EA040075B1 - PIPERIDINE SUBSTITUTE MNK INHIBITORS AND RELATED METHODS - Google Patents

Description

The technical field to which the invention belongs

The present invention generally relates to compounds having activity as MAP kinase (Mnk)-interacting kinase (Mnk) inhibitors, as well as related compositions and methods containing or using such compounds. Such compounds are useful in various therapeutic applications, including the treatment of cancer.

Prerequisites for the creation of the invention

Eukaryotic initiation factor 4E (eIF4E) is a general translation factor, but it has the potential to enhance predominantly the translation of messenger RNAs (mRNAs) that lead to the production of cancer-associated proteins. This selectivity may be related to the increased requirement for eIF4E and its binding partners for translation of mRNAs containing an extensive secondary structure in their 5' untranslated regions (5'UTRs). These mRNAs include those that encode certain proteins that control cell cycle progression and tumorigenesis. Under normal cellular conditions, translation of these cancer-associated mRNAs is suppressed because the availability of active eIF4E is limited; however, their levels may increase when eIF4E is overexpressed or hyperactivated. Elevated levels of eIF4E have been found in many tumor types and cancer cell lines, including colon, breast, bladder, lung, prostate, gastrointestinal, head and neck, Hodgkin's lymphoma, and neuroblastoma.

The initiation of cap-dependent translation is believed to depend on the assembly of eIF4F, a complex of initiation factors including eIF4E, the eIF4G scaffold protein, and the eIF4A RNA helicase. Since eIF4E is the only one of these proteins that directly binds to the mRNA cap structure, it is a key factor for the assembly of eIF4F at the 5' cap. The scaffold protein, eIF4G, also recruits the 40S ribosomal subunit to mRNA through its interaction with eIF3 and binds eIF4B, a protein that promotes the function of the eIF4A RNA helicase, thus facilitating the translation of mRNAs that contain structured 5'-UTRs. The availability of eIF4E as part of the eIF4F complex is a limiting factor in the control of translational rate, and hence eIF4E is an important regulator of mRNA translation.

The regulation of eIF4E activity forms a node of convergence of the PI3K/Akt/mTOR and Ras/Raf/MAPK signaling pathways. The PI3K (phosphoinositide 3-kinase)/PTEN (phosphatase and tensin homologue deleted on chromosome ten)/Akt/mTOR (mammalian target of rapamycin) pathway is frequently involved in oncogenesis and in susceptibility and resistance to cancer therapy. Deregulated signaling through the PI3K/PTEN/Akt/mTOR pathway is often the result of genetic changes in critical components of this pathway and/or mutations in growth factor receptors or signaling components in the upstream signaling pathway. PI3K initiates a cascade of events when activated e.g. by extracellular growth factors, mitogens, cytokines and/or receptors, PDK1 activates Akt which in turn phosphorylates and inactivates the tumor suppressor complex including TSC1 and 2 (tuberous sclerosis complex 1/2 ), which leads to the activation of mTORC1 (target of rapamycin complex 1) by Rheb-GTP. Activation of PDK1 and Akt PI3Ks is negatively regulated by PTEN.

PTEN is a critical tumor suppressor gene and is frequently mutated or downregulated in human cancer. Its loss results in Akt activation and increases downstream mTORC1 signaling. The involvement of the mTOR1 complex (mTORC1) in neoplastic transformation seems to depend on its regulatory role for the eIF4F complex; overexpression of eIF4E can confer resistance to rapamycin. mTORC1 regulates the assembly of the eIF4F complex, which is critical for mRNA translation associated with cell growth, apoptosis prevention, and transformation. mTORC1 achieves this by phosphorylation and inactivation of 4E-BP and subsequent dissociation of 4E-BP from eIF4E. This then allows eIF4E to interact with the eIF4G scaffold protein, allowing assembly of the eIF4F complex to translate structured mRNAs. mTORC1 also promotes the activation of the translational activator, S6K, which phosphorylates the S6 ribosomal protein and other substrates, including eIF4B. mTORC1 signaling is inhibited by rapamycin and its analogs (rapalogues), although these compounds act allosterically and do not directly inhibit mTOR kinase activity.

Given the importance of the PI3K/Akt/mTOR pathway in the regulation of mRNA translation of genes that encode pro-oncogenic proteins and the activated mTORC1 signaling pathway in a high proportion of cancer, these kinases are actively used as targets for oncological drugs. A number of pharmacological inhibitors have been identified, some of which have reached advanced clinical stages. However, it has recently become clear that the mTOR pathway is involved in a complex feedback loop that may impair Akt activation. Long-term treatment of cancer cells or patients with mTOR inhibitors has been shown to induce increased PI3K activity, which results in Akt and eIF4E phosphorylation and promotes cancer cell survival. eIF4E, acting downstream of Akt and mTOR, mimics the action of Akt in oncogenesis and drug resistance, and Akt signaling through eIF4E is an important mechanism of oncogenesis and drug resistance in vivo.

In addition to the PI3K/Akt/mTOR pathway, eIF4E is also a target for the signaling cascade

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Ras/Raf/MAP, which is activated by growth factors, and for the p38 MAP kinase stress-activated pathway. Erk1/2 and p38 then phosphorylate MAP kinase-interacting kinase 1 (Mnk1) and MAP kinase-interacting kinase 2 (Mnk2). The Erk pathway is also upregulated in many cancers, reflecting, for example, activating mutations in Ras (found in about 20% of tumors) or loss of function of the Ras GTPase activator protein NF1. Mnk1 and Mnk2 are threonine/serine protein kinases and specifically phosphorylate the 209 (Ser209) series of eIF4E in the eIF4F complex through an interaction between eIF4E and Mnks that serves to recruit Mnks to act on eIF4E. Mice with mutated eIF4E, in which Ser209 is substituted with alanine, show no eIF4E phosphorylation and significantly reduced tumor growth. It is important to note that although Mnk activity is required for eIF4E-mediated oncogenic transformation, it is not required for normal development. Thus, pharmacological inhibition of Mnks represents an attractive therapeutic strategy for the treatment of cancer.

Despite an increasing understanding of the structure and function of Mnk, there has been little progress in the discovery of pharmacological inhibitors of Mnk, and a relatively small number of Mnk inhibitors have been reported: CGP052088 (Tschopp et al., Mol Cell Biol Res Commun. 3(4 ):205-211, 2000); CGP57380 (Rowlett et al., Am J Physiol Gastrointest Liver Physiol. 294(2): G452-459, 2008); and Cercosporamid (Konicek et al., Cancer Res. 71(5):1849-1857, 2011). These compounds, however, are predominantly used to validate the Mnk target. More recently, additional compounds have been proposed by investigators for the treatment of diseases affected by inhibition of Mnk1 and/or Mnk2 kinase activity, including, for example, compounds disclosed in International Patent Application Publication WO 2014/044691 and various patent documents cited therein, 4(dihydropyridinone -3-yl)amino-5-methylthieno[2,3,-d]pyrimidines disclosed in Yu et al., European Journal of Med. Chem., 95: 116-126, 2015, and 6'-((6-aminopyrimidin-4-yl)amino)-8'-methyl-2Ήspiro[cyclohexane-1,3'-imidazo[1,5-a] pyridine]-1',5'-dione; and various compounds disclosed in International Patent Application Publication WO 2015/200481.

Accordingly, although progress has been made in this area, there remains a significant need for compounds having improved solubility while still retaining activity directed at specific inhibition of Mnk and other receptor activities. The present invention satisfies this need and provides other related benefits.

The essence of the invention

The present invention is directed to compounds that inhibit or modulate Mnk activity, as well as stereoisomers, tautomers, and pharmaceutically acceptable salts of such compounds, as therapeutic candidates. The present invention is also directed to compositions containing such compounds and corresponding methods for the treatment of conditions for which Mnk inhibition would be beneficial, such as cancer.

In one embodiment, the invention is directed to compounds that correspond to formula (I), as well as a stereoisomer, tautomer or pharmaceutically acceptable salt of such compounds:

(I) where the compound of formula (I) is selected from the group consisting of

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In another embodiment, compositions are disclosed comprising a compound of formula (I) in combination with a pharmaceutically acceptable carrier, diluent, or excipient.

In yet another embodiment, methods are provided for treating an Mnk dependent condition in a mammal in need thereof. Such methods include administering to the mammal an effective amount of a compound of formula (I) or compositions comprising such a compound. Such conditions include, but are not limited to, various forms of cancer discussed in more detail below.

These and other aspects of the invention will become apparent with reference to the following detailed description. To this end, the present invention specifies various reference documents that describe in more detail certain general reference information, procedures, compounds and/or compositions, and each of which is incorporated into this application by reference in its entirety.

Detailed description of the invention

In the following description, some specific details are set forth in order to provide a thorough understanding of various embodiments of the invention. However, one skilled in the art will appreciate that the invention may be practiced without these details. Unless the context otherwise requires, in the present description and claims, the word include and its variants, such as includes and including, should be interpreted in an open, inclusive sense (ie, including, but not limited to).

Reference throughout the specification to one embodiment or embodiment means that a particular feature, structure, or characteristic described in connection with an embodiment is included in at least one embodiment of the present invention. Thus, phrases in one embodiment or in an embodiment at various places in this specification do not necessarily refer to the same embodiment. In addition, specific features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

Definitions

In the context of the present description, and unless otherwise indicated, the following terms and phrases have the meanings given below.

The phrase MAP kinase interacting kinase or the term Mnk refers to all isoforms of the MAP kinase interacting kinase protein, including Mnk-1 and Mnk-2.

The compound of the invention may exist in various isomeric forms, as well as in one or more tautomeric forms, including both individual tautomers and mixtures of tautomers. The term isomer is intended to encompass all isomeric forms of the compound of the present invention, including tautomeric forms of the compound.

Some of the compounds described in the present invention may have asymmetric centers and therefore exist in various enantiomeric and diastereomeric forms. The compound of the invention may be in the form of an optical isomer or diastereomer. Accordingly, the invention encompasses the compounds of the invention and their uses as described herein, in the form of their optical isomers, diastereoisomers and mixtures thereof, including a racemic mixture. Optical isomers of the compounds of the invention can be obtained by known methods such as asymmetric synthesis, chiral chromatography, or by chemical resolution of stereoisomers using optically active resolving agents.

Unless otherwise indicated, stereoisomer means one stereoisomer of a compound that is substantially free of other stereoisomers of that compound. Thus, a stereomerically pure compound having one chiral center will be substantially free of the compound's opposite enantiomer. A stereomerically pure compound having two chiral centers will essentially contain no other diastereomers of the compound. A typical stereomerically pure compound contains more than about 80 wt. % of one stereoisomer of the compound and less than about 20 wt. % of other stereoisomers of the compound, for example, more than about 90 wt. % of one stereoisomer of the compound and less than about 10 wt. , or more than about 95 wt.% of one compound stereoisomer and less than about 5 wt.% of other compound stereoisomers, or more than about 97 wt.% of one compound stereoisomer and less than about 3 wt.% of other compound stereoisomers.

If there is a discrepancy between a depicted structure and a name given to that structure, then the depicted structure takes precedence. In addition, if the stereochemistry of a structure or part of a structure is not indicated, for example, by bold or dotted lines, the structure or part of a structure should be considered to encompass all of its stereoisomers. However, in some cases where there is more than one chiral center, the structures and names may be presented as separate enantiomers to help describe the corresponding stereochemistry. Those skilled in the art of organic synthesis will be aware of whether or not the methods used to prepare the compounds result in their production as single enantiomers.

As used herein, a pharmaceutically acceptable salt is a pharmaceutically acceptable salt of a compound of the invention with an organic or inorganic acid or base. Representative pharmaceutically acceptable salts include, for example, alkali metal salts, alkaline earth metal salts, ammonium salts, water-soluble and water-insoluble salts such as acetate, amsonate (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzoate, bicarbonate , bisulfate, bitartrate, borate, bromide, butyrate, calcium, calcium edetate, camsylate, carbonate, chloride, citrate, clavularate, dihydrochloride, edetate, edisylate, estolate, esylate, fiunarate, gluceptate, gluconate, glutamate, glycolyllarsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methyl bromide, methyl nitrate, methyl sulfate, mucinate, napsilate, nitrate, N-methylglucamine, ammonium salt, 3-hydroxy-2-naphthoate , oleate, oxalate, palmitate, pamoate (1,1methene-bis-2-hydroxy-3-naphthoate, einbonate), pantothenate, phosphate/diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate , subacetate, succinate, sulfate, sulfosalicylate, suramate, tannate, tartrate, theoclate, tosylate, triethiodide, and valerate. A pharmaceutically acceptable salt may have more than one charged atom in its structure. In this case, the pharmaceutically acceptable salt may have multiple counterions. Thus, a pharmaceutically acceptable salt can

- 4 040075 have one or more charged atoms and/or one or more counterions.

The terms treat, treating, and treating refer to the improvement or elimination of a disease or symptoms associated with a disease. In some embodiments, such terms refer to minimizing the spread or worsening of a disease by administering one or more prophylactic or therapeutic agents to a patient with such a disease.

The term effective amount refers to an amount of a compound of the invention or other active ingredient sufficient to provide a therapeutic or prophylactic effect in the treatment or prevention of a disease, or to delay the onset or minimize symptoms associated with a disease. In addition, a therapeutically effective amount with respect to a compound of the invention means that amount of a therapeutic agent, used alone or in combination with other therapies, that provides a therapeutic benefit in the treatment or prevention of a disease. Used in connection with the compound of the present invention, this term may include an amount that improves overall therapy, reduces or eliminates the occurrence of symptoms or causes of a disease, or increases therapeutic efficacy or synergy with another therapeutic agent.

The terms modulate, modulate, etc. refer to the ability of a compound to increase or decrease the function or activity of, for example, a MAP kinase (Mnk) interacting kinase. Modulation in its various forms is intended to encompass inhibition, antagonism, partial antagonism, activation, agonism and/or partial agonism of Mnk-associated activity. Mnk inhibitors are compounds that bind, partially or completely block stimulation, reduce, prevent, delay activation, inactivate, desensitize or suppress signal transmission. The ability of a compound to modulate Mnk activity can be demonstrated in an enzymatic assay or a cellular assay.

The patient or subject includes an animal such as a human, cow, horse, sheep, lamb, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit, or guinea pig. The animal may be a mammal, such as a non-primate or primate (eg, ape and human). In one embodiment, the patient is a human, such as an infant, child, adolescent, or adult.

The term prodrug refers to a drug precursor, which is a compound that, when administered to a patient, must undergo chemical transformation as a result of metabolic processes before becoming an active pharmacological agent. Examples of prodrugs of compounds according to formula (I) are esters, acetamides and amides.

Compounds of the invention

The present invention is mainly directed to compounds covered by the genus of formula (I), or their stereoisomers, tautomers or pharmaceutically acceptable salts.

The invention also provides pharmaceutically acceptable salt forms of the compounds of formula (I). The scope of the invention includes acid addition or base addition salts, which are obtained by contacting a pharmaceutically acceptable acid or pharmaceutically acceptable base with a compound of the invention.

To this end, a pharmaceutically acceptable acid addition salt refers to those salts that retain the biological efficacy and free base properties that are not biologically or otherwise undesirable and that form with inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfonic acid, ethane-1,2 -disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfone oic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxoglutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, ptoluenesulfonic acid, trifluoroacetic acid, undecylenes th acid, etc.

Similarly, a pharmaceutically acceptable base addition salt refers to such

- 5 040075 salts that retain the biological effectiveness and properties of free acids that are not biologically or otherwise undesirable. These salts are prepared by adding an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Preferred inorganic salts are the ammonium, sodium, potassium, calcium and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine , diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benetamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines , piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.

A stereoisomer refers to compounds composed of the same atoms, linked by the same bonds, but having different three-dimensional structures that are not interchangeable. The present invention contemplates various stereoisomers and mixtures thereof and includes enantiomers that refer to two stereoisomers whose molecules are non-superimposable mirror images of each other.

The compounds of the invention or their pharmaceutically acceptable salts may contain one or more asymmetric centers and thus enantiomers, diastereomers and other stereoisomeric forms may be formed, which can be defined in terms of absolute stereochemistry as (R) or (S) or as (D ) or (L) for amino acids. The present invention is intended to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R) and (S) or (D) and (L) isomers can be obtained using chiral synthons or chiral reagents or separated using conventional methods such as chromatography and fractional crystallization. Conventional methods for the preparation/isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or racemate of a salt or derivative) using, for example, chiral high performance liquid chromatography (HPLC). When the compounds described in the present invention contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise indicated, the compounds are meant to include both E and Z geometric isomers. Likewise, all tautomeric forms are intended to be included.

The term tautomer refers to the shift of a proton from one atom of a molecule to another atom of the same molecule. Non-limiting examples of tautomers include the enol/keto, lactam/lactim, amide/imide, and amine/imine forms.

Similar tautomers exist for compounds of formula (I). The compounds of the invention are synthesized using conventional synthetic methods and, more specifically, using the general methods described below. Specific synthesis protocols for several compounds in accordance with the present invention are described in the examples.

Pharmaceutical compositions

In one embodiment, a compound of formula (I) is formulated into pharmaceutically acceptable compositions that contain a compound of formula (I) in an amount effective to treat the particular disease or condition of interest when the pharmaceutical composition is administered to a mammal. Pharmaceutical compositions in accordance with the present invention may include a compound of formula (I) in combination with a pharmaceutically acceptable carrier, diluent or excipient.

In this regard, a pharmaceutically acceptable carrier, diluent or excipient includes, without limitation, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, coloring/coloring agent, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, a stabilizer, isotonic agent, diluent or emulsifier that has been approved by the US Food and Drug Administration as being acceptable for use in humans or pets.

Further, mammal includes humans and domestic animals such as laboratory animals and pets (e.g., cats, dogs, pigs, cattle, sheep, goats, horses, rabbits) as well as non-domestic animals such as wild animals and etc.

Pharmaceutical compositions of the invention may be prepared by combining a compound of the invention with a suitable pharmaceutically acceptable carrier, diluent or excipient and may be formulated into solid, semi-solid, liquid or gaseous forms such as tablets, capsules, powders, granules, ointments, solutions , suppositories, injections, dosage forms for inhalation, gels, microspheres and aerosols. Typical routes of administration for such pharmaceuticals

- 6 040075 tic compositions include, without limitation, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal and intranasal. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injections or infusion methods. Pharmaceutical compositions of the invention are formulated so that the active ingredients contained therein can be bioavailable when the composition is administered to a patient. Compositions to be administered to a subject or patient take the form of one or more dosage units, where, for example, a tablet may be a single dosage unit and an aerosolized compound container of the invention may contain multiple dosage units. The actual methods for preparing such dosage forms are known or will be apparent to those skilled in the art; for example, see Remington: The Science and Practice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000). The composition to be administered will in any event contain a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, for the treatment of the disease or condition of interest as described herein.

The pharmaceutical composition of the invention may be in the form of a solid or liquid. In one aspect, the carrier(s) are in the form of solid particles such that the compositions are, for example, in the form of tablets or powder. The carrier(s) may be liquid, with the compositions being, for example, an oral syrup, an injectable liquid or an aerosol, which is useful, for example, for inhalation administration. When the pharmaceutical composition is intended for oral administration, it is preferably in solid or liquid form, with semi-solid, semi-solid, suspension and gel forms included in the forms considered in the present invention, as solid or liquid.

As a solid composition for oral administration, the pharmaceutical composition may be formulated as a powder, granule, compressed tablet, pill, capsule, chewing gum, cachet, or the like. Such a solid composition will typically contain one or more inert diluents or food carriers. In addition, one or more of the following may be present: binders such as carboxymethyl cellulose, ethyl cellulose, microcrystalline cellulose, gum tragacanth, or gelatin; excipients such as starch, lactose or dextrins; disintegrants such as alginic acid, sodium alginate, primogel, corn starch and the like; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; flavoring such as peppermint, methyl salicylate or orange flavoring; and dye.

When the pharmaceutical composition is in the form of a capsule, for example a gelatin capsule, it may contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol or an oil.

The pharmaceutical composition may be in the form of a liquid, such as an elixir, syrup, solution, emulsion or suspension. The liquid may be for oral administration or for delivery by injection, as two examples. When the composition is intended for oral administration, a preferred composition contains, in addition to the compounds of the present invention, one or more of a sweetener, preservatives, coloring/coloring agent and flavor enhancer. One or more of a surfactant, a preservative, a wetting agent, a dispersing agent, a suspending agent, a buffer, a stabilizer, and an isotonic agent may be included in the composition for administration by injection.

Liquid pharmaceutical compositions of the invention, such as solutions, suspensions or other similar forms, may include one or more of the following excipients: sterile diluents such as water for injection, saline, preferably saline, Ringer's solution, isotonic sodium chloride solution, fixed oils such as synthetic mono- or diglycerides which may serve as a solvent or suspending medium, polyethylene glycols, glycerol, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates; and tonicity adjusting agents such as sodium chloride or dextrose. The parenteral preparation may be enclosed in ampoules, disposable syringes or multi-dose glass or plastic vials. Physiological saline is the preferred adjuvant.

The injectable pharmaceutical composition is preferably sterile.

A liquid pharmaceutical composition of the invention intended for parenteral or oral administration should contain such an amount of a compound of the invention that a suitable dosage can be obtained.

The pharmaceutical composition of the invention may be for topical administration, in which case the carrier may suitably comprise a solution, emulsion, ointment or

- 7 040075 gel. The base, for example, may include one or more of the following: petrolatum, lanolin, polyethylene glycols, beeswax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Thickening agents may be present in the topical pharmaceutical composition. If the composition is intended for transdermal administration, it may include a transdermal patch or iontophoresis device.

The pharmaceutical composition of the invention may be for rectal administration, for example in the form of a suppository, which will dissolve in the rectum and release the drug. The composition for rectal administration may contain an oily base as a suitable non-irritating excipient. Such bases include, without limitation, lanolin, cocoa butter, and polyethylene glycol.

The pharmaceutical composition of the invention may include various substances that modify the physical form of a solid or liquid dosage unit. For example, the composition may include substances that form a coating shell around the active ingredients. Substances which form the coating shell are usually inert and may be selected, for example, from sugar, shellac and other enteric substances. Alternatively, the active ingredients may be enclosed in a gelatin capsule.

A pharmaceutical composition of the invention in solid or liquid form may include a substance that binds to a compound of the invention and thus aids in the delivery of the compound. Suitable substances that can act in this capacity include a monoclonal or polyclonal antibody, protein or liposome.

The pharmaceutical composition according to the invention may consist of dosage units that can be administered as an aerosol. The term aerosol is used to refer to a variety of systems, from systems of a colloidal nature to systems consisting of pressurized packages. Delivery can be by liquefied or compressed gas or a suitable pumping system that dispenses the active ingredients. Aerosols of the compounds of the invention can be delivered in single phase, two phase or three phase systems to deliver the active ingredient(s). Delivery of an aerosol includes the necessary container, activators, valves, subcontainers, and the like, which together may form a kit. One skilled in the art will be able to determine preferred aerosols without undue experimentation.

Pharmaceutical compositions according to the invention can be obtained by any of the methods well known in the pharmaceutical art. For example, a pharmaceutical composition intended for administration by injection can be obtained by combining a compound of the invention with sterile distilled water to form a solution. You can add a surfactant to facilitate the formation of a homogeneous solution or suspension. Surfactants are compounds that interact non-covalently with a compound of the invention, thus facilitating the dissolution or homogeneous suspension of the compound in an aqueous delivery system.

In some embodiments, a pharmaceutical composition comprising a compound of Formula I is administered to a mammal in an amount sufficient to inhibit Mnk activity upon administration, and preferably with acceptable toxicity to that target. The Mnk directed activity of the compounds of formula (I) can be determined by one skilled in the art, for example as described in the examples below. Suitable concentrations and dosages will readily be determined by one skilled in the art.

Therapeutic use

The compounds of the invention, or pharmaceutically acceptable salts thereof, are administered in a therapeutically effective amount, which will vary depending on a variety of factors, including the activity of the particular compound used; metabolic stability and duration of action of the compound; age, body weight, general health, sex and diet of the patient; method and time of administration; excretion rate; a combination of drugs; the severity of the particular disorder or condition; and the subject being treated.

An effective amount or therapeutically effective amount refers to that amount of a compound of the invention which, when administered to a mammal, preferably a human, is sufficient to effect treatment, as defined below, of an Mnk-related condition or disease in a mammal, preferably a human. The amount of a compound of the invention that constitutes a therapeutically effective amount will vary depending on the compound, the condition and severity thereof, the route of administration, and the age of the mammal being treated, but will generally be determined by one of ordinary skill in the art based on their own knowledge and of this disclosure.

The compounds of the invention, or pharmaceutically acceptable salts thereof, may also be administered simultaneously with, before, or after administration of one or more other therapeutic agents. Such combination therapy comprises the administration of a single pharmaceutical dosage composition which contains a compound of the invention and one or more additional active agents,

- 8 040075 as well as the introduction of the compounds according to the invention and each active agent in a separate pharmaceutical dosage composition. For example, a compound of the invention and another active agent may be administered to a patient together in a single oral dosage composition such as a tablet or capsule, or each agent may be administered in separate oral dosage compositions. When separate dosage compositions are used, the compounds of the invention and one or more additional active agents may be administered at substantially the same time, i. simultaneously, or at different times, i.e. sequentially; it is assumed that combination therapy includes all of these regimens.

In some embodiments, the disclosed compounds are useful for inhibiting Mnk activity and/or may be useful for assaying Mnk signaling activity in model systems and/or for preventing, treating, or ameliorating symptoms associated with a disease, disorder, or condition involving Mnk, preferably affecting person. A compound that inhibits Mnk activity would be useful in the prevention, treatment, amelioration or reduction of the symptoms or progression of diseases associated with uncontrolled cell growth, proliferation and/or survival, inadequate cellular immune responses or inappropriate cellular inflammatory responses, or diseases that are accompanied by uncontrolled cell growth, proliferation and/or survival, inadequate cellular immune responses, or inappropriate cellular inflammatory responses, in particular when uncontrolled cell growth, proliferation and/or survival, inadequate cellular immune responses, or inadequate cellular inflammatory responses are mediated by Mnk, such as, for example, hematologic tumors, solid tumors and/or their metastases, including leukemias and myelodysplastic syndrome, malignant lymphomas, eg B-cell lymphoma, T-cell lymphoma, hairy cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma and Burkitt's lymphoma, head and neck tumors, including brain tumors and brain metastases, chest tumors, including non-small cell and small cell lung tumors, gastrointestinal tumors, endocrine tumors, breast and other gynecological tumors, urological tumors, including kidney, bladder and prostate tumors, skin tumors and sarcomas, and/or their metastases.

In addition, the compounds of the invention and their pharmaceutical compositions are candidates for use as therapeutic agents for the prevention and/or treatment of diseases associated with cytokines, such as inflammatory diseases, allergies, or other conditions associated with pro-inflammatory cytokines. Typical inflammatory diseases include, without limitation, chronic or acute inflammation, joint inflammation such as chronic inflammatory arthritis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile rheumatoid arthritis, Reiter's syndrome, rheumatoid traumatic arthritis, rubella arthritis, acute synovitis, and gouty arthritis. ; inflammatory skin diseases such as sunburn, psoriasis, erythrodermic psoriasis, pustular psoriasis, eczema, dermatitis, acute or chronic graft formation, atopic dermatitis, contact dermatitis, urticaria and scleroderma; gastrointestinal inflammation such as inflammatory bowel disease, Crohn's disease and related conditions, ulcerative colitis, colitis and diverticulitis; nephritis, urethritis, salpingitis, oophoritis, endomyometritis, spondylitis, systemic lupus erythematosus and related disorders, multiple sclerosis, asthma, meningitis, myelitis, encephalomyelitis, encephalitis, phlebitis, thrombophlebitis, respiratory diseases such as asthma, bronchitis, chronic obstructive disease lung disease (COPD), inflammatory lung disease and adult respiratory distress syndrome, and allergic rhinitis; endocarditis, osteomyelitis, rheumatism, rheumatic pericarditis, rheumatic endocarditis, rheumatic myocarditis, rheumatic mitral valve disease, rheumatic aortic valve disease, prostatitis, prostatocystitis, spondoarthropathy, ankylosing spondylitis, synovitis, tenosynovitis, myositis, pharyngitis, polymyalgia rheumatica, tendonitis, shoulder or bursitis gout, pseudo-gout, vasculitis, inflammatory thyroid diseases selected from the group consisting of granulomatous thyroiditis, lymphocytic thyroiditis, invasive fibrous thyroiditis, acute thyroiditis; Hashimoto's thyroiditis, Kawasaki's disease, Raynaud's phenomenon, Sjögren's syndrome, neuroinflammatory disease, sepsis, conjunctivitis, keratitis, iridocyclitis, optic neuritis, otitis media, lymphadenitis, nasopharyngitis, sinusitis, pharyngitis, tonsillitis, laryngitis, epiglottitis, bronchitis, pneumonitis, stomatitis, gingivitis , esophagitis, gastritis, peritonitis, hepatitis, cholelithiasis, cholecystitis, glomerulonephritis, Goodpasture's disease, crescentic glomerulonephritis, pancreatitis, endomyometritis, myometritis, metritis, cervicitis, endocervicitis, exocervicitis, parametritis, tuberculosis, vaginitis, vulvitis, silicosis, sarcoidosis, pneumoconiosis, heartburn , inflammatory polyarthropathies, psoriatic arthropathy, intestinal fibrosis, bronchiectasis and enteropathic arthropathy.

Although inflammation is the unifying pathogenic process of these diseases, current therapies only treat the symptoms of the disease and not the underlying cause of the inflammation.

The compositions of the present invention are useful for the treatment and/or prevention of inflammatory diseases and their associated complications and disorders.

- 9 040075

Accordingly, some embodiments are directed to a method of treating an Mnk dependent condition in a mammal in need thereof, the method comprising administering to the mammal an effective amount of a pharmaceutical composition as described above (i.e., a pharmaceutical composition comprising any one or more compounds of formula (I)).

Treating or treating, as used herein, encompasses the treatment of a disease or condition of interest in a mammal, preferably a human, having the disease or condition of interest, and includes (i) preventing the occurrence of the disease or condition in the mammal, in particular when such mammal is predisposed to this condition, but this condition has not yet been diagnosed in him;

(ii) inhibition of the disease or condition, ie. stop its development;

(iii) alleviate the disease or condition, i. e. inducing regression of the disease or condition; or (iv) relief of symptoms resulting from the disease or condition, ie. pain relief without affecting the underlying disease or condition.

In the context of the present description, the terms disease and condition may be used interchangeably or may be different because a particular disease or condition may not have a known causal factor (therefore, the etiology is not yet clear) and therefore is not yet recognized as a disease, but only as an undesirable condition or syndrome, where clinicians identified a more or less specific set of symptoms.

As described above, deregulation of protein synthesis is common in human cancer. The key regulator of translational control is eIF4E, whose activity is a key determinant of carcinogenicity. Because eIF4E activation involves phosphorylation of a key serine (Ser209) specifically by MAP kinase (Mnk) interacting kinases, Mnk inhibitors are suitable candidates for use as therapeutic agents for the treatment of cell proliferative disorders such as cancer. Various cancers, including solid tumors, lymphomas, and leukemias, are treatable with the compositions and methods disclosed in the present invention. Types of cancer that can be treated include, but are not limited to, adenocarcinoma of the breast, prostate, and colon; all forms of bronchogenic lung carcinoma; myeloid; melanoma; hepatoma; neuroblastoma; papilloma; apudoma; chorister; branchioma; malignant carcinoid syndrome; carcinoid heart disease; and carcinomas (e.g., Walker, basal cell, basosquamous, Brown-Pierce, ductal, Ehrlich tumor, Krebs 2, Merkel cell, mucinous, non-small cell lung, oat cell, papillary, scirrhous, bronchiolar, bronchogenic, squamous cell, and transitional cellular). Additional cancers that can be treated include histiocytic disorders; leukemia; malignant histiocytosis; Hodgkin's disease; immunoproliferative disease; non-Hodgkin's lymphoma; diffuse large B-cell lymphoma, T-cell lymphoma, B-cell lymphoma, hairy cell lymphoma, Burkitt's lymphoma, plasmacytoma; reticuloendotheliosis; melanoma; chondroblastoma; chondroma; chondrosarcoma; fibroma; fibrosarcoma; giant cell tumors; histiocytoma; lipoma; liposarcoma; mesothelioma; myxoma; myxosarcoma; osteoma; osteosarcoma; chordoma; craniopharyngioma; dysgerminoma; hamartoma; mesenchymal; mesonephroma; myosarcoma; ameloblastoma; cementoma; odontoma; teratoma; thymoma; trophoblastic tumor.

Other cancers that can be treated using the compounds of the invention include, but are not limited to, adenoma; cholangioma; cholesteatoma; cyclindrome; cystadenocarcinoma; cystoadenoma; granulosa cell tumor; ginandroblastoma; hepatoma; hydradenoma; islet cell tumor tumor from Leydig cells; papilloma; tumor from Sertoli cells; thecacellular tumor; leimoma; leiomyosarcoma; myoblastoma; myoma; myosarcoma; rhabdomyoma; rhabdomyosarcoma; ependymoma; ganglioneuroma; glioma; medulloblastoma; meningioma; neurilemma; neuroblastoma; neuroepithelimoma; neurofibroma; neuroma; paraganglioma; nonchromaffin paraganglioma.

In one embodiment, the compounds of the invention are candidates for use as therapeutic agents for the treatment of cancers such as angiokeratoma; angiolymphoid hyperplasia with eosinophilia; sclerosing angioma; angiomatosis; glomangioma; hemangioendothelioma; hemangioma; hemangiopericytoma; hemangiosarcoma; lymphangioma; lymphangiomyoma; lymphangiosarcoma; pinealoma; carcinosarcoma; chondrosarcoma; leaf-shaped cystosarcoma; fibrosarcoma; hemangiosarcoma; leiomyosarcoma; leukosarcoma; liposarcoma; lymphangiosarcoma; myosarcoma; myxosarcoma; ovarian carcinoma; rhabdomyosarcoma; sarcoma; neoplasms; neurofibromatosis; and cervical dysplasia.

In a specific embodiment, the present invention provides methods for treating solid tumor, colon cancer, rectal cancer, colorectal cancer, bladder cancer, gastric cancer, esophageal cancer, head and neck cancer, myelodysplastic syndrome, brain cancer, CNS cancer, malignant glioma. , glioblastoma, hepatocellular carcinoma, hepatocellular carcinoma, thyroid cancer, lung cancer, non-small cell lung cancer, hematologic

- 10 040075 cancer, leukemia, B-cell lymphoma, T-cell lymphoma, hairy cell lymphoma, diffuse large B-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, pancreatic cancer, melanoma, myeloma, multiple myeloma, pancreatic carcinoma , renal cell carcinoma, kidney cancer, cervical cancer, urothelial cancer, prostate cancer, castration-resistant prostate cancer, testicular cancer, breast cancer, or triple negative breast cancer. According to such a method, a therapeutically effective amount of at least one compound according to formula (I), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, may be administered to a subject diagnosed with a cell proliferative disease such as cancer. Alternatively, a pharmaceutical composition comprising at least one compound according to formula (I), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, may be administered to a subject diagnosed with cancer.

In some embodiments, the compounds of the invention are administered to a subject having cancer in combination with other conventional cancer treatments such as radiation therapy or surgery. Radiation therapy is well known in the art and includes x-ray therapy such as gamma irradiation and radiopharmaceutical therapies.

In some embodiments, the Mnk inhibitor compounds of the present invention are used with at least one anticancer agent. Anticancer agents include chemotherapeutic agents. The chemotherapeutic agent includes, but is not limited to, a chromatin function inhibitor, a topoisomerase inhibitor, a microtubule inhibitory drug, a DNA damaging agent, an antimetabolite (such as folate antagonists, pyrimidine analogs, purine analogs, and sugar-modified analogs), a DNA synthesis inhibitor, a DNA interacting agent (such as an intercalating agent); and a DNA repair inhibitor.

Exemplary chemotherapeutic agents include, without limitation, the following groups: antimetabolites/anticancer agents such as pyrimidine analogs (5-fluorouracil, floxuridine, capecitabine, gemcitabine, and cytarabine) and purine analogs, folate antagonists, and related inhibitors (mercaptopurine, thioguanine, pentostatin, and 2 -chlordeoxyadenosine (cladribine)); anti-proliferative/anti-mitotic agents including natural products such as vinca alkaloids (vinblastine, vincristine and vinorelbine), microtubule degrading agents such as taxane (paclitaxel, docetaxel), vincristine, vinblastine, nocodazole, epothilones and navelbine, epidipodophyllotoxins (etoposide, teniposide) DNA damaging agents (actinomycin, amsacrine, anthracyclines, bleomycin, busulfan, camptothecin, carboplatin, chlorambucil, cisplatin, cyclophosphamide, cytoxan, dactinomycin, daunorubicin, doxorubicin, epirubicin, hexatronmethylmelaminexaliplatin, ifosfamide, melphalan, merchlorethamine, mitoksanomicin, mitoksanomicin, plicamycin, procarbazine, taxol, taxotere, temozolamide, teniposide, triethylenethiophosphoramide and etoposide (VP 16)); antibiotics such as dactinomycin (actinomycin D), daunorubicin, doxorubicin (adriamycin), idarubicin, anthracyclines, mitoxantrone, bleomycins, plicamycin (mitramycin), and mitomycin; enzymes (Lasparaginase, which systemically metabolizes L-asparagine and deprives cells that are unable to synthesize their own asparagine); antiplatelet agents; antiproliferative/antimitotic alkylating agents such as nitrogen mustard (mechlorethamine, cyclophosphamide and analogs, melphalan, chlorambucil), ethyleneimines and methylmelamines (hexamethylmelamine and thiotepa), alkyl sulfonates -busulfan, nitrosoureas (carmustine (BCNU) and analogs, streptozocin), trazenes - dacarbazinine (DTIC); antiproliferative/antimitotic antimetabolites such as folic acid analogs (methotrexate); platinum coordination complexes (cisplatin, carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide; hormones, hormone analogues (estrogen, tamoxifen, goserelin, bicalutamide, nilutamide) and aromatase inhibitors (letrozole, anastrozole); anticoagulants (heparin, synthetic heparin salts and other thrombin inhibitors); fibrinolytic agents (such as tissue plasminogen activator, streptokinase and urokinase), aspirin, dipyridamole, ticlopidine, clopidogrel, abciximab; anti-migration means; antisecretory agents (breveldin); immunosuppressants (cyclosporine, tacrolimus (FK-506), sirolimus (rapamycin), azathioprine, mycophenolate mofetil); anti-angiogenic compounds (TNP470, genistein) and growth factor inhibitors (vascular endothelial growth factor (VEGF) inhibitors, fibroblast growth factor (FGF) inhibitors); angiotensin receptor blocker; nitric oxide donors; antisense oligonucleotides; antibodies (trastuzumab, rituximab); chimeric antigen receptors; cell cycle inhibitors and differentiation inducers (tretinoin); mTOR inhibitors, topoisomerase inhibitors (doxorubicin (adriamycin), amsacrine, camptothecin, daunorubicin, dactinomycin, eniposide, epirubicin, etoposide, idarubicin, irinotecan (CPT-11) and mitoxantrone, topotecan, irinotecan), corticosteroids (cortisone, dexamethasone, hydrocortisone, methylprednisolone , prednisone and prednisolone); growth factor signal transduction kinase inhibitors; mitochondrial dysfunction inducers, toxins such as cholera toxin, ricin, Pseudomonas exotoxin, Bordetella pertussis adenylate cyclase toxin, or diphtheria toxin, and caspase activators; and chromatin destroying agents.

In some embodiments, the Mnk inhibitor of the present invention

- 11 040075 are used simultaneously, in the same composition or in separate compositions, or sequentially with additional agent(s) as part of a combination therapy regimen.

Mnk inhibitors according to formula (I), including their respective salts and pharmaceutical compositions of compounds of formula (I), are also effective as therapeutic agents for the treatment or prevention of cytokine-mediated disorders such as inflammation in a patient, preferably a human. In one embodiment, a compound or composition according to the invention is particularly useful for the treatment or prevention of a disease selected from the group consisting of chronic or acute inflammation, chronic inflammatory arthritis, rheumatoid arthritis, psoriasis, COPD, inflammatory bowel disease, septic shock, Crohn's disease , ulcerative colitis, multiple sclerosis and asthma.

In a further aspect of the invention, the compounds of the invention or pharmaceutically acceptable compositions of the compounds of the invention are provided as inhibitors of Mnk activity. Such inhibition is achieved by contacting an Mnk expressing cell with a compound or pharmaceutically acceptable composition to reduce or inhibit Mnk activity to provide therapeutic efficacy against an Mnk dependent condition in a mammal in need thereof.

Therapeutically effective doses of a compound of formula (I) or a composition of a compound of formula (I) are typically in the range of about 1 to 2000 mg/day, about 10 to about 1000 mg/day, about 10 to about 500 mg/day , about 10 to about 250 mg/day, about 10 to about 100 mg/day, or about 10 to about 50 mg/day. Therapeutically effective doses may be administered in one or more doses. However, it should be understood that the specific dosages of the compounds of the invention for any particular patient will depend on a variety of factors such as age, sex, body weight, general health, diet, individual response of the patient being treated, time of administration, severity of the disease being treated. treatment, activity of the particular compound used, dosage form, route of administration, and concomitant treatment. A therapeutically effective amount for a given situation can easily be determined by the ordinary clinician or physician by routine experimentation and skill and judgement. In any event, the compound or composition will be administered at dosages and in a manner that will deliver a therapeutically effective amount depending on the unique condition of the patient.

General synthesis methods

Method 1.

The preparation of intermediate IV, where X=halogen or another leaving group such as OTf, -OTs or -OMs, was carried out by exposing compound II to a ketone III or a ketone equivalent such as IIIa, IIIb or IIIc under acidic conditions. More specifically, by subjecting compound II, where X is Cl or Br, to ketone III in 1,4-dioxane and concentrated sulfuric acid, intermediate compound IV is obtained by heating.

Compounds of formula I were synthesized by Buchwald-Hartwig coupling, type coupling

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Ullmann or nucleophilic aromatic substitution. Thus contacting an intermediate IV where X=halogen or another leaving group such as -OTf, -OTs or -OMs with a compound of formula V or Va under conditions suitable for coupling or nucleophilic aromatic substitution gave compounds of formula I or Ia. Compound Ia can be subjected to a deprotection procedure to give compound I.

Method 2.

N'^N N'^N

I la

In an alternative method, the leaving group X of intermediate IV, where X=halogen or another leaving group such as -OTf, -OTs or -OMs, can be replaced by the corresponding N nucleophile under conditions similar to those described above for the synthesis of compound I, thus obtaining an intermediate V or a protected intermediate Va. Compound Va can be subjected to a deprotection procedure to give Compound V.

Compounds of formula I or Ia were readily synthesized by contacting an intermediate V with a pyrimidine compound VI or VIa, where X=halogen or another leaving group such as -OTf, -OTs or -OMs, under Buchwald-Hartwig coupling conditions, coupling by type Ullmann or nucleophilic aromatic substitution. Compound Ia can be subjected to a deprotection procedure to give compound I.

More specific methods for the synthesis of compounds of formula I are described below. It should be understood that if protecting groups (P) are used during the synthesis of intermediates, or if a compound of formula I contains one or more protecting groups, then such protecting groups are removed by methods known in the art of chemistry.

It should also be understood that the group R ¹ , R ² or R ³ compounds of formula I can be formed at a suitable stage of the synthesis by conventional methods known in chemistry.

Examples

The following examples are presented for purposes of illustration and not limitation.

Example 1 (R)-6-((6-amino-5-methylpyrimidin-4-yl)amino)-1',8-dimethyl-2H-spiro[imidazo[1,5a]pyridine-3,3'- piperidine]-1,5-dione (comp. No. 1Fa) and (S)-6-((6-amino-5-methylpyrimidin-4-yl)amino)-1',8dimethyl-2H-spiro[imidazo[1 ,5-a]pyridine-3,3'-piperidine]-1,5-dione (comp. No. 1Fb).

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Pd ₂ dba ₃ , Pd(OAc) ₂ XPhos, XantPhos

Synthesis of N-(6-((1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo [1,5-a]pyridine-3,3'-piperidine] 6- yl)amino)-5-methylpyrimidin-4-yl)cyclopropanecarboxamide (3).

Suspension of N-(6-amino-5-methylpyrimidin-4-yl)cyclopropanecarboxamide (1, 0.82 g, 4.29 mmol), 6-bromo-1',8-dimethyl-2H-spiro[imidazo[1, 5-a]pyridine-3,3'-piperidine]-1,5-dione (2, 1.40 g, 4.29 mmol) and cesium carbonate (2.79 g, 8.58 mmol) in 1.4 -dioxane (20 ml) was purged with argon for 15 min. To this mixture was added 9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene (0.12 g, 0.21 mmol), 2-(dicyclohexylphosphino)-2',4',6'-triisopropylbiphenyl (0. 10 g, 0.21 mmol), tris(dibenzylideneacetone) dipalladium (0.20 g, 0.21 mmol) and palladium acetate (0.049 g, 0.21 mmol). The mixture was purged for another 5 minutes and the vessel was sealed. The reaction mixture was stirred at 100° C. for 16 hours. After completion, the reaction mass was diluted with 5% methanol in dichloromethane (150 ml) and passed through a pad of celite. The filtrate was concentrated and the crude material was purified by combiflash column chromatography using 3-5% methanol in dichloromethane to give N(6-((1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro [imidazo[1,5-a]pyridine-3,3'-piperidine]-6-yl)amino)-5-methylpyrimidin-4-yl)cyclopropanecarboxamide (3) as a yellow solid. Yield: 0.96 g, 51%; MS (ESI) m/z 438.1 [M+1]+.

(R)-6-((6-amino-5-methylpyrimidin-4-yl)amino)-1',8-dimethyl-2H-spiro[imidazo[1,5-a]pyridine3,3'-piperidine]- 1,5-dione (comp. No. 1Fa) and (S)-6-((6-amino-5-methylpyrimidin-4-yl)amino)-1',8-dimethyl2H-spiro[imidazo[1,5- a]pyridine-3,3'-piperidine]-1,5-dione (comp. No. 1Fb).

To a solution of N-(6-((1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]- 6-yl)amino)-5-methylpyrimidin-4-yl)cyclopropanecarboxamide (3, 0.95 g, 2.17 mmol) in tetrahydrofuran, ethanol and water (1:1:1, 30 ml) was added potassium hydroxide (1 .22 g, 21.71 mmol). The reaction mixture was stirred at 60° C. for 18 hours. After completion, the reaction mixture was cooled to room temperature and extracted with 10% methanol in dichloromethane (3x50 ml). The combined organic layer was washed with water, brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude compound was purified by combiflash column chromatography using 7-8% methanol in dichloromethane to give 6-((6-amino-5-methylpyrimidin-4-yl)amino)-1',8-dimethyl-2Hspiro[imidazo[ 1,5-a]pyridine-3,3'-piperidine]-1,5-dione (rac-1F) as an off-white solid. Yield: 555 mg, 69%; MS (ESI) m/z 370.4 [M+1] ⁺ .

Chiral purification of the racemic compound 6-((6-amino-5-methylpyrimidin-4-yl)amino)-1',8dimethyl-2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]- 1,5-dione (Comp. No. 1F) was carried out by SFC chiral HPLC on a Chiralpak-IA column (250x21 mm, 5 μm) using an isocartoic mixture of 15% methanol/carbon dioxide. Peak-1: R _t =17 min, ee=98.16%;

1H NMR (400 MHz, DMSO-d ₆ ) δ 9.82 (br s, 1H), 8.48 (s, 1H), 8.12 (s, 1H), 7.98 (s, 1H), 6.50 (s, 2H),

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3.33-3.30(m, 1H), 3.02-2.93(m, 1H), 2.81-2.79(m, 1H), 2.46-2.45(m, 1H ), 2.44 (s, 3H), 2.21 (s, 3H), 1.98 (s,

3H), 1.93-1.91(m, 2H), 1.72-1.68(m, 1H), 1.49-1.47(m, 1H). Peak-2: R _t =19.8 min, ee=96.58%; ¹ H NMR (400

MHz, DMSO-d6) δ 9.87 (brs, 1H), 8.48 (s, 1H), 8.11 (s, 1H), 7.98 (s, 1H), 6.48 (s , 2H), 3.33-3.30 (m, 1H),

2.99-2.93 (m, 1H), 2.81-2.79 (m, 1H), 2.49-2.48 (m, 1H), 2.44 (s, 3H), 2, 21 (s, 3H), 1.98 (s, 3H), 1.93-1.91 (m,

2H), 1.71-1.68 (m, 1H), 1.47 (d, J=12 Hz, 1H).

Example 2 (R)-4-amino-6-((1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo[1,5-a]pyridine3,3' -piperidin]-6-yl)amino)pyrimidine-5-carbonitrile (comp. No. 2Fa) and (S)-4-amino-6-((1',8-dimethyl-1,5dioxo-1,5-dihydro -2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-6-yl)amino)pyrimidine-5-carbonitrile (Comp. No. 2Fb)

Fa2Fb

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Synthesis of 6-bromo-8-methyl-2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-1,5-dione hydrochloride (3).

5-bromo-3-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (1, 10.0 g, 43.2 mmol), tert-butyl 3-oxopiperidine-1-carboxylate (2 , 10.31 g, 51.8 mmol) and a 4 M solution of hydrogen chloride in 1,4-dioxane (100 ml) in 1,4-dioxane (100 ml). The reaction mixture was heated at 110°C for 16 hours. After completion, the reaction mixture was evaporated under reduced pressure and the solid residue was washed with simple ether to obtain 6-bromo-8-methyl-2Hspiro[imidazo[1,5-a]pyridine-3, 3'-piperidine]-1,5-dione hydrochloride (3) as a light yellow solid. Yield: 20.0 g, 93%; MS (ESI) m/z 312.27 [M+1]+.

Synthesis of 6-bromo-1',8-dimethyl-2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-1,5-dione (4).

To a solution of 6-bromo-8-methyl-2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-1,5-dione hydrochloride (3, 10.0 g, 28.8 mmol ) in N,N-dimethylformamide (75 mL) was added N,N-diisopropylethylamine (9.29 g, 72.04 mmol) and methyl iodide (4.50 g, 31.7 mmol) at room temperature. reaction mixture

- 16 040075 stirred at room temperature for 3 hours Then the reaction mixture was quenched with ice water and the solid precipitate was isolated by filtration. The solid was washed with diethyl ether and dried under reduced pressure to give 6-bromo-1',8-dimethyl-2H-cnuro[umidazo[1,5-a]nurudin3,3'-piperidine]-1,5-dione (4 ) as a light brown solid. Yield: 5.3 g, 57%; MS (ESI) m/z 326.2 [M+1]+ .

Synthesis of tert-butyl(1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo[1,5-a]πiridin-3,3'-πiperidin]-6-yl) carbamate (5).

To a solution of 6-bromo-1',8-dimethyl-2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-1,5-dione (4, 15.0 g, 46, 1 mmol) in 1,4-dioxane (150 ml) in a sealed tube were added tert-butyl carbamate (7.01 g, 59.0 mmol) and cesium carbonate (30.0 g, 92.0 mmol) under argon. The reaction mixture was purged with argon for 15 min followed by the addition of 9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene (2.66 g, 4.6 mmol) and tris(dibenzylideneacetone)dipalladium (4.1 g, 4 .60 mmol). The mixture was purged with argon for an additional 5 min and the reaction mixture was heated at 100° C. for 16 h. After completion, the reaction mass was concentrated in vacuo to give the crude compound, which was purified by silica gel column chromatography (3-5% methanol in dichloromethane) to give tert-butyl (1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-6-yl)carbamate (5) as a dark brown solid which was used in the next step without any further purification. Yield: 9.0 g, 54%; MS (ESI) m/z 363.14 [M+1] ⁺ .

Chiral resolution of tert-butyl (R)-(1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo[1,5a]pyridine-3,3'-piperidine]-6 -yl) carbamate (5A) and tert-butyl (S)-(1',8-dimethyl-1,5-dioxo-1,5-dihydro2H-spiro[imidazo[1,5-a]pyridine-3,3 '-piperidine]-6-yl) carbamate (5B).

Racemic compound tert-butyl(1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo[1,5a]pyridine-3,3'-piperidine]-6-yl)carbamate (5, 8.0 g) was purified with chiral SFC using a Chiralpak-IG column (250 x 21 mm, 5 μm) using an iso-map gradient of 0.2% triethylamine in methanol/carbon dioxide (40:60). After purification, peak-1 was isolated at Rt=8.0 min (2.4 g, ei=99.82%) and peak-2 was isolated at Rt=12.8 min (3.4 g, ei=99.60 %). The absolute stereochemistry of peak 1 and peak 2 was not determined. For ease of illustration, peak-1 was randomly assigned the name tert-butyl(R)-(1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo[1,5-a]pyridine- 3,3'-piperidine]-6yl)carbamate (5A), and peak-2 was randomly assigned the name tert-butyl (S)-(1',8-dimethyl-1,5dioxo-1,5-dihydro-2H- spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-6-yl)carbamate (5B).

Synthesis of (R)-6-amino-1',8-dimethyl-2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-1,5-dione (6A).

To a solution of tert-butyl (R)-(1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo[1,5-a]pyridine3,3'-piperidine]-6 -yl) carbamate (5A, 2.40 g, 6.6 mmol) in dichloromethane (20 ml) was added a 4 M solution of hydrogen chloride in 1,4-dioxane (20 ml). The reaction mixture was stirred at room temperature for 3 hours. After completion, the reaction mass was concentrated in vacuo and the crude compound was suspended in saturated sodium bicarbonate solution. The aqueous layer was extracted with dichloromethane; the organic layer was dried over sodium sulfate, filtered and concentrated. The residue was washed with diethyl ether to give the desired compound (R)-6-amino-1',8-dimethyl2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-1,5-dione (6A ) as a light brown solid. Yield: 1.7 g, 98%; MS (ESI) m/z 263.3 [M+1] ⁺

Synthesis of (R)-4-amino-6-((1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo[1,5-a]pyridine-3,3' piperidine]-6-yl)amino)pyrimidine-5-carbonitrile (Comp. No. 2Fa).

To a solution of (R)-6-amino-1',8-dimethyl-2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-1,5-dione (6A, 1.70 g, 6.4 mmol) in tert-butanol (25 ml) was added 4-amino-6-chloropyrimidine-5-carbonitrile (7, 1.0 g, 6.4 mmol) and p-toluenesulfonic acid (1.10 g , 6.4 mmol). The reaction mixture was heated at 120°C for 15 h in a hermetically sealed tube. Then the reaction mixture was concentrated and the residue was dissolved in a mixture of 20% isopropanol/chloroform (100 ml). The organic layer was washed with saturated sodium bicarbonate solution, dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography using combiflash (5-7% methanol in dichloromethane) to give (R)-4-amino-6-((1',8-dimethyl-1,5-dioxo-1,5-dihydro -2Hspiro[imidazo[1,5-a]pyridine-3,3'-piperidin]-6-yl)amino)pyrimidine-5-carbonitrile (Comp. No. 2Fa) as a white solid. Yield: 0.61 g, 25%. MS (ESI) m/z 381.13 [M+1] ⁺ ; ee=97.94%; R _t =19.96 min, Chiralpak IC (4.6x250 mm, 5 μm) using a methanol/carbon dioxide isomap gradient (40:60). 1H NMR (400 MHz, DMSO-d6) δ 10.02 (s, 1H), 8.58 (s, 1H), 8.49 (s, 1H), 8.34 (s, 1H), 7.79 (brs, 2H), 3.31-3.28 (m, 1H), 2.98-2.94 (m, 1H), 2.82-2.81 (m, 1H), 2.45 (s, 3H), 2.23 (s, 3H), 1.99-1.94 (m, 2H), 1.72-1.70 (m, 1H), 1.50 (d, J=12 .76 Hz, 1H).

Synthesis of (S)-6-amino-1',8-dimethyl-2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-1,5-dione (6B).

To a solution of tert-butyl (S)-(1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo[1,5-a]pyridine3,3'-piperidine]-6 -yl) carbamate (5B, 3.40 g, 9.3 mmol) in dichloromethane (30 ml) was added a 4 M solution of hydrogen chloride in 1,4-dioxane (30 ml). The reaction mixture was stirred at room temperature for 3 hours. After completion, the reaction mass was concentrated in vacuo and the crude

- 17 040075 the compound was suspended in a saturated sodium bicarbonate solution. The aqueous layer was extracted with dichloromethane; the organic layer was dried over sodium sulfate and concentrated. The residue was washed with diethyl ether to give the desired compound (S)-6-amino-1',8-dimethyl-2Hspiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-1,5-dione (6B ) as a light brown solid. Yield: 2.1 g, 85%; MS (ESI) m/z 263.3 [M+1]+.

Synthesis of (S)-4-amino-6-((1',8-dimethyl-1,5-dioxo-1,5-dihydro-2H-spiro[imidazo[1,5-a]pyridine-3,3' piperidine]-6-yl)amino)pyrimidine-5-carbonitrile (Comp. No. 2Fb).

To a solution of (S)-6-amino-1',8-dimethyl-2H-spiro[imidazo[1,5-a]pyridine-3,3'-piperidine]-1,5-dione (6B, 2.0 g, 7.6 mmol) in tert-butanol (30 ml) was added 4-amino-6-chloropyrimidine-5-carbonitrile (7, 1.17 g, 7.6 mmol) and p-toluenesulfonic acid (1.30 g , 7.6 mmol). The reaction mixture was heated at 120°C for 15 h in a hermetically sealed tube. Then the reaction mixture was concentrated and the residue was dissolved in a mixture of 20% isopropanol/chloroform (100 ml). The organic layer was washed with saturated sodium bicarbonate solution, dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography using combiflash (5-7% methanol in dichloromethane) to give (S)-4-amino-6-((1',8-dimethyl-1,5-dioxo-1,5-dihydro -2Hspiro[imidazo[1,5-a]pyridine-3,3'-piperidin]-6-yl)amino)pyrimidin-5-carbonitrile (Comp. No. 2Fb) as a white solid. Yield: 0.49 g, 18%. MS (ESI) m/z 381.17[M+1]+; ee=95.58%, Rt=8.89 min, Chiralpak IC (4.6x250 mm, 5 µm) using a methanol/carbon dioxide isomap gradient (40:60). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.02 (s, 1H), 8.58 (s, 1H), 8.49 (s, 1H), 8.34 (s, 1H), 7 .79 (brs, 2H), 3.29-3.28 (m, 1H), 2.94-2.91 (m, 1H), 2.81-2.80 (m, 1H), 2 .45 (s, 3H), 2.22 (s, 3H), 1.93-1.88 (m, 2H), 1.72-1.70 (m, 1H), 1.49 (d, J =12.76 Hz, 1H).

Example 3

Using appropriate reactants, the following compounds can be prepared according to general methods 1 or 2.

- 18 040075

WE 0 vAnh nW N Y- NH 0 h ₂ n \ J 3K O VAAh - _V W N p—NH O H ₂ N Cl 3L 0 \_Anh N 7 - NH O Pj H ₂ N Cl 3R 0 \_Λ-ΝΗ N E—NH °__J^ H ₂ N Cl ZM O \Anh N E—NH 0 h ₂ n ci 3S N 2“NH O |q H ₂ N Cl 3N o \Ανη NH O h ₂ n 3T N e—NH 0 P| _H2N ' thirty Xt9 N 2"NH O h ₂ n \ 3U O A ^NH QX NfVNH bf| N ZR ex _H2NCl rac-2F O \ Anh N ? - NH O H ₂ N \ N 3Q 0 \Anh rt w N^ NH H ₂ N 3V O \ Λ-ΝΗ rt.Qt) N p— NH O j ⁴ H ₂ N \ ' 3W 0 \__Anh ^XxX N 2“NH O 2Fb o \_Anh N y—NH O J ⁴ _h2nci ' h ₂ n\ N

- 19 040075

sx 0 A ^nh N NH 0 H ₂ N Cl 3Z O \AnH db N E-NH O H ₃ N Cl 3Y 0 \Anh QD N 3 - NH О H ₂ N Cl 3AA O yA ^NH NV—NH 0 ''--F H _? ^N'FF lFa L h ₂ n \ 3BB O y7% ⁿ rt w N y- NH 0 S h ₂ n ci )<_ ^f lFb o \Anh ,-00 N 2~NH O ^N H ₂ N 3CC O \γΝΗ rt QA N 7—NH 0 H ₂ N Cl 2 Fa 0 \Anh N QnH O J ¹¹ HiN^V N 3DD N NH 0 H ₂ N Cl 3EE O OChi N 7-NH 0 L H ₂ N Cl 3LL O \ Z^NH ^F V ^F Qtr Nfj-NHb H ₂ N \ 3FF N 7 - NH 0 ⁴ H ₂ N Cl 3MM L N Qnh O y ¹ h ₂ n \ N 3GG : V D , NV - NH 0 h ₂ n \ 3NN O vAnh N E—NH O J ⁴ h ₂ n \ N

- 20 040075

Example 4 6'-((6-amino-5-chloropyrimidin-4-yl)amino)-8'-methyl-2Ή-spiro[cyclohexane-1,3'-imidazo[1,5-a]pyridine]- 1',5'-dione hydrochloride (comp. No. 4A).

Compound 4a can be prepared according to the procedure described in Example 223 of US Pat. No. 9,382,248.

Biological research

Cellular analysis of peIF4E signaling activity.

Phosphorylated eIF4E was analyzed using the CisBio peIF4E HTRF® Assay Kit (CisBio, cat# 64EF4PEG). Cells were seeded in a 96-well tissue culture plate in the appropriate growth medium (90 μl). Compounds (10X) were diluted using 3-fold serial dilutions in cell culture medium and added to cells. The plates were incubated for 2 hours at 37°C. The cell supernatant was carefully removed either by aspiration of the supernatant or by tapping the plate. 50 μl of supplemented lysis buffer (1X) was immediately added and incubated for at least 30 min at room temperature with shaking. After homogenization by up and down pipetting, 16 μl of cell lysate was transferred from a 96-well cell culture plate to a small volume 384-well white plate. 4 μl of premixed antibody solutions (v/v) were prepared in detection buffer and added. The plate was covered with a plate sealer and incubated overnight at room temperature. Fluorescence emission at two different wavelengths was read (665 and 620 nm) on a Wallac Victor2. Emission ratios were converted to percent inhibition and imported into GraphPad Prism software. The compound concentration required to achieve 50% inhibition of enzyme activity (IC ₅₀ ) was calculated using concentrations ranging from 20 μM to 0.1 nM (12 point curve). IC ₅₀ values were determined using a non-linear regression model available in GraphPad Prism 5.

The results of these analyzes are presented in the table below. For this IC ₅₀ , values less than 0.001 μM are indicated as +++, from 0.001 to 0.01 μM are indicated as ++, and greater than 0.01 μM are indicated as + (NA means not known).

MNK Inhibition Decreases Expression of Immune Checkpoint Receptors and Ligands

Upon activation of the T cell receptor (TCR) signaling pathway, T cells proliferate, producing

- 21 040075 cytokines (eg IL-2) and induce expression of immune checkpoint receptors. Programmed cell death receptor 1 (PD-1) is an inhibitory checkpoint receptor expressed on the surface of activated T cells as well as on myeloid cells. The ligand for PD-1, programmed cell death ligand-1 (PD-L1, B7-H1/CD274), is not expressed by T cells or normal epithelial cells, but is expressed by antigen presenting cells and is overexpressed in some types of cancer. The interaction of PD-1 with PD-L1 leads to an antiproliferative effect on T cells and ultimately to T cell depletion and apoptosis. To investigate the role of MNK in activated T cells and tumor cells, the effect of an MNK inhibitor on immune checkpoint molecules was investigated.

Expression of PD-1 (CD279).

To study the effect of MNK inhibitors on PD-1 expression, Jurkat cells (Clone E6.1, ATCC, transformed T cells) were used, which express PD-1 when activated via T cell receptor (TCR) signaling. Briefly, Jurkat cells were grown in 1X RPMI with 1X penicillin/streptomycin and 10% FBS, then about 3x106 Jurkat cells were activated in the presence of 1 μg/ml PHA (Sigma) and 50 ng/ml PMA (Sigma). Test cells were treated simultaneously with various concentrations of MNK inhibitor (0, 0.01, 0.1, 1, 3 and 10 μM). After 48 hours, culture supernatants were harvested and tested in a sandwich ELISA for the presence of IL-2 using a DuoSet ELISA containing human IL-2 (R&D Systems, Minneapolis, MN). PD-1 levels on Jurkat cells were assayed by incubation with human FcR block followed by contact with allophycocyanin (APC)-conjugated anti-PD-1 antibody (5 µl per 100 µl assay volume, Biolegend, San Diego, CA) for 25 min at 4°C, without washing the cells, fixed dead cell stain (1:10,000; BD Biosciences, San Jose, CA) was added and incubated again for 10 min at 4°C. The cells were washed twice with running buffer and finally the cells were fixed with fixing buffer for 15 min at 4°C. After fixation, cells were washed twice with flow buffer, resuspended in flow buffer, and evaluated for fluorescence using a BD Accuri C6 flow cytometer. Data were analyzed using C6 cytometer software (BD Biosciences, San Jose, CA) or Attune Nxt Cytometer (Invitrogen, Carlsbad, CA).

Activation of Jurkat T cells by PHA and PMA induced cell surface expression of PD-1 in approximately 25-30% of stimulated Jurkat cells compared to non-induced cells (Unstim) and induced a 1000-fold increase in IL-2 cytokine production, respectively. Treatment of PHA/PMA-activated Jurkat T cells with an MNK inhibitor resulted in a concentration-dependent decrease in the expression of the immunoinhibitory PD-1 receptor, a decrease of up to 50% at the highest concentration compared to control. Furthermore, this decrease in PD-1 was not the result of blocking Jurkat T-cell activation per se, since inhibition of MNK by an MNK inhibitor did not alter cytokine production as measured by IL-2 levels. Finally, inhibition of MNK by various other MNK inhibitors had no effect on cell viability. Indeed, various other MNK inhibitors in the Jurkat T-cell assay have shown the ability to down-regulate immune checkpoint inhibitors without affecting cell viability. The results of these analyzes are presented in the table below. For this, the percentage inhibition of PD-1 positive cells (10 μM) greater than 50% is indicated as +++, from 10% to 50% is indicated as ++, and less than 10% is indicated as + (NA means not known).

Water solubility.

Solubility, the phenomenon of the dissolution of a solute in a solvent to form a homogeneous system, is one of the important parameters to achieve the desired concentration of a compound in the systemic circulation for the desired (expected) pharmacological response. Compounds having good water solubility are desirable because they result in good in vivo bioavailability due to their high dissolution rate after administration to a subject. Compounds having good water solubility also facilitate the development of compositions, the manufacture of compositions, and contribute to the stability of the compositions.

High performance thermodynamic dissolution procedure.

In a 96-well plate, 10 mM DMSO stock solutions (50-100 μl) of each compound were dried in a stream of heated nitrogen using a SPE-96 plate dryer (upstream flow rate=50 L/min, temperature=60°C, bottom flow rate =20 l/min, temperature=80°C). After DMSO was completely removed, the remaining substances were dissolved in test solvents, including deionized water, simulated fasting intestinal juice (FaSSIF, pH 6.5) and simulated fasting gastric juice (FaSSGF, pH 1.6). Compounds prepared in free base form were evaluated under conditions with or without one equivalent of TFA. The theoretical maximum concentrations of aqueous solutions were 10 mM. Each well was capped and incubated for 18 hours at room temperature or 37°C. Stirring of the solutions during incubation was accomplished by shaking the plate at 750 rpm or adding StirStix (stainless steel capillaries) and stirring the mixture using a rotary magnetic stirrer. After the incubation period

- 22 040075 Aliquots from each well were filtered. The solubility of all samples was quantified after comparison with known concentration standards using HPLC-UV.

The results of the water solubility analysis are presented in the table below.

pEIF4E signaling activity, PD-1 inhibitory activity and water solubility

Comm . No. pEIF4E signaling activity PD-1 inhibitory activity Water solubility deionized H ₂ O FaSSGF (pH 1.2) FaSSIF (pH 6.5) 3-piperidine-substituted compounds BEHIND + NA SW + AP ++ + + + 2, 6 4, 1 0.256 3D ++ 3.3 4.4 0.11 gas- 1F ++ ++ 4.3 9, 8 0.24 WE + WE ++ 0.24 0.7 <1 3G ++ ZN + 31 +++ 3J ++ ZK + + + + 1.53 1, 68 0.158 3L ++ + + + 3.2 3, 1 0.07 ZM 3N ++ ZO +++

- 23 040075

ZR + + + 3Q ++ 3R +++ >2.09 0.068 3S ++ +++ 4.2 10 0.6 ST + ++ 10 10 0.5 3U + gas- 2F ++ ++ 1, 84 0.079 3V + 1, 82 0.107 3W ++ 1.4 0.593 ZX +++ 3Y + + + 1Ea + ++ >1.85 1.53 0.125 IFb ++ ++ >1.85 >1.85 0.088 2 Fa ++ ++ 1.84 1.32 0.077 2Fb + ++ 1.77 1.77 0.151 311 3JJ ZKK 3LL ZMM 3NN ZOO ZRR 3QQ 3RR 3SS ZTT Cycloalkyl-substituted compounds 4A +++ +++ 0.0003 0.26 0.045 4-piperidine-substituted compounds 3Z ++ ZAA ++ + 4.4 3.2 0.03 ZVV +++ + ZSS ++ ++ 0.77 1.9 0.04 3DD +++ +++ 2.1 3 0.13 SEE ++ +++ 3FF +++ ++ 3GG ++ 3, 6 ZNN ++ + 2.4 0.018

The inventors unexpectedly found that the dissolution rate of the Mnk inhibitor compound can be significantly improved compared to comparative compound 4a by: (i)

-24040075 displacement of imidazopyridine in the compound with 3- or 4-piperidine; and (ii) substitution of the pyrimidine in the compound with a lower alkyl, halogen or cyano. Such substitutions or structural features cause chirality in molecules.

The improvement in the water solubility of the compounds of the invention is particularly noticeable in low pH environments, such as gastric fluid. For example, as shown in the table, compounds 3D, 3K, 3L, 3X, 3Y, rac-1F, 1Fa, 1Fb, rac-2F, 2Fa and 2Fb showed excellent solubility in simulated fasting gastric juice (FaSSGF) pH 1.6.

Importantly, in addition to improved water solubility, the 3- and 4-piperidine-substituted compounds also retained their anti-cancer activity in the form of their ability to inhibit pEIF4E signaling. In some cases, the anti-cancer activity is further manifested in the ability of these 3- and 4-piperidine-substituted compounds to inhibit PD-1, as can be seen in the table.

The various embodiments described above may be combined to provide additional embodiments. All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-patent publications cited in the present invention and/or listed in the application information sheet are incorporated herein by reference in their entirety. Aspects of the embodiments may be modified, if necessary, to take advantage of the concept of various patents, applications, and publications to provide other additional embodiments.

These and other variations of the embodiments are possible in light of the above detailed description. As a general rule, in the following claims, the terms used should not be construed as limiting the claims to the specific embodiments disclosed in the specification and claims, but should be construed as including all possible embodiments together with the full scope of equivalents covered by the claims. Accordingly, the claims are not limited to the disclosure.

Claims (8)

CLAIM 1. The compound or its stereoisomer, tautomer or pharmaceutically acceptable salt selected from the group consisting of - 25 040075 2. A compound or stereoisomer, tautomer or pharmaceutically acceptable salt thereof according to claim 1, selected from the group consisting of - 26 040075 3. A compound having a structure about or a pharmaceutically acceptable salt thereof. 4. A compound having a structure or a pharmaceutically acceptable salt thereof. 5. A compound according to any one of claims 1 to 4, wherein the pharmaceutically acceptable salt is an organic or inorganic acid salt selected from the group consisting of acetate, mesylate, sulfate, citrate, oxalate, hydrochloride, dihydrochloride, isethionate, lactate, and laurate. 6. A pharmaceutical composition for the treatment of an Mnk-dependent condition, comprising (i) a therapeutically effective amount of at least one compound or its stereoisomer, tautomer or pharmaceutically acceptable salt according to any one of claims 1 to 4; (ii) in combination with a pharmaceutically acceptable carrier, diluent or excipient. 7. A method of treating an Mnk-dependent condition in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of at least one compound according to any one of claims 1 to 4, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the Mnk-dependent condition is is cancer or inflammation. 8. Method for treating solid tumor, colorectal cancer, bladder cancer, gastric cancer, esophageal cancer, head and neck cancer, CNS cancer, malignant glioma, glioblastoma, hepatocellular cancers, thyroid cancer, lung cancer, non-small cell cancer, small cell lung cancer, melanoma, myeloma, pancreatic cancer, pancreatic carcinoma, renal cell carcinoma, cervical cancer, urothelial cancer, prostate cancer, castration-resistant prostate cancer, ovarian cancer, breast cancer, triple negative breast cancer, leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, B-cell lymphoma, T-cell lymphoma, hairy cell lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, multiple myeloma, and myelodysplastic syndrome in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of a compound according to any one of claims. -4 or its stereoisomer, a tautomer; or a pharmaceutically acceptable salt.