WO2020157188A1

WO2020157188A1 - Dihydrooxadiazinones for the treatment of hyperproliferative diseases

Info

Publication number: WO2020157188A1
Application number: PCT/EP2020/052276
Authority: WO
Inventors: Manuel ELLERMANN; Stefan Nikolaus GRADL; Charlotte Christine KOPITZ; Martin Lange; Adrian Tersteegen; Philip Lienau; Simon Anthony HERBERT; Detlev Sülzle; Timothy A. Lewis; Heidi GREULICH; Xiaoyun Wu
Original assignee: Bayer Aktiengesellschaft; The Broad Institute, Inc.
Priority date: 2019-02-01
Filing date: 2020-01-30
Publication date: 2020-08-06

Abstract

The present invention provides dihydrooxydiazinone compounds of general formula (I) : (I), in which R1, R2, R3, and R4, are as defined herein, methods of preparing said compounds, intermediate compounds useful for preparing said compounds, pharmaceutical compositions and combinations comprising said compounds and the use of said compounds for manufacturing pharmaceutical compositions for the treatment or prophylaxis of diseases, in particular of hyperproliferative diseases, as a sole agent or in combination with other active ingredients.

Description

DIHYDROOXADIAZINONES FOR THE TREATMENT OF HYPERPROLIFERATIVE DISEASES

The present invention provides dihydrooxadiazinone compounds of general formula (I) as described and defined herein, methods of preparing said compounds, pharmaceutical compositions and the use of said compounds for the treatment or prophylaxis of diseases, in particular of hyperproliferative diseases.

BACKGROUND

Cancer kills over 550,000 people in the United States and over 8 million people world-wide each year. New agents, including small molecules, molecules that impact tissue-specific growth requirements, and immunomodulatory agents, have been shown to benefit a subset of patients whose cancers have unique genomic mutations or other characteristics. Unfortunately, many cancer patients are still left without effective therapeutic options.

One approach to identify new anti-cancer agents is phenotypic screening to discover novel small molecules displaying strong selectivity between cancer cell lines, followed by predictive chemogenomics to identify the cell features associated with drug response. In the 1990s, Weinstein and colleagues demonstrated that the cytotoxic profile of a compound can be used to identify cellular characteristics, such as gene-expression profiles and DNA copy number, which correlate with drug sensitivity. The ability to identify the features of cancer cell lines that mediate their response to small molecules has strongly increased in recent years with automated high-throughput chemosensitivity testing of large panels of cell lines coupled with comprehensive genomic and phenotypic characterization of the cell lines. Phenotypic observations of small molecule sensitivity can be linked to expression patterns or somatic alterations, as in the case of trastuzumab-sensitive HER2- amplified breast cancer or erlotinib-sensitive EGFR-mutant lung cancer.

Phenotypic screening identified some of the compounds known in the literature to be PDE3 inhibitors to be useful for the treatment of certain cancers. Co-expression of PDE3A and/or PDE3B and Schlafen 12 (SLFN12) polynucleotides or polypeptides are typically required for cells to be sensitive. PDE3A and/or B inhibitors which cause drug sensitivity have been found to stabilze the formation of a complex between PDE3A and/or PDE3B and SLFN12. PDE3A and/or B inhibitors which do not cause inhibition of tumor cell proliferation typically do not stabilize the PDE3A- and/or PDE3B - SLFN12 complex.

From US 4,489,074, US 4,495,185 (EP0080296), JP59062578 are some oxazinones known a cardiatonic agents. Especially the cardiac mode of action mediated unwanted effects of PDE-3 inhibitors (Movsesian &Kukreja, S.H. Francis et al. (eds.), Phosphodiesterases as Drug Targets, Handbook of Experimental Pharmacology 204, 2011 ; p 237ff) may limit their therapeutic use when PDE3-inhibiting agents are used on a short- or/and long term basis, e.g. in cancer patients and a suitable therapeutic window is needed.

Some dihydrooxydiazinones are known, however, the state of the art does not describe the dihydrooxadiazinone compounds of general formula (I) of the present invention as described and defined herein.

SUMMARY

It has now been found, and this constitutes at least in part one basis of the present invention, that the compounds of the present invention have stil surprising and advantageous properties.

In particular, the compounds of the present invention have surprisingly been found to inhibit tumor cell proliferation with IC₅₀ values of < 100 nM in e.g. HeLa cells. Additionally, the compounds do not inhibit enzymatic PDE3A and/or PDE3B at the concentration at which they inhibit tumor cell proliferation but at concentrations where IC₅₀ values for enzymatic PDE3A and/or PDE3B inhibition may be > 10 times higher than IC₅₀ values for tumor cell proliferation. Without wishing to be bound by theory, this distinction in inhibitory properties may be associated with PDE3A and/or PDE3B-SLFN12 complex induction and/or improved pharmacokinetic parameters in vitro or in vivo and/or improved physicochemical properties and/or improved safety pharmacological properties. With these advantageous properties, the compounds described herein may therefore be used for the treatment or prophylaxis of hyperproliferative diseases, such as cancer diseases.

The present invention provides compounds of general formula (I) which modulate formation of a PDE3A-SLFN12 complex and/or PDE3B-SLFN12 complex, methods for their preparation, pharmaceutical composition and the use thereof and methods of treatment or prophylaxis of diseases, in particular of hyperproliferative diseases more particularly of cancer diseases. These and other features of the present teachings are set forth herein.

In accordance with a first aspect, the present invention provides compounds of general formula (I):

where

R¹ is selected from a hydrogen atom, a halogen atom, a cyano group, a CrC3-alkyl group, a CrC3-haloalkyl group, and a CrC3-haloalkoxy group;

R² is selected from a hydrogen atom and a halogen atom,

with the proviso that both, R¹ and R² can not be a hydrogen atom at the same time;

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from C1-C3- haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom or a CrC3-alkyl group;

or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

DETAILED DESCRIPTION

DEFINITIONS

Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this invention belongs. The following references provide one of skill with a general definition of many of the terms used in this invention: Singleton et al. , Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings ascribed to them below, unless specified otherwise.

Structures drawn include all permissible rotations about bonds. The term“substituted” means that one or more hydrogen atoms on the designated atom or group are replaced with a selection from the indicated group, provided that the designated atom's normal valency under the existing circumstances is not exceeded. Combinations of substituents and/or variables are permissible.

The term“optionally substituted” means that the number of substituents can be equal to or different from zero. Unless otherwise indicated, it is possible that optionally substituted groups are substituted with as many optional substituents as can be accommodated by replacing a hydrogen atom with a non-hydrogen substituent on any available carbon or nitrogen atom. Commonly, it is possible for the number of optional substituents, when present, to be 1 , 2, 3, in particular 1 , or 2.

As used herein, the term“one or more”, e.g. in the definition of the substituents of the compounds of general formula (I) of the present invention, means“1 , 2, 3, 4 or 5, particularly 1 , 2, 3 or 4, more particularly 1 , 2 or 3, even more particularly 1 or 2”.

When residues in the compounds according to the invention are substituted, it is possible for said groups to be mono-substituted or poly-substituted with substituent(s), unless otherwise specified. Within the scope of the present invention, the meanings of all groups which occur repeatedly are independent from one another. It is possible that groups in the compounds according to the invention are substituted with one, two or three identical or different substituents, particularly with one substituent.

The terms“oxo”, “an oxo group” or“an oxo substituent” mean an oxygen atom attached by a double bond =0. Oxo may be attached to atoms of suitable valency, for example to a saturated carbon atom or to a sulfur atom. For example, but without limitation, one oxo group can be attached to a carbon atom, resulting in the formation of a carbonyl group C(=0) , or two oxo groups can be attached to one sulfur atom, resulting in the formation of a sulfonyl group -S(=0)₂.

The term “ring substituent” means a substituent attached to an aromatic or nonaromatic ring which replaces an available hydrogen atom on the ring.

Should a composite substituent be composed of more than one parts, e.g. (Ci-C₄-alkyl)-0-(CrC₄-alkyl)-, a hyphen at the beginning or at the end of such a composite substituent indicates the point of attachment of said composite substituent to the rest of the molecule. Should the composite substituent be substituted said substitutent may be bound at any suitable carbon atom of the composite substitutent.

Should a ring, comprising carbon atoms and optionally one or more heteroatoms, such as nitrogen, oxygen or sulfur atoms for example, be substituted with a substituent, it is possible for said substituent to be bound at any suitable position of said ring, be it bound to a suitable carbon atom and/or to a suitable heteroatom.

The term“comprising” when used in the specification includes“consisting of’.

If within the present text any item is referred to as“as mentioned herein”, it means that it may be mentioned anywhere in the present text.

If within the present text any item is referred to as“supra” within the description it indicates any of the respective disclosures made within the specification in any of the preceding pages, or above on the same page.

If within the present text any item is referred to as“infra” within the description it indicates any of the respective disclosures made within the specification in any of the subsequent pages, or below on the same page.

The term “halogen atom” means a fluorine, chlorine, bromine or iodine atom, particularly a fluorine, chlorine or bromine atom.

The term“Ci-C3-alkyl-” means a linear or branched, saturated hydrocarbon group having 1 , 2, or 3, carbon atoms, such as, for example, a methyl-, ethyl-, propyl-, /so-propyl-, or 1 or 2 carbon atoms (“CrC2-alkyl-”), e.g., a methyl group or an ethyl group.

The same definitions can be applied should the alkyl group be placed within a chain as a bivalent“CrC3-alkylene” moiety. All names as mentioned above then will bear an“ene” added to the end, thus e.g., a“pentyl” becomes a bivalent“pentylene” group. In addition, the term“CrC3-heteroalkyl” refers to a CrC3-alkyl group in which one or more of the carbon atoms have been replaced with an atom selected from N, O, S, or P, which are substituted as mentioned herein to satisfy atom valency requirements.

The term“CrC3-hydroxyalkyl” means a linear or branched, saturated, monovalent hydrocarbon group in which the term“CrC33-alkyl” is defined supra, and in which 1 , 2 or 3 hydrogen atoms are replaced with a hydroxy group, such as, for example, a hydroxymethyl-, 1-hydroxyethyl-, 2-hydroxyethyl-, 1 ,2-dihydroxyethyl-, 3-hydroxypropyl-, 2-hydroxypropyl-, 1-hydroxypropyl-, 1-hydroxypropan-2-yl-, 2-hydroxypropan-2-yl-, 2,3- dihydroxypropyl-, or a 1 ,3-dihydroxypropan-2-yl-.

The term “Ci-C3-haloalkyl” means a linear or branched, saturated, monovalent hydrocarbon group in which the term“CrC3-alkyl” is as defined supra, and in which one or more of the hydrogen atoms are replaced, identically or differently, with a halogen atom. Particularly, said halogen atom is a fluorine atom. Said CrC3-haloalkyl group is, for example, a fluoromethyl-, difluoromethyl-, trifluoromethyl-, 1-fluoroethyl-, 2-fluoroethyl-, 1 , 1-difluoroethyl-, 2,2-difluoroethyl-, 2,2,2-trifluoroethyl-, 2-fluoro-propyl, pentafluoroethyl- , 3,3,3-trifluoropropyl- or a 1 ,3-difluoropropan-2-yl group. Particularly haloalkyl is trifluoromethyl or difluoromethyl.

The term“CrC₆-alkoxy” means a linear or branched, saturated, monovalent group of formula (CrC₆-alkyl)-0-, in which the term“CrC₆-alkyl” is as defined supra, such as, for example, a methoxy-, ethoxy-, n-propoxy-, isopropoxy-, n-butoxy-, sec-butoxy-, isobutoxy- , tert-butoxy-, pentyloxy-, isopentyloxy or a n-hexyloxy group, or an isomer thereof.

The term“CrC3-haloalkoxy” means a linear or branched, saturated, monovalent CrC3-alkoxy group, as defined supra, in which one or more of the hydrogen atoms is replaced, identically or differently, with a halogen atom. Particularly, said halogen atom is a fluorine atom. Said CrC3-haloalkoxy group is, for example, a fluoromethoxy-, difluoromethoxy-, trifluoromethoxy-, 2,2,2-trifluoroethoxy- or a pentafluoroethoxy group.

The same definitions can be applied should the alkenyl group be placed within a chain as a bivalent“CrC₆-alkenylene” moiety. All names as mentioned above then will bear a“ene” added to their end, thus e.g., a“pentenyl” becomes a bivalent“pentenylene” group.

The term“C4-C6-cycloalkyl-” means a saturated monocyclic or bicyclic hydrocarbon ring which contains 4, 5, or 6, carbon atoms (“C4-C6-cycloalkyl-”). Said C4-C6-cycloalkyl- group may be, for example, a monocyclic hydrocarbon ring, such as, for example, cyclobutyl-, cyclopentyl-, cyclohexyl- ring. A cycloalkyl group may be optionally substituted as defined at the respective part wherein such term is used.

The term“4- to 6-membered heterocycloalkyl” means a saturated or a partially unsaturated heterocycle with 4, 5, or 6 ring atoms respectively, 4, 5 or 6 ring atoms in total, which contains one or two identical or different ring heteroatoms selected from the series N, O, and S, said heterocycloalkyl group being attached to the rest of the molecule via any one of the carbon atoms or if present, a nitrogen atom.

Said heterocycloalkyl group, without being limited thereto, can be a 4-membered ring, such as, for example, a azetidinyl-, oxetanyl- or thietanyl group; or a 5-membered ring, such as a tetrahydrofuranyl-, 1 ,3-dioxolanyl-, thiolanyl-, pyrrolidinyl-, imidazolidinyl-, pyrazolidinyl-, 1 , 1-dioxidothiolanyl-, 1 ,2-oxazolidinyl-, 1 ,3-oxazolidinyl- or a 1 ,3-thiazolidinyl group, for example; or a 6-membered ring, such as, for example, a tetrahydropyranyl-, tetrahydrothiopyranyl-, piperidinyl-, morpholinyl-, dithianyl-, thiomorpholinyl-, piperazinyl-, 1 ,3-dioxanyl-, 1 ,4-dioxanyl- or a 1 ,2-oxazinanyl group, for example. Particularly, “4- to 6-membered heterocycloalkyl” means a 4- to 6-membered heterocycloalkyl as defined supra containing one ring nitrogen atom or an oxygen atom or a sulfur atom and if it contains a nitrogen atom it may optionally contain one further ring heteroatom from the series: N, O, S. More particularly, “5- or 6-membered heterocycloalkyl” means a monocyclic, saturated heterocycle with 5 or 6 ring atoms in total, containing one ring nitrogen atom and optionally one further ring heteroatom from the series: N, O. Said heterocycloalkyl group is being attached to the rest of the molecule via any carbon atom or where applicable via any nitrogen atom.

The term“heteroaryl” means a monovalent, monocyclic or bicyclic aromatic ring having 5, or 6 ring atoms (a“5- to 6-membered heteroaryl” group), which contains at least one ring heteroatom and optionally one, two or three further ring heteroatoms from the series: N, O and/or S, and which is bound via a ring carbon atom or a heteroatom to the rest of the molecule (if allowed by valency).

Said heteroaryl group can be a 5-membered heteroaryl group, such as, for example, a thienyl-, furanyl-, pyrrolyl-, oxazolyl-, thiazolyl-, imidazolyl-, pyrazolyl-, isoxazolyl-, isothiazolyl-, oxadiazolyl-, triazolyl-, thiadiazolyl- or a tetrazolyl group; or a 6-membered heteroaryl group, such as, for example, a pyridyl-, pyridazinyl-, pyrimidyl-, pyrazinyl- or a triazinyl group;

In general, and unless otherwise mentioned, the heteroaryl or heteroarylene groups include all possible isomeric forms thereof, e.g.: tautomers and positional isomers with respect to the point of linkage to the rest of the molecule. Thus, for some illustrative non-restricting examples, the term pyridinyl includes pyridin-2-yl, pyridin-3-yl and pyridin-4-yl; or the term thienyl includes thien-2-yl and thien-3-yl.

The term“Oi-Ob”, as used in the present text, e.g. in the context of the definition of “CrC₆-alkyl”,“CrC₆-haloalkyl”,“CrC₆-alkoxy” or“CrC₆-haloalkoxy” is to be understood as meaning an alkyl group having a whole number of carbon atoms of 1 to 6, i.e. 1 , 2, 3, 4, 5 or 6 carbon atoms. It is to be understood further that said term “CrCe” is to be interpreted as disclosing any sub-range comprised therein, e.g. Ci-Ce _, C₂-C_{5 ,} C₃-C_{4 ,} Cr C2 , C1-C3 , C1-C4 , C1-C5 , C1-C6 ; particularly C1-C2 , C1-C3 , C1-C4 , C1-C5 , C1-C6 , more particularly C₁-C₄ ; in the case of “CrC₆-haloalkyl-” or“CrC₆-haloalkoxy-” even more particularly C₁-C₂.

Similarly, as used herein, the term“C2-C6”, as used throughout this text, e.g., in the context of the definitions of “C2-C6-alkenyl-” and“C2-C6-alkynyl-”, is to be understood as meaning an alkenyl- group or an alkynyl group having a whole number of carbon atoms from 2 to 6, i.e., 2, 3, 4, 5 or 6 carbon atoms. It is to be understood further that said term “C₂-C₆” is to be interpreted as disclosing any sub-range comprised therein, e.g., C₂-C_{6 ,} C₃- C5 , C3-C4 , C2-C3 , C2-C4 , C2-C5 ; particularly C2-C3.

Further, as used herein, the term“C₃-C₈”, as used in the present text, e.g. in the context of the definition of “Cs-Cs-cycloalkyl”, means a cycloalkyl group having a finite number of carbon atoms of 3 to 8, i.e. 3, 4, 5, 6, 7 or 8 carbon atoms.

Unless specifically stated or obvious from context, as used herein, the term“about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. About can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1 %, 0.5%, 0.1 %, 0.05%, or 0.01 % of the stated value. Unless otherwise clear from context, all numerical values provided herein are modified by the term about.

By "agent" is meant any small molecule chemical compound, antibody, nucleic acid molecule, or polypeptide, or fragments thereof.

By “ameliorate” is meant decrease, suppress, attenuate, diminish, arrest, or stabilize the development or progression of a disease.

By "analog" is meant a molecule that is not identical, but has analogous functional or structural features. For example, a polypeptide analog retains the biological activity of a corresponding naturally-occurring polypeptide, while having certain biochemical modifications that enhance the analog's function relative to a naturally occurring polypeptide. Such biochemical modifications could increase the analog's protease resistance, membrane permeability, or half-life, without altering, for example, ligand binding. An analog may include an unnatural amino acid.

In this disclosure, "comprises," "comprising," "containing" and "having" and the like can have the meaning ascribed to them in U.S. Patent law and can mean " includes," "including," and the like; "consisting essentially of" or "consists essentially" likewise has the meaning ascribed in U.S. Patent law and the term is open-ended, allowing for the presence of more than that which is recited so long as basic or novel characteristics of that which is recited is not changed by the presence of more than that which is recited, but excludes prior art embodiments.

“Detect” refers to identifying the presence, absence or amount of the analyte to be detected. In particular embodiments, the analyte is a PDE3A and/or PDE3B or SLFN12 polypeptide.

By“disease” is meant any condition or disease that damages or interferes with the normal function of a cell, tissue, or organ. Examples of diseases include hyperproliferatiotive disorder, cancer types such as e.g., adenocarcinoma, breast cancer, cervical cancer, liver cancer, lung cancer and melanoma.

By "effective amount" is meant the amount of a compound described herein required to ameliorate the symptoms of a disease relative to an untreated patient. The effective amount of active compound(s) used to practice the present invention for therapeutic treatment of a disease varies depending upon the manner of administration, the age, body weight, and general health of the subject. Ultimately, the attending physician or veterinarian will decide the appropriate amount and dosage regimen. Such amount is referred to as an "effective" amount.

In still other embodiments, the PDE3A and/or PDE3B modulator is a compound of formula (I).

As used herein, the term“leaving group” means an atom or a group of atoms that is displaced in a chemical reaction as stable species taking with it the bonding electrons. In particular, such a leaving group is selected from the group comprising: halide, in particular a chloro-, bromo- or iodo group, a (methylsulfonyl)oxy-, [(4- methylphenyl)sulfonyl]oxy-, [(trifluoromethyl)sulfonyl]oxy-, [(nonafluorobutyl)sulfonyl]oxy-, [(4-bromophenyl)sulfonyl]oxy-, [(4-nitrophenyl)sulfonyl]oxy-, [(2-nitrophenyl)sulfonyl]oxy-, [(4-isopropylphenyl)sulfonyl]oxy-, [(2,4,6-triisopropylphenyl)sulfonyl]oxy-, [(2,4,6- trimethylphenyl)sulfonyl]oxy-, [(4-te/f-butylphenyl)sulfonyl]oxy-, (phenylsulfonyl)oxy- and a [(4-methoxyphenyl)sulfonyl]oxy group.

By “modulator” is meant any agent that binds to a polypeptide and alters a biological function or activity of the polypeptide. A modulator includes, without limitation, agents that reduce or eliminate a biological function or activity of a polypeptide (e.g., an “inhibitor”). For example, a modulator may inhibit a catalytic activity of a polypeptide. A modulator includes, without limitation, agents that increase or decrease binding of a polypeptide to another agent. For example, a modulator may promote binding of a polypeptide to another polypeptide.

Unless specifically stated or obvious from context, as used herein, the term "or" is understood to be inclusive. Unless specifically stated or obvious from context, as used herein, the terms "a", "an", and "the" are understood to be singular or plural.

As used herein, the term“protective group” is a protective group attached to an oxygen or nitrogen atom in intermediates used for the preparation of compounds of the general formula (I). Such groups are introduced e.g., by chemical modification of the respective hydroxy or amino group in order to obtain chemoselectivity in a subsequent chemical reaction. Protective groups for hydroxy and amino groups are descibed for example in T.W. Greene and P.G.M. Wuts in Protective Groups in Organic Synthesis, 4th edition, Wiley 2006; more specifically, protective groups for amino groups can be selected from substituted sulfonyl groups, such as a mesyl-, tosyl- or a phenylsulfonyl group, acyl groups such as a benzoyl-, acetyl- or a tetrahydropyranoyl group, or carbamate based groups, such as a tert-butoxycarbonyl group (Boc). Protective groups for hydroxy groups can be selected from acyl groups such as a benzoyl-, acetyl, pivaloyl- or a tetrahydropyranoyl group, or can include silicon, as in e.g., a tert-butyldimethylsilyl-, tert- butyldiphenylsilyl-, triethylsilyl- or a triisopropylsilyl group.

Where the plural form of the word compounds, salts, polymorphs, hydrates, solvates and the like, is used herein, this is taken to mean also include a single compound, salt, polymorph, isomer, hydrate, solvate or the like.

The term“prodrugs” or“prodrug” designates compounds which themselves can be biologically active or inactive, but are converted (for example metabolically or hydrolytically) into compounds according to the invention during their residence time in the body. Derivatives of the compound 6 and the salts thereof which are converted into compound 6 or a salt thereof in a biological system (bioprecursors or pro-drugs) are covered by the invention. Said biological system may be, for example, a mammalian organism, particularly a human subject. The bioprecursor is, for example, converted into a compound of formula (I) or a salt thereof by metabolic processes.

The term“pharmaceutically acceptable salt(s)” of the compounds of formula (I) include those derived from pharmaceutically acceptable inorganic and organic acids and bases. For example, see S. M. Berge, et al.“Pharmaceutical Salts,” J. Pharm. Sci. 1977, 66, 1-19.

As used herein, the term“pharmaceutically acceptable salt” refers to a salt formed by the addition of a pharmaceutically acceptable acid or base to a compound disclosed herein.

As used herein, the phrase“pharmaceutically acceptable” refers to a substance that is acceptable for use in pharmaceutical applications from a toxicological perspective and does not adversely interact with the active ingredient.

Unless specifically stated or obvious from context, as used herein, if a range is provided, the upper and lower limit are always meant to be included. Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub- range from the group consisting 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50.

By“reference” is meant a standard or control condition.

By "stable compound' or "stable structure" is meant a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.

The term“substituent” refers to a group“substituted” on, e.g., an alkyl, haloalkyl, cycloalkyl, heterocyclyl, heterocycloalkenyl, cycloalkenyl, aryl, or heteroaryl group at any atom of that group, replacing one or more hydrogen atoms therein. In one aspect, the substituent(s) on a group are independently any one single, or any combination of two or more of the permissible atoms or groups of atoms delineated for that substituent. In another aspect, a substituent may itself be substituted with any one of the above substituents. Further, as used herein, the phrase “optionally substituted” means unsubstituted (e.g., substituted with an H) or substituted.

The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

It will be understood that the description of compounds herein is limited by principles of chemical bonding known to those skilled in the art. Accordingly, where a group may be substituted by one or more of a number of substituents, such substitutions are selected so as to comply with principles of chemical bonding with regard to valencies, etc. and to give compounds which are not inherently unstable. For example, any carbon atom will be bonded to two, three, or four other atoms, consistent with the four valence electrons of carbon.

Compound Forms and Salts

It is possible for the compounds of formula (I) to exist in free form, e.g. as a free base, or as a free acid, or as a zwitterion, or to exist in the form of a salt. Said salt may be any salt, either an organic or inorganic addition salt, particularly any pharmaceutically acceptable organic or inorganic addition salt, which is customarily used in pharmacy, or which is used, for example, for isolating or purifying the compounds of the present invention.

A salt, for example, can be formed between an anion and a positively charged substituent (e.g., amino) on a compound described herein. Suitable anions include chloride, bromide, iodide, sulfate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, and acetate. Likewise, a salt can also be formed between a cation and a negatively charged substituent (e.g., carboxylate) on a compound described herein. Suitable cations include sodium ion, potassium ion, magnesium ion, calcium ion, and an ammonium cation such as tetramethylammonium ion. Examples of prodrugs include Ci-e alkyl esters of carboxylic acid groups, which, upon administration to a subject, are capable of providing active compounds.

A suitable pharmaceutically acceptable salt of the compounds of the present invention may be, for example, an acid-addition salt of a compound of the present invention bearing a nitrogen atom, in a chain or in a ring, for example, which is sufficiently basic, such as an acid-addition salt with an inorganic acid, or “mineral acid”, such as hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfamic, bisulfuric, phosphoric, or nitric acid, for example, or with an organic acid, such as formic, acetic, acetoacetic, pyruvic, trifluoroacetic, propionic, butyric, hexanoic, heptanoic, undecanoic, lauric, benzoic, salicylic, 2-(4-hydroxybenzoyl)-benzoic, camphoric, cinnamic, cyclopentanepropionic, digluconic, 3-hydroxy-2-naphthoic, nicotinic, pamoic, pectinic, 3-phenylpropionic, pivalic, 2-hydroxyethanesulfonic, itaconic, trifluoromethanesulfonic, dodecylsulfuric, ethanesulfonic, benzenesulfonic, para-toluenesulfonic, methanesulfonic, 2- naphthalenesulfonic, naphthalinedisulfonic, camphorsulfonic acid, citric, tartaric, stearic, lactic, oxalic, malonic, succinic, malic, adipic, alginic, maleic, fumaric, D-gluconic, mandelic, ascorbic, glucoheptanoic, glycerophosphoric, aspartic, sulfosalicylic, or thiocyanic acid, for example.

Further examples of suitable acid salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, succinate, sulfate, tartrate, thiocyanate, tosylate and undecanoate. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the present invention and their pharmaceutically acceptable acid addition salts.

Further, another suitably pharmaceutically acceptable salt of a compound of formula (I), which is sufficiently acidic, is an alkali metal salt, for example a sodium or potassium salt, an alkaline earth metal salt, for example a calcium, magnesium or strontium salt, or an aluminium or a zinc salt, or an ammonium salt derived from ammonia or from an organic primary, secondary or tertiary amine having 1 to 20 carbon atoms, such as ethylamine, diethylamine, triethylamine, ethyldiisopropylamine, monoethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, dimethylaminoethanol, diethylaminoethanol, tris(hydroxymethyl)aminomethane, procaine, dibenzylamine, N- methylmorpholine, arginine, lysine, 1 ,2-ethylenediamine, N-methylpiperidine, N-methyl- glucamine, N,N-dimethyl-glucamine, N-ethyl-glucamine, 1 ,6-hexanediamine, glucosamine, sarcosine, serinol, 2-amino-1 , 3-propanediol, 3-amino-1 , 2-propanediol, 4- amino-1 ,2,3-butanetriol, or a salt with a quarternary ammonium ion having 1 to 20 carbon atoms, such as tetramethylammonium, tetraethylammonium, tetra(n-propyl)ammonium, tetra(n-butyl)ammonium, N-benzyl-N,N,N-trimethylammonium, choline or benzalkonium.

In certain embodiments salts are derived from appropriate bases include alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium and N-(alkyl)4⁺ salts. The present invention also envisions the quaternization of any basic nitrogen- containing groups of the compounds disclosed herein. Water or oil-soluble or dispersible products may be obtained by such quaternization. Salt forms of the compounds of any of the formulae herein can be amino acid salts of carboxyl groups (e.g., L-arginine, -lysine, - histidine salts).

Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418; Journal of Pharmaceutical Science, 66, 2 (1977); and "Pharmaceutical Salts: Properties, Selection, and Use A Handbook; Wermuth, C. G. and Stahl, P. H. (eds.) Verlag Helvetica Chimica Acta, Zurich, 2002 [ISBN 3-906390-26-8] each of which is incorporated herein by reference in their entireties.

Those skilled in the art will further recognise that it is possible for acid addition salts of the claimed compounds to be prepared by reaction of the compounds with the appropriate inorganic or organic acid via any of a number of known methods. Alternatively, alkali and alkaline earth metal salts of acidic compounds of the present invention are prepared by reacting the compounds of the present invention with the appropriate base via a variety of known methods.

The present invention includes all possible salts of the compounds of the present invention as single salts, or as any mixture of said salts, in any ratio.

The neutral forms of the compounds may be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise the salts are equivalent to the parent form of the compound for the purposes of the present invention.

In the present text, in particular in the Experimental Section, for the synthesis of intermediates and of examples of the present invention, when a compound is mentioned as a salt form with the corresponding base or acid, the exact stoichiometric composition of said salt form, as obtained by the respective preparation and/or purification process, is, in most cases, unknown.

Unless specified otherwise, suffixes to chemical names or structural formulae relating to salts, such as "hydrochloride", "trifluoroacetate", "sodium salt", or "x HCI", "x CF₃COOH", "x Na⁺", for example, mean a salt form, the stoichiometry of which salt form not being specified.

This applies analogously to cases in which synthesis intermediates or example compounds or salts thereof have been obtained, by the preparation and/or purification processes described, as solvates, such as hydrates, with (if defined) unknown stoichiometric composition.

In addition to salt forms, the present invention provides compounds which are in a prodrug form. Prodrugs of the compounds described herein are those compounds that undergo chemical changes under physiological conditions to provide the compounds of the present invention. Additionally, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to the compounds of the present invention when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent. Prodrugs are often useful because, in some situations, they may be easier to administer than the parent drug. They may, for instance, be more bioavailable by oral administration than the parent drug. The prodrug may also have improved solubility in pharmacological compositions over the parent drug. A wide variety of prodrug derivatives are known in the art, such as those that rely on hydrolytic cleavage or oxidative activation of the prodrug. An example, without limitation, of a prodrug would be a compound of the present invention which is administered as an ester (the "prodrug"), but then is metabolically hydrolyzed to the carboxylic acid, the active entity. Additional examples include peptidyl derivatives of a compound of the present invention. The compounds of the present invention can exist as a hydrate, or as a solvate, wherein the compounds of the present invention contain polar solvents, in particular water, methanol or ethanol for example, as structural element of the crystal lattice of the compounds. It is possible for the amount of polar solvents, in particular water, to exist in a stoichiometric or non-stoichiometric ratio. In the case of stoichiometric solvates, e.g. a hydrate, hemi-, (semi-), mono-, sesqui-, di-, tri-, tetra-, penta- etc. solvates or hydrates, respectively, are possible. The present invention includes all such hydrates or solvates.

The present invention also includes various hydrate and solvate forms of the compounds.

It is possible for the compounds of general formula (I) to exist as isotopic variants. The invention therefore includes one or more isotopic variant(s) of the compounds of general formula (I), particularly deuterium-containing compounds of general formula (I).

The term“Isotopic variant” of a compound or a reagent is defined as a compound exhibiting an unnatural proportion of one or more of the isotopes that constitute such a compound.

The term“Isotopic variant of the compound of general formula (I)” is defined as a compound of general formula (I) exhibiting an unnatural proportion of one or more of the isotopes that constitute such a compound.

The expression“unnatural proportion” means a proportion of such isotope which is higher than its natural abundance. The natural abundances of isotopes to be applied in this context are described in“Isotopic Compositions of the Elements 1997”, Pure Appl. Chem., 70(1), 217-235, 1998. Examples of such isotopes include stable and radioactive isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, bromine and iodine, such as ²H (deuterium), ³H (tritium), ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁷0, ¹⁸0, ³²P, ³³P, ³³S, ³⁴S, ³⁵S, ³⁶S, ¹⁸F, ³⁶CI, ⁸² Br, ¹²³l, ¹²⁴l, ¹²⁵l, ¹²⁹l and ¹³¹1, respectively.

The compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may be radiolabeled with radioactive isotopes, such as for example tritium (³H), iodine-125 (¹²⁵l) or carbon-14 (¹⁴C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention particularly deuterium-containing compounds of formula

(I)·

formula (la)

Formula (la) shows the positions D¹, D², D³ and D⁴ in which anyone of the hydrogen atoms may be exchanged by a deuterium atom. Additionally in residues R¹-R⁴ if these residues contain a heteroatom-H or carbon-H bond accessible for a chemical reaction such an exchange may be possible. Hydrogen atoms can be replaced by deuterium atoms using methods known to those with ordinary skill in the art to obtain a heteroatom- D or carbon-D bond. Anyone of R¹, R², or R⁴ themselves can also be deuterium instead of hydrogen.

Thus one aspct of the invention are those compounds wherein independently anyone of R¹, R², or R⁴ is deuterium and/or anyone of the hydrogen atoms as shown in formula (I) are replaced by a deuterium atom and or anyone of R¹, R², R³,or R⁴ bears a deuterium atom at a chemically accessible position or any combination of positions being deuterated at the same time.

With respect to the treatment and/or prophylaxis of the diseases specified herein the isotopic variant(s) of the compounds of general formula (I) may contain deuterium (“deuterium-containing compounds of general formula (I)”). Isotopic variants of the compounds of general formula (I) in which one or more radioactive isotopes, such as ³H or ¹⁴C, are incorporated are useful e.g. in drug and/or substrate tissue distribution studies. These isotopes are particularly preferred for the ease of their incorporation and detectability. Positron emitting isotopes such as ¹⁸F or ¹¹C may be incorporated into a compound of general formula (I). These isotopic variants of the compounds of general formula (I) are useful for in vivo imaging applications. Deuterium-containing and ¹³C- containing compounds of general formula (I) can be used in mass spectrometry analyses in the context of preclinical or clinical studies.

Isotopic variants of the compounds of general formula (I) can generally be prepared by methods known to a person skilled in the art, such as those described in the schemes and/or examples herein, by substituting a reagent for an isotopic variant of said reagent, particularly for a deuterium-containing reagent. Depending on the desired sites of deuteration, in some cases deuterium from D2O can be incorporated either directly into the compounds or into reagents that are useful for synthesizing such compounds. Deuterium gas is also a useful reagent for incorporating deuterium into molecules. Catalytic deuteration of olefinic bonds and acetylenic bonds is a direct route for incorporation of deuterium. Metal catalysts (i.e. Pd, Pt, and Rh) in the presence of deuterium gas can be used to directly exchange deuterium for hydrogen in functional groups containing hydrocarbons. A variety of deuterated reagents and synthetic building blocks are commercially available from companies such as for example C/D/N Isotopes, Quebec, Canada; Cambridge Isotope Laboratories Inc., Andover, MA, USA; and CombiPhos Catalysts, Inc., Princeton, NJ, USA.

The term“deuterium-containing compound of general formula (I)” is defined as a compound of general formula (I), in which one or more hydrogen atom(s) is/are replaced by one or more deuterium atom(s) and in which the abundance of deuterium at each deuterated position of the compound of general formula (I) is higher than the natural abundance of deuterium, which is about 0.015%. Particularly, in a deuterium-containing compound of general formula (I) the abundance of deuterium at each deuterated position of the compound of general formula (I) is higher than 10%, 20%, 30%, 40%, 50%, 60%, 70% or 80%, particularly higher than 90%, 95%, 96% or 97%, even more particularly higher than 98% or 99% at said position(s). It is understood that the abundance of deuterium at each deuterated position is independent of the abundance of deuterium at other deuterated position(s).

The selective incorporation of one or more deuterium atom(s) into a compound of general formula (I) may alter the physicochemical properties (such as for example acidity [C. L. Perrin, et al. , J. Am. Chem. Soc., 2007, 129, 4490], basicity [C. L. Perrin et al., J. Am. Chem. Soc., 2005, 127, 9641], lipophilicity [B. Testa et al., Int. J. Pharm., 1984, 19(3), 271]) and/or the metabolic profile of the molecule and may result in changes in the ratio of parent compound to metabolites or in the amounts of metabolites formed. Such changes may result in certain therapeutic advantages and hence may be preferred in some circumstances. Reduced rates of metabolism and metabolic switching, where the ratio of metabolites is changed, have been reported (A. E. Mutlib et al., Toxicol. Appl. Pharmacol., 2000, 169, 102). These changes in the exposure to parent drug and metabolites can have important consequences with respect to the pharmacodynamics, tolerability and efficacy of a deuterium-containing compound of general formula (I). In some cases deuterium substitution reduces or eliminates the formation of an undesired or toxic metabolite and enhances the formation of a desired metabolite (e.g. Nevirapine: A. M. Sharma et al., Chem. Res. Toxicol., 2013, 26, 410; Efavirenz: A. E. Mutlib et al., Toxicol. Appl. Pharmacol., 2000, 169, 102). In other cases the major effect of deuteration is to reduce the rate of systemic clearance. As a result, the biological half-life of the compound is increased. The potential clinical benefits would include the ability to maintain similar systemic exposure with decreased peak levels and increased trough levels. This could result in lower side effects and enhanced efficacy, depending on the particular compound’s pharmacokinetic/ pharmacodynamic relationship. ML-337 (C. J. Wenthur et al., J. Med. Chem., 2013, 56, 5208) and Odanacatib (K. Kassahun et al., WO2012/112363) are examples for this deuterium effect. Still other cases have been reported in which reduced rates of metabolism result in an increase in exposure of the drug without changing the rate of systemic clearance (e.g. Rofecoxib: F. Schneider et al., Arzneim. Forsch. / Drug. Res., 2006, 56, 295; Telaprevir: F. Maltais et al., J. Med. Chem., 2009, 52, 7993). Deuterated drugs showing this effect may have reduced dosing requirements (e.g. lower number of doses or lower dosage to achieve the desired effect) and/or may produce lower metabolite loads.

A compound of general formula (I) may have multiple potential sites of attack for metabolism. To optimize the above-described effects on physicochemical properties and metabolic profile, deuterium-containing compounds of general formula (I) having a certain pattern of one or more deuterium-hydrogen exchange(s) can be selected. Particularly, the deuterium atom(s) of deuterium-containing compound(s) of general formula (I) is/are attached to a carbon atom and/or is/are located at those positions of the compound of general formula (I), which are sites of attack for metabolizing enzymes such as e.g. cytochrome P450.

In some aspects, the compounds of formula (I) may be isomers. "Isomers" are different compounds that have the same molecular formula. "Stereoisomers" are isomers that differ only in the way the atoms are arranged in space. As used herein, the term "isomer" includes any and all geometric isomers and stereoisomers. For example, "isomers" include geometric double bond cis- and trans-isomers, also termed E- and Z- isomers; R- and S-enantiomers; diastereomers, (d)-isomers and (l)-isomers, racemic mixtures thereof; and other mixtures thereof, as falling within the scope of this invention The symbol— denotes a bond that can be a single or a double bond as described herein. Provided herein are various geometric isomers and mixtures thereof resulting from the arrangement of substituents around a carbon-carbon double bond or arrangement of substituents around a carbocyclic ring. Substituents around a carbon-carbon double bond are designated as being in the "Z" or "E" configuration wherein the terms "Z" and "E" are used in accordance with lUPAC standards. Unless otherwise specified, structures depicting double bonds encompass both the "E" and "Z" isomers.

Substituents around a carbon-carbon double bond alternatively can be referred to as "cis" or "trans," where "cis" represents substituents on the same side of the double bond and "trans" represents substituents on opposite sides of the double bond. The arrangement of substituents around a carbocyclic ring can also be designated as "cis" or "trans." The term "cis" represents substituents on the same side of the plane of the ring, and the term "trans" represents substituents on opposite sides of the plane of the ring. Mixtures of compounds wherein the substituents are disposed on both the same and opposite sides of plane of the ring are designated "cis/trans."

The compounds of the present invention optionally contain one or more asymmetric centres, depending upon the location and nature of the various substituents desired, which are e.g. carbon atoms having four different substituents. It is possible that one or more asymmetric carbon atoms are present in the (R) or (S) configuration, which can result in racemic mixtures in the case of a single asymmetric centre, and in diastereomeric mixtures in the case of multiple asymmetric centres.

The term "(±)" is used to designate a racemic mixture where appropriate.

In certain instances, it is possible that asymmetry also be present due to restricted rotation about a given bond, for example, the central bond adjoining two substituted aromatic rings of the specified compounds.

When a compound is an enantiomer, the stereochemistry at each chiral carbon can be specified by either (R) or (S). Resolved compounds whose absolute configuration is unknown can be designated (+) or (-) depending on the direction (dextro- or levorotatory) which they rotate plane polarized light at the wavelength of the sodium D line. Certain of the compounds described herein contain one or more asymmetric centers and can thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined, in terms of absolute stereochemistry at each asymmetric atom, as (R)- or (S)-. The present chemical entities, pharmaceutical compositions and methods are meant to include all such possible isomers, including racemic mixtures, optically substantially pure forms and intermediate mixtures.

Preferred compounds are those which produce the more desirable biological activity should they be different for each of the isomers. Separated, pure or partially purified isomers and stereoisomers or racemic or diastereomeric mixtures of the compounds of the present invention are also included within the scope of the present invention. The purification and the separation of such materials can be accomplished by standard techniques known in the art including chiral high pressure liquid chromatography (HPLC), the formation and crystallization of chiral salts, or prepared by asymmetric syntheses.

The optical isomers can be obtained by resolution of the racemic mixtures according to conventional processes, for example, by the formation of diastereoisomeric salts using an optically active acid or base or formation of covalent diastereomers. Examples of appropriate acids are tartaric, diacetyltartaric, ditoluoyltartaric and camphorsulfonic acid. Mixtures of diastereoisomers can be separated into their individual diastereomers on the basis of their physical and/or chemical differences by methods known in the art, for example, by chromatography or fractional crystallisation. The optically active bases or acids are then liberated from the separated diastereomeric salts. A different process for separation of optical isomers involves the use of chiral chromatography (e.g., HPLC columns using a chiral phase), with or without conventional derivatisation, optimally chosen to maximise the separation of the enantiomers. Suitable HPLC columns using a chiral phase are commercially available, such as those manufactured by Daicel, e.g., Chiracel OD and Chiracel OJ, for example, among many others, which are all routinely selectable. Enzymatic separations, with or without derivatisation, are also useful. The optically active compounds of the present invention can likewise be obtained by chiral syntheses utilizing optically active starting materials.

In order to distinguish different types of isomers from each other reference is made to lUPAC Rules Section E (Pure Appl Chem 45, 11-30, 1976).

The present invention includes all possible stereoisomers of the compounds of the present invention as single stereoisomers, or as any mixture of said stereoisomers, e.g. (R)- or (S)- isomers, in any ratio. Preferred is the stereoisomer which shows the desired effect. For compounds of formula (I) wherein R⁴=methyl it is discovered that the compounds having said methyl group in the S-configuration do have a significantly better pharmacological effect.

Thus as one aspect of the invention for the configuration of the alkyl group in R⁴ the invention particularly inlcudes those compounds in which R⁴=Ci-C3-alkyl, more particularly R⁴=methyl with S-configuration as indicated in formula (la)

formula (la)

Isolation of a single stereoisomer, e.g. a single enantiomer or a single diastereomer, of a compound of the present invention is achieved by any suitable state of the art method, such as chromatography, especially chiral chromatography, for example.

Further, it is possible for the compounds of the present invention to exist as tautomers. For example, any compound of the present invention which contains an pyrazol moiety as a heteroaryl group for example can exist as a 1 H tautomer, or a 2H tautomer, or even a mixture in any amount of the two tautomers, namely :

2H tautomer 1 H tautomer

The present invention includes all possible tautomers of the compounds of the present invention as single tautomers, or as any mixture of said tautomers, in any ratio.

Further, the compounds of the present invention can potential exist as N-oxides, which are defined in that at least one nitrogen of the compounds of the present invention is oxidised. The present invention includes all such chemically possible N-oxides.

Furthermore, the present invention includes all possible crystalline forms, or polymorphs, of the compounds of the present invention, either as single polymorph, or as a mixture of more than one polymorph, in any ratio.

Moreover, the present invention also includes prodrugs of the compounds according to the invention. The term “prodrugs” here designates compounds which themselves can be biologically active or inactive, but are converted (for example metabolically or hydrolytically) into compounds according to the invention during their residence time in the body.

Thus the present invention includes prodrugs of the compounds of formula (I). In yet another embodiment the present invention includes for a compound according to any of the claims or embodimetns herein a stereoisomer, a tautomer, an N- oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

In yet another embodiment the present invention includes stereoisomers, tautomes, an N-oxides, hydrates, solvates, or a salts, or a mixture of same of a compounds of formula (I).

In another embodiment the present invention includes stereoisomers, tautomes, hydrates, solvates, or a salts, or a mixture of same of a compounds of formula (I).

In a further embodiment the present invention includes stereoisomers, tautomes, solvates, or a salts, or a mixture of same of a compounds of formula (I).

In yet a further embodiment the present invention includes stereoisomers, tautomes, solvates, or a salts, or a mixture of same of a compounds of formula (I).

In yet another embodiment the present invention includes stereoisomers, tautomes, or a salts, or a mixture of same of a compounds of formula (I).

DESCRIPTION In accordance with a first aspect, the present invention provides compounds of general formula (I):

where

R² is selected from a hydrogen atom and a halogen atom;

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from C1-C3- haloalkyl group, a hydroxy group, a CrC3-alkoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom or a CrC3-alkyl group; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

In accordance with an embodiment of the first aspect, the present invention provides compounds of general formula (I):

where

R¹ is selected from a hydrogen atom, a halogen atom, a cyano group, a CrC₃-alkyl group, a CrC₃-haloalkyl group, and a CrC₃-haloalkoxy group;

R² is selected from a hydrogen atomand a halogen atom;

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from Cr C₃-haloalkyl group, a hydroxy group, a CrC₃-alkoxy group, a C₄-C₆-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

R² is selected from a hydrogen atom and a halogen atom;

R⁴ is a CrC₃-alkyl group;

where R¹ is selected from a hydrogen atom, a fluorine atom, a chlorine atom, a cyano group, a methyl group, a trifluoromethyl group, and a trifluoromethoxy group;

R² is selected from a hydrogen atom and a fluorine atom, a chlorine atom;

R³ is a CrC6-alkoxy group which is optionally substituted with a group selected from C₁-C₃- haloalkyl group, a hydroxy group, a CrC₃-alkoxy group, a C₄-C₆-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom or a methyl group;

where

R¹ is selected from a hydrogen atom, a fluorine atom, a chlorine atom, a cyano group, a methyl group, a trifluoromethyl group, and a trifluoromethoxy group;

R² is selected from a hydrogen atom and a fluorine atom, a chlorine atom;

R⁴ is a hydrogen atom;

where

R² is selected from a hydrogen atom and a fluorine atom, a chlorine atom;

R³ is a CrC6-alkoxy group which is optionally substituted with a group selected from Cr

C₃-haloalkyl group, a hydroxy group, a CrC₃-alkoxy group, a C₄-C₆-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a methyl group;

where

R¹ is selected from a hydrogen atom, a fluorine atom, a chlorine atom, a methyl group, and a trifluoromethyl group;

R² is selected from a hydrogen atom and a fluorine atom, a chlorine atom;

R³ is a Ci-C₆-alkoxy group which is optionally substituted with a group selected from Cr C3-haloalkyl group, a hydroxy group, a Ci-C3-alkoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom or a methyl group;

where

R² is selected from a hydrogen atom and a fluorine atom, a chlorine atom;

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from Cr C3-haloalkyl group, a hydroxy group, a Ci-C3-alkoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom;

or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same. In accordance with an embodiment of the first aspect, the present invention provides compounds of general formula (I):

where

R² is selected from a hydrogen atom and a fluorine atom, a chlorine atom; with the proviso that both, R¹ and R² can not be a hydrogen atom at the same time;

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from Cr C3-haloalkyl group, a hydroxy group, a CrC3-alkoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a methyl group;

where

R¹ is selected from a hydrogen atom, a fluorine atom, a chlorine atom, and a trifluoromethyl group;

R² is selected from a hydrogen atom and a fluorine atom;

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluoroethyl group, a hydroxy group, methoxy group, an ethoxy group, a C4-C6-cycloalkyl group, a 4- to 6- membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom or a methyl group;

where

R¹ is selected from a hydrogen atom, a fluorine atom, a chlorine atom, and a trifluoromethyl group; R² is selected from a hydrogen atom and a fluorine atom;

R⁴ is a hydrogen atom;

where

R² is selected from a hydrogen atomand a a fluorine atom;

R⁴ is a methyl group;

where

R¹ is selected from a fluorine atom, and a trifluoromethyl group;

R² is a hydrogen atom;

R⁴ is a hydrogen atom or a methyl group;

where

R¹ is selected from a fluorine atom, and a trifluoromethyl group;

R² is a hydrogen atom;

R³ is a Ci-C₆-alkoxy group which is optionally substituted with a group selected from trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluoroethyl group, a hydroxy group, methoxy group, an ethoxy group, a C4-C6-cycloalkyl group, a 4- to 6- membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom;

where

R¹ is selected from a fluorine atom, and a trifluoromethyl group;

R² is a hydrogen atom;

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluoroethyl group, a hydroxy group, methoxy group, a, ethoxy group, a C4-C6-cycloalkyl group, a 4- to 6- membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a methyl group;

where

R¹ is a trifluoromethyl group;

R² a hydrogen atom;

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluoroethyl group, a hydroxy group, methoxy group, an ethoxy group, a cyclobutyl group, a tetrahydrofuryl group, a pyrazol-3-yl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom or a methyl group;

where

R¹ is a trifluoromethyl group;

R² a hydrogen atom;

R⁴ is a hydrogen atom;

where

R¹ is a trifluoromethyl group;

R² a hydrogen atom;

R⁴ is a methyl group;

where

R¹ is a trifluoromethyl group;

R² a hydrogen atom;

R³ is a CrC4-alkoxy group which is optionally substituted with a group selected from trifluoromethyl group, a difluoroethyl group, a fluoroethyl group, methoxy group, a with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom or a methyl group;

where

R¹ is a trifluoromethyl group;

R² a hydrogen atom;

R³ is a CrC4-alkoxy group which is optionally substituted with a group selected from trifluoromethyl group, a difluoroethyl group, a fluoroethyl group, and a methoxy group, with the proviso that an unsubstituted methoxy group is excluded;

where

R¹ is a trifluoromethyl group;

R² a hydrogen atom; R³ is a CrC4-alkoxy group which is optionally substituted with a group selected from trifluoromethyl group, a difluoroethyl group, a fluoroethyl group, methoxy group, a with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a methyl group; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

In accordance with an embodiment of the first aspect, the present invention provides compounds of general formula (I) selected from

1 5-[4-(2-methoxyethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

2 (6S)-6-methyl-5-[3-(trifluoromethyl)-4-(3,3,3-trifluoropropoxy)phenyl]-3,6- dihydro-2H-1 ,3,4-oxadiazin-2-one,

3 (6S)-6-methyl-5-[4-propoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

4 (6S)-5-{4-[(2R)-2-fluoropropoxy]-3-(trifluoromethyl)phenyl}-6-methyl-3, 6-dihydro- 2H-1 ,3,4-oxadiazin-2-one,

5 (6S)-5-[4-butoxy-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

6 (6S)-5-[4-(2,2-difluoropropoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3, 6-dihydro- 2H-1 ,3,4-oxadiazin-2-one,

7 (6S)-5-[4-ethoxy-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

8 (6S)-5-[4-(2-methoxyethoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H- 1 ,3,4-oxadiazin-2-one and

9 (6S)-6-methyl-5-[4-(2, 2, 2-trifluoroethoxy)-3-(trifluoromethyl)phenyl]-3, 6-dihydro- 2H-1 ,3,4-oxadiazin-2-one, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

2 (6S)-6-methyl-5-[3-(trifluoromethyl)-4-(3,3,3-trifluoropropoxy)phenyl]-3,6- dihydro-2H-1 ,3,4-oxadiazin-2-one, 3 (6S)-6-methyl-5-[4-propoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

1 5-[4-(2-methoxyethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

In accordance with a further embodiment of the first aspect, the present invention provides compounds of general formula (I):

wherein the compound is selected from the group

5-{4-[(2R)-2-hydroxypropoxy]-3-(trifluoromethyl)phenyl}-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

5-[4-(2-hydroxy-2-methylpropoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

5-{4-[(2S)-2-hydroxypropoxy]-3-(trifluoromethyl)phenyl}-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

5-[4-(1 H-pyrazol-3-ylmethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one, 5-[4-(2-methoxyethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one,

5-[4-(cyclobutylmethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one,

5-{4-[(2S)-tetrahydrofuran-2-ylmethoxy]-3-(trifluoromethyl)phenyl}-3,6-dihydro-2H-

1 ,3,4-oxadiazin-2-one,

5-{4-[(2R)-tetrahydrofuran-2-ylmethoxy]-3-(trifluoromethyl)phenyl}-3,6-dihydro-2H-

1 ,3,4-oxadiazin-2-one,

5-[3-(trifluoromethyl)-4-(3,3,3-trifluoropropoxy)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-

2-one,

5-[4-butoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one,

5-[4-propoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one and

5-[4-(3,3-dimethylbutoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one

or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same.

In accordance with a further embodiment of the first aspect, the present invention provides compounds of general formula (I): wherein the compound is selected from the group (6S)-5-{4-[(2R)-2-hydroxypropoxy]-3-(trifluoromethyl)phenyl}-6-methyl-3,6-dihydro-2H-

1 ,3,4-oxadiazin-2-one,

(6S)-5-[4-(2-hydroxy-2-methylpropoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3, 6-dihydro- 2H-1 ,3,4-oxadiazin-2-one,

(6S)-5-{4-[(2S)-2-hydroxypropoxy]-3-(trifluoromethyl)phenyl}-6-methyl-3,6-dihydro-2H-

1 ,3,4-oxadiazin-2-one,

(6S)-6-methyl-5-[4-(1 H-pyrazol-3-ylmethoxy)-3-(trifluoromethyl)phenyl]-3, 6-dihydro- 2H-1 ,3,4-oxadiazin-2-one,

(6S)-5-[4-(2-methoxyethoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

(6S)-5-[4-(cyclobutylmethoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-

1 ,3,4-oxadiazin-2-one,

(6S)-6-methyl-5-{4-[(2S)-tetrahydrofuran-2-ylmethoxy]-3-(trifluoromethyl)phenyl}-3,6- dihydro-2H-1 ,3,4-oxadiazin-2-one,

(6S)-6-methyl-5-{4-[(2R)-tetrahydrofuran-2-ylmethoxy]-3-(trifluoromethyl)phenyl}-3,6- dihydro-2H-1 ,3,4-oxadiazin-2-one, (6S)-6-methyl-5-[3-(trifluoromethyl)-4-(3,3,3-trifluoropropoxy)phenyl]-3,6-dihydro-2H-

1.3.4-oxadiazin-2-one,

(6S)-5-[4-butoxy-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one,

(6S)-6-methyl-5-[4-propoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin- 2-one and

(6S)-5-[4-(3,3-dimethylbutoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-

1.3.4-oxadiazin-2-one

Further embodiments of the present invention

where

R¹ is selected from a hydrogen atom, a halogen atom, a cyano group, a CrC3-alkyl group, a CrC3-haloalkyl group, and a CrC3-haloalkoxy group; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

where

R¹ is selected from a hydrogen atom, a fluorine atom, a chlorine atom, a cyano group, a methyl group, a trifluoromethyl group, and a trifluoromethoxy group; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same

where

R¹ is selected from a hydrogen atom, a fluorine atom, a chlorine atom, a methyl group, and a trifluoromethyl group; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same In accordance with an embodiment of the first aspect, the present invention provides compounds of general formula (I):

where

R¹ is selected from a hydrogen atom, a fluorine atom, a chlorine atom, and a trifluoromethyl group; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same

where

R¹ is selected from a fluorine atom, and a trifluoromethyl group or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

where

R¹ is a trifluoromethyl group or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N- oxide or a mixture of same.

where

R² is selected from a hydrogen atom and a halogen atom; with the proviso that both, R¹ and R² can not be a hydrogen atom at the same time;

where

R² is selected from a hydrogen atom and a halogen atom;

where

R² is selected from a hydrogen atom and a fluorine atom; with the proviso that both, R¹ and R² can not be a hydrogen atom at the same time;

where

R² is a hydrogen atom; with the proviso that both, R¹ and R² can not be a hydrogen atom at the same time;

where R² is a fluorine atom; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

where

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from C1-C3- haloalkyl group, a hydroxy group, a CrC3-alkoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

wherein

In accordance with an embodiment of the first aspect, the present invention provides compounds of general formula (I): wherein

R³ is a Ci-C₆-alkoxy group which is optionally substituted with a group selected from trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluoroethyl group, a hydroxy group, methoxy group, a ethoxy group, a C4-C6-cycloalkyl group, a 4- to 6- membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

wherein

R³ is a Ci-C₆-alkoxy group which is optionally substituted with a group selected from trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluoroethyl group, a hydroxy group, methoxy group, a ethoxy group, a C4-C6-cycloalkyl group, a 4- to 6- membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

wherein

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluoroethyl group, a hydroxy group, methoxy group, a ethoxy group, a cyclobutyl group, a tetrahydrofuran- 2-yl group, a pyrazol-3-yl group, with the proviso that an unsubstituted methoxy group is excluded;

wherein

R³ is a Ci-C₆-alkoxy group which is optionally substituted with a group selected from hydroxy, trifluoromethyl group, a difluoroethyl group, a fluoroethyl group, a methoxy group, a cyclobutyl group, a pyrazol-3-yl group, a tetrahydrofuran-2-yl group,

wherein

R³ is a CrC4-alkoxy group which is optionally substituted with a group selected from hydroxy, trifluoromethyl group, a difluoroethyl group, a fluoroethyl group, a methoxy group, a cyclobutyl group, a pyrazol-3-yl group, a tetrahydrofuran-2-yl group,

wherein

In accordance with a further embodiment of the first aspect, the present invention wherein

In accordance with a further embodiment of the first aspect, the present invention wherein R³ is a CrC₆-alkoxy group which is substituted with a group selected from CrC3-haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C6-cycloalkyl group, a 4- to 6- membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

In accordance with yet another embodiment of the first aspect, the present invention provides compounds of general formula (I):

wherein

R³ is a CrC₆-alkoxy group with the proviso that an unsubstituted methoxy group is excluded;

wherein

R³ is a CrC4-alkoxy group which is optionally substituted with a group selected from C1-C3- haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

wherein

R³ is a CrC4-alkoxy group which is substituted with a group selected from CrC3-haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C6-cycloalkyl group, a 4- to 6- membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

wherein R³ is a CrC4-alkoxy group which is optionally substituted with a group selected from a trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluoroethyl group a hydroxy group, a methoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

wherein

R³ is a CrC4-alkoxy group which is optionally substituted with a group selected from a trifluoromethyl group, a difluoroethyl group, a fluoroethyl group, a methoxy group, with the proviso that an unsubstituted methoxy group is excluded;

wherein

R³ is a CrC3-alkoxy group which is optionally substituted with a group selected from a trifluoromethyl group, a difluoroethyl group, a fluoroethyl group, a methoxy group, with the proviso that an unsubstituted methoxy group is excluded;

wherein

R⁴ is a hydrogen atom or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

In accordance with yet another embodiment of the first aspect, the present invention provides compounds of general formula (I): wherein

R⁴ is a CrC3-alkyl group or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

wherein

R⁴ is a methyl group or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

In accordance with a further embodiment of the first aspect, the present invention provides the compounds as specifically exemplified in the experimental section or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

In another aspect, the invention provides a pharmaceutical composition containing one or more pharmaceutically acceptable carriers or excipients and a compound of formula (I)

In yet another aspect, the invention provides a pharmaceutical composition containing one or more pharmaceutically acceptable carriers or excipients and a compound of formula (I)

a pharmaceutically acceptable salt or prodrug thereof.

“Reference” in the context of this invention means an average expression in a representative panel of tumor cells or tumor cell lines.

In various embodiments of any aspect delineated herein, the cancer is responsive to a PDE3A- and/or PDE3B-SLFN12 complex modulator.

In various embodiments, the subject has been diagnosed with a cancer responsive to a PDE3A- and/or PDE3B-SLFN12 complex modulator. In various embodiments of any aspect delineated herein, the cancer is such as e.g., tumors of the anus, the brain, the breast, the bones, the central and peripheral nervous system, the colon, the eye, the kidney, the endocrine glands (e.g., thyroid and adrenal cortex), the endometrium, the esophagus, the gastrointestinal tract (including gastrointestinal stromal tumors), the germ cells, the head and the neck, the kidney, the liver, the larynx and hypopharynx, the lung, the mesothelioma, the pancreas, the prostate, the rectum, the reproductive organs (e.g., cervix, ovary, prostate), the respiratory tract, the small intestine, the skin, the soft tissue, the stomach, the testis, the thyroid gland, the parathyroid gland, ureter, the urogenital tract, vagina and vulva and the connective tissue and metastases of these tumors. Malignant neoplasias include inherited cancers exemplified by Retinoblastoma and Wilms tumor etc.).

In various embodiments, the cancer is a skin cancer, especially melanoma, lung adenocarcinoma or a cervical cancer.

In various embodiments of any aspect delineated herein, the PDE3A- and/or PDE3B-SLFN12 complex modulator is administered orally.

In various embodiments of any aspect delineated herein, the PDE3A- and/or PDE3B-SLFN12 complex modulator is administered by intravenous injection.

More particularly the compounds of formula (I) are suitable for the treatment of a patient having a cancer that is sensitive to treatment with a phosphodiesterase 3A/B (PDE3A and/or B)-SLF12 complex modulator by detecting co-expression of PDE3A and / or PDE3B and Schlafen 12 ( SLFN12 ) and / or SLFN12L mRNA, polynucleotides or polypeptides and/or a lack of decrease in expression of CREB3L1 mRNA, polynucleotides or polypeptides in a cancer cell derived from such patients. The compounds of formula (I) are selective for cancer cell killing while minimizing enzymatic inhibition of PDE3A and PDE3B

Compositions and articles defined by the invention were isolated or otherwise manufactured in connection with the examples provided below. Other features and advantages of the invention will be apparent from the detailed description, and from the claims.

In a particular further embodiment of the first aspect, the present invention provides combinations of two or more of the above mentioned embodiments under the heading “further embodiments of the first aspect of the present invention”.

The present invention provides any sub-combination within any embodiment or aspect of the present invention of compounds of general formula (I), supra. The present invention provides any sub-combination within any embodiment or aspect of the present invention of intermediate compounds of general formula (I).

The present invention provides the compounds of general formula (I) which are disclosed in the Example Section of this text, infra.

Kits

The invention further provides kits comprising a compound of formula (I) and/or means for characterizing the responsiveness or resistance of a subject to PDE3A- and/ or PDE3B-SLFN12 complex modulator, especially to compounds of formula (I) treatment.

Also provided herein are kits that can include the compound of formula (I) in form of a therapeutic composition containing an effective amount of said compound in e.g., a unit dosage form.

In some embodiments, the kit comprises a sterile container which includes a therapeutic or diagnostic composition; such containers can be boxes, ampoules, bottles, vials, tubes, bags, pouches, blister-packs, or other suitable container forms known in the art. Such containers can be made of plastic, glass, laminated paper, metal foil, or other materials suitable for holding medicaments.

In one embodiment, if desired, the kit further comprises instructions for measuring PDE3A and/or PDE3B and SLFN12 and/or instructions for administering the PDE3A- and/or PDE3B-SLFN12 complex modulator to a subject having a malignancy, e.g., a malignancy selected as responsive to PDE3A- and/or PDE3B-SLFN12 complex modulator treatment.

In particular embodiments, the instructions include at least one of the following: description of the therapeutic agent; dosage schedule and administration for treatment or prevention of malignancy or symptoms thereof; precautions; warnings; indications; counter-indications; over dosage information; adverse reactions; animal pharmacology; clinical studies; and/or references. The instructions may be printed directly on the container (when present), or as a label applied to the container, or as a separate sheet, pamphlet, card, or folder supplied in or with the container.

The practice of the present invention employs, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are well within the purview of the skilled artisan. Such techniques are explained fully in the literature, such as, “Molecular Cloning: A Laboratory Manual”, second edition (Sambrook, 1989); “Oligonucleotide Synthesis” (Gait, 1984); “Animal Cell Culture” (Freshney, 1987); “Methods in Enzymology”“Handbook of Experimental Immunology” (Weir, 1996);“Gene Transfer Vectors for Mammalian Cells” (Miller and Calos, 1987);“Current Protocols in Molecular Biology” (Ausubel, 1987);“PCR: The Polymerase Chain Reaction”, (Mullis, 1994); “Current Protocols in Immunology” (Coligan, 1991). These techniques are applicable to the production of the polynucleotides and polypeptides of the invention, and, as such, may be considered in making and practicing the invention. Particularly useful techniques for particular embodiments will be discussed in the sections that follow.

The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the assay, screening, and therapeutic methods of the invention, and are not intended to limit the scope of the invention.

General synthesis of the compounds of formula (I)

The compounds according to the invention of general formula (I) can be prepared according to the following schemes 1 through 6. The schemes and procedures described below illustrate synthetic routes to the compounds of general formula (I) of the invention and are not intended to be limiting. It is clear to the person skilled in the art that the order of transformations as exemplified in schemes 1 through 6 can be modified in various ways. The order of transformations exemplified in these schemes is therefore not intended to be limiting. In addition, interconversion of any of the substituents, R¹, R², R³, or R⁴ can be achieved before and/or after the exemplified transformations. These modifications can be such as the introduction of protecting groups, cleavage of protecting groups, reduction or oxidation of functional groups, halogenation, metallation, substitution or other reactions known to the person skilled in the art. These transformations include those which introduce a functionality which allows for further interconversion of substituents. Appropriate protecting groups and their introduction and cleavage are well-known to the person skilled in the art (see for example T.W. Greene and P.G.M. Wuts in Protective Groups in Organic Synthesis, 3^rd edition, Wiley 1999). Specific examples are described in the subsequent paragraphs.

Five routes for the preparation of compounds of general formula (I) are described in schemes 1 , 2, 3, 4, 5 and 6. Each alternative route is considered a separate aspect of the invention. Synthesis Routes

Route 1 : Synthesis of compounds of formula (I) by nucleophilic aromatic substitution

Part I: Synthesis of intermediate compounds of formula (ID

Scheme 1

Scheme 1: Route for the preparation of intermediate compounds of formula (II), in which R¹, R² and R⁴ have the meaning as defined supra or in anyone of the claims; the meaning of X is as defined below in context of Scheme 2 and the paragraphs (i), 0, (k) for compounds of formula (II).

(a) Hydroxy(tosyloxy)iodo)benzene, DMSO, water, RT, 18 h; or iodine, DMSO, 60 °C, 18h. For the preparation of the starting material for steps (a) and (h) see e.g. Scheme 3.

(b) H2NNHCOOCH₃, HCI (aq), MeOH, RT, 5h;

(c) potassium carbonate, acetonitrile, 60 °C, 18h or NaOEt/EtOH, 0 °C, 10 min, or NaH, EtOH, 0 °C 10 min;

(d) sodium formate, sodium hydrogencarbonate, CH3CN, water, 65°C, 24h;

(e) potassium acetate, potassium iodide, 18h, RT;

(f) H₂NNHCOOCH₃, HCI (aq), MeOH, RT, 5-18h;

(g) potassium carbonate, CH₃CN, 60 °C, 18h or NaOEt/EtOH, 0 °C 10 min, or NaH, EtOH, 0 °C 10 min;

(h) acetic acid, bromine, hydrogen bromide, 18h, RT; for the preparation of the starting material for steps (a) and (h) see e.g. Scheme 3.

Compounds of formula (II) containing chiral centers can be optionally separated by methods known to the person skilled in the art, such as e.g. chiral chromatography, to obtain individual enantiomers or diastereomers. Part II: Conversion of the intermediate compounds of formula (ID into compounds of general formula (I):

(i), 0), (k): R³ = OR⁵,

R⁵ = a CrC₆-alkyl group which is optionally substituted with a group independently selected from CrC3-haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C₆- cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methyl group is excluded; and Pg2 is a protecting group suitable for hydroxy groups, e.g. a benzyl group.

Scheme 2

Scheme 2: Route for the preparation of compounds of general formula (I) via formula (II) in which R¹, R² and R⁴ have the meaning as defined supra,

(i) Route 1 via nucleophilic aromatic substitution to introduce R³ substituents

Compounds of general formula (I), can be obtained by reacting intermediate compounds of formula (II), in which

R⁴ has the meaning as defined supra,

X is F or Cl (as reflected in scheme 2), and if X is Cl, R¹ or R²can not be F; particularly X is F;

R¹ and R² have the meaning as defined supra, with the proviso that if X is Cl, R¹ or

R²can not be F, and with the proviso that at least one of R¹ and R² exerts an electron withdrawing effect; particularly, R¹ is selected from fluorine, and -CF₃, and R² is hydrogen or fluorine; more particularly, R¹ is -CF₃ and R² is hydrogen; generally, substituents having an electron withdrawing effect can be selected from halogen, cyano, and Ci-C3-haloalkyl group (this definition applicable also below); with a corresponding alcohol R⁵-OH, optionally as the alcohol or as an alkoxide salt, such as a sodium or potassium salt,

optionally in the presence of a strong base, such as potassium tert-butoxide, or sodium hydride, or sodium metal,

optionally in an inert solvent, such as THF or DMF,

optionally in the presence of a base, such as cesium carbonate, in a solvent, such as DMSO, optionally at elevated temperatures ranging from RT to 160 °C, particularly from 60° C to 150 °C, for 2h-7d;

whereby

R⁵ is a Ci-C₆-alkyl group which is optionally substituted with a group independently selected from CrC3-haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4- C₆-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methyl group is excluded.

(j) Alternative Route via transition metal catalyzed, particularly palladium catalyzed amination, to introduce R³ substituents

X is Cl, Br, I, (as reflected in scheme 2), or a group selected from (CrC4-alkylsulfonyl)oxy, (Ci-C4-fluoroalkylsulfonyl)oxy and (phenylsulfonyl)oxy, the phenyl present in (phenylsulfonyl)oxy being optionally substituted with one, two, three, four or five substituents, each of them independently selected from halogen, nitro, cyano, Cr C4-alkyl and CrC4-alkoxy;

R¹ or R² are as defined supra but are different from Cl, Br, I; particularly, R¹ is -CF₃ and R² is hydrogen;

R⁴ has the meaning as defined supra,

with a corresponding alcohol R⁵-OH,

in the presence of a base, such as potassium phosphate or cesium carbonate, a palladium catalyst, such as [(2-Di-terf-butylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1 , 1 '- biphenyl)-2-(2'-amino-1 , 1 '-biphenyl)]palladium(ll) methanesulfonate or [(2-Di -tert- butylphosphino-3-methoxy-6-methyl-2',4',6'-triisopropyl-1 , 1 '-biphenyl)-2-(2- aminobiphenyl)]palladium(ll) methanesulfonate, optionally a ligand, such as 2-Di(tert- butyl)phosphino-2',4',6'-triisopropyl-3-methoxy-6-methylbiphenyl or 2-(Di-terf- butylphosphino)-2',4',6'- triisopropyl-3, 6-dimethoxy-1 , 1 '-biphenyl, in an inert solvent, such as dioxane or toluene, at elevated temperatures ranging from 60-160 °C, for 2h-7d;

whereby

R⁵ = is a defined supra.

The thus obtained compounds of formula (I) containing chiral centers can be optionally separated by chiral chromatography to obtain individual enantiomers or diastereomers.

(k) Alternative Route via Mitsunobu reaction, to introduce R³ substituents

Compounds of general formula (I), can be obtained in a two step procedure (k-1) (Removal of the protecting group Pg2), followed by (k-2) (Mitsunobu reaction)).

(k-1 )

Compounds of general formula (II), where X is -OH, can be obtained by reacting intermediate compounds of formula (II), in which

X is OPg2, where Pg2 is a protecting group suitable for hydroxy groups, e.g. a benzyl group;

R¹ or R² are as defined supra ; R⁴ has the meaning as defined supra,

with hydrogen gas, in the presence of a palladium catalyst, such as Palladium on carbon, in a solvent, such as ethanol, at temperatures ranging from room temperature to 60 °C, for 2h-3d, at hydrogen pressure ranging from 1 bar to 10 bar, particularly 1 bar.

(k-2)

Compounds of general formula (I), can be obtained by reacting intermediate compounds of formula (II), in which X is -OH,

R¹ or R² are as defined supra, R⁴ has the meaning as defined supra ;

with a corresponding alcohol R⁵-OH,

in the presence of an dialkylazodicarboxylate, particularly diisopropylazodicarboxylate, a phosphine, such as triphenylphosphine, in an inert solvent, such as THF, at temperatures ranging from 0 °C to room temperature, for 2h-3d;

whereby

R⁵ = is as defined supra.

Route 2

Part I: Synthesis of intermediate compounds of formula (III) from compounds of formula ilYli

Scheme 3

Scheme 3: Route for the preparation of intermediate compounds of formula (III) from compounds of formula (IV), in which R¹ and R² have the meaning as defined supra. Benzoic acids of formula (IV) are well known to the person skilled in the art and are commercially available in considerable structural variety.

Step(a) (IV) to (IV-1)

A compound of formula (IV) and oxalyl chloride, in a solvent such as e.g. DMF/DCM, are reacted at a temperature range of 0 °C to RT, for 1h-20 h, then HN(OCH3)CH3 _*HCI, Et3N/CH2Cl2, is added at RT, and the mixture is subsequently reacted for 1h- 3d;

Step (b) (IV-1) to (III)

A compound (IV-1) and R⁴CH₂MgBr, in which R⁴ is as defined supra, in a solvent such as e.g. THF, are reacted at 0 °C to RT, for 1h - 20 h, in order to obtain an intermediate compound of formula (III); Route 3: Stereoselective synthesis of intermediate compounds of formula (lla), in which R⁴ is different from hydrogen

Scheme 4 Scheme 4: Stereoselective route for the preparation of intermediate compounds of formula (I la), constituting a sub-set of the intermediate compounds of formula (II), from intermediate compounds of formula (III) in which R¹, R², and R⁴ have the meaning as defined supra, with the proviso that R⁴ is different from hydrogen.

(c) LiHMDS/THF, 1 h, -78 °C, then tBDMSCI (terf-butyldimethylchloro silane), -78 °C to

RT, 15h - 2d;

(d) AD-Mix-a, CH₃S0₂NH₂/tBu0H/water, 0 °C to RT 15 h - 2d;

(e) 1. H ₂NNHCOOCH₃, HCI, MeOH; 2. NaOEt/EtOH or NaOMe/MeOH or NaH/EtOH;

Route 4: Alternative synthesis of intermediate compounds of formula (II)

Scheme 5

Scheme 5: Route for the preparation of intermediate compounds of formula (II) from compounds of formulae (V) and (VI), in which R¹, R² and R⁴ have the meaning as defined supra and M is a metal-containing group, such as Li, or MgBr, or MgCI; and the meaning of X is as defined above in context of Scheme 2 and the paragraphs (i), (j), (k) for compounds of formula (II), and DG is a group displacable from compounds of formula (VI) with reactands of formula (V), selected from morpholinyl or N(OCH₃)CH₃ (Weinreb amide); and Pg is a protecting group suitable for hydroxy groups, e.g. a tri-(Ci-C4-alkyl)-silyl group such as tert-butyl-dimethylsilyl. Compounds of formulae (V) and (VI) are known to the person skilled in the art and can be readily prepared from commercially available precursors by known methods.

(a) THF, -20 °C - 20 °C, 1h - 24h, (b) 1. H₂NNHCOOCH₃, HCI, MeOH; 2. TBAF (tetrabutylammonium fluoride), THF; 3. NaOEt/EtOH or NaOMe/MeOH;

Compounds of formula (II) can be converted to compounds of general formula (I) as described above in context of Scheme 2.

Scheme 6

Scheme 6: Route for the preparation of compounds of general formula (I) from compounds of formulae (VII) and (VI), in which R¹, R², R³ (potentially containing functional groups suitably protected), and R⁴ have the meaning as defined supra and M is a metal-containing group, such as Li, or MgBr, or MgCI; and DG is a group displacable from compounds of formula (VII) by reactands of formula (VI), particularly but not limited to N(OCH3)CH3 (Weinreb amide); and Pg is a protecting group suitable for hydroxy groups, e.g. a tri-(Ci- C4-alkyl)-silyl group such as tert-butyl-dimethylsilyl. Compounds of formulae (VII) and (VI) are known to the person skilled in the art and can be readily prepared from commercially available precursors by known methods.

(a) THF, -20 °C - 20 °C, 1 h - 24h, (b) 1.) H₂NNHCOOCH₃, HCI, MeOH; 2.) TFA, DCM, 0

°C - 20 °C; 3.) NaOEt/EtOH or NaOMe/MeOH, 20 °C;

Compounds of formula (I) containing chiral centers can be optionally separated by methods known to the person skilled in the art, such as e.g. chiral chromatography, to obtain individual enantiomers or diastereomers.

Compounds (VII) are either commercially available or can be prepared according to procedures available from the public domain, as understandable to the person skilled in the art.

Some further aspects of the invention are the synthesis routes according to the schemes 1-6 above whereby each alternative route stands as a single aspect.

The present invention futher provides methods of preparing compounds of the present invention of general formula (I), said methods comprising the steps as described in the Experimental Section herein.

The present invention in particular provides a method of preparing a compound of general formula (I) as defined supra, said method comprising starting from a compound of formula

(II)

formula (II)

EITHER

wherein X is a fluorine atom or a chlorine atom

R¹ is selected from a hydrogen atom, a halogen atom, a cyano group, a Ci-C3-alkyl group, a Ci-C3-haloalkyl group, and a Ci-C3-haloalkoxy group,

R² is a hydrogen atom or a halogen atom;

with the proviso that at least one of R¹ and R² exerts an electron withdrawing effect and

R⁴ is a hydrogen atom or a Ci-C3-alkyl group

with the proviso that if X is a chlorine atom, R¹ and R² are not a fluorine atom, to react with a corresponding alcohol R⁵-OH, optionally as the alcohol or as an alkoxide salt, such as a sodium or potassium salt,

whereby

R⁵ is a CrC₆-alkyl group which is optionally substituted with a group selected from Ci-C3-haloalkyl group, a hydroxy group, a Ci-C3-alkyoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methyl group is excluded comprising the following conditions:

optionally in the presence of a strong base,

optionally in an inert solvent,

optionally in the presence of a further base, in a solvent,

optionally at elevated temperatures ranging from room temperature to 160 °C; in order to obtain a compound of formula (I)

formula (I)

OR

comprising starting from a compound of formula (II)

formula (II)

wherein

X is Cl, Br, I, or a group selected from (Ci-C4-alkylsulfonyl)oxy, (C1-C4- fluoroalkylsulfonyl)oxy and (phenylsulfonyl)oxy, the phenyl present in (phenylsulfonyl)oxy being optionally substituted with one, two, three, four or five substituents, each of them independently selected from halogen, nitro, cyano, Cr C4-alkyl and CrC4-alkoxy,

R¹ is selected from a hydrogen atom, a halogen atom, a cyano group, a CrC3-alkyl group, a CrC3-haloalkyl group, and a CrC3-haloalkoxy group,

R² is a hydrogen atom or a halogen atom;

with the proviso, that both, R¹ and R² are different from a chlorine atom, a bromine atom or a iodine atom;

R⁴ ia a hydrogen atom or a CrC3-alkyl group

to react with a corresponding alcohol R⁵-OH, whereby R⁵ is as defined supra,

in the presence of a base, a palladium catalyst, optionally a ligand,

in an inert solvent,

at elevated temperatures ranging form 60-160°C in order to obtain a compound of formula (I)

formula (II)

wherein X is OPg2, whereby

Pg2 is a suitable protecting group of a hydroxy group, particularly it could be a protecting group which can be cleaved by the contact with hydrogen, such as a benzyl group,

R¹ is selected from a hydrogen atom, a halogen atom, a cyano group, a CrC3-alkyl group, a CrC3-haloalkyl group, and a CrC3-haloalkoxy group, and

R² is a hydrogen atom or a halogen atom;

which is removed by a suitable method depending on the protecting group used, if the proteting group is a benzyl group, the reaction mixture is contacted with hydrogen gas in the presense of a palladium catalyst in a solvent at temperatures ranging from room temperature to 60°C under hydrogen pressure ranging from 1 bar to 10 bar resulting in a compound of formula (II), wherein X=OH, and

subsequently reacting the the so obtained compound of formula (II) with a corresponding alcohol R⁵-OH

whereby R⁵ is defined as mentioned supra,

in the presence of a dialkylazodicarboxylate, a phosphine, in an inert solvent at lower temperatures ranging from 0°C to room temperature

in order to obtain a compound of formula (I)

formula (I)

The present invention in particular provides a method of preparing a compound of general formula (I) as defined supra, said method comprising starting from a compound of formula (II)

formula (II)

EITHER

wherein X is a fluorine atom or a chlorine atom

R¹ is selected from a hydrogen atom, a fluorine atom, a cyano group, a methyl group, a trifluoromethyl group, a difluormethyl group, a difluoroethyl group, a fluroroethyl group, and a trifluoromethoxy group;

R² is a hydrogen atom or a fluorine atom;

R⁴ ia a hydrogen atom or a methyl group

with the proviso that if X is a chlorine atom, R¹ and R² are not a fluorine atom, to react with a corresponding alcohol R⁵-OH, optionally as the alcohol or as a sodium salt or a potassium alkoxide salt,

whereby

R⁵ is a CrC₆-alkyl group which is optionally substituted with a group selected from CrC3-haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methyl group is excluded comprising the following conditions:

optionally in the presence of potassium tert-butoxide, or sodium hydride, or sodium metal, optionally in tetrahydrofuran or dimethylformamid,

optionally in the presence of cesium carbonate, optionally in dimethylsulfoxid,

optionally at elevated temperatures ranging from 60°C to 150 °C; in order to obtain a compound of formula (I)

formula (I) OR

comprising starting from a compound of formula (II)

formula (II)

wherein

X is Cl, Br, I, or a group selected from (Ci-C4-alkylsulfonyl)oxy, (C1-C4- fluoroalkylsulfonyl)oxy and (phenylsulfonyl)oxy, the phenyl present in

(phenylsulfonyl)oxy being optionally substituted with one, two, three, four or five substituents, each of them independently selected from halogen, nitro, cyano, Cr C4-alkyl and CrC4-alkoxy,

R¹ is selected from a hydrogen atom, a fluorine atom, a cyano group, a methyl group, a trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluroroethyl group, and trifluoromethoxy group,

R² is a hydrogen atom or a fluorine atom;

R⁴ is a hydrogen atom or a methyl group to react with a corresponding alcohol R⁵-OH, whereby R⁵ is as defined supra,

in the presence of potassium phosphate or cesium carbonate,

in the rpesence of a palladium catalyst [(2-Di-te/f-butylphosphino-3,6-dimethoxy-2',4',6'- triisopropyl-1 , 1 '-biphenyl)-2-(2'-amino-1 , 1 '-biphenyl)]palladium(l I) methanesulfonate or [(2-Di-te/f-butylphosphino-3-methoxy-6-methyl-2',4',6'-triisopropyl-1 , 1 '-biphenyl)-2-(2- aminobiphenyl)]palladium(ll) methanesulfonate,

optionally a ligand, selected from 2-Di(tert-butyl)phosphino-2',4',6'-triisopropyl-3-methoxy- 6-methylbiphenyl and 2-(Di-te/f-butylphosphino)-2',4',6'- triisopropyl-3, 6-dimethoxy- 1 , 1 '- biphenyl,

in dioxane or toluene,

formula (I)

OR

comprising starting from a compound of formula (II)

formula (II)

wherein X is OPg, whereby

Pg is a benzyl group,

R¹ is selected from a hydrogen atom, a fluorine atom, a cyano group, a methyl group, a trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluroroethyl group, and trifluoromethoxy group, R² is a hydrogen atom or a fluorine atom;

R⁴ is a hydrogen atom or a methyl group which is contacted with hydrogen gas in the presense of a palladium on carbon in an alcohol at temperatures ranging from room temperature to 60°C under hydrogen pressure ranging from 1 bar to 10 bar resulting in a compound of formula (II), wherein X=OH, and subsequently reacting the the so obtained compound of formula (II) with a corresponding alcohol R⁵-OH whereby R⁵ is defined as mentioned supra,

in the presence of a diisopropylazodicarboxylate, triphenylphosphine, in tetrahydrofurane at lower temperatures ranging from 0°C to room temperature

in order to obtain a compound of formula (I)

formula (I)

The present invention more particularly provides a method of preparing a compound of general formula (I) as defined supra, said method comprising starting from a compound of formula (II)

formula (II)

wherein X is a fluorine atom or a chlorine atom

R¹ is selected from a fluorine atom, and a -CF₃ group, and R² is a hydrogen atom or a fluorine atom;

R⁴ is a CrC3-alkyl group, to react with a corresponding alcohol R⁵-OH, optionally as the alcohol or its sodium or potassium salt, comprising the following conditions:

optionally in the presence of potassium tert-butoxide or sodium hydride, in THF or DMF, optionally in the presence of cesium carbonate, at optionally elevated temperatures ranging from from 60°C to 150 °C;

whereby

R⁵ is a CrC₆-alkyl group which is optionally substituted with a group selected from CrC3-haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methyl group is excluded. in order to obtain a compound of formula (I)

formula (I)

The present invention alternatively provides a method of preparing a compound of general formula (I) as defined supra, said method comprising starting from a compound of formula

(II)

formula (II)

wherein X is Cl, Br, I, or a group selected from a (CrC₄-alkylsulfonyl)oxy group, a (C₁-C₄- fluoroalkylsulfonyl)oxy group and a (phenylsulfonyl)oxy group, the phenyl moiety present in said (phenylsulfonyl)oxy group being optionally substituted with one, two, three, four or five substituents, each of them independently selected from halogen, nitro, cyano, a CrC₄-alkyl group and a CrC₄-alkoxy group;

R¹ is selected from a hydrogen atom, a halogen atom, a cyano group, a CrC₃-alkyl group, a CrC₃-haloalkyl group, and a CrC₃-haloalkoxy group; but is different from Cl, Br, I;

R² is a hydrogen atom or a fluorine atom;

particularly, R¹ is -CF₃ and R² is hydrogen

R⁴ is a hydrogen atom or a CrC₃-alkyl group; to react with a corresponding alcohol R⁵-OH,

in which R⁵ is a CrC₆-alkyl group which is optionally substituted with a group selected from CrC₃-haloalkyl group, a hydroxy group, a CrC₃-alkyoxy group, a C₄-C₆-cycloalkyl group, a 4 to 6-membered heterocycloalkyl group, a 5 to 6-membered heteroaryl group, with the proviso that an unsubstituted methyl group is excluded, comprising the following conditions:

in the presence of a base, and

a palladium catalyst, optionally a ligand,

in an inert solvent, at elevated temperatures;

in order to obtain a compound of formula (I)

formula (I)

The present invention alternatively provides a method of preparing a compound of general formula (I) as defined supra , said method comprising starting from a compound of formula

(II)

formula (II)

wherein

X is Cl, Br, I, or a group selected from a (Ci-C₄-alkylsulfonyl)oxy group, a (C₁-C₄- fluoroalkylsulfonyl)oxy group and a (phenylsulfonyl)oxy group, the phenyl moiety present in said (phenylsulfonyl)oxy group being optionally substituted with one, two, three, four or five substituents, each of them independently selected from halogen, nitro, cyano, a Ci-C₄-alkyl group and a Ci-C₄-alkoxy group;

R¹ is -CF₃;

R² is a hydrogen atom or a fluorine atom;

R⁴ is a hydrogen atom or a Ci-C₃-alkyl group; to react with a corresponding alcohol R⁵-OH,

in the presence of a base, and

a palladium catalyst,

optionally a ligand,

in an inert solvent,

at elevated temperatures;

in order to obtain a compound of formula (I)

formula (I) The present invention alternatively provides a method of preparing a compound of general formula (I) as defined supra, said method comprising starting from a compound of formula

(II)

formula (II)

wherein

X is Cl, Br, I, or a group selected from a (CrC4-alkylsulfonyl)oxy group, a (C1-C4- fluoroalkylsulfonyl)oxy group and a (phenylsulfonyl)oxy group, the phenyl moiety present in said (phenylsulfonyl)oxy group being optionally substituted with one, two, three, four or five substituents, each of them independently selected from halogen, nitro, cyano, a CrC4-alkyl group and a CrC4-alkoxy group;

R¹ is -CF₃;

R² is a hydrogen atom;

R⁴ is a hydrogen atom or a CrC3-alkyl group; to react with a corresponding alcohol R⁵-OH,

in which R⁵ is a CrC₆-alkyl group which is optionally substituted with a group selected from CrC3-haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C6-cycloalkyl group, a 4 to 6-membered heterocycloalkyl group, a 5 to 6-membered heteroaryl group, with the proviso that an unsubstituted methyl group is excluded, comprising the following conditions:

in the presence of a base, and

a palladium catalyst,

optionally a ligand,

in an inert solvent,

at elevated temperatures;

in order to obtain a compound of formula (I)

formula (I)

(II)

formula (II)

wherein

R¹ is selected from a hydrogen atom, a halogen atom, a cyano group, a CrC3-alkyl group, a CrC3-haloalkyl group, and a CrC3-haloalkoxy group; but is different from Cl, Br, I;

R² is selected from a hydrogen atomand a halogen atom; but is different from Cl, Br, I; particularly, R¹ is -CF3 and R² is hydrogen;

in which R⁵ is a CrC₆-alkyl group which is optionally substituted with a group selected from CrC3-haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methyl group is excluded. comprising the following conditions:

in the presence of potassium phosphate or cesium carbonate, a palladium catalyst selected from [(2-Di-te/f-butylphosphino-3,6-dimethoxy-2',4',6'- triisopropyl-1 , 1 '-biphenyl)-2-(2'-amino- 1 , 1 '-biphenyl)]palladium(l I) methanesulfonate and [(2-Di-te/f-butylphosphino-3-methoxy-6-methyl-2',4',6'-triisopropyl-1 , 1 '-biphenyl)-2-(2- aminobiphenyl)]palladium(ll) methanesulfonate,

optionally a ligand, selected from 2-Di(tert-butyl)phosphino-2',4',6'-triisopropyl-3-methoxy- 6-methylbiphenyl and 2-(Di-te/f-butylphosphino)-2',4',6'- triisopropyl-3, 6-dimethoxy-1 , T- biphenyl,

in dioxane or toluene,

at elevated temperatures ranging from 60-160 °C;

in order to obtain a compound of formula (I)

formula (I)

The present invention also provides a method of preparing a compound of general formula (I) as defined supra, said method starting from a compound of formula (II)

formula (II) to obtain a compound of formula (I) in any of the process alternatives as described supra, further comprising reactions steps in order to obtain the starting compound of formula (II), by

Step A:

wherein a compound of formula (V)

(V)

wherein

R¹ is selected from a hydrogen atom, a halogen atom, a cyano group, a Ci-C3-alkyl group, a Ci-C3-haloalkyl group, and a Ci-C3-haloalkoxy group;

R² is selected from a hydrogen atomand a halogen atom;

M is a methyl containing group, selected from lithium, -MgBr, -MgCI

X is as defined in any of the alternative route above, is reacted with a compound of formula (VI)

wherein Pg is a protecting group suitable for hydroxy groups, such as e.g. tri-(Ci-C4-alkyl)- silyl group such as tert-butyl-dimethylsilyl and

DG is a displacable group selected from morpholiny and N(OCH3)CH3 (Weinreb amide) in an inert solvent, such as e.g. tetrahydrofuran,

at a temparature range from -20°C to room temperature in order to obain a compound of formula (VII)

which then subsequently is further converted into a compound of formula (II)

formula (II)

in a three step reaction by reacting the compound of formula (VII) a) with H₂N-NH-C(0)0CH₃ under acidic conditions in an alcohol and further reacting the mixture

b) with tetrabutylammoniumfluoride in tetrahydrofuran followed by

c) addition of a base such as sodiumethanolate in ethanol or sodiummethanolate in methanol.

In some aspects the invention provides a method as anyone outlined above

wherein

R¹ is trifluoromethyl and R² is a hydrogen atom.

In accordance with a further aspect, the present invention provides intermediate compounds which are useful for the preparation of the compounds of general formula (I), supra.

Even more particularly, the invention provides the intermediate compounds of general formula (II) in particular intermediates as used in the experimental section.

In accordance with another aspect, the present invention provides the use of said intermediate compounds for the preparation of a compound of general formula (I) as defined supra.

The present invention provides the intermediate compounds which are disclosed in the Example Section of this text, infra.

The present invention provides any sub-combination within any embodiment or aspect of the present invention of intermediate compounds of general formula (I), supra.

The compounds of general formula (I) of the present invention can be converted to any salt, particularly pharmaceutically acceptable salts, as described herein, by any method which is known to the person skilled in the art. Similarly, any salt of a compound of general formula (I) of the present invention can be converted into the free compound, by any method which is known to the person skilled in the art. Methods and Administration

Compounds of general formula (I) of the present invention demonstrate a valuable pharmacological spectrum of action, which could not have been predicted. Compounds of the present invention have surprisingly been found to effectively modulate PDE3A and/or PDE3B and it is likely therefore that said compounds may be used for the treatment or prophylaxis of diseases, more particularly hyperproliferative diseases, even more particularly cancer diseases in humans and animals.

More particularly the compounds of formula (I) are suitable for the treatment of a patient having a cancer that is sensitive to treatment with a phosphodiesterase 3A/B, (PDE3A and/or PDE3B)-SLF12 complex modulator by detecting co-expression of PDE3A and / or PDE3B and Schlafen 12 ( SLFN12 ) and / or SLFN12L mRNA, polynucleotides or polypeptides and/or a lack of decrease in expression of CREB3L1 mRNA, polynucleotides or polypeptides in a cancer cell derived from such patients. The compounds of formula (I) are selective for cancer cell killing while minimizing enzymatic inhibition of PDE3A and PDE3B.

Compounds of the present invention can be utilized to inhibit tumor growth by inducing a SLFN12 complex formation. This method comprises administering to a mammal in need thereof, including a human, an amount of a compound of general formula (I), or a pharmaceutically acceptable salt, isomer, polymorph, metabolite, hydrate, solvate or ester thereof, which is effective to treat the disease.

The present compounds of formula (I) may additionally show improved physicochemical properties and/or improved safety pharmacological properties.

Thus a further aspect of the invention are compounds if formula (I) which show improved physicochemical properties compared to compounds of the state of the art.

Another aspect of the invention are those compounds of formula (I) which show an improved safety pharmacological properties.

Further Definitions

By "alteration" is meant a change (increase or decrease) in the expression levels, structure or activity of a gene or polypeptide as detected by standard art known methods such as those described herein. As used herein, in one embodiment an alteration includes an about 10% change in expression levels, particularly an about 25% change, more particularly an about 40% change, and most particularly an about 50% or greater (e.g., ,

60% or greater, 70% or greater, 80% or greater, 90% or greater) change in expression levels. In certain embodiments an alteration includes a 10% or less (including 10 %) change in expression levels, particularly a 25% or less (including 25%) change, more particularly a 40% or less (including 40%) change, and most particularly a 50% or less (including 50%) or greater change in expression levels. In other embodiments an alteration includes a 9% - 1 1 % (including 9% and 1 1 %) change in expression levels, particularly a 10%-25% (including 10% and 25%) change, more particularly a 25% - 40% (including 25% and 40%) change, and most particularly a 40%-50% (including 40% - 50%) or greater than 50% (including 50%) change in expression levels. In other certain embodiments an alteration includes a 9% - 11 % (including 9% and 1 1 %) change in expression levels, particularly a 22%-28% (including 22% and 28%) change, more particularly a 35% - 45% (including 35% and 45%) change, and most particularly a 45%-55% (including 45% - 55%) or a greater or equal to 55% change in expression levels

By "fragment" is meant a portion of a polypeptide or nucleic acid molecule. This portion contains, particularly, at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% of the entire length of the reference nucleic acid molecule or polypeptide. In certain embodiments this portion contains, particularly, at least 9%-1 1 % (including 9% and 1 1 %), 18%-22% (including 18% ands 22%), 27%-33% (including 27% and 33%), 36%-44% (including 36% and 44%), 45%-55% (including 45% and 55%), 54%-66% (including 54% and 66%), 63%-77% (including 63% and 77%), 72%-88%(including 72%and 88%), or 81 %-99% (including 81 % and 99%) of the entire length of the reference nucleic acid molecule or polypeptide A fragment may contain about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 200, about 300, about 400, about 500, about 600, about 700, about 800, about 900, or about 1000 nucleotides or amino acids. In certain embodiments a fragment may contain 9-11 , about 18-22, 27-33, 36-44, 45-55, 54-66, 63-77, 72-88, 81- 99, 90-1 10, 180-220, 270-330, 360-440, 450-550, 540-660, 630-770, 720-880, 810-990, or 900-1 100 nucleotides or amino acids (including for each the mentioned limitation e.g. for“9-11” means including 9 and 11.

By“modulator” is meant any agent that binds to a polypeptide and alters a biological function or activity of the polypeptide. A modulator includes, without limitation, agents that reduce or eliminate a biological function or activity of a polypeptide (e.g., an“inhibitor”). For example, a modulator may inhibit a catalytic activity of a polypeptide. A modulator includes, without limitation, agents that increase or decrease binding of a polypeptide to another agent. For example, a modulator may promote binding of a polypeptide to another polypeptide. In some embodiments, the modulator of PDE3A and/or PDE3B polypeptide is a compound of formula (I).

“Hematopoietic hyperproliferative diseases” also known as myoproliferative diseases include e.g. polycythemia vera, essential thrombocytosis, thrombocytosis, primary myelofibrosis, and others.

“Hyperproliferative diseases” include, but are not limited to, for example: psoriasis, keloids, and other hyperplasias affecting the skin, benign hyperproliferative diseases, hematopoietic hyperproliferative diseases (including polycythemia vera, essential thrombocytosis, primary myelofibrosis), benign prostate hyperplasia (BPH), cancer (especially metastatic or malignant tumors, more specifically solid tumors and haematological tumors).

“Benign hyperproliferative diseases” include for example, endometriosis, leiomyoma and benign prostate hyperplasia.

By“marker” or“biomarker” is meant any protein or polynucleotide having an alteration in expression level or activity (e.g., at the protein or mRNA level) that is associated with a disease or disease. In particular embodiments, a marker of the invention is PDE3A and/or PBE3B or SLFN12 or CREB3L1 polypeptide or polynucleotide.

“Solid tumours” are such as e.g. cancers of the breast, brain, digestive tract, eye, head and neck, liver, parathyroid, reproductive organs, respiratory tract, skin, thyroid, urinary tract, and their distant metastases. Those diseases also include lymphomas, sarcomas, and leukaemias.

Examples of breast cancers include, but are not limited to, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, and lobular carcinoma in situ.

Examples of“brain cancers” include, but are not limited to, brain stem and hypophtalmic glioma, cerebellar and cerebral astrocytoma, medulloblastoma, ependymoma, as well as neuroectodermal and pineal tumour. Tumours of the“digestive tract” include, but are not limited to, anal, colon, colorectal, oesophageal, gallbladder, gastric, pancreatic, rectal, small-intestine, and salivary gland cancers.

“Eye cancers” include, but are not limited to, intraocular melanoma and retinoblastoma.

“Head-and-neck cancers” include, but are not limited to, laryngeal, hypopharyngeal, nasopharyngeal, oropharyngeal cancer, lip and oral cavity cancer and squamous cell.

Examples of “liver cancers” include, but are not limited to, hepatocellular carcinoma (liver cell carcinomas with or without fibrolamellar variant), cholangiocarcinoma (intrahepatic bile duct carcinoma), and mixed hepatocellular cholangiocarcinoma.

Examples of cancers of the“respiratory tract” include, but are not limited to, small-cell and non-small-cell lung carcinoma, as well as bronchial adenoma and pleuropulmonary blastoma.

“Reproductive organs” include female- and male reproductive organs.

“T umours of the female reproductive organs” include, but are not limited to, endometrial, cervical, ovarian, vaginal, and vulvar cancer, as well as sarcoma of the uterus. T“umours of the male reproductive organs” include, but are not limited to, prostate and testicular cancer.

“Skin cancers” include, but are not limited to, squamous cell carcinoma, Kaposi’s sarcoma, malignant melanoma (melanoma), Merkel cell skin cancer, and non melanoma skin cancer.

“Tumours of the urinary tract” include, but are not limited to, bladder, penile, kidney, renal pelvis, ureter, urethral and human papillary renal cancers.

“Lymphomas” include, but are not limited to, AIDS-related lymphoma, non-Hodgkin’s lymphoma, cutaneous T-cell lymphoma, Burkitt lymphoma, Hodgkin’s disease, and lymphoma of the central nervous system.

“Sarcomas” include, but are not limited to, sarcoma of the soft tissue, osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma, and rhabdomyosarcoma.

“Leukemias” include, but are not limited to, acute myeloid leukemia (AML), acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia.

By "subject" is meant a mammal, including, but not limited to, a human or non-human mammal, such as e.g. a bovine, equine, canine, ovine, or feline. As used herein, the terms“treat,” treating,”“treatment,” and the like refer to reducing or ameliorating a disease and/or symptoms associated therewith. It will be appreciated that, although not precluded, treating a disease or condition does not require that the disease, condition or symptoms associated therewith be completely eliminated.

Any compositions or methods provided herein can be combined with one or more of any of the other compositions and methods provided herein.

The term “treating” or “treatment” as stated throughout this document is used conventionally, and includes for example the management or care of a subject for the purpose of combating, alleviating, reducing, relieving, and/or improving the condition of a disease or disease, such as e.g. a carcinoma. These diseases have been well characterized in humans, but also exist with a similar etiology in other mammals, and can be treated by administering pharmaceutical compositions of the present invention.

The compounds of the present invention can be used in particular in therapy and prevention, i.e. prophylaxis, of hyperproliferative diseases, more particularly of cancer diseases, e.g. heamatological cancer diseases and tumour growth and metastasis, especially in solid tumours and heamatological cancer diseases of all indications and stages with or without pre-treatment of the tumour.

Generally, the use of chemotherapeutic agents and/or anti-cancer agents in combination with a compound or pharmaceutical composition of the present invention will serve to:

1. yield better efficacy in reducing the growth of a tumour or even eliminate the tumour as compared to administration of either agent alone,

2. provide for the administration of lesser amounts of the administered chemo therapeutic agents,

3. provide for a chemotherapeutic treatment that is well tolerated in the patient with fewer deleterious pharmacological complications than observed with single agent chemotherapies and certain other combined therapies,

4. provide for treating a broader spectrum of different cancer types in mammals, especially humans,

5. provide for a higher response rate among treated patients,

6. provide for a longer survival time among treated patients compared to standard chemotherapy treatments,

7. provide a longer time for tumour progression, and/or 8. yield efficacy and tolerability results at least as good as those of the agents used alone, compared to known instances where other cancer agent combinations produce antagonistic effects.

In another aspect, the cell is in vitro. In another embodiment, the cell is in vivo.

Optionally, an anti-neoplasia therapeutic (e.g., compounds of general formula (I)) may be administered in combination with any other standard anti-neoplasia therapy or conventional chemotherapeutic agent, such as e.g. an alkylating agent; such methods are known to the skilled artisan and described in Remington's Pharmaceutical Sciences by E. W. Martin. If desired, agents of the invention are administered in combination with any conventional anti-neoplastic therapy, including but not limited to, surgery, radiation therapy, or chemotherapy for the treatment of a neoplasia (e.g., melanoma, lung adenocarcinoma or a cervical cancer).

The present invention also provides compounds of formula (I) for methods of treating hyperproliferative diseases, more particularly cancer diseases including hematological cancer diseases and solid tumors.

In one embodiment the invention provides methods of treatment mentioned above where tumors are selected from list given above, more particularly the tumors are: tumors of the anus, the brain, the breast, the bones, the central and peripheral nervous system, the colon, the eye, the kidney, the endocrine glands (e.g., thyroid and adrenal cortex), the endometrium, the esophagus, the gastrointestinal tract (including gastrointestinal stromal tumors), the germ cells, the head and the neck, the kidney, the liver, the larynx and hypopharynx, the lung, the mesothelioma, the pancreas, the prostate, the rectum, the reproductive organs (e.g., cervix, ovary, prostate), the respiratory tract, the small intestine, the skin, the soft tissue, the stomach, the testis, the thyroid gland, the parathyroid gland, ureter, the urogenital tract, vagina and vulva and the connective tissue and metastases of these tumors. Malignant neoplasias include inherited cancers exemplified by Retinoblastoma and Wilms tumor.

In one embodiment the invention provides methods of treatment of cancer or the use of the compounds formula (I) for the treatment of a cancer disease, where said cancer disease is selected from melanoma and cervical cancer.

In one embodiment the invention provides methods of treatment of cancer or the use of the compounds formula (I) for the treatment of a cancer disease, where said cancer disease is melanoma. In one embodiment the invention provides methods of treatment of cancer or the use of the compounds formula (I) for the treatment of a cancer disease, where said cancer disease is cervical cancer.

These diseases have been well characterized in humans, but also exist with a similar etiology in other mammals, and can be treated by administering pharmaceutical compositions of the present invention.

The term“treating” or“treatment” as used in the present text is used conventionally, e.g., the management or care of a subject for the purpose of combating, alleviating, reducing, relieving, improving the condition of a disease or disorder, such as e.g. cancer.

The compounds of the present invention can be used in particular in therapy and prevention, i.e. prophylaxis, of hyperproliferative diseases, more particularly cancer diseases.

In accordance with a further aspect, the present invention provides compounds of general formula (I), as described supra, or stereoisomers, tautomers, N-oxides, hydrates, solvates, and salts thereof, particularly pharmaceutically acceptable salts thereof, or mixtures of same, for use in the treatment or prophylaxis of diseases, in particular hyperproliferative diseases.

The pharmaceutical activity of the compounds according to the invention can be explained by their activity by modulation of phosphodiesterase 3A/B (PDE3A and/or B) .

Thus a further aspect of the invention is a method of treatment comprising administering a compound of formula (I) or a pharmaceutical composition thereof to a patient suffering from a cancer disease being sensitive to a treatment with a PDE3A and/or PDE3B modulator.

Another aspect of the invention is a method of treatment comprising the steps of

deriving cancer cells from a patient,

detecting co-expression of PDE3A and/or

detecting co-expression of PDE3B

and detecting co-expression of Schlafen 12 ( SLFN12 ) and / or SLFN12L mRNA, polynucleotides or polypeptides

and/or a lack of decrease in expression of CREB3L1 mRNA, polynucleotides or polypeptides in said cancer cells, summarizing the overall results whether the data collected indicate that said cancer cells are sensitive to the treatment with a compound of formula (I), and administering a compound of formula (I) to said patient.

In accordance with a further aspect, the present invention provides the use of compounds of general formula (I), as described supra, or stereoisomers, tautomers, N-oxides, hydrates, solvates, and salts thereof, particularly pharmaceutically acceptable salts thereof, or mixtures of same, for the treatment or prophylaxis of diseases, in particular hyperprol iterative diseases, particularly cancer diseases.

In accordance with a further aspect, the present invention provides the a compound of formula (I), described supra, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, particularly a pharmaceutically acceptable salt thereof, or a mixture of same, for use in the prophylaxis or treatment of diseases, in particular hyperprol iterative diseases, particularly cancer diseases.

In accordance with a further aspect, the present invention provides the use of compounds of general formula (I), as described supra, or stereoisomers, tautomers, N-oxides, hydrates, solvates, and salts thereof, particularly pharmaceutically acceptable salts thereof, or mixtures of same, in a method of treatment or prophylaxis of diseases, in particular hyperproliferative diseases, particularly cancer diseases.

In accordance with a further aspect, the present invention provides a method of treating a hyperproliferative disease such as cancer in a subject, the method comprising administering to the subject a compound of any of claims 1-9, thereby treating the hyperproliferative disease

In accordance with a further aspect, the present invention provides use of a compound of general formula (I), as described supra, or stereoisomers, tautomers, N-oxides, hydrates, solvates, and salts thereof, particularly pharmaceutically acceptable salts thereof, or mixtures of same, for the preparation of a pharmaceutical composition, particularly a medicament, for the prophylaxis or treatment of diseases, in particular hyperproliferative diseases, particularly cancer diseases.

In accordance with a further aspect, the present invention provides a method of treatment or prophylaxis of diseases, in particular hyperproliferative diseases, particularly cancer diseases, comprising administering an effective amount of a compound of general formula (I), as described supra, or stereoisomers, tautomers, N-oxides, hydrates, solvates, and salts thereof, particularly pharmaceutically acceptable salts thereof, or mixtures of same to a patient in need thereof PHARMACEUTICAL COMPOSITION

In accordance with a further aspect, the present invention provides pharmaceutical compositions, in particular a medicament, comprising a compound of general formula (I), as described supra, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, a salt thereof, particularly a pharmaceutically acceptable salt, or a mixture of same, and one or more excipients), in particular one or more pharmaceutically acceptable excipient(s). Conventional procedures for preparing such pharmaceutical compositions in appropriate dosage forms can be utilized.

The present invention furthermore provides pharmaceutical compositions, in particular medicaments, which comprise at least one compound according to the invention, conventionally together with one or more pharmaceutically suitable excipients, and for their use for the above mentioned purposes.

It is possible for the compounds according to the invention to have systemic and/or local activity. For this purpose, they can be administered in a suitable manner, such as, for example, via the oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, vaginal, dermal, transdermal, conjunctival, otic route or as an implant or stent.

For these administration routes, it is possible for the compounds according to the invention to be administered in suitable administration forms.

For oral administration, it is possible to formulate the compounds according to the invention to dosage forms known in the art that deliver the compounds of the invention rapidly and/or in a modified manner, such as, for example, tablets (uncoated or coated tablets, for example with enteric or controlled release coatings that dissolve with a delay or are insoluble), orally-disintegrating tablets, films/wafers, films/lyophylisates, capsules (for example hard or soft gelatine capsules), sugar-coated tablets, granules, pellets, powders, emulsions, suspensions, aerosols or solutions. It is possible to incorporate the compounds according to the invention in crystalline and/or amorphised and/or dissolved form into said dosage forms.

Parenteral administration can be effected with avoidance of an absorption step (for example intravenous, intraarterial, intracardial, intraspinal or intralumbal) or with inclusion of absorption (for example intramuscular, subcutaneous, intracutaneous, percutaneous or intraperitoneal). Administration forms which are suitable for parenteral administration are, inter alia, preparations for injection and infusion in the form of solutions, suspensions, emulsions, lyophylisates or sterile powders. Examples which are suitable for other administration routes are pharmaceutical forms for inhalation [inter alia powder inhalers, nebulizers], nasal drops, nasal solutions, nasal sprays; tablets/films/wafers/capsules for lingual, sublingual or buccal administration; suppositories; eye drops, eye ointments, eye baths, ocular inserts, ear drops, ear sprays, ear powders, ear-rinses, ear tampons; vaginal capsules, aqueous suspensions (lotions, mixturae agitandae), lipophilic suspensions, emulsions, ointments, creams, transdermal therapeutic systems (such as, for example, patches), milk, pastes, foams, dusting powders, implants or stents.

The compounds according to the invention can be incorporated into the stated administration forms. This can be effected in a manner known per se by mixing with pharmaceutically suitable excipients. Pharmaceutically suitable excipients include, inter alia,

• fillers and carriers (for example cellulose, microcrystalline cellulose (such as, for example, Avicel^®), lactose, mannitol, starch, calcium phosphate (such as, for example, Di-Cafos^®)),

• ointment bases (for example petroleum jelly, paraffins, triglycerides, waxes, wool wax, wool wax alcohols, lanolin, hydrophilic ointment, polyethylene glycols),

• bases for suppositories (for example polyethylene glycols, cacao butter, hard fat),

• solvents (for example water, ethanol, isopropanol, glycerol, propylene glycol, medium chain-length triglycerides, fatty oils, liquid polyethylene glycols, paraffins),

• surfactants, emulsifiers, dispersants or wetters (for example sodium dodecyl sulfate), lecithin, phospholipids, fatty alcohols (such as, for example, Lanette^®), sorbitan fatty acid esters (such as, for example, Span^®), polyoxyethylene sorbitan fatty acid esters (such as, for example, Tween^®), polyoxyethylene fatty acid glycerides (such as, for example, Cremophor^®), polyoxethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, glycerol fatty acid esters, poloxamers (such as, for example, Pluronic^®),

• buffers, acids and bases (for example phosphates, carbonates, citric acid, acetic acid, hydrochloric acid, sodium hydroxide solution, ammonium carbonate, trometamol, triethanolamine),

• isotonicity agents (for example glucose, sodium chloride),

• adsorbents (for example highly-disperse silicas), • viscosity-increasing agents, gel formers, thickeners and/or binders (for example polyvinylpyrrolidone, methylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, carboxymethylcellulose-sodium, starch, carbomers, polyacrylic acids (such as, for example, Carbopol^®); alginates, gelatine),

• disintegrants (for example modified starch, carboxymethylcellulose-sodium, sodium starch glycolate (such as, for example, Explotab^®), cross- linked polyvinylpyrrolidone, croscarmellose-sodium (such as, for example, AcDiSol^®)),

• flow regulators, lubricants, glidants and mould release agents (for example magnesium stearate, stearic acid, talc, highly-disperse silicas (such as, for example, Aerosil^®)),

• coating materials (for example sugar, shellac) and film formers for films or diffusion membranes which dissolve rapidly or in a modified manner (for example polyvinylpyrrolidones (such as, for example, Kollidon^®), polyvinyl alcohol, hydroxypropylmethylcellulose, hydroxypropylcellulose, ethylcellulose, hydroxypropylmethylcellulose phthalate, cellulose acetate, cellulose acetate phthalate, polyacrylates, polymethacrylates such as, for example, Eudragit^®)),

• capsule materials (for example gelatine, hydroxypropylmethylcellulose),

• synthetic polymers (for example polylactides, polyglycolides, polyacrylates, polymethacrylates (such as, for example, Eudragit^®), polyvinylpyrrolidones (such as, for example, Kollidon^®), polyvinyl alcohols, polyvinyl acetates, polyethylene oxides, polyethylene glycols and their copolymers and blockcopolymers),

• plasticizers (for example polyethylene glycols, propylene glycol, glycerol, triacetine, triacetyl citrate, dibutyl phthalate),

• penetration enhancers,

• stabilisers (for example antioxidants such as, for example, ascorbic acid, ascorbyl palmitate, sodium ascorbate, butylhydroxyanisole, butylhydroxytoluene, propyl gallate),

• preservatives (for example parabens, sorbic acid, thiomersal, benzalkonium chloride, chlorhexidine acetate, sodium benzoate),

• colourants (for example inorganic pigments such as, for example, iron oxides, titanium dioxide), • flavourings, sweeteners, flavour- and/or odour-masking agents.

The present invention furthermore relates to a pharmaceutical composition which comprise at least one compound according to the invention, conventionally together with one or more pharmaceutically suitable excipient(s), and to their use according to the present invention.

COMBINATIONS

In accordance with another aspect, the present invention provides pharmaceutical combinations, in particular medicaments, comprising at least one compound of general formula (I) of the present invention and at least one or more further active ingredients, in particular for the treatment and/or prophylaxis of a hyperproliferative disease, a cancer disease.

Particularly, the present invention provides a pharmaceutical combination, which comprises:

• one or more first active ingredients, in particular compounds of general formula (I) as defined supra, and

• one or more further active ingredients, in particular a hyperproliferative disease, a cancer disease.

The term“combination” in the present invention is used as known to persons skilled in the art, it being possible for said combination to be a fixed combination, a non-fixed combination or a kit-of-parts.

A“fixed combination” in the present invention is used as known to persons skilled in the art and is defined as a combination wherein, for example, a first active ingredient, such as one or more compounds of general formula (I) of the present invention, and a further active ingredient are present together in one unit dosage or in one single entity. One example of a“fixed combination” is a pharmaceutical composition wherein a first active ingredient and a further active ingredient are present in admixture for simultaneous administration, such as e.g. in a formulation. Another example of a“fixed combination” is a pharmaceutical combination wherein a first active ingredient and a further active ingredient are present in one unit without being in admixture.

A non-fixed combination or“kit-of-parts” in the present invention is used as known to persons skilled in the art and is defined as a combination wherein a first active ingredient and a further active ingredient are present in more than one unit. One example of a non- fixed combination or kit-of-parts is a combination wherein the first active ingredient and the further active ingredient are present separately. It is possible for the components of the non-fixed combination or kit-of-parts to be administered separately, sequentially, simultaneously, concurrently or chronologically staggered.

The compounds of the present invention can be administered as the sole pharmaceutical agent or in combination with one or more other pharmaceutically active ingredients where the combination causes no unacceptable adverse effects. The present invention also provides such pharmaceutical combinations. For example, the compounds of the present invention can be combined with known anti-cancer-agents, said anti-cancer agents including but not limited to:

1311-chTNT, abarelix, abemaciclib, abiraterone, acalabrutinib, aclarubicin, adalimumab, ado-trastuzumab emtansine, afatinib, aflibercept, aldesleukin, alectinib, alemtuzumab, alendronic acid, alitretinoin, alpharadin, altretamine, amifostine, aminoglutethimide, hexyl aminolevulinate, amrubicin, amsacrine, anastrozole, ancestim, anethole dithiolethione, anetumab ravtansine, angiotensin II, antithrombin III, apalutamide, aprepitant, arcitumomab, arglabin, arsenic trioxide, asparaginase, atezolizumab, avelumab, axicabtagene ciloleucel, axitinib, azacitidine, basiliximab, belotecan, bendamustine, besilesomab, belinostat, bevacizumab, bexarotene, bicalutamide, bisantrene, bleomycin, blinatumomab, bortezomib, bosutinib, buserelin, brentuximab vedotin, brigatinib, busulfan, cabazitaxel, cabozantinib, calcitonine, calcium folinate, calcium levofolinate, capecitabine, capromab, carbamazepine carboplatin, carboquone, carfilzomib, carmofur, carmustine, catumaxomab, celecoxib, celmoleukin, cemiplimab, ceritinib, cetuximab, chlorambucil, chlormadinone, chlormethine, cidofovir, cinacalcet, cisplatin, cladribine, clodronic acid, clofarabine, cobimetinib, copanlisib, crisantaspase, crizotinib, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, daratumumab, darbepoetin alfa, dabrafenib, dasatinib, daunorubicin, decitabine, degarelix, denileukin diftitox, denosumab, depreotide, deslorelin, dianhydrogalactitol, dexrazoxane, dibrospidium chloride, dianhydrogalactitol, diclofenac, dinutuximab, docetaxel, dolasetron, doxifluridine, doxorubicin, doxorubicin + estrone, dronabinol, durvalumab, eculizumab, edrecolomab, elliptinium acetate, elotuzumab, eltrombopag, enasidenib, endostatin, enocitabine, enzalutamide, epirubicin, epitiostanol, epoetin alfa, epoetin beta, epoetin zeta, eptaplatin, eribulin, erlotinib, esomeprazole, estradiol, estramustine, ethinylestradiol, etoposide, everolimus, exemestane, fadrozole, fentanyl, filgrastim, fluoxymesterone, floxuridine, fludarabine, fluorouracil, flutamide, folinic acid, formestane, fosaprepitant, fotemustine, fulvestrant, gadobutrol, gadoteridol, gadoteric acid meglumine, gadoversetamide, gadoxetic acid, gallium nitrate, ganirelix, gefitinib, gemcitabine, gemtuzumab, Glucarpidase, glutoxim, GM-CSF, goserelin, granisetron, granulocyte colony stimulating factor, histamine dihydrochloride, histrelin, hydroxycarbamide, 1-125 seeds, lansoprazole, ibandronic acid, ibritumomab tiuxetan, ibrutinib, idarubicin, ifosfamide, imatinib, imiquimod, improsulfan, indisetron, incadronic acid, ingenol mebutate, inotuzumab ozogamicin, interferon alfa, interferon beta, interferon gamma, iobitridol, iobenguane (1231), iomeprol, ipilimumab, irinotecan, Itraconazole, ixabepilone, ixazomib, lanreotide, lansoprazole, lapatinib, lasocholine, lenalidomide, lenvatinib, lenograstim, lentinan, letrozole, leuprorelin, levamisole, levonorgestrel, levothyroxine sodium, lisuride, lobaplatin, lomustine, lonidamine, lutetium Lu 177 dotatate, masoprocol, medroxyprogesterone, megestrol, melarsoprol, melphalan, mepitiostane, mercaptopurine, mesna, methadone, methotrexate, methoxsalen, methylaminolevulinate, methylprednisolone, methyltestosterone, metirosine, midostaurin, mifamurtide, miltefosine, miriplatin, mitobronitol, mitoguazone, mitolactol, mitomycin, mitotane, mitoxantrone, mogamulizumab, molgramostim, mopidamol, morphine hydrochloride, morphine sulfate, mvasi, nabilone, nabiximols, nafarelin, naloxone + pentazocine, naltrexone, nartograstim, necitumumab, nedaplatin, nelarabine, neratinib, neridronic acid, netupitant/palonosetron, nivolumab, pentetreotide, nilotinib, nilutamide, nimorazole, nimotuzumab, nimustine, nintedanib, niraparib, nitracrine, nivolumab, obinutuzumab, octreotide, ofatumumab, olaparib, olaratumab, omacetaxine mepesuccinate, omeprazole, ondansetron, oprelvekin, orgotein, orilotimod, osimertinib, oxaliplatin, oxycodone, oxymetholone, ozogamicine, p53 gene therapy, paclitaxel, palbociclib, palifermin, palladium-103 seed, palonosetron, pamidronic acid, panitumumab, panobinostat, pantoprazole, pazopanib, pegaspargase, PEG-epoetin beta (methoxy PEG-epoetin beta), pembrolizumab, pegfilgrastim, peginterferon alfa-2b, pembrolizumab, pemetrexed, pentazocine, pentostatin, peplomycin, Perflubutane, perfosfamide, Pertuzumab, picibanil, pilocarpine, pirarubicin, pixantrone, plerixafor, plicamycin, poliglusam, polyestradiol phosphate, polyvinylpyrrolidone + sodium hyaluronate, polysaccharide-K, pomalidomide, ponatinib, porfimer sodium, pralatrexate, prednimustine, prednisone, procarbazine, procodazole, propranolol, quinagolide, rabeprazole, racotumomab, radium-223 chloride, radotinib, raloxifene, raltitrexed, ramosetron, ramucirumab, ranimustine, rasburicase, razoxane, refametinib, regorafenib, ribociclib, risedronic acid, rhenium-186 etidronate, rituximab, rolapitant, romidepsin, romiplostim, romurtide, rucaparib, samarium (153Sm) lexidronam, sargramostim, sarilumab, satumomab, secretin, siltuximab, sipuleucel-T, sizofiran, sobuzoxane, sodium glycididazole, sonidegib, sorafenib, stanozolol, streptozocin, sunitinib, talaporfin, talimogene laherparepvec, tamibarotene, tamoxifen, tapentadol, tasonermin, teceleukin, technetium (99mTc) nofetumomab merpentan, 99mTc-HYNIC-[Tyr3]-octreotide, tegafur, tegafur + gimeracil + oteracil, temoporfin, temozolomide, temsirolimus, teniposide, testosterone, tetrofosmin, thalidomide, thiotepa, thymalfasin, thyrotropin alfa, tioguanine, tisagenlecleucel, tislelizumab, tocilizumab, topotecan, toremifene, tositumomab, trabectedin, trametinib, tramadol, trastuzumab, trastuzumab emtansine, treosulfan, tretinoin, trifluridine + tipiracil, trilostane, triptorelin, trametinib, trofosfamide, thrombopoietin, tryptophan, ubenimex, valatinib, valrubicin, vandetanib, vapreotide, vemurafenib, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, vismodegib, vorinostat, vorozole, yttrium-90 glass microspheres, zinostatin, zinostatin stimalamer, zoledronic acid, zorubicin.

Based upon standard laboratory techniques known to evaluate compounds useful for the treatment of hyperproliferative diseases, more particularly cancer diseases by standard toxicity tests and by standard pharmacological assays for the determination of treatment of the conditions identified above in mammals, and by comparison of these results with the results of known active ingredients or medicaments that are used to treat these conditions, the effective dosage of the compounds of the present invention can readily be determined for treatment of each desired indication. The amount of the active ingredient to be administered in the treatment of one of these conditions can vary widely according to such considerations as the particular compound and dosage unit employed, the mode of administration, the period of treatment, the age and sex of the patient treated, and the nature and extent of the condition treated.

The total amount of the active ingredient to be administered will generally range from about 0.001 mg/kg to about 500 mg/kg body weight per day, particularly about 0.001 mg/kg to about 200 mg/kg body weight per day, and more particularly from about 0.01 mg/kg to about 50 mg/kg body weight per day. Clinically useful dosing schedules will range from one to three times a day dosing to once every four weeks dosing. In addition, it is possible for "drug holidays", in which a patient is not dosed with a drug for a certain period of time, to be beneficial to the overall balance between pharmacological effect and tolerability. It is possible for a unit dosage to contain from about 0.5 mg to about 1500 mg of active ingredient, and can be administered one or more times per day or less than once a day. The average daily dosage for administration by injection, including intravenous, intramuscular, subcutaneous and parenteral injections, and use of infusion techniques will particularly be from 0.01 to 200 mg/kg of total body weight. The average daily rectal dosage regimen will particularly be from 0.01 to 200 mg/kg of total body weight. The average daily vaginal dosage regimen will particularly be from 0.01 to 200 mg/kg of total body weight. The average daily topical dosage regimen will particularly be from 0.1 to 200 mg administered between one to four times daily. The transdermal concentration will particularly be that required to maintain a daily dose of from 0.01 to 200 mg/kg. The average daily inhalation dosage regimen will particularly be from 0.01 to 100 mg/kg of total body weight. For oral administration the dosing schedule maybe once or two time or three times daily and a dose range as referred to above for general dosing is possible.

Of course the specific initial and continuing dosage regimen for each patient will vary according to the nature and severity of the condition as determined by the attending diagnostician, the activity of the specific compound employed, the age and general condition of the patient, time of administration, route of administration, rate of excretion of the drug, drug combinations, and the like. The desired mode of treatment and number of doses of a compound of the present invention or a pharmaceutically acceptable salt or ester or composition thereof can be ascertained by those skilled in the art using conventional treatment tests.

EXAMPLES

EXPERIMENTAL SECTION

NMR peak forms are stated as they appear in the spectra, possible higher order effects have not been considered.

Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker (300 or 400 MHz ¹H, 75 or 101 MHz ¹³C) spectrometer. Proton, fluorine, and carbon chemical shifts are reported in ppm (d) referenced to the NMR solvent. Data are reported as follows: chemical shifts, multiplicity (br = broad, s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet; coupling constant(s) in Hz).

The ¹H-NMR data of selected compounds are listed in the form of ¹H-NMR peaklists. Therein, for each signal peak the d value in ppm is given, followed by the signal intensity, reported in round brackets. The d value-signal intensity pairs from different peaks are separated by commas. Therefore, a peaklist is described by the general form: di (intensityi), 62 (intens^), ... , d, (intensity,), ... , d_h (intensity_n).

The intensity of a sharp signal correlates with the height (in cm) of the signal in a printed NMR spectrum. When compared with other signals, this data can be correlated to the real ratios of the signal intensities. In the case of broad signals, more than one peak, or the center of the signal along with their relative intensity, compared to the most intense signal displayed in the spectrum, are shown. A ¹ H-NMR peaklist is similar to a classical ¹ H-NMR readout, and thus usually contains all the peaks listed in a classical NMR interpretation. Moreover, similar to classical ¹H-NMR printouts, peaklists can show solvent signals, signals derived from stereoisomers of the particular target compound, peaks of impurities, ¹³C satellite peaks, and/or spinning sidebands. The peaks of stereoisomers, and/or peaks of impurities are typically displayed with a lower intensity compared to the peaks of the target compound (e.g., with a purity of >90%). Such stereoisomers and/or impurities may be typical for the particular manufacturing process, and therefore their peaks may help to identify a reproduction of the manufacturing process on the basis of "by-product fingerprints". An expert who calculates the peaks of the target compound by known methods (MestReC, ACD simulation, or by use of empirically evaluated expectation values), can isolate the peaks of the target compound as required, optionally using additional intensity filters. Such an operation would be similar to peak-picking in classical ¹H-NMR interpretation. A detailed description of the reporting of NMR data in the form of peaklists can be found in the publication "Citation of NMR Peaklist Data within Patent Applications" (cf. http://www.researchdisclosure.com/searching-disclosures, Research Disclosure Database Number 605005, 2014, 01 Aug 2014). In the peak picking routine, as described in the Research Disclosure Database Number 605005, the parameter "MinimumHeight" can be adjusted between 1% and 4%. However, depending on the chemical structure and/or depending on the concentration of the measured compound it may be reasonable to set the parameter "MinimumHeight" <1 %.

Chemical names were generated using the ACD/Name software from ACD/Labs. In some cases generally accepted names of commercially available reagents were used in place of ACD/Name generated names.

The following table 1 lists the abbreviations used in this paragraph and in the Examples section as far as they are not explained within the text body. Other abbreviations have their meanings customary perse to the skilled person.

Table 1 : Abbreviations

The various aspects of the invention described in this application are illustrated by the following examples which are not meant to limit the invention in any way.

The example testing experiments described herein serve to illustrate the present invention and the invention is not limited to the examples given.

EXPERIMENTAL SECTION - GENERAL PART

All reagents, for which the synthesis is not described in the experimental part, are either commercially available, or are known compounds or may be formed from known compounds by known methods by a person skilled in the art.

The compounds and intermediates produced according to the methods of the invention may require purification. Purification of organic compounds is well known to the person skilled in the art and there may be several ways of purifying the same compound. In some cases, no purification may be necessary. In some cases, the compounds may be purified by crystallization. In some cases, impurities may be stirred out using a suitable solvent. In some cases, the compounds may be purified by chromatography, particularly flash column chromatography, using for example prepacked silica gel cartridges, e.g. Biotage SNAP cartidges KP-Sil^® or KP-NH^® in combination with a Biotage autopurifier system (SP4^® or Isolera Four^®) and eluents such as e.g. gradients of hexane/ethyl acetate or DCM/methanol. In some cases, the compounds may be purified by preparative HPLC using for example a Waters autopurifier equipped with a diode array detector and/or on line electrospray ionization mass spectrometer in combination with a suitable prepacked reverse phase column and eluents such as e.g. gradients of water and acetonitrile which may contain additives such as e.g. trifluoroacetic acid, formic acid or aqueous ammonia.

In some cases, purification methods as described above can provide those compounds of the present invention which possess a sufficiently basic or acidic functionality in the form of a salt, such as e.g., in the case of a compound of the present invention which is sufficiently basic, a trifluoroacetate or formate salt for example, or, in the case of a compound of the present invention which is sufficiently acidic, an ammonium salt for example. A salt of this type can either be transformed into its free base or free acid form, respectively, by various methods known to the person skilled in the art, or be used as salts in subsequent biological assays. It is to be understood that the specific form (e.g. salt, free base etc.) of a compound of the present invention as isolated and as described herein is not necessarily the only form in which said compound can be applied to a biological assay in order to quantify the specific biological activity.

UPLC-MS Standard Procedures

Analytical UPLC-MS was performed as described below. The masses (m/z) are reported from the positive mode electrospray ionisation unless the negative mode is indicated (ESI- ). In most of the cases method 1 is used. If not, it is indicated.

EXPERIMENTAL SECTION - GENERAL PROCEDURES

Analytical LC-MS Methods:

Method 1 :

Instrument: Waters Acquity UPLC-MS SingleQuad; Column: Acquity UPLC BEH C18 1.7 pm, 50x2.1 mm; eluent A: water + 0.1 vol % formic acid (99%), eluent B: acetonitrile; gradient: 0-1.6 min 1-99% B, 1.6-2.0 min 99% B; flow 0.8 ml/min; temperature: 60 °C; DAD scan: 210-400 nm.

Method 2:

Instrument: Waters Acquity UPLC-MS SingleQuad; Column: Acquity UPLC BEH C18 1.7 pm, 50x2.1 mm; eluent A: water + 0.2 vol % aqueous ammonia (32%), eluent B: acetonitrile; gradient: 0-1.6 min 1-99% B, 1.6-2.0 min 99% B; flow 0.8 ml/min; temperature: 60 °C; DAD scan: 210-400 nm.

Method 3:

Instrument: SHIMADZU LCMS - UFLC 20-AD - LCMS 2020 MS detector; Column: Waters

Atlantis dC18 3pm, 2.1 x 100 mm; eluent A: water + 0.1 % formic acid (v/v), eluent B: acetonitrile + 0.1% formic acid (v/v); gradient: 0-5.00 min 5-100% B 5.00-5.40 min 100% B; flow: 0.6 mL/min; temperature: 40°C; PDA scan: 210 - 420 nm.

Preparative LC-MS Methods:

Method 4:

Instrument: Waters Acquity UPLC-MS SingleQuad; Colum: Acquity UPLC BEH C18 1.7 50x2.1mm; eluent A: water + 0.2 vol % aqueous ammonia (32%), eluent B: acetonitrile; gradient: 0-1.6 min 1-99% B, 1.6-2.0 min 99% B; flow 0.8 ml/min; temperature: 60 °C; DAD scan: 210-400 nm

Method 5:

Instrument: Waters Acquity UPLC-MS SingleQuad; Column: Acquity UPLC BEH C18 1.7 50x2.1mm; eluent A: water + 0.1 vol % formic acid (99%), eluent B: acetonitrile; gradient: 0-1.6 min 1-99% B, 1.6-2.0 min 99% B; flow 0.8 ml/min; temperature: 60 °C; DAD scan: 210-400 nm

EXPERIMENTAL SECTION - GENERAL PROCEDURES

General details

All reactions were carried out under nitrogen (N2) atmosphere. All reagents and solvents were purchased from commercial vendors and used as received. Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker (300 or 400 MHz ¹H, 75 or 101 MHz ¹³C) spectrometer. Proton and carbon chemical shifts are reported in ppm (d) referenced to the NMR solvent. Data are reported as follows: chemical shifts, multiplicity (br = broad, s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet; coupling constant(s) in Hz). Flash chromatography was performed using 40-60 pm Silica Gel (60 A mesh) on a Teledyne Isco Combiflash Rf or a Biotage Isolera. Analytical thin layer chromatography (TLC) was performed on EM Reagent 0.25 mm silica gel 60-F plates.

EXPERIMENTAL SECTION - INTERMEDIATES

Intermediate 1

2-bromo-1-[4-fluoro-3-(trifluoromethyl)phenyl]ethan-1-one

1-[4-fluoro-3-(trifluoromethyl)phenyl]ethanone (15 ml, 97 mmol, commercial) was dissolved in acetic acid (120 ml_) at room temperature. Bromine (5.0 ml, 100 % purity, 97 mmol) was dissolved in acetic acid (10 ml_) and the solution was added dropwise into the reaction mixture, which was stirred overnight, its colour turned from brown to orange. The mixture was concentrated under reduced pressure and used as crude material (27.7 g) in the next step.

Intermediate 2

2-[4-fluoro-3-(trifluoromethyl)phenyl]-2-oxoethyl acetate

2-bromo-1-[4-fluoro-3-(trifluoromethyl)phenyl]ethanone (27.7 g, 97.0 mmol, Intermediate 1) was dissolved under nitrogen in DMF (100 ml_), potassium acetate (28.6 g, 291 mmol) and potassium iodide (16.1 g, 97.0 mmol) were added and the mixture was stirred at room temperature overnight. The mixture was diluted with water and extracted with ethyl acetate three times. The combined organic layers were dried over anhydrous sodium sulfate and the filtrate was concentrated under reduced pressure. The crude material (25.7 g) was used without further purification.

LC-MS (Method 1): R_t = 1.14 min; MS (ESIneg): m/z = 263 [M-H]-

Intermediate 3

methyl (2Z)-2-{2-(acetyloxy)-1-[4-fluoro-3-(trifluoromethyl)phenyl]ethylidene}hydrazine-1- carboxylate

2-[4-fluoro-3-(trifluoromethyl)phenyl]-2-oxoethyl acetate (25.7 g, 97.3 mmol, Intermediate 2) was dissolved in methanol (170 ml_) and acidified to pH 5 using aqueous hydrochloric acid (1M). Then methyl hydrazinecarboxylate (9.64 g, 107 mmol) was added and the mixture was stirred for 16 hours at room temperature. The resulting precipitate was collected by filtration and the solids were washed with methanol. The filtrates were combined and concentrated. The resulting solids were triturated in a mixture of methanol and DCM (1 :1 v/v) amd the white precipitate was collected. The solids from both filtrations were combined to yield the title product (15.6 g) which was used without further purification.

LC-MS (Method 1): R_t = 1.15 min; MS (ESIneg): m/z = 335 [M-H]-

Intermediate 4

5-[4-fluoro-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one

Methyl (2)-2-{2-(acetyloxy)-1-[4-fluoro-3-(trifluoromethyl)phenyl]ethylidenhydrazine- carboxylate (3.30 g, 9.81 mmol, Intermediate 3) was suspended in ethanol (83 ml_) under nitrogen and sodium ethylate solution in ethanol (5.5 ml_, 21 % purity, 15 mmol) was added. It was stirred at room temperature for 30 min. The reaction mixture was diluted with aqueous saturated ammonium chloride solution and water and stirred for 1 hour. Precipitated product was filtered off. The filter cake was washed with water and dried under vacuo to give 2.50 g (95 % purity) of the titled compound.

LC-MS (Method 1): R_t = 1.03 min; MS (ESIneg): m/z = 261 [M-H]- Ή-NMR (400 MHz, DMSO-d6) d [ppm]: 1.228 (0.61), 1.246 (1.18), 1.264 (0.57), 2.327

(0.49), 2.669 (0.50), 4.164 (0.49), 4.181 (0.47), 4.631 (1.19), 5.421 (16.00), 7.617 (1.38),

7.640 (2.05), 7.665 (1.50), 8.028 (2.22), 8.045 (2.02), 8.060 (1.16), 8.066 (1.06), 8.072

(1.26), 8.081 (1.28), 8.088 (1.01), 8.094 (1.06), 11.215 (0.91). Intermediate 5

1-[4-chloro-3-(trifluoromethyl)phenyl]propan-1-one

To a cooled solution of 4-chloro-N-methoxy-N-methyl-3-(trifluoromethyl)benzamide (118.1 g, 441 mmol) in THF (833 ml_) at 0 °C, was added a solution of ethylmagnesium bromide (3M in diethyl ether, 309 ml_, 927mmol) dropwise over a period of 3 hours via cannula.

After this time, the reaction mixture was partitioned between EtOAc and 1M aqueous hydrogen chloride solution, passed through a Celite pad, with the organic layer separated, the aqueous layer back-extracted with further aliquots of EtOAc and the combined orgnaic layers washed with saturated aquoeus sodium chloride solution, dried (MgS04), filtered and concentrated in vacuo. The residual material was dry loaded and purified by dry flash chromatography (silica gel, eluting with heptanes- EtOAc, 95:5) to afford 70.10 g the title compound (67%) as a yellow solid. LCMS (Method 3, 2 min) 100% @ Rt 1.25 mins, MS (ESIpos) m/z no ionisation observed

¹H NMR (250 MHz, Chloroform-d) d = 1.27 (t, J = 7.2 Hz, 3H), 3.03 (q, J = 7.2 Hz, 2H), 7.64 (d, J = 8.4 Hz, 1 H), 8.08 (dd, J = 8.3, 2.0 Hz, 1 H), 8.30 (d, J = 1.9 Hz, 1 H).

Intermediate 6

tert-Butyl({(1 E)-1-[4-chloro-3-(trifluoromethyl)phenyl]prop-1-en-1-yl}oxy)dimethylsilane

To a cooled solution of 1-[4-chloro-3-(trifluoromethyl)phenyl]propan-1-one (70.10 g, 296.25 mmol, Intermediate 5) in tetrahydrofuran (550 ml_) at -78 °C, was added a solution of LHMDS (1M in THF, 296 mL, 296.25 mmol) dropwise over a period of 2 hours via cannula. After 1 h, a solution of tBDMSCI (44.65 g, 296.25 mmol) in THF (70 mL) was added dropwise via cannula. The reaction mixture was allowed to warm to RT and was stirred at RT for 72 h and partitioned between EtOAc and 1M aqueous hydrogen chloride solution. The organic layer was separated, with the aqueous layer back-extracted with a further aliquot of EtOAc and the combined organic layer was washed with saturated aquoeus sodium chloride solution, dried (MgS04), filtered and concentrated in vacuo. The residue was purified via dry flash chromatography (silica gel, eluting with heptanes-EtOAc, 1 :0, then 7:3) to afford 93.78 g the title compound ( 90%) as a pale yellow solid. LCMS (Method 3, 2 min) 100% @ Rt 2.03 mins, MS (ESIpos) m/z no ionisation observed

¹H NMR (500 MHz, Chloroform-d) d = -0.00 (s, 6H), 1.02 (s, 9H), 1.77 (d, J = 6.9 Hz, 3H), 5.33 (q, J = 6.9 Hz, 1 H), 7.43 (d, J = 8.4 Hz, 1 H), 7.55 (dd, J = 8.4, 2.0 Hz, 1 H), 7.80 (d, J

= 2.0 Hz, 1 H).

Intermediate 7

tert-Butyl({(1 E)-1-[4-fluoro-3-(trifluoromethyl)phenyl]prop-1-en-1-yl}oxy)dimethylsilane

A solution of 4-fluoro-3-trifluoromethylpropiophenone (12 g, 55 mmol, CAS 239107-27-8) was dissolved in 60 ml_ of THF and cooled in a dry ice bath before addition of 60 ml_ (60 mmol) of 1 N LHMDS (in THF). After 1 h, a solution of 9.0 g of tBDMSCI (60 mmol) in 15 ml_ THF was added dropwise and the reaction was stirred, warming to room temperature overnight. The next day, the reaction mixture was concentrated and stirred in 500 ml_ of hexane for 30 min before filtering and concentrating. Chromatography with hexane on silica gel pretreated with Et₃N yielded 15.3 g product (84%).

¹H NMR (400 MHz, CDCh) d 7.71 (dd, J = 6.9, 2.1 Hz, 1 H), 7.62 (ddd, J = 7.3, 4.7, 2.2 Hz, 1 H), 7.13 (t, J = 9.4 Hz, 1 H), 5.26 (q, J = 6.9 Hz, 1 H), 1.76 (d, J = 6.9 Hz, 3H), 1.02 (s, 9H), -0.00 (s, 6H). ¹⁹F NMR (376 MHz, CDCI₃) d -61.49 (d, J = 12.7 Hz), -116.85 (q, J = 12.8 Hz).

Intermediate 8

(2S)-1-[4-Chloro-3-(trifluoromethyl)phenyl]-2-hydroxypropan-1-one

Following a literature procedure (J. Org. Chem. 1992, 57, 5067), to tert-butanol: water (1 :1 v:v 0.83 L) was added methanesulfonamide (8.40 g, 88.35 mmol), and AD-mix-a (124 g, CAS-Nr 4039-32-1). The mixture was cooled to 0 °C before addition of tert-butyl({(1 E)-1- [4-chloro-3-(trifluoromethyl)phenyl]prop-1-en-1-yl}oxy)dimethylsilane (31.00 g, 88.35 mmol, Intermediate 6) portionwise over the course of 30 minutes. The mixture was kept at 0 °C for 5 hours and then was allowed to warm to room temperature overnight. After this time, sodium sulfite (75 g) was added and stirred for 30 minutes, with the mixture was filtered through a pad of Celite, washing with water and EtOAc. The organic layer was separated washed with saturated aqueous sodium chloride solution, dried (MgS04), filtered and concentrated in vacuo. The residual material was purified via dry flash chromatography (silica gel, eluting with heptanes-EtOAc, 4:1) to afford 18.58 g the title compound (78%, 98.16 %ee) as a pale yellow solid. LCMS (Method 3, 2 min) 94% @ Rt 1.10 mins, MS (ESIpos) m/z no ionisation observed

¹H NMR (250 MHz, Chloroform-d) d = 1.36 (d, J = 7.0 Hz, 3H), 3.46 (d, J = 6.5 Hz, 1 H), 5.03 (p, J = 7.0 Hz, 1 H), 7.57 (d, J = 8.3 Hz, 1 H), 7.92 (dd, J = 8.3, 2.0 Hz, 1 H), 8.16 (d, J

= 1.9 Hz, 1 H). Chiral Analysis: Column: Amylose-2 25cm Mobile phase: 95:5 Heptane:

I PA Flow rate: 1 ml/min UV at 250nm Runtime: 25mins Enantiomer 1 retention time = 8.57 mins @ 0.92% Enantiomer 2 retention time = 16.09 mins @ 99.08% %ee = 98.16 Intermediate 9

(S)-1-(4-Fluoro-3-(trifluoromethyl)phenyl)-2-hydroxypropan-1-one

Following a literature procedure (J. Org. Chem. 1992, 57, 5067), to 90 ml_ of tert- butanol and 90 ml_ water was added 1.8 g methanesulfonamide, and 26 g AD-mix-a (Aldrich) and the mixture was cooled on an ice bath before addition of the 6.0 g of tert-butyl({(1 E)-1-[4- fluoro-3-(trifluoromethyl)phenyl]prop-1-en-1-yl}oxy)dimethylsilane (18 mmol, Intermediate 7). The mixture was kept cold for several hours and warmed to room temperature overnight. The next day the mixture was cooled on an ice bath, 18 g of sodium sulfite was added and stirred 30 min. Water and CH2CI2 were added and the CF^Chwas separated, dried, and concentrated before chromatography with 0-30% EtOAc yielded 3.5 g of product as an oil (82%).

¹H NMR (400 MHz, CDCI₃) d 8.25 (dd, J = 6.7, 1.7 Hz, 1 H), 8.17 (ddd, J = 8.3, 4.6, 2.2 Hz, 1 H), 7.38 (t, J = 9.2 Hz, 1 H), 5.15 (q, J = 6.5 Hz, 1 H), 3.62 (s, 1 H), 1.48 (d, J = 7.0 Hz, 3H). ¹⁹F NMR (376 MHz, CDC ) d -61.78 (d, J = 12.5 Hz), -104.89 (q, J = 12.5 Hz). Mass 237

(M + 1)⁺.

Intermediate 10

(6S)-5-[4-Chloro-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one

To a solution of (S)-1-(4-chloro-3-(trifluoromethyl)phenyl)-2-hydroxypropan-1-one (18.6 g, 69.14 mmol) in MeOH (70 ml_) was added methyl hydrazinecarboxylate (6.85 g, 76.05 mmol) and 0.1 M aqueous hydrogen chloride solution (0.69 ml_, 0.69 mmol), with the resulting mixture heated at reflux temperature for 1 h. After this time, the reaction was concentrated in vacuo, with the residual material azeotroped with further aliquots of MeOH (twice). To this material was added freshly prepared methanolic NaOMe solution (6.36 g Na (276.54 mmol) consumed in 180 ml_ MeOH). After 80 mins, AcOH (15.83 ml_, 276.54 mmol) was added and the solution was concentrated, with the residual material partitioned between EtOAc and water. The organic layer was isolated, washed with saturated aqueous sodium chloride solution, dried (MgS04), filtered and concentrated in vacuo. The residual material was purified by Biotage Isolera™ chromatography (340 g KP-Sil, eluting with DCM, 100%), with the desired fractions combined and concentrated in vacuo. This resulting material was re-dissolved in EtOAc and washed with 0.1 M aqueous hydrogen chloride solution, then saturated aqueous sodium chloride solution, dried (MgSO filtered and concentrated in vacuo to afford the title compound (10.62 g, 48%, 96.85%ee) as a pale yellow solid

LCMS (Method 3, 2 min) 97% @ Rt = 1.13 mins, MS (ESIpos) m/z no ionisation observed ¹H NMR (250 MHz, Chloroform-d) d = 1.65 (d, J = 7.0 Hz, 3H), 5.55 (q, J = 7.0 Hz, 1 H), 7.61 (d, J = 8.5 Hz, 1 H), 7.75 (dd, J = 8.4, 2.0 Hz, 1 H), 8.02 (d, J = 2.1 Hz, 1 H), 8.27 (s, 1 H). Column: Chiralcel OJ-H 25cm Mobile phase: 70:30 Heptane: I PA Flow rate: 1ml/min UV at 280nm Runtime: 20mins; Enantiomer 1 (minor) retention time: 7.47 mins; Enantiomer 2 (major) retention time: 11.01 mins; 96.85%ee Intermediate 11

(6S)-5-(-[(4-fluoro-3-(trifluoromethyl)phenyl)-)]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-

2-one

To 3.4 g of (S)-1-(4-fluoro-3-(trifluoromethyl)phenyl)-2-hydroxypropan-1-one (14 mmol, Intermediate 9) in 15 ml_ of MeOH and 1.41 g of methyl hydrazinecarboxylate (15.7 mmol) was added 9 drops of 0.1 N HCI solution (J. Med. Chem. 1992, 35, 163) and the mixture was heated at reflux 1 h. After cooling, the reaction was concentrated, MeOH was added and concentrated to remove HCI and water (twice). To this was added a NaOMe solution (1.64 g Na (71.5 mmol) consumed in 45 ml_ MeOH). After 2 h, 4.3 ml_ of HOAc (72 mmol) was added and the solution was concentrated and partitioned between EtOAc and water. The EtOAc was removed, dried, and concentrated. Chromatography with 10-40% EtOAc in hexane yielded the product as an oil which solidified upon standing overnight, 2.25 g (57%). ¹H NMR (400 MHz, CDCh) d 8.57 (s, 1 H), 7.95 (dd, J = 6.6, 2.0 Hz, 1 H), 7.85 (ddd, J = 8.3, 4.4, 2.3 Hz, 1 H), 7.31 (t, J = 9.2 Hz, 1 H), 5.56 (q, J = 7.0 Hz, 1 H), 1.65 (d, J = 7.0 Hz, 4H). ¹⁹F NMR (376 MHz, CDCI₃) d -61.68 (d, J = 12.6 Hz), -110.74 (q, J = 12.7 Hz). LC-MS (Method 5): Mass 277 (M+H)⁺.

EXPERIMENTAL SECTION - EXAMPLES

Example 1

5-[4-(2-methoxyethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one

2-methoxyethan-1-ol (300 pi, 3.8 mmol) was dissolved in THF (16 mL). The solution was cooled to 0 °C and potassium tert-butoxide (1.07 g, 9.54 mmol) was added and the resulting mixture was stirred for 15 min. 5-[4-fluoro-3-(trifluoromethyl)phenyl]-3,6-dihydro- 2H-1 ,3,4-oxadiazin-2-one (1.00 g, 3.81 mmol, Intermediate 4) was added in several portions and the resulting mixture was stirred at RT for 16h. Water was added and the mixture was extracted with ethyl acetate (3x). The combined organic layers were dried over anhydrous sodium sulfate and the filtrate was concentrated under reduced pressure. The crude material was purified using mass triggered prep HPLC to yield the title product (143 mg, 95 % purity, 11 % yield).

LC-MS (Method 1): R_t = 0.97 min; MS (ESIpos): m/z = 319 [M+H]⁺

Ή-NMR (400 MHz, DMSO-d6) d [ppm]: 1.107 (1.75), 2.518 (0.71), 2.523 (0.46), 3.313 (16.00), 3.330 (7.65), 3.675 (1.38), 3.684 (1.14), 3.687 (1.53), 3.691 (1.16), 3.698 (1.49), 4.292 (1.38), 4.299 (1.05), 4.303 (1.51), 4.314 (1.32), 5.378 (7.03), 7.357 (0.91), 7.378 (0.97), 7.923 (2.32), 7.946 (0.73), 7.951 (0.52), 11.063 (2.04).

Example 2

(6S)-6-methyl-5-[3-(trifluoromethyl)-4-(3,3,3-trifluoropropoxy)phenyl]-3,6-dihydro-2H- 1 ,3,4-oxadiazin-2-one

(6S)-5-[4-Chloro-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one (60.0 g, 205 pmol, Intermediate 8), [(2-di-tert-butylphosphino-3-methoxy-6-methyl- 2',4',6'-triisopropyl-1 ,T-biphenyl)-2-(2-aminobiphenyl)]palladium(ll) methanesulfonate (8.6 mg, 10.3 pmol), 2-di(tert-butyl)phosphino-2',4',6'-triisopropyl-3-methoxy-6-methylbiphenyl (4.8 mg, 10.3 pmol) and caesiumcarbonate (93.5 mg, 287 pmol) were added to a 5 ml reaction vessel, the vessel was sealed and flushed with argon. Degassed toluene (1000 mI) and 3, 3, 3-trifluoro-1 -propanol (89.6 mg 1.0 mmol) were added and the mixture was stirred at 95 °C overnight. The mixture was filtered through a 2 g silica column, the column flushed with a DCM:MeOH mixture (9: 1), and the solvent removed under reduced pressure, with further purification achieved using mass triggered prep HPLC to give the title compound (1 1.5 mg, 90 % purity, 14 % yield).

LC-MS (Method 1): R_t = 1.30 min; MS (ESIpos): m/z = 371.07 [M+H]⁺

¹H NMR (400 MHz, ACETONITRILE-ds) d ppm 1.49 (d, 3 H) 2.67 - 2.79 (m, 2 H) 4.36 (t, 2 H) 5.61 (q, 1 H) 7.21 (d, 1 H) 7.86 (dd, 1 H) 7.93 (d, 1 H) 9.09 (br. S, 1 H)

The following compounds can be made according to the procedure as described in the synthesis of examples 1 or 2 above:

5-{4-[(2R)-2-hydroxypropoxy]-3-(trifluoromethyl)phenyl}-3,6-dihydro-2H-1 ,3,4-oxadiazin-

2-one,

5-[4-(2-hydroxy-2-methylpropoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one, 5-{4-[(2S)-2-hydroxypropoxy]-3-(trifluoromethyl)phenyl}-3,6-dihydro-2H-1 ,3,4-oxadiazin-

2-one,

5-[4-(1 H-pyrazol-3-ylmethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-

2-one,

5-[4-(2-methoxyethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one,

5-{4-[(2S)-tetrahydrofuran-2-ylmethoxy]-3-(trifluoromethyl)phenyl}-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

5-{4-[(2R)-tetrahydrofuran-2-ylmethoxy]-3-(trifluoromethyl)phenyl}-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

5-[3-(trifluoromethyl)-4-(3,3,3-trifluoropropoxy)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one,

5-[4-butoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one,

5-[4-propoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one,

5-[4-(3,3-dimethylbutoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one,

(6S)-5-{4-[(2R)-2-hydroxypropoxy]-3-(trifluoromethyl)phenyl}-6-methyl-3,6-dihydro-2H-

1 ,3,4-oxadiazin-2-one,

(6S)-5-[4-(2-hydroxy-2-methylpropoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro- 2H-1 ,3,4-oxadiazin-2-one,

1 ,3,4-oxadiazin-2-one,

(6S)-6-methyl-5-[4-(1 H-pyrazol-3-ylmethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-

1 ,3,4-oxadiazin-2-one,

(6S)-5-[4-(cyclobutylmethoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

(6S)-6-methyl-5-{4-[(2R)-tetrahydrofuran-2-ylmethoxy]-3-(trifluoromethyl)phenyl}-3,6- dihydro-2H-1 ,3,4-oxadiazin-2-one and

(6S)-5-[4-(3,3-dimethylbutoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one. Example 3

(6S)-6-methyl-5-[4-propoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one

(6S)-6-Methyl-5-[4-propoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one was produced using an analogous procedure to Example 2 from (6S)-5-[4-chloro-3- (trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one (Intermediate 8) and 1-propanol affording the title compound in 19 % yield.

LC-MS (Method 1): R_t = 1.27 min; MS (ESIpos): m/z = 317.04 [M+H]⁺

1H NMR (400 MHz, ACETONITRILE-ds) d ppm 1.02 (t, 3 H) 1.49 (d, 3 H) 1.74 - 1.86 (m, 2 H) 4.10 (t, 2 H) 5.60 (q, 1 H) 7.18 (d, 1 H) 7.83 (dd, 1 H) 7.91 (d, 1 H) 9.06 (br s, 1 H)

Example 4

(6S)-5-{4-[(2R)-2-fluoropropoxy]-3-(trifluoromethyl)phenyl}-6-methyl-3,6-dihydro-2H-

1 ,3,4-oxadiazin-2-one

1 ,3,4-oxadiazin-2-one was produced using an analogous procedure used to produce Example 2 from (6S)-5-[4-chloro-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-

1 ,3,4-oxadiazin-2-one (Intermediate 8) and 2-fluoropropan-1-ol, affording the title compound in 9% yield.

LC-MS (Method 2): R_t = 1.07 min; MS (ESIneg): m/z = 333.13 [M-H]- ¹H NMR (400 MHz, ACETONITRILE-ds) d ppm 1.37 - 1.47 (m, 3 H) 1.49 (d, 3 H) 4.14 - 4.33 ( , 2 H) 4.91 - 5.00 ( , 1 H) 5.04 - 5.12 ( , 1 H) 5.60 (q, 1 H) 7.19 (d, 1 H) 7.85 (dd, 1 H) 7.93 (d, 1 H) 9.09 (s, 1 H)

Example 5

(6S)-5-[4-butoxy-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one

(6S)-5-[4-butoxy-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one was produced using an analogous protocol used to produce Example 2 from (6S)-5- [4-chloro-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one (Intermediate 8) and 1-butanol affording the title compound in 14 % yield.

LC-MS (Method 1): R_t = 1.37 min; MS (ESIpos): m/z = 331.05 [M+H]⁺

¹H NMR (400 MHz, ACETONITRILE-ds) d ppm 0.95 (t, 3 H) 1.43 - 1.53 (m, 5 H) 1.70 - 1.83 (m, 2 H) 4.14 (t, 2 H) 5.60 (q, 1 H) 7.19 (d, 1 H) 7.83 (dd, 1 H) 7.91 (d, 1 H) 9.06 (s, 1 H)

Example 6

(6S)-5-[4-(2,2-difluoropropoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one

(6S)-5-[4-(2,2-difluoropropoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one was produced using an analogous protocol used to produce Example 2 from (6S)-5-[4-chloro-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin- 2-one (Intermediate 8) and 2,2 difluoropropanol affording the title compound in 6 % yield.

LC-MS (Method 1): R_t = 1.14 min; MS (ESIpos): m/z = 353.13 [M+H]⁺

¹H NMR (400 MHz, ACETONITRILE-ds) d ppm 1.49 (d, 3 H) 1.75 (t, 3 H) 4.35 (t, 2 H) 5.61 (q, 1 H) 7.21 (d, 1 H) 7.87 (dd, 1 H) 7.96 (d, 1 H) 9.1 1 (s, 1 H)

Example 7

(6S)-5-[4-ethoxy-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one

(6S)-5-[4-ethoxy-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one was produced using an analogous protocol used to produce Example 2 from (6S)-5- [4-chloro-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one (Intermediate 8) and ethanol affording the title compound in 36 % yield.

LC-MS (Method 1): R_t = 1.18 min; MS (ESIpos): m/z = 303.06 [M+H]⁺

1H NMR (400 MHz, ACETONITRILE-ds) d ppm 1.39 (t, 3 H) 1.49 (d, 3 H) 4.20 (q, 2 H) 5.60 (q, 1 H) 7.18 (d, 1 H) 7.84 (dd, 1 H) 7.91 (d, 1 H) 9.06 (br s, 1 H)

Example 8

(6S)-5-[4-(2-methoxyethoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one

To 1 mL of 2-methoxyethanol was added 10 mg of sodium metal (0.43 mmol) and the mixture was stirred until the metal was consumed. To the basic solution was added (S)- 5-(4-fluoro-3-(trifluoromethyl)phenyl)-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one (100 mg, 0.36 mmol, Intermediate 11) of and the solution was heated at 60 °C for 3 h. After cooling, NH4CI (aq) was added and was rinsed with EtOAc. The combined EtOAc was dried and concentrated, chromatography with 0-50% EtOAc in hexane isolated 14 mg of product (11 %) as a white solid. ¹H NMR (400 MHz, CDCh) d 8.28 (s, 1 H), 7.90 (s, 1 H), 7.77 (d, J = 8.5 Hz, 1 H), 7.10 (d, J = 8.7 Hz, 1 H), 5.54 (q, J = 6.9 Hz, 1 H), 4.34 - 4.21 (m, 2H), 3.89 - 3.74 (m, 2H), 3.48 (s, 3H), 1.63 (d, J = 6.9 Hz, 4H). ¹⁹F NMR (376 MHz, CDCh) d -62.72. Mass 333 (M + 1). Chiral SCF HPLC analysis (AS-H column, 10 min run, 0-50% MeOH) indicated an 87.5: 12.5 ratio of enantiomers.

Example 9

(6S)-6-methyl-5-[4-(2,2,2-trifluoroethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one

To 4.0 mL of DMF was added 2.6 mL of trifluoroethanol (36 mmol), 1.2 g of CS2CO3 (3.7 mmol) and 1.0 g of (S)-5-(4-fluoro-3-(trifluoromethyl)phenyl)-6-methyl-3,6-dihydro-2H- 1 ,3,4-oxadiazin-2-one (3.6 mmol, Intermediate 1 1) and the mixture was heated at 70 °C for 6 h. After cooling, water and EtOAc were added, the layers were separated and the water rinsed twice with EtOAc. The combined EtOAc layers were rinsed with brine, dried, and concentrated. Chromatography with 10-50% EtOAc in hexanes yielded 1.02 g of off- white solid (80%). ¹H NMR (400 MHz, CDCh) d 8.38 (s, 1 H), 7.96 (d, J = 1.7 Hz, 1 H), 7.83 (dd, J = 8.7, 2.1 Hz, 1 H), 7.08 (d, J = 8.8 Hz, 1 H), 5.55 (q, J = 7.0 Hz, 1 H), 4.52 (q, J = 7.8 Hz, 2H), 1.64 (d, J = 7.0 Hz, 3H). ¹⁹F NMR (376 MHz, CDCh) d -62.73, -73.86. Mass 357 (M + 1)⁺. EXPERIMENTAL SECTION - BIOLOGICAL ASSAYS

Examples were tested in selected biological assays one or more times. When tested more than once, data are reported as either average values or as median values, wherein

• the average value, also referred to as the arithmetic mean value, represents the sum of the values obtained divided by the number of times tested, and

• the median value represents the middle number of the group of values when ranked in ascending or descending order. If the number of values in the data set is odd, the median is the middle value. If the number of values in the data set is even, the median is the arithmetic mean of the two middle values.

Examples were synthesized one or more times. When synthesized more than once, data from biological assays represent average values or median values calculated utilizing data sets obtained from testing of one or more synthetic batch.

The in vitro activity of the compounds of the present invention can be demonstrated in the following assays:

Assay 1

Cell proliferation measurement

The antiproliferative activity of the compounds of the general formula (I) was examined in vitro in human cancer cells. For this purpose, the appropriate number of cells (Hela: 800; SK-MEL-3: 1000; IGR37: 2000) were plated in 384-well plates with appropriate growth medium (Hela: DMEM/Ham's F12 (Biochrom; # FG 4815 with stabile Glutamine), FCS 10% final (Biochrom; # S 0415); SK-MEL-3: McCoy's 5A (Biochrom; # F 1015), FCS 10% final (Biochrom; # S 0415), L-Alanyl-L-Glutamine final: 2mM, (Biochrom; # K 0302) ;IGR37 DMEM; (Biochrom; # FG 0445, high glucose; + FCS 10% final (Biochrom; # S 0415)) and incubated at 37°C overnight. After 24 h, cells on one plate (0 h plate) were treated with 30 mI/cavity of CTG solution (Promega Cell Titer Glo (catalogue # G755B and G756B)) and incubated at room temperature for 10 min, and luminescence was measured by means of a VICTOR V (Perkin Elmer), in order to determine cell viability on commencement of treatment. The cells on the test plate were treated with the compounds of the general formula (I) as and incubated at 37°C for 72 h. The compounds were added to the cells by means of an HP D300 digital dispenser in a 10-step 2.5-fold dilution series generally starting at a maximum final drug concentration of 100nM . As control, the cells were treated with vehicle (DMSO at 0.3% final concentration). After 72 h, the cells were treated with 30 mI/cavity of CTG solution (Promega Cell Titer Glo (catalogue # G755B and G756B)) and incubated at room temperature for 10 min, and luminescence was measured by means of a VICTOR V (Perkin Elmer), in order to determine cell viability at the end of treatment. The percentage effect on cell growth and the IC5₀ derived therefrom were determined for each test substance using the values from the 0 h plate (= maximum inhibition) and the DMSO control (= minimum inhibition). The IC5₀ values were calculated using a 4-parameter fit.

Table 2: Anti-proliferation IC₅o values of several examples in vitro in different cell lines

Thus one aspect of the invention is the use of the compounds of formula (I) for the treatment of cervical cancer.

Another aspect of the invention is the use of the compounds of formula (I) for the treatment of skin cancer, especially melanoma.

Yet another aspect of the invention is the use of compounds of formula (I), for the treatment of skin cancer, especially melanoma, and cervical cancer.

Another aspect are compounds of formula (I) which effectively inhibit tumor cell proliferation ( e.g . in HeLa cells) with IC5₀ values of < 100 nM.

Another aspect are compounds of formula (I) which effectively inhibit tumor cell proliferation (e.g. in HeLa cells) with IC5₀ values of < 10 nM. Another aspect are compounds of formula (I) which effectively inhibit tumor cell proliferation (e.g. in HeLa cells) with IC50 values of < 1 nM.

Assay 2

Cell proliferation measurement

The antiproliferative activity of the compounds of the general formula (I) is examined in vitro in human cancer cells. For this purpose, 500Hel_a cells, A2058 cells, DU145 cells, HMCB cells, IGR37 cells, NCIH1734 cells, OSRC2 cells, or 750 CAL51 cells, C0L0741 cells, DBTRG05MG cells, DKMG cells, G292CLONEA141 B1 cells, GB1 cells, HEL cells, HEL9217 cells, JHUEM1 cells, L3.3 cells, LI7 cells, TE4 cells, or 1000 8505C cells, HUT78 cells, NCI H 1563 cells, NCIH2122 cells, NCIH2172 cells, RVH421 cells, SKMEL3 cells, or 1500 C32 cells, HS578T cells, JHOM1 cells, NCIH196 cells, OVKATE cells, are plated in 384-well plates with appropriate growth medium and incubated at 37°C overnight. After 24 h, the cells on the test plate are treated with the compounds of the general formula (I) as and incubated at 37°C for 72 h. The compounds are added to the cells by means of an HP D300 digital dispenser in a 10 (or more) -step dilution series. As control, the cells are treated with vehicle (DMSO at 0.3% final concentration). After 72 h, the cells are treated with 20 mI/well of 50% CTG solution in PBS (Promega Cell Titer Glo (catalogue # G755B and G756B)) and incubated at room temperature for 10 min, and luminescence is measured by means of a VICTOR V (Perkin Elmer), in order to determine cell viability at the end of treatment. The percentage effect on cell growth and the IC50 derived therefrom are determined for each test substance using the values from untreated wells (= percent viability). The IC50 values are calculated using a 4-parameter fit.

Assay 3

Method for PDE3A enzyme inhibition

The commercially available 3H-cAMP Scintillation Proximity Assay (SPA, Perkin Elmer) system was used for enzyme inhibition studies. For the determination of the in vitro effect of example compounds on the PDE3A reactions 2 mI of the respective example compound solution in DMSO (serial dilutions) were placed in wells of microtiter plates (lsoplate-96/200W; Perkin Elmer). 50 mI of a dilution of PDE3A cell extract from Sf9 cells overexpressing human full length PDE3A (SB Drug Discovery, UK) in buffer A (50 mM Tris/HCI pH 7.5, 8.3 mM MgCI₂, 1.7 mM EDTA, 0.2% BSA) was added. The dilution of the PDE3A cell extract was chosen such that the reaction kinetics was linear and less than 70% of the substrate was consumed (typical dilution 1 :5000). The reaction was started by addition of 50 pi (0.025 pCi) of 1 :2000 in buffer A w/o BSA diluted substrate [8-3H] adenosine 3', 5'-cyclic phosphate (1 pCi/mI; Perkin Elmer). After incubation at room temperature for 60 min, the reaction was stopped by addition of 25 pi of a suspension containing 18 mg/ml yttrium scintillation proximity beads (Perkin Elmer) in water. The microtiter plates were sealed and measured in a Microbeta scintillation counter (PerkinElmer Wallac). IC50 values were determined from sigmoidal curves by plotting percentage PDE3A activity vs log compound concentration.

Assay 4

PDE3B enzyme inhibition

The commercially available 3H-cAMP Scintillation Proximity Assay (SPA, Perkin Elmer) system was used for enzyme inhibition studies. For the determination of the in vitro effect of example compounds on the PDE3B reactions 2 mI of the respective example compound solution in DMSO (serial dilutions) were placed in wells of microtiter plates (lsoplate-96/200W; Perkin Elmer). 50 mI of a dilution of PDE3B cell extract from Sf9 cells overexpressing human full length PDE3B (SB Drug Discovery, UK) in buffer A (50 mM Tris/HCI pH 7.5, 8.3 mM MgCI₂, 1.7 mM EDTA, 0.2% BSA) was added. The dilution of the PDE3B cell extract was chosen such that the reaction kinetics was linear and less than 70% of the substrate was consumed (typical dilution 1 :6000). The reaction was started by addition of 50 mI (0.025 pCi) of 1 :2000 in buffer A w/o BSA diluted substrate [8-3H] adenosine 3', 5'-cyclic phosphate (1 pCi/mI; Perkin Elmer). After incubation at room temperature for 60 min, the reaction was stopped by addition of 25 mI of a suspension containing 18 mg/ml yttrium scintillation proximity beads (Perkin Elmer) in water. The microtiter plates were sealed and measured in a Microbeta scintillation counter (PerkinElmer Wallac). IC50 values were determined from sigmoidal curves by plotting percentage PDE3B activity vs log compound concentration.

Another aspect of the invention are compounds of formula (I) which effectively inhibit tumor cell proliferation with IC50 values of < 100 nM in e.g. HeLa cells while IC50 values for enzymatic PDE3A or PDE3B inhibition are often > 10 times higher than IC50 values for tumor cell proliferation.

One aspect of the invention are compounds of formula (I) which effectively inhibit tumor cell proliferation with IC50 values of < 100 nM in e.g. HeLa cells while IC50 values for enzymatic PDE3A or PDE3B inhibition are often > 30 times higher than IC50 values for tumor cell proliferation.

One aspect of the invention are compounds of formula (I) which effectively inhibit tumor cell proliferation with IC50 values of < 1 nM in e.g. HeLa cells while IC50 values for enzymatic PDE3A or PDE3B inhibition are often > 10 times higher than IC50 values for tumor cell proliferation.

Table 6: Inhibition of PDE3A and PDE3B

Assay 5

In vivo pharmacokinetics in rodents (e.g. mice)

The housing and handling of animals was performed in strict compliance with the European and German Guidelines for Laboratory Animal Welfare. Animals received food and water ad libitum. For the quantification of circulating compounds in plasma, a certain dose (1 - 100 mg/kg) was orally administered to female NMRInu/nu mice at the age of 6- 8 weeks in a solubilized form (n=3 mice per time point).

Blood was collected into Lithium-Heparin tubes (Monovetten®, Sarstedt) and centrifuged for 15 min at 3000 rpm. A small aliquot (e.g. 100 pL) from the supernatant (plasma) was taken and precipitated by addition of an aliquot ice cold acetonitrile (e.g. of 400 pL) and frozen at -20 °C over night. Samples were subsequently thawed and centrifuged at 3000 rpm, 4°C for 20 minutes. Aliquots of the supernatants were taken for analytical testing using an Agilent HPLC-system with LCMS/MS detection. PK parameters were calculated by non-compartmental analysis using a PK calculation software. Assay 6

Validation of PDE3A modulator-induced PDE3A protein interactions using immunoprecipitation and immunoblotting

HeLa cells are transfected with ORF overexpression constructs expressing V5- tagged SLFN12, or V5-tagged GFP. ORF expression constructs are obtained from the TRC (clone IDs: TRCN0000468231 , TRCN0000476272, ccsbBroad304_99997). At 72 hours post transfection, cells are treated with 10 mM DNMDP or trequinsin for 4 hours followed by lysis using the ModRipa lysis buffer and immunoprecipitation of PDE3A. For each condition, 2 mg total protein lysate is incubated with 1 pg of anti-PDE3A antibody at 4° C overnight, after which 7.5 mI each of Protein A- and Protein G- Dynabeads (Life Technologies 10001 D and 10003D) are added and incubated for another 1 hour. Beads are washed and bound proteins are eluted with 30 mI of LDS PAGE gel loading buffer. Input (~60 pg total protein lysate) and IP products wearere resolved on 4-12% Tris-Glycine PAGE gels and immunoblotted with an anti-V5 antibody (Life Technologies R96205, 1 :5000), the Bethyl anti-PDE3A antibody (1 :1000), and secondary antibodies from LiCOR Biosciences (Cat.# 926-32210 and 926068021 , each at 1 :10,000). Blots were washed and imaged using a LiCOR Odyssey infrared imager.

Assay 7

In vivo xenotransplantation models

The anti-tumor activities of Compounds of the invention can be examined in murine xenotransplantation models of human cancer. For this purpose, mice can be implanted subcutaneously with tumor cells. At a mean tumor size of 20-40 mm² animals can be randomized into treatment and control groups (at least n=10 animals/group) and treatment can be started with vehicle only or respective Compound. The oral application volume can be 10 ml/kg. In the case of twice daily treatments, the time interval between two applications per day can be 6-7h. The tumor size and the body weight can be determined at least weekly. The tumor area can be detected by means of an electronic caliper [length (mm) x width (mm)]. The experiment can be ended when the study reaches the pre determined ethical endpoint based on German and European animal welfare regulations. In vivo anti-tumor efficacy can be presented as T/C ratio at study end (Treatment/Control; mean tumor weight of treatment group / mean tumor weight of control group). A compound having a T/C below 0.5 is defined as active (i.e., effective). Statistical analysis can be assessed using SigmaStat software. A one-way analysis of variance can be performed and differences to the control can be compared by a pair-wise comparison procedure (Dunn’s method).

Assay 8

Effects on cardiovascular function in conscious telemetered rats

The measurement of cardiovascular parameters in small laboratory animals such as the rat by telemetry is an integral part of cardiac safety assessment. The telemetry technology provides precise measurements while avoiding stress artifacts inherent with the use of physical or chemical restraint and is suitable to detect potential drug-induced alterations of cardiovascular parameters.

Conscious telemetered normotensive Wistar rats (n=4-6/group) are treated with single doses of a compound (three dose groups and a vehicle control group). Cardiovascular parameters such as arterial systolic and diastolic blood pressure, heart rate, left ventricular systolic and end diastolic pressure, left ventricular contractility, as well as body temperature and locomotor activity are continuously monitored before and over 24 hours after administration. For statistical analysis of cardiovascular parameters a mixed model approach and a Dunnett’s t-test procedure is used.

Other Embodiments

From the foregoing description, it will be apparent that variations and modifications may be made to the invention described herein to adopt it to various usages and conditions. Such embodiments are also within the scope of the following claims.

The recitation of a listing of elements in any definition of a variable herein includes definitions of that variable as any single element or combination (or subcombination) of listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

Claims

1. A compound of formula (I)

where

R² is selected from a hydrogen atom and a halogen atom;

R³ is a Ci-C₆-alkoxy group which is optionally substituted with a group selected from a CrC3-haloalkyl group, a hydroxy group, a CrC3-alkoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, and a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom or a Ci-C3-alkyl group;

2. The compound according to claim 1 ,

where

R² is selected from a hydrogen atom, a fluorine atom, and a chlorine atom;

R⁴ is a hydrogen atom or a methyl group; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a salt of a stereoisomer, a salt of a tautomer, a salt of an N-oxide or a mixture of same.

3. The compound according to claim 1 ,

where

R² is selected from a hydrogen atom, a fluorine atom, and a chlorine atom;

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from a CrC3-haloalkyl group, a hydroxy group, a CrC3-alkoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, and a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom or a methyl group;

4. The compound according to claim 1 ,

where

R² is selected from a hydrogen atom and a fluorine atom;

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from a trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluoroethyl group, a hydroxy group, methoxy group, an ethoxy group, a C4-C6-cycloalkyl group, a 4- to 6- membered heterocycloalkyl group, and a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom or a methyl group;

5. The compound according to claim 1 ,

where R¹ is selected from a fluorine atom, and a trifluoromethyl group;

R² is a hydrogen atom;

R⁴ is a hydrogen atom or a methyl group;

6. The compound according to claim 1 ,

where

R¹ is a trifluoromethyl group;

R² a hydrogen atom;

R³ is a CrC₆-alkoxy group which is optionally substituted with a group selected from a trifluoromethyl group, a difluoromethyl group, a difluoroethyl group, a fluoroethyl group, a hydroxy group, methoxy group, an ethoxy group, a cyclobutyl group, a tetrahydrofuryl group, and a pyrazol-3-yl group, with the proviso that an unsubstituted methoxy group is excluded;

R⁴ is a hydrogen atom or a methyl group;

7. The compound according to claim 1 ,

where

R¹ is a trifluoromethyl group;

R² a hydrogen atom;

R³ is a CrC4-alkoxy group which is optionally substituted with a group selected from a trifluoromethyl group, a difluoroethyl group, a fluoroethyl group, and a methoxy group, with the proviso that an unsubstituted methoxy group is excluded;

8. The compound according to claim 1 , which is selected from the group:

(6S)-6-methyl-5-[3-(trifluoromethyl)-4-(3,3,3-trifluoropropoxy)phenyl]-3,6-dihydro-2H- 1 ,3,4-oxadiazin-2-one,

(6S)-6-methyl-5-[4-propoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one,

(6S)-5-{4-[(2R)-2-fluoropropoxy]-3-(trifluoromethyl)phenyl}-6-methyl-3,6-dihydro-2H- 1 ,3,4-oxadiazin-2-one,

(6S)-5-[4-(2,2-difluoropropoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

(6S)-5-[4-ethoxy-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one,

(6S)-6-methyl-5-[4-(2,2,2-trifluoroethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

2-one,

5-{4-[(2S)-2-hydroxypropoxy]-3-(trifluoromethyl)phenyl}-3,6-dihydro-2H-1 ,3,4-oxadiazin-

2-one,

5-[4-(2-methoxyethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one, 5-[4-(cyclobutylmethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2- one, 5-{4-[(2S)-tetrahydrofuran-2-ylmethoxy]-3-(trifluoromethyl)phenyl}-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one,

5-[4-butoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one,

5-[4-propoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4-oxadiazin-2-one,

1 ,3,4-oxadiazin-2-one,

(6S)-5-[4-(3,3-dimethylbutoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one.

9. The compound according to claim 1 , which is selected from the group:

5-[4-(2-methoxyethoxy)-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one

(6S)-6-methyl-5-[3-(trifluoromethyl)-4-(3,3,3-trifluoropropoxy)phenyl]-3,6- dihydro-2H-1 ,3,4-oxadiazin-2-one (6S)-6-methyl-5-[4-propoxy-3-(trifluoromethyl)phenyl]-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one

(6S)-5-{4-[(2R)-2-fluoropropoxy]-3-(trifluoromethyl)phenyl}-6-methyl-3, 6-dihydro- 2H-1 ,3,4-oxadiazin-2-one

(6S)-5-[4-butoxy-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one

(6S)-5-[4-(2,2-difluoropropoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3, 6-dihydro- 2H-1 ,3,4-oxadiazin-2-one

(6S)-5-[4-ethoxy-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H-1 ,3,4- oxadiazin-2-one

(6S)-5-[4-(2-methoxyethoxy)-3-(trifluoromethyl)phenyl]-6-methyl-3,6-dihydro-2H- 1 ,3,4-oxadiazin-2-one

(6S)-6-methyl-5-[4-(2, 2, 2-trifluoroethoxy)-3-(trifluoromethyl)phenyl]-3, 6-dihydro- 2H-1 ,3,4-oxadiazin-2-one

10. A method of preparing a compound of general formula (I) according to any one of claims 1 to 9, said method comprising

starting from a compound of formula (II)

formula (II)

wherein X is a fluorine atom or a chlorine atom

R² is a hydrogen atom or a halogen atom;

with the proviso that particularly at least one of R¹ and R² exerts an electron withdrawing effect and

R⁴ is a hydrogen atom or a CrC3-alkyl group

with the proviso that if X is a chlorine atom, R¹ and R² are not a fluorine atom, and reacting said compound with a corresponding alcohol R⁵-OH, or alkoxide salts thereof, such as a sodium or potassium salt thereof,

whereby

R⁵ is a Ci-C₆-alkyl group which is optionally substituted with a group selected from

CrC3-haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C6-cycloalkyl group, a 4 to 6-membered heterocycloalkyl group, a 5 to 6-membered heteroaryl group, with the proviso that an unsubstituted methyl group is excluded wherein the reaction occurs with the following conditions:

optionally in the presence of a strong base,

optionally in an inert solvent,

optionally in the presence of a further base, in a solvent,

formula (I)

wherein R³ is -OR⁵.

11. A method of preparing a compound of general formula (I) according to any one of claims 1 to 9, said method comprising

starting from a compound of formula (II)

formula (II) wherein

X is Cl, Br, I, or a group selected from (Ci-C4-alkylsulfonyl)oxy, (C1-C4- fluoroalkylsulfonyl)oxy and (phenylsulfonyl)oxy, the phenyl present in (phenylsulfonyl)oxy being optionally substituted with one, two, three, four or five substituents, each of them independently selected from halogen, nitro, cyano, Cr

C4-alkyl and CrC4-alkoxy,

R² is a hydrogen atom or a halogen atom;

R⁴ ia a hydrogen atom or a CrC3-alkyl group

and reacting said compound with a corresponding alcohol R⁵-OH, whereby R⁵ is as defined for the first alternative,

in the presence of a base, a palladium catalyst,

in an inert solvent,

at elevated temperatures ranging form 60°-160°C in order to obtain a compound of formula (I)

formula (I)

wherein R³ is -OR⁵.

12. A method of preparing a compound of general formula (I) according to any one of claims 1 to 9, said method comprising

starting from a compound of formula (II)

formula (II)

wherein X is OPg2, whereby

Pg2 is a protecting group of a hydroxy group which can be cleaved by a suitable cleavage method for said protecting group,

R² is a hydrogen atom or a halogen atom;

and subsequently removing the protecting group, resulting in a compound of formula (II), wherein X=OH, and

whereby R⁵ is defined as for the first alternative,

in order to obtain a compound of formula (I)

formula (I)

13. The methods of preparing a compound of general formula (I) according to claims 10, 11 and 12,

said methods comprising

for compound (II): R¹ is trifluoromethyl and R² is a hydrogen atom,

the following conditions: according to claim 10:

the strong base is selected from potassium tert-butoxide, sodium hydride, and sodium metal,

for the optionally used further base cesium chloride is selected,

optionally in a temperature range from 60° to 150°C;

according to claim 1 1 :

the base is selected from potassium phosphate or cesium carbonate,

the palladium catalyst is selcted from [(2-Di-te/f-butylphosphino-3,6-dimethoxy-

2',4',6'-triisopropyl-1 , 1 '-biphenyl)-2-(2'-amino-1 , 1 '-biphenyl)]palladium(ll) methanesulfonate and [(2-Di-te/f-butylphosphino-3-methoxy-6-methyl-2',4',6'- triisopropyl-1 , 1 '-biphenyl)-2-(2-aminobiphenyl)]palladium(ll) methanesulfonate, the optionally used ligand is selected from2-Di(tert-butyl)phosphino-2',4',6'- triisopropyl-3-methoxy-6-methylbiphenyl or 2-(Di-te/f-butylphosphino)-2',4',6'- triisopropyl-3,6-dimethoxy-1 ,1 '-biphenyl,

according to claim 12:

the protecting group is a benzyl group,

the palladium ctalyst is palladium on carbon, the solvent is selected from ethanol and methanol,

the transfer of the so obtained compound of formula (II) with X=oH into a compound of formula (I) with X=OR⁵ being conducted

in the presence of diisopropylazodicarboxylate and

14. A compound of general formula (I) according to any one of claims 1 to 9 for use in the treatment or prophylaxis of a hyperproliferative disease.

15. A pharmaceutical composition comprising a compound of general formula (I) according to any one of claims 1 to 9 and one or more pharmaceutically acceptable excipients.

16. A pharmaceutical combination comprising:

· one or more first active ingredients, in particular compounds of general formula (I) according to any one of claims 1 to 9, and

• one or more further active ingredients.

17. The use of a compound of general formula (I) according to any one of claims 1 to 9 for the treatment or prophylaxis of a disease.

18. The use according to claim 14, or 17, wherein the disease is a hyperprol iterative disease.

19. The use according to claim 18, wherein the hyperproliferative disease is a cancer disease.

20. The use according to claim 19, wherein the cancer disease is cervical cancer or melanoma.

21. A method of treating a hyperproliferative disease such as cancer in a subject, the method comprising administering to the subject a compound of any of the preceding claims 1-9, thereby treating the hyperproliferative disease.

22. A compound having the structure of general formula (II):

(I I),

in which R¹, R² and R⁴ have the meaning as defined for the compound of general formula (I) according to anyone of claims 1-9 and X is a OR⁵ group whereby

R⁵ is a CrC₆-alkyl group which is optionally substituted with a group selected from C1-C3- haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methyl group is excluded.

23. Use of a compound of general formula (II) according to claim 22,

R⁵ is a CrC₆-alkyl group which is optionally substituted with a group selected from C1-C3- haloalkyl group, a hydroxy group, a CrC3-alkyoxy group, a C4-C6-cycloalkyl group, a 4- to 6-membered heterocycloalkyl group, a 5- to 6-membered heteroaryl group, with the proviso that an unsubstituted methyl group is excluded,

for the manufacture of a compound of formula (I).