dosage regimen. Alternatively, one or more other therapeutic agents may be part of a single dosage form, mixed together with a compound of this invention in a single composition. If administered as a multiple dosage regime, one or more other therapeutic agent and a compound or composition of the invention may be administered simultaneously, sequentially or within a period of time from one another, for example within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 18, 20, 21, 22, 23, or 24 hours from one another. In some embodiments, one or more other therapeutic agent and a compound or composition of the invention are administered as a multiple dosage regimen within greater than 24 hours apart.

As used herein, the term “combination,” “combined,” and related terms refers to the simultaneous or sequential administration of therapeutic agents in accordance with this invention. For example, a compound of the present invention may be administered with one or more other therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a single unit dosage form. Accordingly, the present invention provides a single unit dosage form comprising a compound of the current invention, one or more other therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

The amount of a compound of the invention and one or more other therapeutic agent (in those compositions which comprise an additional therapeutic agent as described above) that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. Preferably, a composition of the invention should be formulated so that a dosage of between 0.01-100 mg/kg body weight/day of a compound of the invention can be administered.

In those compositions which comprise one or more other therapeutic agent, the one or more other therapeutic agent and a compound of the invention may act synergistically. Therefore, the amount of the one or more other therapeutic agent in such compositions may be less than that required in a monotherapy utilizing only that therapeutic agent. In such compositions a dosage of between 0.01-1,000 μg/kg body weight/day of the one or more other therapeutic agent can be administered.

The amount of one or more other therapeutic agent present in the compositions of this invention may be no more than the amount that would normally be administered in a composition comprising that therapeutic agent as the only active agent. Preferably the amount of one or more other therapeutic agent in the presently disclosed compositions will range from about 50% to 100% of the amount normally present in a composition comprising that agent as the only therapeutically active agent. In some embodiments, one or more other therapeutic agent is administered at a dosage of about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% of the amount normally administered for that agent. As used herein, the phrase “normally administered” means the amount an FDA approved therapeutic agent is approved for dosing per the FDA label insert.

The compounds of this invention, or pharmaceutical compositions thereof, may also be incorporated into compositions for coating an implantable medical device, such as prostheses, artificial valves, vascular grafts, stents and catheters. Vascular stents, for example, have been used to overcome restenosis (re-narrowing of the vessel wall after injury). However, patients using stents or other implantable devices risk clot formation or platelet activation. These unwanted effects may be prevented or mitigated by pre-coating the device with a pharmaceutically acceptable composition comprising a kinase inhibitor. Implantable devices coated with a compound of this invention are another embodiment of the present invention.

4.2.1. Exemplary Other Therapeutic Agents

In some embodiments, one or more other therapeutic agent is a CDK inhibitor, such as a CDK4/CDK6 inhibitor. In some embodiments, a CDK 4/6 inhibitor is selected from palbociclib (Ibrance®, Pfizer); ribociclib (Kisqali®, Novartis); abemaciclib (Ly2835219, Eli Lilly); and trilaciclib (G1T28, G1 Therapeutics).

In some embodiments, one or more other therapeutic agent is a platinum-based therapeutic, also referred to as platins. Platins cause cross-linking of DNA, such that they inhibit DNA repair and/or DNA synthesis, mostly in rapidly reproducing cells, such as cancer cells. In some embodiments, a platinum-based therapeutic is selected from cisplatin (Platinol®, Bristol-Myers Squibb); carboplatin (Paraplatin®, Bristol-Myers Squibb; also, Teva; Pfizer); oxaliplatin (Eloxitin® Sanofi-Aventis); nedaplatin (Aqupla®, Shionogi), picoplatin (Poniard Pharmaceuticals); and satraplatin (JM-216, Agennix).

In some embodiments, one or more other therapeutic agent is a taxane compound, which causes disruption of microtubules, which are essential for cell division. In some embodiments, a taxane compound is selected from paclitaxel (Taxol®, Bristol-Myers Squibb), docetaxel (Taxotere®, Sanofi-Aventis; Docefrez®, Sun Pharmaceutical), albumin-bound paclitaxel (Abraxane®; Abraxis/Celgene), cabazitaxel (Jevtana®, Sanofi-Aventis), and SID530 (SK Chemicals, Co.) (NCT00931008).

In some embodiments, one or more other therapeutic agent is a nucleoside inhibitor, or a therapeutic agent that interferes with normal DNA synthesis, protein synthesis, cell replication, or will otherwise inhibit rapidly proliferating cells.

In some embodiments, one or more other therapeutic agent is an mTOR inhibitor, which inhibits cell proliferation, angiogenesis and glucose uptake. In some embodiments, an mTOR inhibitor is everolimus (Afinitor®, Novartis); temsirolimus (Torisel®, Pfizer); and sirolimus (Rapamune®, Pfizer).

In some embodiments, one or more other therapeutic agent is a proteasome inhibitor. Approved proteasome inhibitors useful in the present invention include bortezomib (Velcade®, Takeda); carfilzomib (Kyprolis®, Amgen); and ixazomib (Ninlaro®, Takeda).

In some embodiments, one or more other therapeutic agent is a growth factor antagonist, such as an antagonist of platelet-derived growth factor (PDGF), or epidermal growth factor (EGF) or its receptor (EGFR). Approved PDGF antagonists which may be used in the present invention include olaratumab (Lartruvo®; Eli Lilly). Approved EGFR antagonists which may be used in the present invention include cetuximab (Erbitux®, Eli Lilly); necitumumab (Portrazza®, Eli Lilly), panitumumab (Vectibix®, Amgen); and osimertinib (targeting activated EGFR, Tagrisso®, AstraZeneca).

In some embodiments, one or more other therapeutic agent is an aromatase inhibitor. In some embodiments, an aromatase inhibitor is selected from exemestane (Aromasin®, Pfizer); anastazole (Arimidex®, AstraZeneca) and letrozole (Femara®, Novartis).

In some embodiments, one or more other therapeutic agent is an antagonist of the hedgehog pathway. Approved hedgehog pathway inhibitors which may be used in the present invention include sonidegib (Odomzo®, Sun Pharmaceuticals); and vismodegib (Erivedge®, Genentech), both for treatment of basal cell carcinoma.

In some embodiments, one or more other therapeutic agent is a folic acid inhibitor. Approved folic acid inhibitors useful in the present invention include pemetrexed (Alimta®, Eli Lilly).

In some embodiments, one or more other therapeutic agent is a CC chemokine receptor 4 (CCR4) inhibitor. CCR4 inhibitors being studied that may be useful in the present invention include mogamulizumab (Poteligeo®, Kyowa Hakko Kirin, Japan).

In some embodiments, one or more other therapeutic agent is an isocitrate dehydrogenase (IDH) inhibitor. IDH inhibitors being studied which may be used in the present invention include AG120 (Celgene; NCT02677922); AG221 (Celgene, NCT02677922; NCT02577406); BAY1436032 (Bayer, NCT02746081); IDH305 (Novartis, NCT02987010).

In some embodiments, one or more other therapeutic agent is an arginase inhibitor. Arginase inhibitors being studied which may be used in the present invention include AEB1102 (pegylated recombinant arginase, Aeglea Biotherapeutics), which is being studied in Phase 1 clinical trials for acute myeloid leukemia and myelodysplastic syndrome (NCT02732184) and solid tumors (NCT02561234); and CB-1158 (Calithera Biosciences).

In some embodiments, one or more other therapeutic agent is a glutaminase inhibitor. Glutaminase inhibitors being studied which may be used in the present invention include CB-839 (Calithera Biosciences).

In some embodiments, one or more other therapeutic agent is a topoisomerase inhibitor. Approved topoisomerase inhibitors useful in the present invention include irinotecan (Onivyde®, Merrimack Pharmaceuticals); topotecan (Hycamtin®, GlaxoSmithKline). Topoisomerase inhibitors being studied which may be used in the present invention include pixantrone (Pixuvri®, CTI Biopharma).

In some embodiments, one or more other therapeutic agent is an inhibitor of anti-apoptotic proteins, such as BCL-2. Approved anti-apoptotics which may be used in the present invention include venetoclax (Venclexta®, AbbVie/Genentech); and blinatumomab (Blincyto®, Amgen). Other therapeutic agents targeting apoptotic proteins which have undergone clinical testing and may be used in the present invention include navitoclax (ABT-263, Abbott), a BCL-2 inhibitor (NCT02079740).

In some embodiments, one or more other therapeutic agent is an androgen receptor inhibitor. Approved androgen receptor inhibitors useful in the present invention include enzalutamide (Xtandi®, Astellas/Medivation); approved inhibitors of androgen synthesis include abiraterone (Zytiga®, Centocor/Ortho); approved antagonist of gonadotropin-releasing hormone (GnRH) receptor (degaralix, Firmagon®, Ferring Pharmaceuticals).

In some embodiments, one or more other therapeutic agent is a selective estrogen receptor modulator (SERM), which interferes with the synthesis or activity of estrogens. Approved SERMs useful in the present invention include raloxifene (Evista®, Eli Lilly).

In some embodiments, one or more other therapeutic agent is an inhibitor of bone resorption. An approved therapeutic which inhibits bone resorption is Denosumab (Xgeva®, Amgen), an antibody that binds to RANKL, prevents binding to its receptor RANK, found on the surface of osteoclasts, their precursors, and osteoclast-like giant cells, which mediates bone pathology in solid tumors with osseous metastases. Other approved therapeutics that inhibit bone resorption include bisphosphonates, such as zoledronic acid (Zometa®, Novartis).

In some embodiments, one or more other therapeutic agent is an inhibitor of interaction between the two primary p53 suppressor proteins, MDMX and MDM2. Inhibitors of p53 suppression proteins being studied which may be used in the present invention include ALRN-6924 (Aileron), a stapled peptide that equipotently binds to and disrupts the interaction of MDMX and MDM2 with p53. ALRN-6924 is currently being evaluated in clinical trials for the treatment of AML, advanced myelodysplastic syndrome (MDS) and peripheral T-cell lymphoma (PTCL) (NCT02909972; NCT02264613).

In some embodiments, one or more other therapeutic agent is an inhibitor of transforming growth factor-beta (TGF-beta or TGFß). Inhibitors of TGF-beta proteins being studied which may be used in the present invention include NIS793 (Novartis), an anti-TGF-beta antibody being tested in the clinic for treatment of various cancers, including breast, lung, hepatocellular, colorectal, pancreatic, prostate and renal cancer (NCT02947165). In some embodiments, the inhibitor of TGF-beta proteins is fresolimumab (GC1008; Sanofi-Genzyme), which is being studied for melanoma (NCT00923169); renal cell carcinoma (NCT00356460); and non-small cell lung cancer (NCT02581787). Additionally, in some embodiments, the additional therapeutic agent is a TGF-beta trap, such as described in Connolly et al. (2012) Int'l J. Biological Sciences 8:964-978. One therapeutic compound currently in clinical trials for treatment of solid tumors is M7824 (Merck KgaA—formerly MSB0011459X), which is a bispecific, anti-PD-L1/TGFß trap compound (NCT02699515); and (NCT02517398). M7824 is comprised of a fully human IgG1 antibody against PD-L1 fused to the extracellular domain of human TGF-beta receptor II, which functions as a TGFß “trap.”

In some embodiments, one or more other therapeutic agent is selected from glembatumumab vedotin-monomethyl auristatin E (MMAE) (Celldex), an anti-glycoprotein NMB (gpNMB) antibody (CR011) linked to the cytotoxic MMAE. gpNMB is a protein overexpressed by multiple tumor types associated with cancer cells' ability to metastasize.

In some embodiments, one or more other therapeutic agent is an antiproliferative compound. Such antiproliferative compounds include, but are not limited to aromatase inhibitors; antiestrogens; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule active compounds; alkylating compounds; histone deacetylase inhibitors; compounds which induce cell differentiation processes; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors; antineoplastic antimetabolites; platin compounds; compounds targeting/decreasing a protein or lipid kinase activity and further anti-angiogenic compounds; compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase; gonadorelin agonists; anti-androgens; methionine aminopeptidase inhibitors; matrix metalloproteinase inhibitors; bisphosphonates; biological response modifiers; antiproliferative antibodies; heparanase inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds used in the treatment of hematologic malignancies; compounds which target, decrease or inhibit the activity of Flt-3; Hsp90 inhibitors such as 17-AAG (17-allylaminogeldanamycin, NSC330507), 17-DMAG (17-dimethylaminoethylamino-17-demethoxy-geldanamycin, NSC707545), IPI-504, CNF1010, CNF2024, CNF1010 from Conforma Therapeutics; temozolomide (Temodal©); kinesin spindle protein inhibitors, such as SB715992 or SB743921 from GlaxoSmithKline, or pentamidine/chlorpromazine from CombinatoRx; MEK inhibitors such as ARRY142886 from Array BioPharma, AZd6244 from AstraZeneca, PD181461 from Pfizer and leucovorin.

The term “aromatase inhibitor” as used herein relates to a compound which inhibits estrogen production, for instance, the conversion of the substrates androstenedione and testosterone to estrone and estradiol, respectively. The term includes, but is not limited to steroids, especially atamestane, exemestane and formestane and, in particular, non-steroids, especially aminoglutethimide, roglethimide, pyridoglutethimide, trilostane, testolactone, ketokonazole, vorozole, fadrozole, anastrozole and letrozole. Exemestane is marketed under the trade name Aromasin™. Formestane is marketed under the trade name Lentaron™. Fadrozole is marketed under the trade name Afema™. Anastrozole is marketed under the trade name Arimidex™ Letrozole is marketed under the trade names Femara™ or Femar™. Aminoglutethimide is marketed under the trade name Orimeten™. A combination of the invention comprising a chemotherapeutic agent which is an aromatase inhibitor is particularly useful for the treatment of hormone receptor positive tumors, such as breast tumors.

The term “antiestrogen” as used herein relates to a compound which antagonizes the effect of estrogens at the estrogen receptor level. The term includes, but is not limited to tamoxifen, fulvestrant, raloxifene and raloxifene hydrochloride. Tamoxifen is marketed under the trade name Nolvadex™. Raloxifene hydrochloride is marketed under the trade name Evista™. Fulvestrant can be administered under the trade name Faslodex™. A combination of the invention comprising a chemotherapeutic agent which is an antiestrogen is particularly useful for the treatment of estrogen receptor positive tumors, such as breast tumors.

The term “anti-androgen” as used herein relates to any substance which is capable of inhibiting the biological effects of androgenic hormones and includes, but is not limited to, bicalutamide (Casodex™). The term “gonadorelin agonist” as used herein includes, but is not limited to abarelix, goserelin and goserelin acetate. Goserelin can be administered under the trade name Zoladex™.

The term “topoisomerase I inhibitor” as used herein includes, but is not limited to topotecan, gimatecan, irinotecan, camptothecian and its analogues, 9-nitrocamptothecin and the macromolecular camptothecin conjugate PNU-166148. Irinotecan can be administered, e.g. in the form as it is marketed, e.g. under the trademark Camptosar™. Topotecan is marketed under the trade name Hycamptin™.

The term “topoisomerase II inhibitor” as used herein includes, but is not limited to the anthracyclines such as doxorubicin (including liposomal formulation, such as Caelyx™) daunorubicin, epirubicin, idarubicin and nemorubicin, the anthraquinones mitoxantrone and losoxantrone, and the podophillotoxines etoposide and teniposide. Etoposide is marketed under the trade name Etopophos™. Teniposide is marketed under the trade name VM 26-Bristol Doxorubicin is marketed under the trade name Acriblastin™ or Adriamycin™. Epirubicin is marketed under the trade name Farmorubicin™. Idarubicin is marketed. under the trade name Zavedos™. Mitoxantrone is marketed under the trade name Novantron.

The term “microtubule active agent” relates to microtubule stabilizing, microtubule destabilizing compounds and microtublin polymerization inhibitors including, but not limited to taxanes, such as paclitaxel and docetaxel; vinca alkaloids, such as vinblastine or vinblastine sulfate, vincristine or vincristine sulfate, and vinorelbine; discodermolides; cochicine and epothilones and derivatives thereof. Paclitaxel is marketed under the trade name Taxol™ Docetaxel is marketed under the trade name Taxotere™. Vinblastine sulfate is marketed under the trade name Vinblastin R.P™. Vincristine sulfate is marketed under the trade name Farmistin™.

The term “alkylating agent” as used herein includes, but is not limited to, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNU or Gliadel). Cyclophosphamide is marketed under the trade name Cyclostin™. Ifosfamide is marketed under the trade name Holoxan™.

The term “histone deacetylase inhibitors” or “HDAC inhibitors” relates to compounds which inhibit the histone deacetylase and which possess antiproliferative activity. This includes, but is not limited to, suberoylanilide hydroxamic acid (SAHA).

The term “antineoplastic antimetabolite” includes, but is not limited to, 5-fluorouracil or 5-FU, capecitabine, gemcitabine, DNA demethylating compounds, such as 5-azacytidine and decitabine, methotrexate and edatrexate, and folic acid antagonists such as pemetrexed. Capecitabine is marketed under the trade name Xeloda™. Gemcitabine is marketed under the trade name Gemzar™.

The term “platin compound” as used herein includes, but is not limited to, carboplatin, cis-platin, cisplatinum and oxaliplatin. Carboplatin can be administered, e.g., in the form as it is marketed, e.g. under the trademark Carboplat™. Oxaliplatin can be administered, e.g., in the form as it is marketed, e.g. under the trademark Eloxatin™.

The term “compounds targeting/decreasing a protein or lipid kinase activity; or a protein or lipid phosphatase activity; or further anti-angiogenic compounds” as used herein includes, but is not limited to, protein tyrosine kinase and/or serine and/or threonine kinase inhibitors or lipid kinase inhibitors, such as a) compounds targeting, decreasing or inhibiting the activity of the platelet-derived growth factor-receptors (PDGFR), such as compounds which target, decrease or inhibit the activity of PDGFR, especially compounds which inhibit the PDGF receptor, such as an N-phenyl-2-pyrimidine-amine derivative, such as imatinib, SU101, SU6668 and GFB-111; b) compounds targeting, decreasing or inhibiting the activity of the fibroblast growth factor-receptors (FGFR); c) compounds targeting, decreasing or inhibiting the activity of the insulin-like growth factor receptor I (IGF-IR), such as compounds which target, decrease or inhibit the activity of IGF-IR, especially compounds which inhibit the kinase activity of IGF-I receptor, or antibodies that target the extracellular domain of IGF-I receptor or its growth factors; d) compounds targeting, decreasing or inhibiting the activity of the Trk receptor tyrosine kinase family, or ephrin B4 inhibitors; e) compounds targeting, decreasing or inhibiting the activity of the AxI receptor tyrosine kinase family; f) compounds targeting, decreasing or inhibiting the activity of the Ret receptor tyrosine kinase; g) compounds targeting, decreasing or inhibiting the activity of the Kit/SCFR receptor tyrosine kinase, such as imatinib; h) compounds targeting, decreasing or inhibiting the activity of the C-kit receptor tyrosine kinases, which are part of the PDGFR family, such as compounds which target, decrease or inhibit the activity of the c-Kit receptor tyrosine kinase family, especially compounds which inhibit the c-Kit receptor, such as imatinib; i) compounds targeting, decreasing or inhibiting the activity of members of the c-Abl family, their gene-fusion products (e.g. BCR-Abl kinase) and mutants, such as compounds which target decrease or inhibit the activity of c-Abl family members and their gene fusion products, such as an N-phenyl-2-pyrimidine-amine derivative, such as imatinib or nilotinib (AMN107); PD180970; AG957; NSC 680410; PD173955 from ParkeDavis; or dasatinib (BMS-354825); j) compounds targeting, decreasing or inhibiting the activity of members of the protein kinase C (PKC) and Raf family of serine/threonine kinases, members of the MEK, SRC, JAK/pan-JAK, FAK, PDK1, PKB/Akt, Ras/MAPK, PI3K, SYK, TYK2, BTK and TEC family, and/or members of the cyclin-dependent kinase family (CDK) including staurosporine derivatives, such as midostaurin; examples of further compounds include UCN-01, safingol, BAY 43-9006, Bryostatin 1, Perifosine; llmofosine; RO 318220 and RO 320432; GO 6976; lsis 3521; LY333531/LY379196; isochinoline compounds; FTIs; PD184352 or QAN697 (a P13K inhibitor) or AT7519 (CDK inhibitor); k) compounds targeting, decreasing or inhibiting the activity of protein-tyrosine kinase inhibitors, such as compounds which target, decrease or inhibit the activity of protein-tyrosine kinase inhibitors include imatinib mesylate (Gleevec™) or tyrphostin such as Tyrphostin A23/RG-50810; AG 99; Tyrphostin AG 213; Tyrphostin AG 1748; Tyrphostin AG 490; Tyrphostin B44; Tyrphostin B44 (+) enantiomer; Tyrphostin AG 555; AG 494; Tyrphostin AG 556, AG957 and adaphostin (4-{[(2,5-dihydroxyphenyl)methyl]amino}-benzoic acid adamantyl ester; NSC 680410, adaphostin); 1) compounds targeting, decreasing or inhibiting the activity of the epidermal growth factor family of receptor tyrosine kinases (EGFR1ErbB2, ErbB3, ErbB4 as homo- or heterodimers) and their mutants, such as compounds which target, decrease or inhibit the activity of the epidermal growth factor receptor family are especially compounds, proteins or antibodies which inhibit members of the EGF receptor tyrosine kinase family, such as EGF receptor, ErbB2, ErbB3 and ErbB4 or bind to EGF or EGF related ligands, CP 358774, ZD 1839, ZM 105180; trastuzumab (Herceptin™), cetuximab (Erbitux™), Iressa, Tarceva, OSI-774, Cl-1033, EKB-569, GW-2016, E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3 or E7.6.3, and 7H-pyrrolo-[2,3-d]pyrimidine derivatives; m) compounds targeting, decreasing or inhibiting the activity of the c-Met receptor, such as compounds which target, decrease or inhibit the activity of c-Met, especially compounds which inhibit the kinase activity of c-Met receptor, or antibodies that target the extracellular domain of c-Met or bind to HGF, n) compounds targeting, decreasing or inhibiting the kinase activity of one or more JAK family members (JAK1/JAK2/JAK3/TYK2 and/or pan-JAK), including but not limited to PRT-062070, SB-1578, baricitinib, pacritinib, momelotinib, VX-509, AZD-1480, TG-101348, tofacitinib, and ruxolitinib; o) compounds targeting, decreasing or inhibiting the kinase activity of PI3 kinase (PI3K) including but not limited to ATU-027, SF-1126, DS—7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib, pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, and idelalisib; and; and q) compounds targeting, decreasing or inhibiting the signaling effects of hedgehog protein (Hh) or smoothened receptor (SMO) pathways, including but not limited to cyclopamine, vismodegib, itraconazole, erismodegib, and IPI-926 (saridegib).

The term “PI3K inhibitor” as used herein includes, but is not limited to compounds having inhibitory activity against one or more enzymes in the phosphatidylinositol-3-kinase family, including, but not limited to PI3Kα, PI3Kγ, PI3Kδ, PI3Kβ, PI3K-C2α, PI3K-C2β, PI3K-C2γ, Vps34, p110-α, p110-β, p110-γ, p110-δ, p85-α, p85-β, p55-γ, p150, p101, and p87. Examples of PI3K inhibitors useful in this invention include but are not limited to ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib, pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, and idelalisib.

The term “BTK inhibitor” as used herein includes, but is not limited to compounds having inhibitory activity against Bruton's Tyrosine Kinase (BTK), including, but not limited to AVL-292 and ibrutinib.

The term “SYK inhibitor” as used herein includes, but is not limited to compounds having inhibitory activity against spleen tyrosine kinase (SYK), including but not limited to PRT-062070, R-343, R-333, Excellair, PRT-062607, and fostamatinib.

Further examples of BTK inhibitory compounds, and conditions treatable by such compounds in combination with compounds of this invention can be found in WO2008039218 and WO2011090760, the entirety of which are incorporated herein by reference.

Further examples of SYK inhibitory compounds, and conditions treatable by such compounds in combination with compounds of this invention can be found in WO2003063794, WO2005007623, and WO2006078846, the entirety of which are incorporated herein by reference.

Further examples of PI3K inhibitory compounds, and conditions treatable by such compounds in combination with compounds of this invention can be found in WO2004019973, WO2004089925, WO2007016176, U.S. Pat. No. 8,138,347, WO2002088112, WO2007084786, WO2007129161, WO2006122806, WO2005113554, and WO2007044729 the entirety of which are incorporated herein by reference.

Further examples of JAK inhibitory compounds, and conditions treatable by such compounds in combination with compounds of this invention can be found in WO2009114512, WO2008109943, WO2007053452, WO2000142246, and WO2007070514, the entirety of which are incorporated herein by reference.

Further anti-angiogenic compounds include compounds having another mechanism for their activity, e.g. unrelated to protein or lipid kinase inhibition e.g. thalidomide (Thalomid™) and TNP-470.

Examples of proteasome inhibitors useful for use in combination with compounds of the invention include, but are not limited to bortezomib, disulfiram, epigallocatechin-3-gallate (EGCG), salinosporamide A, carfilzomib, ONX-0912, CEP-18770, and MLN9708.

Compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase are e.g. inhibitors of phosphatase 1, phosphatase 2A, or CDC25, such as okadaic acid or a derivative thereof.

Compounds which induce cell differentiation processes include, but are not limited to, retinoic acid, α- γ- or δ-tocopherol or α- γ- or δ-tocotrienol.

The term cyclooxygenase inhibitor as used herein includes, but is not limited to, Cox-2 inhibitors, 5-alkyl substituted 2-arylaminophenylacetic acid and derivatives, such as celecoxib (Celebrex™), rofecoxib (Vioxx™), etoricoxib, valdecoxib or a 5-alkyl-2-arylaminophenylacetic acid, such as 5-methyl-2-(2′-chloro-6′-fluoroanilino)phenyl acetic acid, lumiracoxib.

The term “bisphosphonates” as used herein includes, but is not limited to, etridonic, clodronic, tiludronic, pamidronic, alendronic, ibandronic, risedronic and zoledronic acid. Etridonic acid is marketed under the trade name Didronel™. Clodronic acid is marketed under the trade name Bonefos™. Tiludronic acid is marketed under the trade name Skelid™. Pamidronic acid is marketed under the trade name Aredia™. Alendronic acid is marketed under the trade name Fosamax™. Ibandronic acid is marketed under the trade name Bondranat™. Risedronic acid is marketed under the trade name Actonel™. Zoledronic acid is marketed under the trade name Zometa™. The term “mTOR inhibitors” relates to compounds which inhibit the mammalian target of rapamycin (mTOR) and which possess antiproliferative activity such as sirolimus (Rapamune®), everolimus (Certican™), CCI-779 and ABT578.

The term “heparanase inhibitor” as used herein refers to compounds which target, decrease or inhibit heparin sulfate degradation. The term includes, but is not limited to, PI-88. The term “biological response modifier” as used herein refers to a lymphokine or interferons.

The term “inhibitor of Ras oncogenic isoforms”, such as H-Ras, K-Ras, or N-Ras, as used herein refers to compounds which target, decrease or inhibit the oncogenic activity of Ras; for example, a “farnesyl transferase inhibitor” such as L-744832, DK8G557 or R115777 (Zarnestra™). The term “telomerase inhibitor” as used herein refers to compounds which target, decrease or inhibit the activity of telomerase. Compounds which target, decrease or inhibit the activity of telomerase are especially compounds which inhibit the telomerase receptor, such as telomestatin.

The term “methionine aminopeptidase inhibitor” as used herein refers to compounds which target, decrease or inhibit the activity of methionine aminopeptidase. Compounds which target, decrease or inhibit the activity of methionine aminopeptidase include, but are not limited to, bengamide or a derivative thereof.

The term “proteasome inhibitor” as used herein refers to compounds which target, decrease or inhibit the activity of the proteasome. Compounds which target, decrease or inhibit the activity of the proteasome include, but are not limited to, Bortezomib (Velcade™) and MLN 341.

The term “compounds used in the treatment of hematologic malignancies” as used herein includes, but is not limited to, FMS-like tyrosine kinase inhibitors, which are compounds targeting, decreasing or inhibiting the activity of FMS-like tyrosine kinase receptors (Flt-3R); interferon, 1-β-D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors, which are compounds which target, decrease or inhibit anaplastic lymphoma kinase.

Compounds which target, decrease or inhibit the activity of FMS-like tyrosine kinase receptors (Flt-3R) are especially compounds, proteins or antibodies which inhibit members of the Flt-3R receptor kinase family, such as PKC412, midostaurin, a staurosporine derivative, SU11248 and MLN518.

The term “antiproliferative antibodies” as used herein includes, but is not limited to, trastuzumab (Herceptin™), Trastuzumab-DM1, erbitux, bevacizumab (Avastin™), rituximab (Rituxan®), PRO64553 (anti-CD40) and 2C4 Antibody. By antibodies is meant intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies formed from at least 2 intact antibodies, and antibodies fragments so long as they exhibit the desired biological activity.

For the treatment of acute myeloid leukemia (AML), compounds of the current invention can be used in combination with standard leukemia therapies, especially in combination with therapies used for the treatment of AML. In particular, compounds of the current invention can be administered in combination with, for example, farnesyl transferase inhibitors and/or other drugs useful for the treatment of AML, such as Daunorubicin, Adriamycin, Ara-C, VP-16, Teniposide, Mitoxantrone, Idarubicin, Carboplatinum and PKC412.

Other anti-leukemic compounds include, for example, Ara-C, a pyrimidine analog, which is the 2′-alpha-hydroxy ribose (arabinoside) derivative of deoxycytidine. Also included is the purine analog of hypoxanthine, 6-mercaptopurine (6-MP) and fludarabine phosphate. Compounds which target, decrease or inhibit activity of histone deacetylase (HDAC) inhibitors such as sodium butyrate and suberoylanilide hydroxamic acid (SAHA) inhibit the activity of the enzymes known as histone deacetylases. Specific HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), Trichostatin A and compounds disclosed in U.S. Pat. No. 6,552,065 including, but not limited to, N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof and N-hydroxy-3-[4-[(2-hydroxyethyl){2-(1H-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof, especially the lactate salt. Somatostatin receptor antagonists as used herein refer to compounds which target, treat or inhibit the somatostatin receptor such as octreotide, and SOM230. Tumor cell damaging approaches refer to approaches such as ionizing radiation. The term “ionizing radiation” referred to above and hereinafter means ionizing radiation that occurs as either electromagnetic rays (such as X-rays and gamma rays) or particles (such as alpha and beta particles). Ionizing radiation is provided in, but not limited to, radiation therapy and is known in the art. See Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, Devita et al., Eds., 4thEdition, Vol. 1, pp. 248-275 (1993).

Also included are EDG binders and ribonucleotide reductase inhibitors. The term “EDG binders” as used herein refers to a class of immunosuppressants that modulates lymphocyte recirculation, such as FTY720. The term “ribonucleotide reductase inhibitors” refers to pyrimidine or purine nucleoside analogs including, but not limited to, fludarabine and/or cytosine arabinoside (ara-C), 6-thioguanine, 5-fluorouracil, cladribine, 6-mercaptopurine (especially in combination with ara-C against ALL) and/or pentostatin. Ribonucleotide reductase inhibitors are especially hydroxyurea or 2-hydroxy-1H-isoindole-1,3-dione derivatives.

Photodynamic therapy as used herein refers to therapy which uses certain chemicals known as photosensitizing compounds to treat or prevent cancers. Examples of photodynamic therapy include treatment with compounds, such as Visudyne™ and porfimer sodium.

Implants containing corticosteroids refers to compounds, such as fluocinolone and dexamethasone.

Other chemotherapeutic compounds include, but are not limited to, plant alkaloids, hormonal compounds and antagonists; biological response modifiers, preferably lymphokines or interferons; antisense oligonucleotides or oligonucleotide derivatives; shRNA or siRNA; or miscellaneous compounds or compounds with other or unknown mechanism of action.

The structure of the active compounds identified by code numbers, generic or trade names may be taken from the actual edition of the standard compendium “The Merck Index” or from databases, e.g. Patents International (e.g. IMS World Publications).

In some embodiments, one or more other therapeutic agent is an immuno-oncology agent. As used herein, the term “an immuno-oncology agent” refers to an agent which is effective to enhance, stimulate, and/or up-regulate immune responses in a subject. In some embodiments, the administration of an immuno-oncology agent with a compound of the invention has a synergic effect in treating a cancer.

An immuno-oncology agent can be, for example, a small molecule drug, an antibody, or a biologic or small molecule. Examples of biologic immuno-oncology agents include, but are not limited to, cancer vaccines, antibodies, and cytokines. In some embodiments, an antibody is a monoclonal antibody. In some embodiments, a monoclonal antibody is humanized or human.

In some embodiments, an immuno-oncology agent is (i) an agonist of a stimulatory (including a co-stimulatory) receptor or (ii) an antagonist of an inhibitory (including a co-inhibitory) signal on T cells, both of which result in amplifying antigen-specific T cell responses.

In some embodiments, an immuno-oncology agent is a cytokine that inhibits T cell activation (e.g., IL-6, IL-10, TGF-β, VEGF, and other immunosuppressive cytokines) or a cytokine that stimulates T cell activation, for stimulating an immune response.

In some embodiments, an immuno-oncology agent is an antagonist of inhibitory receptors on NK cells or an agonists of activating receptors on NK cells. In some embodiments, an immuno-oncology agent is an antagonists of KIR, such as lirilumab.

In some embodiments, an immuno-oncology agent is an agent that inhibits or depletes macrophages or monocytes, including but not limited to CSF-1R antagonists such as CSF-1R antagonist antibodies including RG7155 (WO11/70024, WO11/107553, WO11/131407, WO13/87699, WO13/119716, WO13/132044) or FPA-008 (WO11/140249; WO13169264; WO14/036357).

In some embodiments, an immuno-oncology agent is selected from agonistic agents that ligate positive costimulatory receptors, blocking agents that attenuate signaling through inhibitory receptors, antagonists, and one or more agents that increase systemically the frequency of anti-tumor T cells, agents that overcome distinct immune suppressive pathways within the tumor microenvironment (e.g., block inhibitory receptor engagement (e.g., PD-L1/PD-1 interactions), deplete or inhibit Tregs (e.g., using an anti-CD25 monoclonal antibody (e.g., daclizumab) or by ex vivo anti-CD25 bead depletion), inhibit metabolic enzymes such as IDO, or reverse/prevent T cell energy or exhaustion) and agents that trigger innate immune activation and/or inflammation at tumor sites.

In some embodiments, an immuno-oncology agent is a CTLA-4 antagonist. In some embodiments, a CTLA-4 antagonist is an antagonistic CTLA-4 antibody. In some embodiments, an antagonistic CTLA-4 antibody is YERVOY (ipilimumab) or tremelimumab.

In some embodiments, an immuno-oncology agent is a PD-1 antagonist. In some embodiments, a PD-1 antagonist is administered by infusion. In some embodiments, an immuno-oncology agent is an antibody or an antigen-binding portion thereof that binds specifically to a Programmed Death-1 (PD-1) receptor and inhibits PD-1 activity. In some embodiments, a PD-1 antagonist is an antagonistic PD-1 antibody. In some embodiments, an antagonistic PD-1 antibody is OPDIVO (nivolumab), KEYTRUDA (pembrolizumab), or MEDI-0680 (AMP-514; WO2012/145493). In some embodiments, an immuno-oncology agent may be pidilizumab (CT-011). In some embodiments, an immuno-oncology agent is a recombinant protein composed of the extracellular domain of PD-L2 (B7-DC) fused to the Fc portion of IgG1, called AMP-224.

In some embodiments, an immuno-oncology agent is a PD-L1 antagonist. In some embodiments, a PD-L1 antagonist is an antagonistic PD-L1 antibody. In some embodiments, a PD-L1 antibody is MPDL3280A (RG7446; WO2010/077634), durvalumab (MEDI4736), BMS-936559 (WO2007/005874), and MSB0010718C (WO2013/79174).

In some embodiments, an immuno-oncology agent is a LAG-3 antagonist. In some embodiments, a LAG-3 antagonist is an antagonistic LAG-3 antibody. In some embodiments, a LAG3 antibody is BMS-986016 (WO10/19570, WO14/08218), or IMP-731 or IMP-321 (WO08/132601, WO009/44273).

In some embodiments, an immuno-oncology agent is a CD137 (4-1BB) agonist. In some embodiments, a CD137 (4-1BB) agonist is an agonistic CD137 antibody. In some embodiments, a CD137 antibody is urelumab or PF-05082566 (WO12/32433).

In some embodiments, an immuno-oncology agent is a GITR agonist. In some embodiments, a GITR agonist is an agonistic GITR antibody. In some embodiments, a GITR antibody is BMS-986153, BMS-986156, TRX-518 (WO006/105021, WO009/009116), or MK-4166 (WO11/028683).

In some embodiments, an immuno-oncology agent is an OX40 agonist. In some embodiments, an OX40 agonist is an agonistic OX40 antibody. In some embodiments, an OX40 antibody is MEDI-6383 or MEDI-6469.

In some embodiments, an immuno-oncology agent is an OX40L antagonist. In some embodiments, an OX40L antagonist is an antagonistic OX40 antibody. In some embodiments, an OX40L antagonist is RG-7888 (WO06/029879).

In some embodiments, an immuno-oncology agent is a CD40 agonist. In some embodiments, a CD40 agonist is an agonistic CD40 antibody. In some embodiments, an immuno-oncology agent is a CD40 antagonist. In some embodiments, a CD40 antagonist is an antagonistic CD40 antibody. In some embodiments, a CD40 antibody is lucatumumab or dacetuzumab.

In some embodiments, an immuno-oncology agent is a CD27 agonist. In some embodiments, a CD27 agonist is an agonistic CD27 antibody. In some embodiments, a CD27 antibody is varlilumab.

In some embodiments, an immuno-oncology agent is an immunostimulatory agent. For example, antibodies blocking the PD-1 and PD-L1 inhibitory axis can unleash activated tumor-reactive T cells and have been shown in clinical trials to induce durable anti-tumor responses in increasing numbers of tumor histologies, including some tumor types that conventionally have not been considered immunotherapy sensitive. See, e.g., Okazaki, T. et al. (2013) Nat. Immunol. 14, 1212-1218; Zou et al. (2016) Sci. Transl. Med. 8. The anti-PD-1 antibody nivolumab (Opdivo®, Bristol-Myers Squibb, also known as ONO-4538, MDX1106 and BMS-936558), has shown potential to improve the overall survival in patients with RCC who had experienced disease progression during or after prior anti-angiogenic therapy.

In some embodiments, the immunomodulatory therapeutic specifically induces apoptosis of tumor cells. Approved immunomodulatory therapeutics which may be used in the present invention include pomalidomide (Pomalyst®, Celgene); lenalidomide (Revlimid®, Celgene); ingenol mebutate (Picato®, LEO Pharma).

In some embodiments, an immuno-oncology agent is a cancer vaccine. In some embodiments, the cancer vaccine is selected from sipuleucel-T (Provenge®, Dendreon/Valeant Pharmaceuticals), which has been approved for treatment of asymptomatic, or minimally symptomatic metastatic castrate-resistant (hormone-refractory) prostate cancer; and talimogene laherparepvec (Imlygic®, BioVex/Amgen, previously known as T-VEC), a genetically modified oncolytic viral therapy approved for treatment of unresectable cutaneous, subcutaneous and nodal lesions in melanoma. In some embodiments, an immuno-oncology agent is selected from an oncolytic viral therapy such as pexastimogene devacirepvec (PexaVec/JX-594, SillaJen/formerly Jennerex Biotherapeutics), a thymidine kinase- (TK-) deficient vaccinia virus engineered to express GM-CSF, for hepatocellular carcinoma (NCT02562755) and melanoma (NCT00429312); pelareorep (Reolysin®, Oncolytics Biotech), a variant of respiratory enteric orphan virus (reovirus) which does not replicate in cells that are not RAS-activated, in numerous cancers, including colorectal cancer (NCT01622543); prostate cancer (NCT01619813); head and neck squamous cell cancer (NCT01166542); pancreatic adenocarcinoma (NCT00998322); and non-small cell lung cancer (NSCLC) (NCT00861627); enadenotucirev (NG-348, PsiOxus, formerly known as ColoAdl), an adenovirus engineered to express a full length CD80 and an antibody fragment specific for the T-cell receptor CD3 protein, in ovarian cancer (NCT02028117); metastatic or advanced epithelial tumors such as in colorectal cancer, bladder cancer, head and neck squamous cell carcinoma and salivary gland cancer (NCT02636036); ONCOS-102 (Targovax/formerly Oncos), an adenovirus engineered to express GM-CSF, in melanoma (NCT03003676); and peritoneal disease, colorectal cancer or ovarian cancer (NCT02963831); GL-ONC1 (GLV-lh68/GLV-lh153, Genelux GmbH), vaccinia viruses engineered to express beta-galactosidase (beta-gal)/beta-glucoronidase or beta-gal/human sodium iodide symporter (hNIS), respectively, were studied in peritoneal carcinomatosis (NCT01443260); fallopian tube cancer, ovarian cancer (NCT02759588); or CG0070 (Cold Genesys), an adenovirus engineered to express GM-CSF, in bladder cancer (NCT02365818).

In some embodiments, an immuno-oncology agent is selected from JX-929 (SillaJen/formerly Jennerex Biotherapeutics), a TK- and vaccinia growth factor-deficient vaccinia virus engineered to express cytosine deaminase, which is able to convert the prodrug 5-fluorocytosine to the cytotoxic drug 5-fluorouracil; TG01 and TG02 (Targovax/formerly Oncos), peptide-based immunotherapy agents targeted for difficult-to-treat RAS mutations; and TILT-123 (TILT Biotherapeutics), an engineered adenovirus designated: Ad5/3-E2F-delta24-hTNFα-IRES-hIL20; and VSV-GP (ViraTherapeutics) a vesicular stomatitis virus (VSV) engineered to express the glycoprotein (GP) of lymphocytic choriomeningitis virus (LCMV), which can be further engineered to express antigens designed to raise an antigen-specific CD8+T cell response.

In some embodiments, an immuno-oncology agent is a T-cell engineered to express a chimeric antigen receptor, or CAR. The T-cells engineered to express such chimeric antigen receptor are referred to as a CAR-T cells.

CARs have been constructed that consist of binding domains, which may be derived from natural ligands, single chain variable fragments (scFv) derived from monoclonal antibodies specific for cell-surface antigens, fused to endodomains that are the functional end of the T-cell receptor (TCR), such as the CD3-zeta signaling domain from TCRs, which is capable of generating an activation signal in T lymphocytes. Upon antigen binding, such CARs link to endogenous signaling pathways in the effector cell and generate activating signals similar to those initiated by the TCR complex.

For example, in some embodiments the CAR-T cell is one of those described in U.S. Pat. No. 8,906,682 (June; hereby incorporated by reference in its entirety), which discloses CAR-T cells engineered to comprise an extracellular domain having an antigen binding domain (such as a domain that binds to CD19), fused to an intracellular signaling domain of the T cell antigen receptor complex zeta chain (such as CD3 zeta). When expressed in the T cell, the CAR is able to redirect antigen recognition based on the antigen binding specificity. In the case of CD19, the antigen is expressed on malignant B cells. Over 200 clinical trials are currently in progress employing CAR-T in a wide range of indications. [https://clinicaltrials.gov/ct2/results?term=chimeric+antigen+receptors&pg=1].

In some embodiments, an immunostimulatory agent is an activator of retinoic acid receptor-related orphan receptor γ (RORγt). RORγt is a transcription factor with key roles in the differentiation and maintenance of Type 17 effector subsets of CD4+(Th17) and CD8+(Tc17) T cells, as well as the differentiation of IL-17 expressing innate immune cell subpopulations such as NK cells. In some embodiments, an activator of RORγt is LYC-55716 (Lycera), which is currently being evaluated in clinical trials for the treatment of solid tumors (NCT02929862).

In some embodiments, an immunostimulatory agent is an agonist or activator of a toll-like receptor (TLR). Suitable activators of TLRs include an agonist or activator of TLR9 such as SD-101 (Dynavax). SD-101 is an immunostimulatory CpG which is being studied for B-cell, follicular and other lymphomas (NCT02254772). Agonists or activators of TLR8 which may be used in the present invention include motolimod (VTX-2337, VentiRx Pharmaceuticals) which is being studied for squamous cell cancer of the head and neck (NCTO2124850) and ovarian cancer (NCT02431559).

In some embodiments, an immunostimulatory agent is selected from elotuzumab, mifamurtide, an agonist or activator of a toll-like receptor, and an activator of RORγt.

In some embodiments, an immunostimulatory therapeutic is recombinant human interleukin 15 (rhIL-15). rhIL-15 has been tested in the clinic as a therapy for melanoma and renal cell carcinoma (NCT01021059 and NCT01369888) and leukemias (NCT02689453). In some embodiments, an immunostimulatory agent is recombinant human interleukin 12 (rhIL-12). In some embodiments, an IL-15 based immunotherapeutic is heterodimeric IL-15 (hetIL-15, Novartis/Admune), a fusion complex composed of a synthetic form of endogenous IL-15 complexed to the soluble IL-15 binding protein IL-15 receptor alpha chain (IL15:sIL-15RA), which has been tested in Phase 1 clinical trials for melanoma, renal cell carcinoma, non-small cell lung cancer and head and neck squamous cell carcinoma (NCT02452268). In some embodiments, a recombinant human interleukin 12 (rhIL-12) is NM-IL-12 (Neumedicines, Inc.), NCT02544724, or NCT02542124.

In some embodiments, an immuno-oncology agent is selected from those descripted in Jerry L. Adams ET. AL., “Big opportunities for small molecules in immuno-oncology,” Cancer Therapy 2015, Vol. 14, pages 603-622, the content of which is incorporated herein by reference in its entirety. In some embodiments, an immuno-oncology agent is selected from the examples described in Table 1 of Jerry L. Adams ET. AL. In some embodiments, an immuno-oncology agent is a small molecule targeting an immuno-oncology target selected from those listed in Table 2 of Jerry L. Adams ET. AL. In some embodiments, an immuno-oncology agent is a small molecule agent selected from those listed in Table 2 of Jerry L. Adams ET. AL.

In some embodiments, an immuno-oncology agent is selected from the small molecule immuno-oncology agents described in Peter L. Toogood, “Small molecule immuno-oncology therapeutic agents,” Bioorganic & Medicinal Chemistry Letters 2018, Vol. 28, pages 319-329, the content of which is incorporated herein by reference in its entirety. In some embodiments, an immuno-oncology agent is an agent targeting the pathways as described in Peter L. Toogood.

In some embodiments, an immuno-oncology agent is selected from those described in Sandra L. Ross et al., “Bispecific T cell engager (BiTE®) antibody constructs can mediate bystander tumor cell killing”, PLoS ONE 12(8): e0183390, the content of which is incorporated herein by reference in its entirety. In some embodiments, an immuno-oncology agent is a bispecific T cell engager (BiTE®) antibody construct. In some embodiments, a bispecific T cell engager (BiTE®) antibody construct is a CD19/CD3 bispecific antibody construct. In some embodiments, a bispecific T cell engager (BiTE®) antibody construct is an EGFR/CD3 bispecific antibody construct. In some embodiments, a bispecific T cell engager (BiTE®) antibody construct activates T cells. In some embodiments, a bispecific T cell engager (BiTE®) antibody construct activates T cells, which release cytokines inducing upregulation of intercellular adhesion molecule 1 (ICAM-1) and FAS on bystander cells. In some embodiments, a bispecific T cell engager (BiTE®) antibody construct activates T cells which result in induced bystander cell lysis. In some embodiments, the bystander cells are in solid tumors. In some embodiments, the bystander cells being lysed are in proximity to the BiTE®-activated T cells. In some embodiments, the bystander cells comprise tumor-associated antigen (TAA) negative cancer cells. In some embodiments, the bystander cells comprise EGFR-negative cancer cells. In some embodiments, an immuno-oncology agent is an antibody which blocks the PD-L1/PD1 axis and/or CTLA4. In some embodiments, an immuno-oncology agent is an ex-vivo expanded tumor-infiltrating T cell. In some embodiments, an immuno-oncology agent is a bispecific antibody construct or chimeric antigen receptors (CARs) that directly connect T cells with tumor-associated surface antigens (TAAs).

Exemplary Immune Checkpoint Inhibitors

In some embodiments, an immuno-oncology agent is an immune checkpoint inhibitor as described herein.

The term “checkpoint inhibitor” as used herein relates to agents useful in preventing cancer cells from avoiding the immune system of the patient. One of the major mechanisms of anti-tumor immunity subversion is known as “T-cell exhaustion,” which results from chronic exposure to antigens that has led to up-regulation of inhibitory receptors. These inhibitory receptors serve as immune checkpoints in order to prevent uncontrolled immune reactions.

PD-1 and co-inhibitory receptors such as cytotoxic T-lymphocyte antigen 4 (CTLA-4, B and T Lymphocyte Attenuator (BTLA; CD272), T cell Immunoglobulin and Mucin domain-3 (Tim-3), Lymphocyte Activation Gene-3 (Lag-3; CD223), and others are often referred to as a checkpoint regulators. They act as molecular “gatekeepers” that allow extracellular information to dictate whether cell cycle progression and other intracellular signaling processes should proceed.

In some embodiments, an immune checkpoint inhibitor is an antibody to PD-1. PD-1 binds to the programmed cell death 1 receptor (PD-1) to prevent the receptor from binding to the inhibitory ligand PDL-1, thus overriding the ability of tumors to suppress the host anti-tumor immune response.

In one aspect, the checkpoint inhibitor is a biologic therapeutic or a small molecule. In another aspect, the checkpoint inhibitor is a monoclonal antibody, a humanized antibody, a fully human antibody, a fusion protein or a combination thereof. In a further aspect, the checkpoint inhibitor inhibits a checkpoint protein selected from CTLA-4, PDL1, PDL2, PDl, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK 1, CHK2, A2aR, B-7 family ligands or a combination thereof. In an additional aspect, the checkpoint inhibitor interacts with a ligand of a checkpoint protein selected from CTLA-4, PDL1, PDL2, PDl, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK 1, CHK2, A2aR, B-7 family ligands or a combination thereof. In an aspect, the checkpoint inhibitor is an immunostimulatory agent, a T cell growth factor, an interleukin, an antibody, a vaccine or a combination thereof. In a further aspect, the interleukin is IL-7 or IL-15. In a specific aspect, the interleukin is glycosylated IL-7. In an additional aspect, the vaccine is a dendritic cell (DC) vaccine.

Checkpoint inhibitors include any agent that blocks or inhibits in a statistically significant manner, the inhibitory pathways of the immune system. Such inhibitors may include small molecule inhibitors or may include antibodies, or antigen binding fragments thereof, that bind to and block or inhibit immune checkpoint receptors or antibodies that bind to and block or inhibit immune checkpoint receptor ligands. Illustrative checkpoint molecules that may be targeted for blocking or inhibition include, but are not limited to, CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, GAL9, LAG3, TIM3, VISTA, KIR, 2B4 (belongs to the CD2 family of molecules and is expressed on all NK, γδ, and memory CD8+(αβ) T cells), CD160 (also referred to as BY55), CGEN-15049, CHK 1 and CHK2 kinases, A2aR, and various B-7 family ligands. B7 family ligands include, but are not limited to, B7-1, B7-2, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, B7-H5, B7-H6 and B7-H7. Checkpoint inhibitors include antibodies, or antigen binding fragments thereof, other binding proteins, biologic therapeutics, or small molecules, that bind to and block or inhibit the activity of one or more of CTLA-4, PDL1, PDL2, PD1, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD 160 and CGEN-15049. Illustrative immune checkpoint inhibitors include Tremelimumab (CTLA-4 blocking antibody), anti-OX40, PD-L1 monoclonal Antibody (Anti-B7-H1; MEDI4736), MK-3475 (PD-1 blocker), Nivolumab (anti-PDl antibody), CT-011 (anti-PD1 antibody), BY55 monoclonal antibody, AMP224 (anti-PDL1 antibody), BMS-936559 (anti-PDL1 antibody), MPLDL3280A (anti-PDL1 antibody), MSB0010718C (anti-PDL1 antibody), and ipilimumab (anti-CTLA-4 checkpoint inhibitor). Checkpoint protein ligands include, but are not limited to PD-L1, PD-L2, B7-H3, B7-H4, CD28, CD86 and TIM-3.

In some embodiments, an immune checkpoint inhibitor is REGN2810 (Regeneron), an anti-PD-1 antibody tested in patients with basal cell carcinoma (NCT03132636); NSCLC (NCT03088540); cutaneous squamous cell carcinoma (NCT02760498); lymphoma (NCT02651662); and melanoma (NCT03002376); pidilizumab (CureTech), also known as CT-011, an antibody that binds to PD-1, in clinical trials for diffuse large B-cell lymphoma and multiple myeloma; avelumab (Bavencio®, Pfizer/Merck KGaA), also known as MSB0010718C), a fully human IgG1 anti-PD-L1 antibody, in clinical trials for non-small cell lung cancer, Merkel cell carcinoma, mesothelioma, solid tumors, renal cancer, ovarian cancer, bladder cancer, head and neck cancer, and gastric cancer; or PDR001 (Novartis), an inhibitory antibody that binds to PD-1, in clinical trials for non-small cell lung cancer, melanoma, triple negative breast cancer and advanced or metastatic solid tumors. Tremelimumab (CP-675,206; Astrazeneca) is a fully human monoclonal antibody against CTLA-4 that has been in studied in clinical trials for a number of indications, including: mesothelioma, colorectal cancer, kidney cancer, breast cancer, lung cancer and non-small cell lung cancer, pancreatic ductal adenocarcinoma, pancreatic cancer, germ cell cancer, squamous cell cancer of the head and neck, hepatocellular carcinoma, prostate cancer, endometrial cancer, metastatic cancer in the liver, liver cancer, large B-cell lymphoma, ovarian cancer, cervical cancer, metastatic anaplastic thyroid cancer, urothelial cancer, fallopian tube cancer, multiple myeloma, bladder cancer, soft tissue sarcoma, and melanoma. AGEN-1884 (Agenus) is an anti-CTLA4 antibody that is being studied in Phase 1 clinical trials for advanced solid tumors (NCT02694822).

In some embodiments, a checkpoint inhibitor is an inhibitor of T-cell immunoglobulin mucin containing protein-3 (TIM-3). TIM-3 inhibitors that may be used in the present invention include TSR-022, LY3321367 and MBG453. TSR-022 (Tesaro) is an anti-TIM-3 antibody which is being studied in solid tumors (NCT02817633). LY3321367 (Eli Lilly) is an anti-TIM-3 antibody which is being studied in solid tumors (NCT03099109). MBG453 (Novartis) is an anti-TIM-3 antibody which is being studied in advanced malignancies (NCT02608268).

In some embodiments, a checkpoint inhibitor is an inhibitor of T cell immunoreceptor with Ig and ITIM domains, or TIGIT, an immune receptor on certain T cells and NK cells. TIGIT inhibitors that may be used in the present invention include BMS-986207 (Bristol-Myers Squibb), an anti-TIGIT monoclonal antibody (NCT02913313); OMP-313M32 (Oncomed); and anti-TIGIT monoclonal antibody (NCT03119428).

In some embodiments, a checkpoint inhibitor is an inhibitor of Lymphocyte Activation Gene-3 (LAG-3). LAG-3 inhibitors that may be used in the present invention include BMS-986016 and REGN3767 and IMP321. BMS-986016 (Bristol-Myers Squibb), an anti-LAG-3 antibody, is being studied in glioblastoma and gliosarcoma (NCT02658981). REGN3767 (Regeneron), is also an anti-LAG-3 antibody, and is being studied in malignancies (NCT03005782). IP321 (Immutep S.A.) is an LAG-3-Ig fusion protein, being studied in melanoma (NCT02676869); adenocarcinoma (NCT02614833); and metastatic breast cancer (NCT00349934).

Checkpoint inhibitors that may be used in the present invention include OX40 agonists. OX40 agonists that are being studied in clinical trials include PF-04518600/PF-8600 (Pfizer), an agonistic anti-OX40 antibody, in metastatic kidney cancer (NCT03092856) and advanced cancers and neoplasms (NCT02554812; NCT05082566); GSK3174998 (Merck), an agonistic anti-OX40 antibody, in Phase 1 cancer trials (NCT02528357); MEDI0562 (Medimmune/AstraZeneca), an agonistic anti-OX40 antibody, in advanced solid tumors (NCT02318394 and NCT02705482); MEDI6469, an agonistic anti-OX40 antibody (Medimmune/AstraZeneca), in patients with colorectal cancer (NCT02559024), breast cancer (NCT01862900), head and neck cancer (NCT02274155) and metastatic prostate cancer (NCT01303705); and BMS-986178 (Bristol-Myers Squibb) an agonistic anti-OX40 antibody, in advanced cancers (NCT02737475).

Checkpoint inhibitors that may be used in the present invention include CD137 (also called 4-1BB) agonists. CD137 agonists that are being studied in clinical trials include utomilumab (PF-05082566, Pfizer) an agonistic anti-CD137 antibody, in diffuse large B-cell lymphoma (NCT02951156) and in advanced cancers and neoplasms (NCT02554812 and NCT05082566); urelumab (BMS-663513, Bristol-Myers Squibb), an agonistic anti-CD137 antibody, in melanoma and skin cancer (NCT02652455) and glioblastoma and gliosarcoma (NCT02658981).

Checkpoint inhibitors that may be used in the present invention include CD27 agonists. CD27 agonists that are being studied in clinical trials include varlilumab (CDX-1127, Celldex Therapeutics) an agonistic anti-CD27 antibody, in squamous cell head and neck cancer, ovarian carcinoma, colorectal cancer, renal cell cancer, and glioblastoma (NCT02335918); lymphomas (NCT01460134); and glioma and astrocytoma (NCT02924038).

Checkpoint inhibitors that may be used in the present invention include glucocorticoid-induced tumor necrosis factor receptor (GITR) agonists. GITR agonists that are being studied in clinical trials include TRX518 (Leap Therapeutics), an agonistic anti-GITR antibody, in malignant melanoma and other malignant solid tumors (NCT01239134 and NCT02628574); GWN323 (Novartis), an agonistic anti-GITR antibody, in solid tumors and lymphoma (NCT02740270); INCAGN01876 (Incyte/Agenus), an agonistic anti-GITR antibody, in advanced cancers (NCT02697591 and NCT03126110); MK-4166 (Merck), an agonistic anti-GITR antibody, in solid tumors (NCT02132754) and MEDI1873 (Medimmune/AstraZeneca), an agonistic hexameric GITR-ligand molecule with a human IgG1 Fc domain, in advanced solid tumors (NCT02583165).

Checkpoint inhibitors that may be used in the present invention include inducible T-cell co-stimulator (ICOS, also known as CD278) agonists. ICOS agonists that are being studied in clinical trials include MEDI-570 (Medimmune), an agonistic anti-ICOS antibody, in lymphomas (NCT02520791); GSK3359609 (Merck), an agonistic anti-ICOS antibody, in Phase 1 (NCT02723955); JTX-2011 (Jounce Therapeutics), an agonistic anti-ICOS antibody, in Phase 1 (NCT02904226).

Checkpoint inhibitors that may be used in the present invention include killer IgG-like receptor (KIR) inhibitors. KIR inhibitors that are being studied in clinical trials include lirilumab (IPH2102/BMS-986015, Innate Pharma/Bristol-Myers Squibb), an anti-KIR antibody, in leukemias (NCT01687387, NCT02399917, NCT02481297, NCT02599649), multiple myeloma (NCT02252263), and lymphoma (NCT01592370); IPH2101 (1-7F9, Innate Pharma) in myeloma (NCT01222286 and NCT01217203); and IPH4102 (Innate Pharma), an anti-KIR antibody that binds to three domains of the long cytoplasmic tail (KIR3DL2), in lymphoma (NCT02593045).

Checkpoint inhibitors that may be used in the present invention include CD47 inhibitors of interaction between CD47 and signal regulatory protein alpha (SIRPa). CD47/SIRPa inhibitors that are being studied in clinical trials include ALX-148 (Alexo Therapeutics), an antagonistic variant of (SIRPa) that binds to CD47 and prevents CD47/SIRPa-mediated signaling, in phase 1 (NCT03013218); TTI-621 (SIRPa-Fc,TrilliumTherapeutics), a soluble recombinant fusion protein created by linking the N-terminal CD47-binding domain of SIRPa with the Fc domain of human IgG1, acts by binding human CD47, and preventing it from delivering its “do not eat” signal to macrophages, is in clinical trials in Phase 1 (NCT02890368 and NCT02663518); CC-90002 (Celgene), an anti-CD47 antibody, in leukemias (NCT02641002); and Hu5F9-G4 (Forty Seven, Inc.), in colorectal neoplasms and solid tumors (NCT02953782), acute myeloid leukemia (NCT02678338) and lymphoma (NCT02953509).

Checkpoint inhibitors that may be used in the present invention include CD73 inhibitors. CD73 inhibitors that are being studied in clinical trials include MEDI9447 (Medimmune), an anti-CD73 antibody, in solid tumors (NCT02503774); and BMS-986179 (Bristol-Myers Squibb), an anti-CD73 antibody, in solid tumors (NCT02754141).

Checkpoint inhibitors that may be used in the present invention include agonists of stimulator of interferon genes protein (STING, also known as transmembrane protein 173, or TMEM173). Agonists of STING that are being studied in clinical trials include MK-1454 (Merck), an agonistic synthetic cyclic dinucleotide, in lymphoma (NCT03010176); and ADU-S100 (MIW815, Aduro Biotech/Novartis), an agonistic synthetic cyclic dinucleotide, in Phase 1 (NCT02675439 and NCT03172936).

Checkpoint inhibitors that may be used in the present invention include CSF1R inhibitors. CSF1R inhibitors that are being studied in clinical trials include pexidartinib (PLX3397, Plexxikon), a CSF1R small molecule inhibitor, in colorectal cancer, pancreatic cancer, metastatic and advanced cancers (NCT02777710) and melanoma, non-small cell lung cancer, squamous cell head and neck cancer, gastrointestinal stromal tumor (GIST) and ovarian cancer (NCT02452424); and IMC-CS4 (LY3022855, Lilly), an anti-CSF-1R antibody, in pancreatic cancer (NCT03153410), melanoma (NCT03101254), and solid tumors (NCT02718911); and BLZ945 (4-[2((1R,2R)-2-hydroxycyclohexylamino)-benzothiazol-6-yloxyl]-pyridine-2-carboxylic acid methylamide, Novartis), an orally available inhibitor of CSF1R, in advanced solid tumors (NCT02829723).

Checkpoint inhibitors that may be used in the present invention include NKG2A receptor inhibitors. NKG2A receptor inhibitors that are being studied in clinical trials include monalizumab (IPH2201, Innate Pharma), an anti-NKG2A antibody, in head and neck neoplasms (NCT02643550) and chronic lymphocytic leukemia (NCT02557516).

In some embodiments, the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, ipilimumab, avelumab, durvalumab, atezolizumab, or pidilizumab.

Compounds and compositions described herein are generally useful for the inhibition of eIF4E or a mutant thereof.

The activity of a compound utilized in this invention as an inhibitor of eIF4E, or a mutant thereof, may be assayed in vitro, in vivo or in a cell line. In vitro assays include assays that determine inhibition of eIF4E, or a mutant thereof. Alternate in vitro assays quantitate the ability of the inhibitor to bind to eIF4E. Detailed conditions for assaying a compound utilized in this invention as an inhibitor of eIF4E, or a mutant thereof, are set forth in the Examples below.

Provided compounds are inhibitors of eIF4E and are therefore useful for treating one or more disorders associated with activity of eIF4E. Thus, in certain embodiments, the present invention provides a method for treating an eIF4E-mediated disorder comprising the step of administering to a patient in need thereof a compound of the present invention, or pharmaceutically acceptable composition thereof.

As used herein, the terms “eIF4E-mediated” disorders, diseases, and/or conditions as used herein means any disease or other deleterious condition in which eIF4E, or a mutant thereof, is known to play a role, including, but is not limited to, a cellular proliferative disorder. In some embodiments, a cellular proliferative disorder is cancer as described herein.

Cancer

In some embodiments, the cancer is acoustic neuroma, astrocytoma (e.g. Grade I—Pilocytic Astrocytoma, Grade II—Low-grade Astrocytoma, Grade III—Anaplastic Astrocytoma, or Grade IV—Glioblastoma (GBM)), chordoma, CNS lymphoma, craniopharyngioma, brain stem glioma, ependymoma, mixed glioma, optic nerve glioma, subependymoma, medulloblastoma, meningioma, metastatic brain tumor, oligodendroglioma, pituitary tumors, primitive neuroectodermal (PNET) tumor, or schwannoma. In some embodiments, the cancer is a type found more commonly in children than adults, such as brain stem glioma, craniopharyngioma, ependymoma, juvenile pilocytic astrocytoma (JPA), medulloblastoma, optic nerve glioma, pineal tumor, primitive neuroectodermal tumors (PNET), or rhabdoid tumor. In some embodiments, the patient is an adult human. In some embodiments, the patient is a child or pediatric patient.

Cancer includes, in another embodiment, without limitation, mesothelioma, hepatobilliary (hepatic and billiary duct), bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular melanoma, ovarian cancer, colon cancer, rectal cancer, cancer of the anal region, stomach cancer, gastrointestinal (gastric, colorectal, and duodenal), uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, prostate cancer, testicular cancer, chronic or acute leukemia, chronic myeloid leukemia, lymphocytic lymphomas, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, non-Hodgkins's lymphoma, spinal axis tumors, brain stem glioma, pituitary adenoma, adrenocortical cancer, gall bladder cancer, multiple myeloma, cholangiocarcinoma, fibrosarcoma, neuroblastoma, retinoblastoma, or a combination of one or more of the foregoing cancers.

In some embodiments, the cancer is a solid tumor, such as a sarcoma, carcinoma, or lymphoma. Solid tumors generally comprise an abnormal mass of tissue that typically does not include cysts or liquid areas. In some embodiments, the cancer is selected from renal cell carcinoma, or kidney cancer; hepatocellular carcinoma (HCC) or hepatoblastoma, or liver cancer; melanoma; breast cancer; colorectal carcinoma, or colorectal cancer; colon cancer; rectal cancer; anal cancer; lung cancer, such as non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC); ovarian cancer, ovarian epithelial cancer, ovarian carcinoma, or fallopian tube cancer; papillary serous cystadenocarcinoma or uterine papillary serous carcinoma (UPSC); prostate cancer; testicular cancer; gallbladder cancer; hepatocholangiocarcinoma; soft tissue and bone synovial sarcoma; rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewing sarcoma; anaplastic thyroid cancer; adrenocortical carcinoma; pancreatic cancer; pancreatic ductal carcinoma or pancreatic adenocarcinoma; gastrointestinal/stomach (GIST) cancer; lymphoma; squamous cell carcinoma of the head and neck (SCCHN); salivary gland cancer; glioma, or brain cancer; neurofibromatosis-1 associated malignant peripheral nerve sheath tumors (MPNST); Waldenstrom's macroglobulinemia; or medulloblastoma.

In some embodiments, the cancer is hepatocellular carcinoma (HCC). In some embodiments, the cancer is hepatoblastoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is rectal cancer. In some embodiments, the cancer is ovarian cancer, or ovarian carcinoma. In some embodiments, the cancer is ovarian epithelial cancer. In some embodiments, the cancer is fallopian tube cancer. In some embodiments, the cancer is papillary serous cystadenocarcinoma. In some embodiments, the cancer is uterine papillary serous carcinoma (UPSC). In some embodiments, the cancer is hepatocholangiocarcinoma. In some embodiments, the cancer is soft tissue and bone synovial sarcoma. In some embodiments, the cancer is rhabdomyosarcoma. In some embodiments, the cancer is osteosarcoma. In some embodiments, the cancer is anaplastic thyroid cancer. In some embodiments, the cancer is adrenocortical carcinoma. In some embodiments, the cancer is pancreatic cancer, or pancreatic ductal carcinoma. In some embodiments, the cancer is pancreatic adenocarcinoma. In some embodiments, the cancer is glioma. In some embodiments, the cancer is malignant peripheral nerve sheath tumors (MPNST). In some embodiments, the cancer is neurofibromatosis-1 associated MPNST. In some embodiments, the cancer is Waldenstrom's macroglobulinemia. In some embodiments, the cancer is medulloblastoma.

In some embodiments, the cancer is small cell lung cancer, non-small cell lung cancer, colorectal cancer, multiple myeloma, acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), pancreatic cancer, liver cancer, hepatocellular cancer, neuroblastoma, other solid tumors or other hematological cancers.

In some embodiments, the cancer is small cell lung cancer, non-small cell lung cancer, colorectal cancer, multiple myeloma, or AML.

The present invention further features methods and compositions for the diagnosis, prognosis and treatment of viral-associated cancers, including human immunodeficiency virus (HIV) associated solid tumors, human papilloma virus (HPV)-16 positive incurable solid tumors, and adult T-cell leukemia, which is caused by human T-cell leukemia virus type I (HTLV-I) and is a highly aggressive form of CD4+ T-cell leukemia characterized by clonal integration of HTLV-I in leukemic cells (See https://clinicaltrials.gov/ct2/show/study/NCT02631746); as well as virus-associated tumors in gastric cancer, nasopharyngeal carcinoma, cervical cancer, vaginal cancer, vulvar cancer, squamous cell carcinoma of the head and neck, and Merkel cell carcinoma. (See https://clinicaltrials.gov/ct2/show/study/NCT02488759; see also https://clinicaltrials.gov/ct2/show/study/NCT0240886; https://clinicaltrials.gov/ct2/show/NCT02426892)

In some embodiments, the present invention provides a method for treating a tumor in a patient in need thereof, comprising administering to the patient compound II, or a pharmaceutical salt or composition thereof, and an immuno-oncology agent as described herein. In some embodiments, the tumor comprises any of the cancers described herein. In some embodiments, the tumor comprises melanoma cancer. In some embodiments, the tumor comprises breast cancer. In some embodiments, the tumor comprises lung cancer. In some embodiments the tumor comprises small cell lung cancer (SCLC). In some embodiments, the tumor comprises non-small cell lung cancer (NSCLC).

In some embodiments, the tumor is treated by arresting further growth of the tumor. In some embodiments, the tumor is treated by reducing the size (e.g., volume or mass) of the tumor by at least 5%, 10%, 25%, 50%, 75%, 90% or 99% relative to the size of the tumor prior to treatment. In some embodiments, tumors are treated by reducing the quantity of the tumors in the patient by at least 5%, 10%, 25%, 50%, 75%, 90% or 99% relative to the quantity of tumors prior to treatment.

The following examples are provided for illustrative purposes only and are not to be construed as limiting this invention in any manner.

All manipulations were carried out in a glass reaction tube equipped with a magnetic stir bar under a nitrogen atmosphere. Unless otherwise mentioned, solvents and reagents were purchased from commercial sources and used as received. The reaction was monitored by thin-layer chromatography carried out on silica gel plates and visualized under UV light (254 nm). Analytical LC-MS was performed on an Agilent 1200 infinity LC-MS system coupled with Agilent 6110 quadrupole mass spectrometer and a XBRIGE C18chromatography (4.6×50 mm, 3.5 um). The analysis was conducted at 2.0 mL/min flow rate using a linear gradient of 5 to 95% of B over 1.3 min, temperature: 50° C., (Buffer A: 0.01% TFA-containing H2O; Buffer B: 0.01% TFA-containing acetonitrile) or 1.8 mL/min flow rate using a linear gradient of 10 to 95% of B over 1.5 min, temperature: 50° C. (Buffer A: 10 mmol NH4HCO3-containing H2O; Buffer B: acetonitrile). The NMR spectra were recorded in CDCl3or DMSO-d6on a Bruker 400 MHz instrument with tetramethylsilane (TMS) as the internal standard. NMR data are represented as follows: chemical shift (ppm), multiplicity (s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet), coupling constants in hertz (Hz), and integration.

ABBREVIATIONS

Example 1. Synthesis of Certain Exemplary Compounds

Synthesis of 1.1

Synthesis of 1.2

To a mixture of intermediate 1.1 (43.2 g, 231.0 mmol) and TEA (70.0 g, 693.0 mmol) in DCM (1.2 L) with stirring at 0° C. under N2was added dropwise TMSOTf (102.6 g, 462.0 mmol). The mixture was allowed to warm to rt and stirred for 16 h, poured into water (500 mL), extracted with DCM (3×500 mL), dried over Na2SO4, and concentrated to afford a yellow oil. The oil was dissolved in THE (500 mL) followed by addition of NBS (49.3 g, 277.2 mmol) and reaction stirred at rt for 4 h. The mixture was concentrated and purified by flash silica gel chromatography (DCM 100%) to give intermediate 1.2 as a yellow solid (45.9 g, Y: 77%). ESI-MS (M+H)+: 266.0.

Synthesis of 1.3

A solution of intermediate 1.2 (45.9 g, 173.2 mmol) and sodium thiocyanate (16.8 g, 207.8 mmol) in EtOH (340 mL) was stirred at rt for 16 h. After cooling to rt, ice-water was added. The solid was collected by filtration and dried to give intermediate 1.3 (36 g, Y: 85%) as a yellow solid. ESI-MS (M+H)+: 245.0.

Synthesis of 1.4

To a solution of intermediate 1.3 (2.1 g, 8.6 mmol) and 2-pyrrolidineacetic acid, hydrochloride (2.1 g, 12.9 mmol) in t-BuOH (50 mL) was added AcOH (5 mL). The mixture was stirred at 100° C. for 2 d. The mixture was concentrated and dried in vacuo to give intermediate 1.4 (4.2 g) as crude yellow oil. The oil was used in the next step without any purification. ESI-MS (M+H)+: 356.1.

To a solution of intermediate 1.4 (4.2 g, 8.6 mmol) in MeOH (100 mL) was added conc. H2SO4(1 mL) and stirred at reflux for 16 h. The mixture was then concentrated and purified by silica gel chromatography (PE/EA=5:1) to give intermediate 1.5 (600 mg). To a solution of intermediate 1.5 (600 mg, 1.6 mmol) in THE (60 mL) was added NBS (202 mg, 1.1 mmol). The resulting mixture was stirred at rt for 20 min. The mixture was concentrated and purified by silica gel chromatography (PE/EA=10:1) to give intermediate 1.6 (500 mg, Y: 13%, two steps) as a greenish solid. ESI-MS (M+H)+: 448.0.

Synthesis of I-36-I-39 General Procedure for Suzuki Coupling

To a solution of intermediate 1.6 (1.0 eq) and boronic acids (1.5 eq) in dioxane/H2O (10:1) was added Pd(dppf)Cl2(0.1 eq) and K3PO4(3 eq). The mixture was stirred at 80° C. for 16 h under N2atmosphere. The solvent was removed and residue dissolved in MeOH (10 mL). LiOH (10 eq) was added and the mixture stirred at rt for 16 h. The mixture was concentrated and residue purified by preparative-RPHPLC (MeCN/H2O with 0.05% TFA as a mobile phase) to give the desired compounds I-36, I-37, I-38 and I-39.

Synthesis of 2.1

Synthesis of 2.2

To a solution of intermediate 2.1 (14 g, 65.4 mmol), CuBr2(29.2 g, 130 mmol) and TsOH (1.23 g, 6.5 mmol) in EA (200 mL) was stirred at 80° C. for 16 h. The mixture was cooled to rt, filtered, and the precipitate washed with EA (200 mL), and the filtrate was concentrated to dryness. The crude product was purified by silica gel chromatography (PE/EA=200/1) to give compound 2.2 as a yellow solid (7 g, Y: 36%). ESI-MS (M+H)+: 293.1.

Synthesis of 2.3

A solution of 2.2 (7 g, 23.1 mmol) and sodium thiocyanate (9.3 g, 66.4 mmol) in EtOH (60 mL) was stirred at 30° C. for 16 h. After cooling to rt, ice-water was added. The solid was collected and dried to give the compound 2.3 (5 g, Y: 78%) as a yellow solid. ESI-MS (M+H)+: 272.1.

Synthesis of 2.4

To a solution of compound 2.3 (1.7 g, 6.3 mmol) and ethyl 2-(aminomethyl) pentanoate (1.2 g, 7.5 mmol) in t-BuOH (50 mL) was added AcOH (5 mL). The mixture was stirred at 70° C. for 16 h. The solvent was removed. The residue was purified by silica gel chromatography (PE/EA=10/1) to give the desired compound 2.4 (1.7 g, Y: 63%) as a yellow solid. ESI-MS (M+H)+: 413.1.

Synthesis of 2.5

To a solution of compound 2.4 (1.7 g, 4.1 mmol) in AcOH (50 mL) was added bromine (650 mg, 4.1 mmol) portion wise. The resulting mixture was stirred at rt for 5 min. The reaction mixture was quenched with water (100 mL), adjusted to pH 6-7 with NaHCO3(sat.), extracted with EA (3×100 mL). The combined organic layer was washed with brine (100 mL), dried and concentrated. The crude was purified through silica gel chromatography (PE/EA=10/1) to give the desired compound 2.5 (1.2 g, Y: 60%) as a greenish solid. ESI-MS (M+H)+: 491.1.

Synthesis of I-87

To a solution of compound 2.4 (230 mg, 0.56 mmol) in MeOH (10 mL) was added LiOH (134 mg, 5.58 mmol) and water (1 mL). The mixture was stirred at 70° C. for 2 h. The solvent was removed. The residue was dissolved in water (10 mL), adjusted to pH 5-6 with HCl (1N), extracted with DCM (3×10 mL). The combined organic layer was dried and concentrated. The crude was purified by prep-RPHPLC (MeCN/H2O with 0.05% TFA as a mobile phase) to give the desired compound 1-87 (180 mg, Y: 84%) as a white solid. ESI-MS (M+H)+: 385.1.

Synthesis of I-88

To a solution of compound, 1-87 (100 mg, 0.26 mmol) in AcOH (5 mL) was added bromine (40 mg, 0.26 mmol) portion wise. The resulting mixture was stirred at rt for 5 min. The reaction mixture was quenched with water (10 mL), extracted with DCM (100 mL×3). The combined organic layer was dried and concentrated. The crude was purified by prep-HPLC (MeCN/H2O with 0.05% TFA as a mobile phase) to give the desired compound 1-88 (40 mg, Y: 33%) as a white solid. ESI-MS (M+H)+: 464.1.

Synthesis of I-89-I-92 General Procedure for Suzuki Coupling

To a solution of compound, 1-88 (1 eq) and aryl-boronic acid (1.5 eq) in dioxane/H2O (5/1) was added Pd(dppf)Cl2(0.1 eq) and K3PO4(3 eq). The mixture was stirred at 80° C. for 16 h under N2protection. The solvent was removed. The residue was dissolved in MeOH (10 mL), added LiOH (10 eq). The mixture was stirred at 70° C. for 2 h. The solvent was removed. The residue was dissolved in water (10 mL), adjusted pH to 5-6 with HCl (1N), extracted with DCM (10 mL×3). The combined organic layer was dried and concentrated. The crude was purified by prep-HPLC (MeCN/H2O with 0.05 FA as a mobile phase) to give the desired compounds 1-89, I-90, I-91 and I-92.

Synthesis of 3.1

A solution of 1-(4-phenoxyphenyl) ethan-1-one (2.0 g, 9.0 mmol), CuBr2(2.1 g, 9.1 mmol) and TsOH (162 mg, 0.9 mmol) in EA (20 mL) was stirred at 80° C. for 16 h. T mixture was cooled to rt and filtered. The precipitate was washed with EA (200 mL). The filtrate was concentrated and. the crude product was purified by silica gel chromatography (PE/EA=200/1) to give compound 3.1 as a yellow solid (1.2 g, Y: 44%). ESI-MS (M+H)+: 291.1.

Synthesis of 3.2

A solution of 3.1 (1.2 g, 4.1 mmol) and sodium thiocyanate (402 mg, 4.96 mmol) in EtOH (15 mL) was stirred at 30° C. for 16 h. After cooling to rt, ice-water was added. The solid was collected and dried to give the compound 3.2 (1.0 g, Y: 90%) as a yellow solid. ESI-MS (M+H)+: 270.1.

Synthesis of 3.3

To a solution of compound 3.2 (2.5 g, 9.2 mmol) and amine (1.54 g, 9.2 mmol) in EtOH (120 mL) was added NaHCO3(1.56 g, 18.6 mmol). The mixture was stirred at 50° C. for 16 h. The solvent was removed and the residue purified by silica gel chromatography (PE/EA=10/1) to give the desired compound 3.3 (1.7 g, Y: 48%) as a yellow solid. ESI-MS (M+H)+: 381.1.

Synthesis of 3.4

To a solution of compound 3.3 (1.7 g, 4.5 mmol) in THE (85 mL) was added NBS (396 mg, 2.2 mmol) portion wise. The resulting mixture was stirred at rt for 20 min. The reaction mixture was concentrated to give the crude, which was purified by silica gel chromatography (PE/EA=10/1) to give the desired compound 3.4 (1.14 g, Y: 57%) as a yellow solid. ESI-MS (M+H)+: 459.1.

Synthesis of I-196-I-199

To a solution of compound 3.4 (285 mg, 1.0 eq) and aryl-boronic acid (1.5 eq) in dioxane/H2O (10/1) was added Pd(dppf)Cl2(0.1 eq) and K3PO4(3 eq). The mixture was stirred at 80° C. for 16 h under N2atmosphere. The solvent was removed. The residue was dissolved in MeOH/H2O (5:1; 5 mL), and LiOH (5.0 eq) added. The mixture was stirred at rt for 16 h. The solvent was removed. The residue was dissolved in water (10 mL), pH adjusted to 5-6 with aq. HCl (1N), extracted with EA (10 mL×3). The combined organic layer was dried and concentrated. The crude was purified by prep-RPHPLC (MeCN/H2O with 0.05% TFA as mobile phase) to give the desired compounds I-196, I-197, I-198 & I-199.

1.2 Synthesis of Compounds I-229 to I-240

General information: All evaporations were carried out in vacuo with a rotary evaporator. Analytical samples were dried in vacuo (1-5 mmHg) at rt. Thin layer chromatography (TLC) was performed on silica gel plates, spots were visualized by UV light (214 and 254 nm). Purification by column and flash chromatography was carried out using silica gel (200-300 mesh). Solvent systems are reported as mixtures by volume. All NMR spectra were recorded on a Bruker 400 (400 MHz) spectrometer. 1H chemical shifts are reported in 6 values in ppm with the deuterated solvent as the internal standard. Data are reported as follows: chemical shift, multiplicity (s=singlet, d=doublet, t=triplet, q=quartet, br=broad, m=multiplet), coupling constant (Hz), integration.

LCMS spectra were obtained on an Agilent 1200 series 6110 or 6120 mass spectrometer with electrospray ionization and excepted as otherwise indicated, the general LCMS condition was as follows:

Synthetic Scheme of Compound I-240

Synthesis of 240-01

To a solution of 2-bromo-1-phenylethanone (240-0, 1.98 g, 10.0 mmol) in EtOH (40 mL) was added sodium thiocyanate (0.82 g, 10.0 mmol), then the mixture was stirred at 80° C. overnight. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to give 1-phenyl-2-thiocyanatoethanone (240-01, 1.2 g, 68% yield) as yellow solid. MS Calcd.: 177.0; MS Found: 178.2 [M+H]+.

Synthesis of 240-02

To a solution of 1-phenyl-2-thiocyanatoethanone (240-01, 354.0 mg, 2.0 mmol) in EtOH (10 mL) was added methyl morpholine-2-carboxylate (292.5 mg, 2.0 mmol), then the mixture was stirred at 60° C. overnight. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to give 240-02: methyl 4-(4-phenylthiazol-2-yl)morpholine-2-carboxylate (400.5 mg, 66% yield) as yellow solid. MS Calcd.: 304.1; MS Found: 305.1 [M+H]+.

Synthesis of 240-03

To a solution of methyl 4-(4-phenylthiazol-2-yl) morpholine-2-carboxylate 240-02, (304.0 mg, 1.0 mmol) in DCM (10 mL) was added NBS (267.5 mg, 1.5 mmol), then the mixture was stirred at −60° C. for 0.3 hr. After completion of the reaction indicated by LCMS, the reaction mixture was quenched with saturated Na2S2O3(10 mL), diluted with water (10 mL) and extracted with DCM (20 mL×3). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to give methyl 4-(5-bromo-4-phenylthiazol-2-yl) morpholine-2-carboxylate 240-03, (350.5 mg, 92% yield) as yellow solid. MS Calcd.: 382.0; MS Found: 383.0 [M+H].

Synthesis of 240-04

To a solution of methyl 4-(5-bromo-4-phenylthiazol-2-yl) morpholine-2-carboxylate 240-03 (191.0 mg, 0.5 mmol), 4′-fluorobiphenyl-4-ylboronic acid (324.2 mg, 1.5 mmol) and K3PO4(318.9 mg, 1.5 mmol) in dioxane (10 mL) was added 1,1′-Bis (di-t-butylphosphino)ferrocene palladium dichloride (38.2 mg, 20% w/w), then the mixture was stirred at 60° C. in microwave for 2 hours. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to give methyl 4-(5-(4′-fluorobiphenyl-4-yl)-4-phenylthiazol-2-yl)morpholine-2-carboxylate 240-04 (200.2 mg, 84% yield) as light-yellow solid. MS Calcd.: 474.1; MS Found: 475.0 [M+H]+.

Synthesis of I-240

To a solution of methyl 4-(5-(4′-fluorobiphenyl-4-yl)-4-phenylthiazol-2-yl) morpholine-2-carboxylate (240-04, 118.5 mg, 0.25 mmol) in THE (5 mL) and H2O (1 mL) was added LiOH H2O (21.5 mg, 0.50 mmol), then the mixture was stirred at room temperature for 0.5 hour. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was diluted with 0.05 M HCl solution (10 ml), filtered, and the solid was collected by filtration and washed with water to give 4-(5-(4′-fluorobiphenyl-4-yl)-4-phenylthiazol-2-yl) morpholine-2-carboxylic acid (1-240, 51.2 mg, 44% yield) as off-white solid. MS Calcd.: 460.1; MS Found: 461.2 [M+H]*.

Synthetic Scheme of I-239

Synthesis of 239-01

To a solution of 239-0 (24 g, 121 mmol), 01-R (18.6 g, 133 mmol) and K2CO3(33.6 g, 242 mmol) in dioxane (400 mL) dioxane (400 mL) mixed with water (100 mL) was added Pd(dppf)Cl2(4.9 g, 6 mmol) under N2atmosphere, the reaction mixture was heated to 80° C. and stirred for 3 hr. After completion of the reaction indicated by LCMS, the reaction mixture was filtered and concentrated under reduced pressure to give the residue, which was dissolved in water (200 mL), and extracted with EA (2×100 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated to give the residue, which was purified by silica gel column chromatography to get 239-01 (20 g, 77.2% yield) as a yellow solid. MS Calcd.: 214.1; MS Found: 215.2 [M+H]+.

Synthesis of 239-02

To a solution of 239-01 (14 g, 65.4 mmol) in DCM (140 mL) was added PyBr3(20.8 g, 65.4 mmol), the reaction mixture was stirred at r.t overnight. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to get 239-02 (15 g, 78.5% yield) as a yellow solid. MS Calcd.: 292.0; MS Found: 293.0 [M+H]+.

Synthesis of 239-03

To a solution of 239-02 (10.4 g, 35.6 mmol) in EtOH (100 mL) was added NaSCN (2.9 g, 35.6 mmol), the reaction mixture was stirred at r.t for 4 hr. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to get 239-03 (7.5 g, 77.7% yield) as a yellow solid. MS Calcd.: 271.0; MS Found: 272.0 [M+H]+.

Synthesis of 239-04

To a solution of 239-03 (4 g, 14.7 mmol) in MeOH (250 mL) was added A (2.7 g, 14.7 mmol), the reaction mixture was heated to 60° C. and stirred overnight. After completion of the reaction indicated by LCMS, the reaction mixture was filtered, the solid was washed with MeOH (5 mL) to get 239-04 (3.4 g, 58.1% yield) as a yellow solid. MS Calcd.: 398.1; MS Found: 399.0 [M+H]+.

Synthesis of 239-05

To a solution of 239-04 (400 mg, 1 mmol) in MeCN (20 mL) mixed with water (5 mL) was added Na2S2O4(174 mg, 1 mmol), NaHCO3(84 mg, 1 mmol) and CF3I (1.96 g, 10 mmol), the reaction mixture was stirred at r.t overnight. After completion of the reaction indicated by LCMS, the reaction mixture was diluted with DCM and washed with water, dried over Na2SO4, filtered and concentrated to give the residue, which was purified by prep-HPLC to get 239-05 (190 g, 40.8% yield) as a white solid. MS Calcd.: 466.1; MS Found: 467.0 [M+H]+.

Synthesis of I-239

To a solution of 239-05 (100 mg, 0.21 mmol) in THE (5 mL) mixed with MeOH (1 mL) was added sat·LiOH (2 M, 1 mL), the reaction mixture was stirred at r.t for 1 hr. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, the mixture was adjust pH to 6 with 2 N HCl, the solid was collected by filtration to get the crude, which was purified by prep-HPLC to get I-239 (40 mg, 42.1% yield) as a white solid. MS Calcd.: 452.1; MS Found: 453.0 [M+H]+.

Synthetic Scheme of I-238

Synthesis of 238-01

To a solution 238-0 (1.0 g, 2.53 mmol) in CH2Cl2(20 mL) at −60° C. was added NBS (434 mg, 2.53 mmol). The mixture was stirred at −60° C. for 1 h and then quenched with saturated Na2S2O3(50 mL), diluted with water (50 mL). The resulting mixture was extracted with DCM (200 mL×3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered and concentrated. The crude product was purified by silica gel column chromatograph to give 238-01 (650 mg, yield: 50.0%) as a yellow solid. MS Calcd.: 474.0; MS Found: 475.0 [M+H]+.

Synthesis of 238-02

Synthesis of 1-238

To a solution of 238-02 (90 mg, 0.22 mmol) in THE (3 mL) mixed with MeOH (1 mL) was added sat·LiOH (2 M, 1 mL), the reaction mixture was stirred at r.t for 1 hr. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, the mixture was adjust pH to 6 with 2 N HCl, the solid was collected by filtration to get the crude, which was purified by prep-HPLC to get I-238 (44.6 mg, 28% yield) as a white solid. MS Calcd.: 396.1; MS Found: 397.0 [M+H]+.

Synthetic Scheme of I-237

Synthesis of 237-02

Synthesis of 237-03

To a solution of methyl 4-(4-methylthiazol-2-yl) morpholine-2-carboxylate 237-02 (242.2 mg, 1.0 mmol) in DCM (10 mL) was added NBS (176.2 mg, 0.99 mmol), then the mixture was stirred at −60° C. for 0.2 hr. After completion of the reaction indicated by LCMS, the reaction mixture was quenched with saturated Na2S2O3(10 mL), diluted with water (10 mL) and extracted with DCM (20 mL×3). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to give methyl 4-(5-bromo-4-methylthiazol-2-yl)morpholine-2-carboxylate 237-03 (140.2 mg, 44% yield) as light-yellow solid. MS Calcd.: 320.0; MS Found: 321.0 [M+H]+.

Synthesis of 237-04

To a solution of methyl 4-(5-bromo-4-methylthiazol-2-yl)morpholine-2-carboxylate 237-03 (140.2 mg, 0.44 mmol), 4′-fluorobiphenyl-4-ylboronic acid (280.8 mg, 1.3 mmol) and K3PO4(275.6 mg, 1.3 mmol) in dioxane (10 mL) was added 1,1′-Bis (di-t-butylphosphino)ferrocene palladium dichloride (28.0 mg, 20% w/w), then the mixture was stirred at 60° C. in microwave for 2 hours. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to give methyl 4-(5-(4′-fluorobiphenyl-4-yl)-4-methylthiazol-2-yl) morpholine-2-carboxylate 237-04, (120.0 mg, 66% yield) as light-yellow solid. MS Calcd.: 412.1; MS Found: 413.0 [M+H]+.

Synthesis of I-237

To a solution of methyl 4-(5-(4′-fluorobiphenyl-4-yl)-4-methylthiazol-2-yl) morpholine-2-carboxylate 237-04 (120.0 mg, 0.29 mmol) in THE (5 mL) and H2O (1 mL) was added LiOH·H2O (24.4 mg, 0.58 mmol), then the mixture was stirred at room temperature for 0.5 hour. After completion of the reaction indicated by LCMS, the reaction mixture was diluted with 0.05 M HCl solution (5 ml) and concentrated under reduced pressure to give the residue, which was purified by prep-HPLC to give 4-(5-(4′-fluorobiphenyl-4-yl)-4-methylthiazol-2-yl) morpholine-2-carboxylic acid 1-237 (26.5 mg, 23% yield) as a white solid. MS Calcd.: 398.1; MS Found: 399.1 [M+H]+.

Synthetic Scheme of I-236

Synthesis of 236-01

To a solution of methyl 4-(5-bromo-4-(4′-fluorobiphenyl-4-yl)thiazol-2-yl)morpholine-2-carboxylate 236-01 (238.0 mg, 0.5 mmol), potassium methyltrifluoroborate (183.0 mg, 1.5 mmol) and K3PO4(318.9 mg, 1.5 mmol) in dioxane (10 mL) was added 1,1′-Bis (di-t-butylphosphino)ferrocene palladium dichloride (47.6 mg, 20% w/w), then the mixture was stirred at 60° C. in microwave for 2 hours. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography and prep-HPLC to give methyl 4-(4-(4′-fluorobiphenyl-4-yl)-5-methylthiazol-2-yl)morpholine-2-carboxylate 236-02 (45.8 mg, 22% yield) as light-yellow solid. MS Calcd.: 412.1; MS Found: 413.0 [M+H]*.

Synthesis of I-236

To a solution of methyl 4-(4-(4′-fluorobiphenyl-4-yl)-5-methylthiazol-2-yl) morpholine-2-carboxylate 236-02 (45.8 mg, 0.11 mmol) in THF (5 mL) and H2O (1 mL) was added LiOH·H2O (10.0 mg, 0.22 mmol), then the mixture was stirred at room temperature for 0.3 hour. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was diluted with 0.05 M HCl solution (5 ml), filtered, and the solid was collected by filtration and washed with water to give 4-(4-(4′-fluorobiphenyl-4-yl)-5-methylthiazol-2-yl) morpholine-2-carboxylic acid I-236 (21.75 mg, 50% yield) as off-white solid. MS Calcd.: 398.1; MS Found: 413.2 [M+H]+.

Synthetic Scheme of I-235

Synthesis of 235-01

A solution of ethyl 4-chloro-4-oxobutanoate 235-0 (10.0 g, 6.1 mmol) was added dropwise to 28% aqueous ammonia solution (100 mL) under ice cooling, and then the mixture was allowed to warm to room temperature and was stirred at this temperature for 0.5 h, the mixture was concentrated in vacuo. The residue was diluted with THF and the mixture was filtered. The filtrate was dried over Na2SO4, concentrated in vacuo to give ethyl 4-amino-4-oxobutanoate 235-01 (4.9 g, 55.4% yield) as light-yellow oil.

Synthesis of 235-02

To a solution of ethyl 4-amino-4-oxobutanoate 235-01, (4.35 g, 30.0 mmol) in THF (150 mL) was added Phosphorus pentasulfide (5.73 g, 30.0 mmol), then the mixture was stirred at room temperature overnight. After completion of the reaction indicated by LCMS, the reaction mixture was filtered and concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to give ethyl 4-amino-4-thioxobutanoate 235-02 (1.63 g, 33.7% yield) as light-yellow oil. MS Calcd.: 161.05; MS Found: 162.2 [M+H]+.

Synthesis of 235-04

Synthesis of 235-05

To a solution of ethyl 3-(4-(4′-fluorobiphenyl-4-yl)thiazol-2-yl)propanoate 235-04 (500.0 mg, 1.41 mmol) in DMSO (7.5 mL) was added CF3I (827.9 mg, 4.22 mmol), then the mixture was degassed by N2. Then dppf (150.0 mg, 0.27 mmol) was added followed by 30% H2O2(3.4 mL). The above mixture was stirred at 45° C. for 1 h, then water (20 mL) was added. The resulting mixture was extracted with EtOAc (10 mL×3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered and concentrated. The crude product was purified by silica gel column chromatograph to give ethyl 3-(4-(4′-fluorobiphenyl-4-yl)-5-(trifluoromethyl)thiazol-2-yl)propanoate 235-05 (260.0 mg, yield: 43.6%) as a pale yellow solid. MS Calcd.: 423.09; MS Found: 424.0 [M+H]*.

Synthesis of I-235

To a solution of ethyl 3-(4-(4′-fluorobiphenyl-4-yl)-5-(trifluoromethyl)thiazol-2-yl)propanoate 235-05 (260.0 mg, 0.61 mmol) in EtOH (5 mL) and H2O (1 mL) was added LiOH·H2O (103.2 mg, 2.45 mmol), then the mixture was stirred at room temperature for 1 hour. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was diluted with 1 N HCl solution (5 ml), the resulting mixture was extracted with EtOAc (5 mL×3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC to give 3-(4-(4′-fluorobiphenyl-4-yl)-5-(trifluoromethyl)-thiazol-2-yl)-propanoic acid 1-235 (63.0 mg, yield: 26.0%) as white solid. MS Calcd.: 395.06; MS Found: 396.0 [M+H]+.

Synthetic Scheme of I-234

Synthesis of 234-01

To a solution of ethyl 3-(4-(4′-fluorobiphenyl-4-yl)thiazol-2-yl)propanoate 234-0 (1.0 g, 2.8 mmol) in DCM (50 mL) was added NBS (602.0 mg, 3.4 mmol). The mixture was stirred at room temperature overnight and then quenched with saturated Na2S2O3(5 mL), diluted with water (10 mL). The resulting mixture was extracted with DCM (20 mL×3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered and concentrated. The crude product was purified by silica gel column chromatograph to give ethyl 3-(5-bromo-4-(4′-fluorobiphenyl-4-yl)thiazol-2-yl)propanoate 234-01 (816.0 mg, yield: 66.9%) as a pale yellow solid. MS Calcd.: 433.01; MS Found: 433.8 [M+H]+.

Synthesis of 234-02

Synthesis of I-234

To a solution of ethyl 3-(4-(4′-fluorobiphenyl-4-yl)-5-phenylthiazol-2-yl)propanoate 234-02 (200.0 mg, 0.46 mmol) in EtOH (4 mL) and H2O (1 mL) was added LiOH H2O (77.8 mg, 1.86 mmol), then the mixture was stirred at room temperature for 1 hour. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was diluted with 1 N HCl solution (5 ml), the resulting mixture was extracted with EtOAc (5 mL×3). The combined organic layers were washed with brine (15 mL), dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC to give 3-(4-(4′-fluorobiphenyl-4-yl)-5-phenylthiazol-2-yl)propanoic acid I-234 (47.1 mg, yield: 25.1%) as off-white solid. MS Calcd.: 403.10; MS Found: 404.1 [M+H]+.

Synthetic Scheme of I-233

Synthesis of 233-01

To a solution of 233-0 (3.1 g, 11.4 mmol) in EtOH (250 mL) was added A (1.6 g, 14.7 mmol), the reaction mixture was heated to 85° C. overnight. After completion of the reaction indicated by LCMS, the solvent was removed in vacuo and purified by CC to obtain 233-01 (2.4 g, 53% yield) as a yellow solid. MS Calcd.: 396.1; MS Found: 397.1 [M+H]+.

Synthesis of 233-02

To a solution of 233-01 (340 mg, 0.86 mmol) in MeCN (20 mL) mixed with water (5 mL) was added Na2S2O4(149 mg, 0.86 mmol), NaHCO3(72 mg, 0.86 mmol) and CF3I (1.7 g, 8.6 mmol), the reaction mixture was stirred at r.t overnight. After completion of the reaction indicated by LCMS, the reaction mixture was diluted with DCM and washed with water, dried over Na2SO4, filtered and concentrated to give the residue, which was purified by prep-HPLC to get 233-02 (300 mg, 75% yield) as a white solid. MS Calcd.: 464.1; MS Found: 465.0 [M+H]+.

Synthesis of 1-233

To a solution of 233-02 (120 mg, 0.26 mmol) in THE (5 mL) mixed with MeOH (1 mL) was added sat·LiOH (2 M, 1 mL), the reaction mixture was stirred at r.t for 1 hr. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, the mixture was adjust pH to 6 with 2 N HCl, the solid was collected by filtration to get the crude, which was purified by prep-HPLC to get I-233 (33 mg, 28% yield) as a white solid. MS Calcd.: 450.1; MS Found: 451.1 [M+H]+.

Synthetic Scheme of I-232

Synthesis of 232-01

Synthesis of 1-232

To a solution of 232-01 (110 mg, 0.25 mmol) in THE (3 mL) mixed with MeOH (1 mL) was added LiOH (2 M, 1 mL), the reaction mixture was stirred at r.t for 1 hr. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, the mixture was adjusted pH to 6 with 2 N HCl, the solid was collected by filtration to get the crude, which was purified by prep-HPLC to get I-232 (40.1 mg, 38% yield) as a white solid. MS Calcd.: 422.1; MS Found: 423.1 [M+H]+.

Synthetic Scheme of I-231

Synthesis of 231-02

To a solution of 231-01 (1.00 g, 3.69 mmol) in EtOH (10 mL) was added methyl morpholine-3-carboxylate (0.54 g, 3.72 mmol), then the mixture was stirred at reflux overnight. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to give 231-02 (0.74 g, 50% yield) as a yellow solid. MS Calcd.: 398.1; MS Found: 399.2 [M+H]+.

Synthesis of 231-03

To a solution of 231-02 (430 mg, 1.08 mmol) in DCM (5 mL) was added NBS (150 mg, 2.64 mmol), then the mixture was stirred at r.t. for 5 h. After completion of the reaction indicated by LCMS, the reaction mixture was quenched with saturated Na2S2O3(5 mL), diluted with water (5 mL) and extracted with DCM (20 mL×3). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to give 231-03 (372 mg, 72% yield) as a yellow solid. MS Calcd.: 476.0; MS Found: 477.0 [M+H]+.

Synthesis of 231-04

To a solution of 231-03 (410 mg, 0.86 mmol), phenylboronic acid (315 mg, 2.58 mmol) and K3PO4(547 mg, 2.58 mmol) in dioxane (6 mL) was added 1,1′-Bis (di-t-butylphosphino)ferrocene palladium dichloride (70 mg, 0.09 mmol), then the mixture was stirred at 50° C. over night. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography to give 231-04 (316 mg, 77% yield) as a light-yellow solid. MS Calcd.: 474.1; MS Found: 475.1 [M+H]+.

Synthesis of I-231

To a solution of 231-04 (156 mg, 0.33 mmol) in THF/MeOH/H2O (4 mL/1 mL/1 mL) was added LiOH—H2O (24 mg, 1.00 mmol), then the mixture was stirred at room temperature for 3 hour. After completion of the reaction indicated by LCMS, the reaction mixture was concentrated under reduced pressure to give a residue which was acidify by 1N HCl to PH=6 and filtrated. The precipitate was purified by prep-HPLC (basic method) to give I-231 (49 mg, 32% yield) as a white solid. MS Calcd.: 460.1; MS Found: 461.2 [M+H]+.

Synthetic Scheme of I-230

Synthesis of 230-1

To a solution of 1-(4′-fluorobiphenyl-4-yl) propan-1-one (1.0 g, 4.38 mmol) in DCM (20 mL) was added Br2(0.77 g, 4.82 mmol) at 0° C. Then the reaction was warmed to room temperature and stirred for 3 hours. The reaction was quenched with NaHCO3solution (30 mL). The organic layer was separated and washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. Desired product was obtained as yellow solid (580 mg, Yield: 43.1%).

Synthesis of 230-2

To a solution of 2-bromo-1-(4′-fluorobiphenyl-4-yl) propan-1-one (580 mg, 1.89 mmol) in EtOH (20 mL) was added NaSCN (306 mg, 3.78 mmol). The reaction was heated to 90° C. and stirred at 90° C. for 3 hours. The solvent was concentrated. The solid was dissolved in EtOAc (50 mL). The mixture was washed with waster (40 mL). The organic layer was washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. Desired product was obtained as yellow solid (570 mg, Crude Yield: ˜100%).

Synthesis of 230-3

To a solution of 1-(4′-fluorobiphenyl-4-yl)-2-thiocyanatopropan-1-one (570 mg, 1.99 mmol) in EtOH (10 mL) was added ethyl 3-amino-2-methylpropanoate (524 mg, 3.99 mmol). The reaction was heated to 90° C. and stirred at 90° C. overnight. The solvent was concentrated. The crude product was purified by column chromatography. Desired product was obtained as yellow solid (77 mg, Yield: 9.7%).

Synthesis of I-230

To ethyl 3-(4-(4′-fluorobiphenyl-4-yl)-5-methylthiazol-2-ylamino)-2-methylpropanoate (77 mg, 0.19 mmol) was added MeOH (4 mL), THE (1 mL) and water (1 mL). The LiOH (9 mg, 0.38 mmol) was added into the reaction. The mixture was stirred at room temperature for 5 hours. The reaction was quenched by 1 N HCl solution (20 mL) and the mixture was stirred for 1 hour. The mixture was filtered. The wet cake was dried. Desired product was obtained as white solid (70 mg, Yield: 97.8%). MS Calcd.: 370.1; MS Found: 371.0 [M+H]+.

Synthetic Scheme of I-229

Synthesis of 229-1

1-(4-bromophenyl) ethanone (5.0 g, 25.12 mmol), 4-fluorophenylboronic acid (4.6 g, 32.66 mmol), K2CO3(6.9 g, 50.24 mmol) Pd (PPh3)4(0.5 g), dioxane (20 mL) and water (2 mL) was charged into a reactor protected with N2. The reaction was heated to 90° C. and stirred for 2 hours. The mixture was filtered via diatomite and washed cake with EtOAc (50 mL). The mixture was washed with water (20 mL). The organic layer was concentrated. The crude product was purified by column chromatography. Desired product was obtained as white solid (5.3 g, Yield: 98.5%).

Synthesis of 229-2

To a solution of 1-(4′-fluorobiphenyl-4-yl) ethanone (500 mg, 2.33 mmol) in EtOAc (20 mL) was added CuBr2(1.04 g, 4.67 mmol). The reaction was heated to reflux and stirred overnight. The mixture was cooled to room temperature. The mixture was filtered and washed cake with EtOAc (20 mL). The filtrate was washed with NH4Cl solution (30 mL*3) and brine (30 mL), dried over sodium sulfate, filtered and concentrated. Desired product was obtained as yellow solid (680 mg, Yield: 99.4%).

Synthesis of 229-3

To a solution of 2-bromo-1-(4′-fluorobiphenyl-4-yl) ethanone (680 mg, 2.32 mmol) in EtOH (20 mL) was added NaSCN (376 mg, 4.64 mmol). The reaction was heated to 90° C. and stirred at 90° C. for 3 hours. The solvent was concentrated. The solid was dissolved in EtOAc (50 mL). The mixture was washed with waster (40 mL). The organic layer was washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. Desired product was obtained as yellow solid (600 mg, Yield: 95.3%).

Synthesis of 229-4

Synthesis of 229-5

To a solution of 3-(4-(4′-fluorobiphenyl-4-yl) thiazol-2-ylamino)-2-methylpropanoic acid (1.3 g, 3.65 mmol) in MeOH (30 mL) was added SOCl2(0.87 g, 7.29 mmol) at 0° C. The mixture was stirred at 0° C. for 3 hours. The solvent was concentrated. The crude product was purified by column chromatography. Desired product was obtained as yellow solid (540 mg, Yield: 39.9%).

Synthesis of 229-6

Methyl 3-(4-(4′-fluorobiphenyl-4-yl) thiazol-2-ylamino)-2-methylpropanoate (540 mg, 1.46 mmol), Na2S2O4(507 mg, 2.91 mmol), NaHCO3(245 mg, 2.91 mmol), MeCN (10 mL) and water (5 mL) was charged into a pressure tank reactor. CF3I (about 5 g) was charged into the mixture. The reaction was stirred at room temperature overnight. The reaction was extracted with EtOAc (30 mL). The mixture was washed with waster (40 mL). The organic layer was washed with brine (30 mL), dried over sodium sulfate, filtered and concentrated. Desired product was obtained as yellow solid (400 mg, Yield: 62.6%).

Synthesis of I-229

To methyl 3-(4-(4′-fluorobiphenyl-4-yl)-5-(trifluoromethyl) thiazol-2-ylamino)-2-methylpropanoate (200 mg, 0.46 mmol) was added THE (2 mL) and water (2 mL). The LiOH (13 mg, 0.55 mmol) was added into the reaction. The mixture was stirred at room temperature until material disappeared in TLC (0.5-1 hour). The reaction was quenched by 1 N HCl solution (20 mL) and the mixture was extracted with EtOAc (20 mL). The solvent was concentrated. The crude product was purified by pre-HPLC. Desired product was obtained as orange red solid (25 mg, Yield: 12.9%). MS Calcd.: 424.1; MS Found: 424.9 [M+H]+.

Example 2. Compound Testing in Human eIF4E/4G2 Binding Assay

Human eIF4E (aa 28-217) with a C-terminal His-tag was expressed inE. coliin inclusion bodies. The protein was solubilized with 8 M urea and purified under denaturing conditions using nickel-charged HisTrap HP columns (GE Healthcare). The purified protein was then refolded by diluting in 20 mM Hepes pH 7.0, 0.5 M NaCl, 1 mM DTT, 1 mM EDTA, 0.5 M arginine plus 6 M urea, and then dialyzing overnight into the same buffer without the urea. The protein was further dialyzed into 20 mM Hepes, pH 6.5, 50 mM NaCl, 1 mM EDTA, 1 mM DTT, and concentrated using Hitrap SP sepharose FF columns (GE Healthcare). The concentrated protein was dialyzed into 20 mM Hepes, pH 7.0, 0.5M NaCl, 5 mM DTT and 10% glycerol, and stored at −80° C. until use.

Test compounds (3.43 mM stock in DMSO) were diluted 2-fold in series in DMSO (10 concentration points). Compound solutions (1.2 μl/well) were added into black 384-well polypropylene microplates (Matrix, Thermal Scientific). Twenty-two microliters per well of Assay Buffer (50 mM NaPi, pH 6.5, 50 mM KCl, 1 mM DTT and 0.5 mg/ml gamma globulin) and eight microliters per well of 82.5 nM purified eIF4E in Assay Buffer were added. The samples were incubated at room temperature (20-23° C.) for 15 min. Biotin labeled 4G2 peptide (Ac-Lys-Gln-Tyr-Asp-Arg-Glu-Phe-Leu-Leu-Asp-Phe-Gln-Phe-Met-Pro-Lys(Aha-Bio)-NH2, 1.75 μM stock in DMSO) was diluted to 0.14 μM in Assay Buffer (without DTT) and 5 μl/well was added. The samples were incubated at room temperature for 15 min. Five microliters per well of 6.4 nM Eu-streptavidin (Eu-SA, Perkin Elmer) and 80 nM Allophycocyanin (APC)-anti His antibody (Columbia Biosciences) in Assay Buffer (without DTT) were then added and the samples were incubated at room temperature for 20 min.

Assay signals were monitored by reading excitation at 340 nm and emission fluorescence at 615 nm and 665 nm on an Envision reader (Perkin Elmer). Normalized TR-FRET (time-resolved fluorescence resonance energy transfer) assay signal (Rn) was calculated by the formula:
Rn═[(A−Ba−C×D)/(D−Bd)]×(Dc−Bd)Where A is the fluorescence intensity of the sample at 665 nm,D is the fluorescence intensity of the sample at 615 nm,Ba and Bd are plate backgrounds at 665 nm and 615 nm, respectively,Dc is the fluorescence intensity of 0.78 nM Eu-SA in the assay buffer at 615 nmThe cross-talk factor (C) is determined by the following formula:
C=(Ac−Ba)/(Dc−Bd)Where Ac is the fluorescence intensity of 0.78 nM Eu-SA in the assay buffer at 665 nm.

IC50 values were calculated using xLFit program (IDBS). Table 2 below lists IC50 of some compounds, wherein A represents IC50<5 uM; B represents IC50 between 5-25 uM; and C represents IC50>25 uM.

TABLE 2IC50 of Certain Exemplary Compounds.Comp. No.IC50I-1CI-2CI-3CI-4CI-5CI-6CI-7CI-8CI-9CI-10CI-11CI-12CI-13CI-14CI-15CI-16CI-17CI-18BI-19CI-20CI-21CI-22CI-23CI-24CI-25CI-26CI-27CI-28CI-29CI-30CI-31CI-32CI-33CI-34CI-35CI-36CI-37CI-38CI-39CI-40CI-41CI-42CI-43CI-44CI-45CI-46CI-47CI-52CI-53CI-54CI-55CI-56CI-57CI-58CI-59CI-60CI-61CI-62CI-63CI-64BI-65BI-66AI-67CI-68CI-69BI-70CI-71BI-72AI-73CI-74BI-75BI-76CI-77BI-78CI-79CI-80BI-81CI-82BI-83AI-84CI-85CI-86BI-87CI-88BI-89AI-90CI-91CI-92BI-93CI-94BI-95BI-96CI-97CI-98BI-99CI-100BI-101BI-102CI-103CI-104BI-105BI-106AI-107CI-108BI-109AI-110CI-111AI-112CI-113CI-114AI-115CI-116AI-117CI-118CI-119BI-120CI-121BI-122AI-123CI-124CI-125AI-126CI-127BI-128AI-129CI-130CI-131BI-136AI-137CI-138BI-139BI-140AI-141CI-142CI-143AI-144AI-145CI-146BI-147AI-148CI-149BI-150CI-151CI-152BI-153CI-154AI-155CI-156CI-157BI-158AI-159CI-160CI-161BI-162CI-163BI-164CI-165CI-166BI-167CI-168AI-169CI-170CI-171BI-172BI-173CI-174CI-175CI-176CI-177BI-178CI-179CI-180CI-181CI-182AI-183CI-184CI-185AI-186AI-187CI-188CI-189BI-190CI-191AI-192CI-193CI-194BI-195CI-196AI-197CI-198CI-199BI-200CI-201AI-202CI-203CI-204BI-209AI-210CI-211CI-212BI-213AI-214CI-215CI-216CI-217AI-218CI-219BI-220AI-221AI-222AI-223AI-224AI-225AI-226AI-227AI-228AI-229AI-230BI-231AI-232AI-233AI-234AI-235BI-236BI-237BI-238BI-239BI-240A