RU2803158C2 - Morpholinyl pyridones - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to new morpholinylpyridone compounds of formula (I), pharmaceutical compositions and methods of using such compounds in the treatment of diseases, including breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, kidney cancer, colon cancer, glioma, prostate cancer, ovarian cancer, melanoma and lung cancer, as well as hypoxic tumors.

EFFECT: new compounds of morpholinylpyridones of formula (I) with properties of Vps34 inhibitors have been obtained.

30 cl, 2 tbl, 37 ex

Description

FIELD OF THE INVENTION

The invention provides new morpholinylpyridone compounds of formula (I), pharmaceutical compositions containing such compounds, and methods of using such compounds in the treatment of diseases, including cancer and type II diabetes.

BACKGROUND ART

Enzymes belonging to the phosphatidylinositide 3-kinase (PI3K) family are regulators of several important cellular events. This family consists of three classes, I, II and III, and while the group belonging to class I has been an interesting drug target for many years, class II and III enzymes have been less exploited. Class III PI3K, vacuolar sorting protein 34 (Vps34, PIK3C3), forms a heterodimer with its regulatory subunit p150 (Vps15), and this dimer is involved in several complexes that regulate events associated with vesicular transport, such as autophagy, endocytosis, exocytosis, and micropinocytosis (Amaravadi et al., Clin. Cancer Res., 2011, 17: 654-666; Carpentier et al., 2013, Traffic). This enzyme is responsible for the phosphorylation of phosphatidylinositol (PI) to form phosphatidylinositol (3)-phosphate (PI3P). Binding of this ligand to the PX and FYVE domains leads to the recruitment and delocalization of these effector proteins, resulting in the formation, elongation and movement of vesicles (Backer et al., J. Biochem., 2008, 410: 1-17).

Autophagy is a catabolic process in which cellular components undergo destruction by encapsulation in bilayer membrane vesicles, autophagosomes, which fuse with protease-containing lysosomes. This is a way for the cell to process damaged organelles and misfolded proteins and thereby maintain the functioning of the cell. This pathway is also a way to recycle cell contents to form new structural units (Boya et al., Nat. Cell Biol., 2013, 15: 713-720). Autophagy is a cellular response to stress-related conditions such as nutrient deprivation, acidosis and hypoxia, as well as drug treatment. In view of this, inhibition of autophagy is a means of enhancing the effects of anticancer drugs and desensitizing drug-resistant tumors (Nagelkerke et al., Semin. Cancer Biol., 2014, 31: 99-105). Tumors in the most advanced stages of development show strong activation of the autophagic flux (Leone et al. Trends in Endocrin. Metab., 2013, 24: 209-217). A common marker for studying autophagic flux is the detection of an “autophagic spot” in the autophagosome in the form of lipidated protein LC3 (light chain of microtubule-associated protein 1 (MAP1)). Inhibition of Vps34 results in inhibition of autophagy as measured by LC3 redistribution in the blot (Dowdle et al., Nat. Cell Biol., 2014, 16: 1069-79).

As recently described, deletion of the p150 regulatory subunit results in increased insulin sensitivity in vivo due to decreased internalization of insulin receptors (Nemazanyy, Nature Commun., 2015, 6: 8283). In an animal model with a heterozygous genotype for the kinase dead mutation, this result is confirmed by increased glucose tolerance and increased insulin sensitivity (WO 2013076501).

Inhibition of Vps34 could be beneficial in a variety of disease states, including cancer, inflammatory diseases, autoimmune diseases, neurodegenerative disorders, cardiovascular disorders, type II diabetes, and viral infections (reviewed in Rubinsztein et al., Nat. Rev., 2012, 11 : 709-730). Cancers that would benefit from Vps34 inhibition include, but are not limited to, breast cancer such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, kidney cancer , colon cancer, glioma, prostate cancer, ovarian cancer, melanoma and lung cancer, as well as hypoxic tumors. Thus, there is a need for new and potent Vps34 inhibitors.

Previous inventions describing Vps34 inhibitors for use in treating diseases include WO 2015150555, WO 2015150557, WO 2015108861, WO 2015108881, WO 2012085815, WO 2012085244, WO 2013190510 , Farkas, J. Biol. Chem., 2011, 286(45): 38904-12.

DESCRIPTION OF THE INVENTION

The purpose of this invention is to develop new and potent Vps34 inhibitors. Another aim of the invention is to develop new and potent Vps34 inhibitors that can be used to treat cancer and other diseases such as type II diabetes.

According to one aspect of the invention there is provided a compound of formula (I)

Where

R ¹ represents C ₁ -C ₃ alkyl or cyclopropyl;

R ² is selected from hydrogen, C ₁ -C ₃ haloalkyl and C ₁ -C ₃ alkyl;

A represents

Where

R ³ is selected from R ⁶ , C ₁ -C ₆ alkyl, amino, N-C ₁ -C ₃ alkylamino, N,N-diC ₁ - C ₃ alkylamino and C ₁ -C ₃ alkoxyC ₁ -C ₃ alkyl, wherein said C ₁ -C ₆ alkyl and said C ₁ -C ₃ alkoxyC ₁ -C ₃ alkyl are optionally substituted by one R ⁶ and/or one or more halogen atoms;

R ⁴ is selected from C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl, C ₃ -C ₆ cycloalkyl and phenyl, wherein said phenyl is optionally and independently substituted with one or more fluorine atoms, chlorine atom, methyl, methoxy, dimethylamino, trifluoromethoxy, trifluoromethyl, cyclopropyl;

R ⁵ is selected from halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkyl and C ₃ -C ₆ cycloalkyl;

R ⁶ is selected from phenyl, monocyclic heteroaryl, C ₃ -C ₆ cycloalkyl, heterocyclyl, each of which is optionally substituted with one or more R ⁷ ;

R ⁷ is selected from halogen, amino, N-C ₁ -C ₃ alkylamino, N,N-diC ₁ -C ₃ alkylamino and C 1 -C ₃ alkoxy, C ₁ -C ₃ alkyl, C _{1 -C 3} alkoxy, C ₁ - C ₃ haloalkoxy, C ₃ -C ₆ cycloalkyl, C ₁ -C ₃ haloalkyl and C ₁ -C ₃ alkyl;

or a pharmaceutically acceptable salt thereof.

According to one embodiment of this aspect of the invention, R ² is selected from a hydrogen atom and a C ₁ -C ₃ alkyl, such as a hydrogen atom.

In one embodiment of this aspect of the invention, R ¹ is methyl.

According to one embodiment of this aspect of the invention, R ⁷ is selected from fluorine, cyclopropyl and methyl.

According to one embodiment of this aspect of the invention, R ⁷ represents a fluorine or methyl atom.

According to one embodiment of this aspect of the invention, R ⁴ is selected from methyl, trifluoromethyl, cyclopropyl and phenyl, wherein said phenyl is optionally substituted in the meta position with one substituent selected from fluorine atom, chlorine atom, methyl, methoxy, dimethylamino, trifluoromethoxy, trifluoromethyl and cyclopropyl; And

R ⁵ is selected from chlorine, cyclopropyl, methyl and trifluoromethyl.

According to one embodiment of this aspect of the invention, R ⁴ and R ⁵ are independently selected from C ₁ -C ₃ haloalkyl, such as C ₁ -C ₃ fluoroalkyl, such as trifluoromethyl.

According to one embodiment of this aspect of the invention, R ⁴ is selected from methyl, trifluoromethyl and cyclopropyl; And

R ⁵ is selected from chlorine, cyclopropyl, methyl and trifluoromethyl.

According to one embodiment of this aspect of the invention, R ³ is selected from R ⁶ , C ₁ -C ₃ alkyl, N,N-diC ₁ -C ₃ alkylamino and methoxyC ₁ -C ₃ alkyl, wherein said C ₁ -C ₃ alkyl is optionally substituted one R ⁶ .

According to one embodiment of this aspect of the invention, R ⁶ is selected from phenyl, pyridyl, morpholinyl, imidazolyl, isoxazolyl, pyrazolyl, oxazolyl, cyclopropyl, cyclopentyl, pyrrolidinyl and tetrahydrofuryl, each of which is optionally substituted with one or more R ⁷ .

According to one embodiment of this aspect of the invention, R ⁶ is selected from phenyl, pyridyl, morpholinyl, imidazolyl, pyrazolyl, cyclopropyl, pyrrolidinyl, piperidinyl and tetrahydrofuryl, each of which is optionally substituted with one or more R ⁷ .

According to one embodiment of this aspect of the invention, R ⁶ is selected from phenyl, pyridyl, pyrrolidinyl, pyrazolyl, tetrahydrofuryl, each of which is optionally substituted with one or more R ⁷ .

According to one embodiment of this aspect of the invention, R ⁶ is selected from

According to one embodiment of this aspect of the invention, R ⁶ is selected from

According to one embodiment of this aspect of the invention, R ⁶ is selected from

According to one embodiment of this aspect of the invention, R ³ is selected from

According to one embodiment of this aspect of the invention, R ³ is selected from

According to one embodiment of this aspect of the invention, R ³ is selected from

According to one embodiment of this aspect of the invention, A is

According to one embodiment of this aspect of the invention, A is

According to one embodiment of this aspect of the invention, R ¹ represents methyl or cyclopropyl;

R ² represents a hydrogen atom;

A represents

and R ³ selected from

According to one embodiment of this aspect of the invention, R ¹ represents methyl;

R ² represents a hydrogen atom;

R ⁴ and R ⁵ are CF ₃ ;

A represents

and R ³ selected from

According to one embodiment of this aspect of the invention, R ¹ represents methyl;

R ² represents a hydrogen atom;

R ⁴ represents CF ₃ ;

A represents

and R ³ selected from

According to one embodiment of this aspect of the invention

R ¹ represents methyl or cyclopropyl;

R ² represents a hydrogen atom;

R ⁴ and R ⁵ are CF ₃ ;

A represents

and R ³ selected from

According to one embodiment of this aspect of the invention, said compound is:

4-(3-methylmorpholin-4-yl)-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;

6-[4-[(4-fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one;

6-[4-[(5-fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one;

4-(3-methylmorpholin-4-yl)-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;

4-(3-methylmorpholin-4-yl)-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;

N,N-dimethyl-4-[4-(3-methylmorpholin-4-yl)-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)piperazine-1-sulfonamide;

6-[4-(2-methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one;

6-[4-(4-fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one;

4-(3-methylmorpholin-4-yl)-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;

6-[4-cyclopropylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one;

4-(3-methylmorpholin-4-yl)-6-[4-(1-piperidylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;

4-(3-methylmorpholin-4-yl)-6-[4-morpholinosulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;

6-[4-(1,2-dimethylimidazol-4-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one;

6-[4-(1-methylcyclopropyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one;

4-(3-methylmorpholin-4-yl)-6-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]-1H-pyridin-2-one;

N,N-dimethyl-4-[4-(3-methylmorpholin-4-yl)-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)benzenesulfonamide;

or a pharmaceutically acceptable salt thereof.

According to one embodiment of this aspect of the invention, said compound is:

4-(3-methylmorpholin-4-yl)-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;

6-[4-[(4-Fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one;

6-[4-[(5-Fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one;

4-(3-methylmorpholin-4-yl)-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;

4-(3-methylmorpholin-4-yl)-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;

N,N-dimethyl-4-[4-(3-methylmorpholin-4-yl)-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)piperazine-1-sulfonamide;

6-[4-(2-methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one;

6-[4-(4-fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one;

4-(3-methylmorpholin-4-yl)-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one;

or a pharmaceutically acceptable salt thereof.

According to one aspect of the invention, a compound of the present invention is provided for use in the treatment or prevention of a disease.

According to one aspect of the invention, a compound of the present invention is provided for use in the treatment of cancer. Typically, said cancer is selected from breast cancer, such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, kidney cancer, colon cancer, glioma, prostate cancer, cancer ovarian, melanoma and lung cancer, as well as hypoxic tumors.

According to one aspect of the invention, a compound of the present invention is provided for use in the treatment of type II diabetes.

According to one aspect of the invention, a compound of the present invention is provided for use in the treatment of a disease selected from inflammatory diseases, autoimmune diseases, neurodegenerative disorders, cardiovascular disorders and viral infections.

According to one aspect of the invention, there is provided the use of a compound of the present invention for the preparation of a medicament for the treatment of cancer. Typically, said cancer is selected from breast cancer, such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, kidney cancer, colon cancer, glioma, prostate cancer, cancer ovarian, melanoma and lung cancer, as well as hypoxic tumors.

According to one aspect of the invention, there is provided the use of a compound of the present invention for the preparation of a medicament for the treatment of type II diabetes.

According to one aspect of the invention, there is provided the use of a compound of the present invention for the preparation of a medicament for the treatment of a disease selected from inflammatory diseases, autoimmune diseases, neurodegenerative disorders, cardiovascular disorders and viral infections.

According to one aspect of the invention, there is provided a method of treating cancer, comprising administering a therapeutically effective amount of a compound of the present invention to a patient in need thereof. Typically, said cancer is selected from breast cancer, such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, kidney cancer, colon cancer, glioma, prostate cancer, cancer ovarian, melanoma and lung cancer, as well as hypoxic tumors.

According to one aspect of the invention, a compound of the present invention is provided for use in the treatment of cancer, wherein said cancer treatment further comprises radiation therapy.

According to one aspect of the invention, there is provided a method of treating cancer, comprising administering a therapeutically effective amount of a compound of the present invention to a patient in need thereof, together with radiation therapy.

The compounds of the present invention can also be used for the treatment of cancer in conjunction with radiation therapy and/or surgery. Typically, the use of cytotoxic and/or cytostatic agents in combination with a compound or composition of the present invention will:

(1) achieving better effectiveness in reducing tumor growth or even eliminating tumors compared with the case of administering each of the agents separately;

(2) ensuring the administration of smaller amounts of chemotherapeutic agents used;

(3) providing chemotherapy treatment that is well tolerated by the patient and has fewer harmful pharmacologic complications than observed with single agent chemotherapy and certain other combination therapies;

(4) providing treatment for a wider range of different types of cancer in mammals, especially in humans;

(5) achieving a higher response rate among treated patients;

(6) providing longer survival times among treated patients compared to standard chemotherapy methods;

(7) ensuring a delay in tumor progression and/or

(8) obtaining at least as satisfactory results in terms of efficacy and tolerability as the agents used alone as compared with known antagonistic effects of other combinations of anticancer agents.

According to one aspect of the invention, there is provided a method of treating type II diabetes, comprising administering a therapeutically effective amount of a compound of the present invention to a patient in need thereof.

According to one aspect of the invention, there is provided a method of treating hypoxic tumors, comprising administering a therapeutically effective amount of a compound of the present invention to a patient in need thereof.

According to one aspect of the invention, there is provided a method of treating a disease selected from inflammatory diseases, autoimmune diseases, neurodegenerative disorders and viral infections, comprising administering a therapeutically effective amount of a compound of the present invention to a patient in need thereof.

According to one aspect of the invention, there is provided a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable diluent, carrier and/or excipient.

According to one aspect of the invention, there is provided a pharmaceutical composition containing a therapeutically effective amount of a compound according to claim 1 and another anticancer agent selected from alkylating agents, antimetabolites, anticancer camptothecin derivatives, anticancer herbal agents, antibiotics, enzymes, platinum coordination complexes, tyrosine kinase inhibitors, hormones, hormone antagonists, monoclonal antibodies, interferons and biological response modifiers.

As used herein, the term “C ₁ -C ₆ alkyl” means saturated hydrocarbon groups, both straight and branched chain, consisting of 1-6 carbon atoms. Examples of C ₁ -C ₆ alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, 4-methyl-butyl, n-hexyl, 2-ethyl -butyl group. Among straight-chain C ₁ -C ₆ alkyl groups, typical ones are methyl, ethyl, n-propyl, n-butyl, n-pentyl and n-hexyl groups. Among the branched alkyl groups, mention may be made of isopropyl, isobutyl, sec-butyl, tert-butyl, 4-methylbutyl and 2-ethylbutyl groups.

As used herein, the term “C ₁ -C ₃ alkyl” means saturated hydrocarbon groups, both straight and branched chain, consisting of 1-3 carbon atoms. Examples of C ₁ -C ₃ alkyl groups include methyl, ethyl, n-propyl and isopropyl groups.

As used herein, the term “C ₁ -C ₆ alkoxy” means an O-alkyl group, and “C ₁ -C ₆ alkyl” is used as described above. Examples of C ₁ -C ₆ alkoxy groups include, but are not limited to, methoxy, ethoxy, isopropoxy, n-propoxy, n-butoxy, n-hexoxy, 3-methyl-butoxy groups.

As used herein, the term “C ₁ -C ₃ alkoxy” means an O-alkyl group, and “C ₁ -C ₃ alkyl” is used as described above. Examples of C ₁ -C ₃ alkoxy groups include, but are not limited to, methoxy, ethoxy, isopropoxy and n-propoxy groups.

As used herein, the term "C ₁ -C ₆ haloalkyl" means saturated hydrocarbon groups, both straight and branched chain, consisting of 1-6 carbon atoms, and in which from 1 to all hydrogen atoms are replaced by halogen atom(s) of various types. or the same type. Examples of C ₁ -C ₆ haloalkyl groups include methyl groups substituted with 1-3 halogen atoms, ethyl groups substituted with 1-5 halogen atoms, n-propyl or isopropyl groups substituted with 1-7 halogen atoms, n-butyl or isobutyl groups, substituted with 1-9 halogen atoms, and sec-butyl or tert-butyl groups substituted with 1-9 halogen atoms.

As used herein, the term "C ₁ -C ₃ haloalkyl" means saturated hydrocarbon groups, both straight and branched chain, consisting of 1 to 3 carbon atoms, and in which from 1 to all hydrogen atoms are replaced by halogen atom(s) of various types. or the same type. Examples of C ₁ -C ₃ haloalkyl groups include methyl substituted with 1-3 halogen atoms, ethyl substituted with 1-5 halogen atoms, and n-propyl or isopropyl substituted with 1-7 halogen atoms.

As used herein, the term "C ₁ -C ₃ haloalkoxy" means saturated alkoxy groups, both straight and branched, consisting of 1 to 3 carbon atoms, and in which from 1 to all hydrogen atoms are replaced by halogen atom(s) of various types. or the same type. Examples of C ₁ -C ₃ haloalkoxy groups include a methoxy group substituted with 1 to 3 halogen atoms, an ethoxy group substituted with 1 to 5 halogen atoms, and an n-propoxy or isopropoxy group substituted with 1 to 7 halogen atoms.

As used herein, the term “C ₁ -C ₃ fluoroalkyl” means saturated hydrocarbon groups, both straight and branched chain, consisting of 1 to 3 carbon atoms, and in which from 1 to all hydrogen atoms are replaced by fluorine atom(s). Examples of C ₁ -C ₃ fluoroalkyl groups include methyl substituted with 1-3 fluorine atoms, ethyl substituted with 1-5 fluorine atoms, and n-propyl or isopropyl substituted with 1-7 fluorine atoms.

As used herein, the term “C ₁ -C ₃ fluoroalkoxy” means saturated alkoxy groups, both straight and branched chain, consisting of 1 to 3 carbon atoms, and in which from 1 to all hydrogen atoms are replaced by fluorine atom(s). Examples of C ₁ -C ₃ fluoroalkoxy groups include a methoxy group substituted with 1 to 3 fluorine atoms, an ethoxy group substituted with 1 to 5 fluorine atoms, and an n-propoxy or isopropoxy group substituted with 1 to 7 fluorine atoms.

As used herein, the term “C ₃ -C ₆ cycloalkyl” means a cyclic saturated hydrocarbon group consisting of 3-6 carbon atoms. Examples of C ₃ -C ₆ cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

As used herein, the term “C ₁ -C ₃ alkoxyC ₁ -C ₃ alkyl” means a saturated hydrocarbon group, either straight or branched chain, consisting of 1-3 carbon atoms, substituted by an alkoxy group of 1-3 carbon atoms. Examples of C ₁ -C ₃ alkoxyC ₁ -C ₃ alkyl groups are given below:

As used herein, the term “C ₁ -C ₃ cyanoalkyl” means a straight- or branched-chain cyano(CN) derivative containing one to three carbon atoms, including a carbon atom that forms part of the cyano group. Examples of C ₁ -C ₃ cyanoalkyl groups are given below:

As used herein, the term "N-C ₁ -C ₃ alkylamino" means an amino-containing substituent bearing one C ₁ -C ₃ alkyl group as defined above. Examples of N-C ₁ -C ₃ alkylamino are given below:

As used herein, the term "N,N-diC ₁ -C ₃ alkylamino" means an amino-containing substituent bearing two C ₁ -C ₃ alkyl groups as defined above. Examples of N,N-diC ₁ -C ₃ alkylamino are given below:

As used herein, the term “halogen” means fluorine, chlorine, bromine or iodine. As used herein, the term “halogen atom” means a fluorine, chlorine, bromine or iodine atom. It should be understood that if the substituent is a halogen, it is always associated with a carbon atom.

As used herein, the term “heteroaryl” means a monocyclic aromatic group consisting of carbon atoms in which one to three carbon atoms have been replaced by one or more heteroatoms independently selected from nitrogen, oxygen or sulfur atoms. In a bicyclic aryl, one of the rings may be partially saturated.

As used herein, the term “monocyclic heteroaryl” means a monocyclic aromatic group consisting of carbon atoms in which one to three carbon atoms have been replaced by one or more heteroatoms independently selected from nitrogen, oxygen or sulfur atoms.

Examples of monocyclic heteroaryl groups include, but are not limited to, furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, oxadiazolyl, thiadiazolyl, pyridyl, triazolyl, triazinyl, pyridazyl, isothiazolyl, isoxazolyl, pyrazinyl, pyrazolyl, and pyrimidinyl.

As used herein, the term “heterocyclyl” means a cyclic group consisting of carbon atoms in which one to three carbon atoms are replaced by one or more heteroatoms independently selected from nitrogen, oxygen or sulfur atoms. Examples of heterocyclyl groups include, but are not limited to, tetrahydrofuryl, tetrahydropyranyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl and dioxanyl.

Depending on the substituents present in the compounds of formula (I), the compounds can form salts which are within the scope of the present invention. Salts of the compounds of formula (I) suitable for medical use are those where the counterion is pharmaceutically acceptable.

Suitable salts of the invention include those formed by reaction with organic or inorganic acids or bases. In particular, suitable salts formed by reaction with acids according to the invention include salts formed by reaction with mineral acids, with strong organic carboxylic acids, such as alkane carboxylic acids consisting of 1-4 carbon atoms, which are unsubstituted or substituted, for example , a halogen atom, such as saturated or unsaturated dicarboxylic acids, such as hydroxycarboxylic acids, such as amino acids, or with organic sulfonic acids, such as (C ₁ -C ₄ )alkyl or arylsulfonic acids, which are unsubstituted or substituted, for example , a halogen atom. Pharmaceutically acceptable acid addition salts include those formed from hydrochloric, hydrobromic, sulfuric, nitric, citric, tartaric, acetic, phosphoric, lactic, pyruvic, acetic, trifluoroacetic, succinic, perchloric, fumaric, maleic, glycolic, lactic, salicylic, oxaloacetic, methanesulfonic, ethanesulfonic, p-toluenesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic, benzenesulfonic, isethionic, ascorbic, malic, phthalic, aspartic and glutamic acids, lysine and arginine.

Pharmaceutically acceptable salts with bases include ammonium salts, alkali metal salts, for example, potassium and sodium salts, alkaline earth metal salts, for example, calcium and magnesium salts, and salts with organic bases, for example, dicyclohexylamine, N-methyl-D-glucamine, morpholine , thiomorpholine, piperidine, pyrrolidine, with lower mono-, di- or trialkylamines, for example, ethyl-, tert-butyl-, diethyl-, diisopropyl-, triethyl-, tributyl- or dimethylpropylamine, or with lower mono-, di- - or a tri-hydroxyalkylamine, for example mono-, di- or tri-ethanolamine. In addition, corresponding internal salts can be formed.

The compounds of the invention can be used for prophylaxis and/or treatment as is or in the form of a pharmaceutical composition. Although it is possible to administer the active ingredient as such, it is also possible for it to be present in a pharmaceutical composition. Accordingly, according to the invention there is provided a pharmaceutical composition containing a compound of formula (I) and a pharmaceutically acceptable diluent, excipient and/or carrier. The pharmaceutical compositions of the invention may be in the form of a pharmaceutical composition, which is described below.

Typical compositions for oral administration include suspensions, which may contain, for example, microcrystalline cellulose for bulking, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweeteners or flavoring agents, such as those known in the art; and immediate release tablets, which may contain, for example, microcrystalline cellulose, dibasic calcium phosphate, starch, magnesium stearate, calcium sulfate, sorbitol, glucose and/or lactose and/or other excipients, binders, dry fillers, disintegrants, diluents and lubricants, such as those known in the art. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as gum acacia, gum tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum and the like. Delivery of the compounds of formula (I) can also be accomplished through the oral cavity by sublingual and/or buccal administration. Molded tablets, compressed tablets or freeze-dried tablets are typical forms that can be used. Typical compositions include compositions containing the compound(s) of the present invention with rapidly dissolving diluents such as mannitol, lactose, sucrose and/or cyclodextrins. High molecular weight excipients such as celluloses (Avicel) or polyethylene glycols (PEG) may also be included in such compositions. Such compositions may also include a mucosal adhesion promoting excipient such as hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC), sodium carboxymethylcellulose (SCMC), maleic anhydride copolymer (e.g., Gantrez), and release control agents. such as a copolymer based on an acrylic acid polymer (for example, Carbopol 934). For ease of manufacture and use, lubricants, glidants, flavors, colorants and stabilizers may also be added. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. For oral administration in liquid form, the components of the oral drug may be combined with any orally suitable non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like.

The compositions of the present invention suitable for oral administration may be presented in the form of discrete units such as capsules, wafers, pills or tablets, each containing a predetermined amount of the active ingredient; in the form of powder or granules; in the form of a solution or suspension in an aqueous liquid or non-aqueous liquid, for example, in the form of elixirs, tinctures, suspensions or syrups; or in the form of an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be presented as a bolus, electuary or paste.

The tablet may be made by compression or molding, possibly with one or more additional ingredients. Compressed tablets can be prepared in a suitable machine by compressing the active ingredient into a free-flowing form such as powder or granules, possibly mixed with a binder, lubricant, inert diluent, surfactant or dispersing agent. Molded tablets can be prepared in a suitable machine by molding a mixture of crushed compound moistened with an inert liquid diluent. It is possible that the tablets may be coated or scored and may be formulated to provide a slow or controlled release of the active ingredient. For example, the compounds of the present invention can be administered in a form suitable for immediate release or sustained release. Immediate release or sustained release can be achieved through the use of suitable pharmaceutical compositions containing the compounds of the present invention, or, especially in the case of sustained release, through the use of devices such as subcutaneous implants or osmotic pumps. The compounds of the present invention can also be administered using liposomes.

Typical compositions in unit dosage form are those that contain the active ingredient at an effective dose as given above, or an appropriate proportion thereof.

It should be understood that in addition to the ingredients, particularly those mentioned above, the compositions of this invention may include other agents conventionally used in the art, taking into account the type of composition in question, for example, those suitable for oral administration may include flavorings.

These compositions may be presented in unit dosage form and may be prepared by any of the methods well known in the pharmaceutical field. Such methods may include the step of combining the active ingredient with a carrier that consists of one or more additional ingredients. The compositions can be prepared by continuously and uniformly combining the active ingredient with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired composition.

The compounds of the present invention can also be administered in liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, 1,2-dipalmitoylphosphatidylcholine, phosphatidylethanolamine (cephalin), phosphatidylserine, phosphatidylinositol, diphosphatidylglycerol (cardiolipin), or phosphatidylcholine (lecithin).

Compositions for parenteral administration include aqueous and non-aqueous sterile injectable solutions, which may contain antioxidants, buffers, bacteriostatic agents and solutes to make the composition isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickening agents. These compositions may be presented in single-dose or multi-dose containers, such as sealed ampoules and hermetically sealed vials, and may be stored in a freeze-dried (lyophilized) state, requiring only the addition of a sterile liquid carrier, such as saline or water for injection, immediately prior to application. Extemporaneous solutions and suspensions for injection can be prepared from sterile powders, granules and tablets of the type described earlier. Typical compositions for parenteral administration include injectable solutions or suspensions, which may contain, for example, suitable non-toxic parenterally acceptable diluents or solvents such as polyethylene glycol, ethanol, 1,3-butanediol, water, Ringer's solution, isotonic sodium chloride solution, or other suitable dispersing or wetting and suspending agents, including synthetic mono- or diglycerides, and fatty acids, including oleic acid or cremophor.

Typical compositions for nasal, aerosol or inhalation administration include solutions in saline, which may contain, for example, benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability and/or other solubilizing or dispersing agents, for example, agents known in the art technology.

Compositions for rectal administration can be presented in the form of a suppository with typical carriers such as cocoa butter, synthetic glycerol esters (glycerides) or polyethylene glycol. Typically, such carriers are solid at normal temperatures but melt and/or dissolve in the rectum to release the drug.

Compositions for topical administration to the oral cavity, for example, buccally or sublingually, include lozenges containing the active ingredient in a flavored base such as sucrose and gum acacia or tragacanth, and lozenges containing the active ingredient in a base such as gelatin and glycerin or sucrose and gum arabic. Typical compositions for topical administration include a topical carrier such as Plastibase® (polyethylene thickened mineral oil).

The compounds of formula (I) may be administered as a single pharmaceutical agent or in combination with one or more additional therapeutic agents without the combination causing any unacceptable adverse effects. Such a pharmaceutical composition involves administering a single pharmaceutical composition that contains a compound of formula (I) and one or more additional therapeutic agents, and administering a compound of formula (I) and each additional therapeutic agent as part of its own separately administered pharmaceutical composition. For example, a compound of Formula (I) and a therapeutic agent may be administered to a patient together in a single orally administered composition, such as a capsule or tablet, or each agent may be administered in separate dosage compositions.

When compositions are used for separate administration, administration of a compound of formula (I) and one or more additional therapeutic agents can be performed substantially in parallel (eg, simultaneously) or spaced apart in time (eg, sequentially).

The amount of active ingredient required to achieve a therapeutic effect will undoubtedly vary depending on the specific compound, the route of administration, the subject being treated, including the type, species, age, weight, sex and health status of the subject and renal and hepatic function. in a given subject, and the particular disorder or disease being treated, as well as its severity. A physician, veterinarian, or normally trained clinician can readily determine and prescribe the effective amount of a drug needed to prevent, counteract, or delay the progression of the condition.

Oral dosages of the present invention, when used to achieve these effects, will range from about 0.01 mg per kg body weight per day (mg/kg/day) to about 100 mg/kg/day, preferably from 0. 01 mg per kg body weight per day (mg/kg/day) to 10 mg/kg/day and most preferably 0.1 to 5.0 mg/kg/day for an adult. If administered orally, the compositions may be formulated as tablets or other presentation forms prepared in discrete units containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0 , 10.0, 15.0, 25.0, 50.0, 100 and 500 milligrams of the active ingredient, to adjust the dosage of the symptomatic therapy of the patient being treated. Typically, the drug contains from about 0.01 mg to about 500 mg of active ingredient, preferably from about 1 mg to about 100 mg of active ingredient. If administered intravenously, the most preferred doses will be in the range of about 0.1 to about 10 mg/kg/minute when infused at a constant rate. The compounds of the present invention can be administered in a single daily dose, or the total daily dosage can be administered in divided doses on a two, three or four times daily basis. In addition, the compounds of the present invention can be administered in intranasal form through topical application together with suitable intranasal excipients or can be administered by transdermal routes using such forms of transdermal skin patches as are well known to those of ordinary skill in the art. If administered in the form of a transdermal delivery system, the dosing will certainly be continuous rather than intermittent throughout the dosing regimen.

Receiving connections

The compounds of the present invention can be prepared as the free base or a pharmaceutically acceptable salt thereof by the method described below. In the following description of such methods, it is intended that, where appropriate, suitable protecting groups will be added to and subsequently removed from the various reagents and intermediates in a manner readily apparent to one skilled in the art of organic synthesis. Traditional techniques for the use of such protecting groups, as well as examples of suitable protecting groups, are described, for example, in Protective Groups in Organic Synthesis, edited by TW Greene, PGM Wutz, ^4th edition, Wiley-Interscience, New York, 2006. It is obvious that for an alternative Microwave radiation can be used to heat reaction mixtures.

According to another aspect of the present invention, there is provided a method for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² and A are, unless otherwise stated, as defined herein. This method includes:

(1) formation of the corresponding compound of formula (I)

A compound of formula (I) can be prepared (Scheme 1) starting from, for example, a compound of formula (II), where R ^x can be F, OCH ₃ , OS(CH ₃ ) ₃ or OSiR'R''R''' (wherein R', R'' and R''' are independently aryl (such as phenyl) or alkyl (such as methyl or t-butyl)). If R ^x represents F, then conversion to compound (I) can be accomplished by, for example, acid hydrolysis using an aqueous HCl solution. If R ^x is OCH ₃ , then conversion to compound (I) can be accomplished by reaction, for example, with trimethylsilyl iodide in a suitable solvent such as chloroform, or by reaction with HBr in a suitable solvent such as acetic acid, or by reaction with BBr ₃ in a suitable solvent such as dichloromethane. If R ^x is OC(CH ₃ ) ₃ , then conversion to compound (I) can be accomplished by reaction with, for example, trifluoroacetic acid in a suitable solvent such as dichloromethane. If R ^x is OSiR'R''R''', then conversion to compound (I) can be accomplished using, for example, HCl in a suitable solvent such as methanol, or using tetrabutylammonium fluoride in tetrahydrofuran. If enantiomerically pure or enriched compound (II) is used in this reaction, enantiomerically pure or enantiomerically enriched compound (I) is obtained.

The compounds of formula (II) are commercially available or known from the literature, or they are prepared by standard methods known in the art. The compound of formula (I) or (II) can be separated into its enantiomers by standard methods known in the art, for example, by chromatography on a chiral stationary phase.

General methods

All solvents used were of analytical grade, and commercially available anhydrous solvents were routinely used to carry out the reactions. Starting materials were obtained from commercial sources or prepared according to published procedures in the literature. Room temperature means +20-25°C. The compositions of solvent mixtures are given in volume percentages or volume ratios.

Microwave heating was performed in a Biotage Initiator microwave cavity irradiating continuously at 2.45 GHz. Obviously, microwave radiation can be used to heat reaction mixtures.

Normal phase chromatography was performed manually on silica gel 60 (0.040-0.063 mm) from Merck or automated using the ISCO Combiflash® Companion™ system using SiliaSep™ normal phase flash chromatography columns and the specified solvent system.

Nuclear magnetic resonance (NMR) spectra were recorded on an NMR spectrometer at 400 MHz (or higher field) equipped with a suitably configured probe. Spectra were recorded at ambient temperature unless otherwise stated. Chemical shifts are given in ^ppm downfield and upfield relative to the TMS (tetramethylsilane) signal (0.00 ^ppm ). The following reference signals were used: the residual signal of the solvent DMSO-d ₆ δ 2.5, CDCl ₃ δ 7.26, or methanol-d ₄ δ 3.31. The multipletities of resonant signals are denoted as s, d, t, q, m and br for singlet, doublet, triplet, quartet, multiplet and broadened signals, respectively.

High pressure liquid chromatography (HPLC) was performed on a reverse phase column. A linear gradient was applied using, for example, mobile phase A (aqueous 0.1% NH ₃ solution, or aqueous 0.1% acetic acid solution, or aqueous 0.1% formic acid solution) and B ( acetonitrile or methanol). Mass spectrometry (MS) analyzes were performed in positive ion mode using electrospray ionization (ES+).

Preparative chromatography was performed on a Gilson-PREP GX271 or GX281 using Trilution Ic as software on a reverse phase column. A linear gradient was applied using, for example, mobile phase A (aqueous 0.1% NH ₃ solution, or aqueous 0.1% acetic acid solution, or aqueous 0.1% formic acid solution) and B ( acetonitrile or methanol).

Preparative chiral chromatography for enantiomer separation was performed on a Thar SFC system using supercritical fluid chromatography (SFC) on a chiral stationary phase. A linear gradient was applied using mobile phase A (carbon dioxide) and B (acetonitrile or methanol or ethanol or 2-propanol or any mixtures thereof). Excipients (such as diethylamine or isopropylamine or ammonia or formic acid or TFA) may be used.

Compound names are given using BIOVIA Draw 16.1.

Abbreviations

Example 1

(3R)-4-(2,6-Dichloro-4-pyridyl)-3-methyl-morpholine

2,6-Dichloro-4-iodopyridine (2 g; 7.3 mmol), (3R)-3-methylmorpholine hydrochloride (1.1 g; 7.99 mmol), PPh ₃ (120 mg; 0.46 mmol), Pd(OAc) ₂ (50 mg; 0.22 mmol) and freshly ground K ₃ PO ₄ (5.5 g; 25.9 mmol) were transferred to DMF (15 ml). The resulting mixture was purged with nitrogen for 5 minutes and stirred at 100°C overnight. After cooling to RT, the mixture was poured into water (50 ml) and EtOAc (15 ml). The organic layer was separated and the aqueous layer was extracted with EtOAc (3×10 ml). The combined organic extracts were washed with brine, treated with Na ₂ SO ₄ , filtered, concentrated and purified on a silica gel column eluting with 10-60% EtOAc in heptane to give the product as a solid (520 mg; 29%). MS ES+ m/z 247 [M+H] ⁺ .

Example 2

(3R)-4-(2-tert-Butoxy-6-chloro-4-pyridyl)-3-methyl-morpholine

(3R)-4-(2,6-Dichloro-4-pyridyl)-3-methyl-morpholine (520 mg; 2.1 mmol), KOtBu (500 mg; 4.46 mmol) and 4 molecular sieves (approximately 10 beads; 4-8 mesh) were transferred to toluene (8 ml) and stirred at 90°C for 2 hours. After cooling to RT, the mixture was diluted with EtOAc (10 ml), brine (5 ml) and water ( 5 ml). The organic layer was separated and the aqueous layer was extracted with EtOAc (10 ml). The combined organic extracts were washed with brine, concentrated and purified on a silica gel column eluting with 0-40% EtOAc in heptane to give the product as an oil. MS ES+ m/z 285 [M+H] ⁺ .

Example 3

tert-Butyl-4-[6-tert-butoxy-4-[(3R)-3-methylmorpholin-4-yl]-2-pyridyl]-3-(trifluoromethyl)piperazine-1-carboxylate

Mixture of tert-butyl 3-(trifluoromethyl)piperazine-1-carboxylate (500 mg; 1.97 mmol), (3R)-4-(2-tert-butoxy-6-chloro-4-pyridyl)-3-methyl -morpholine (670 mg; 2.36 mmol), KOtBu (440 mg; 3.94 mmol) and PEPPSI-iPr (133 mg; 0.2 mmol) in 1,4-dioxane (15 ml) were heated in a microwave reactor at 130°C for 3 hours. The reaction mixture was poured into water (10 ml) and extracted with EtOAc (3 x 20 ml). The combined organic extracts were washed with brine, treated with Na ₂ SO ₄ , filtered, concentrated and purified on a silica gel column eluting with 20-30% EtOAc in heptane to give the product as an oil (420 mg; 42%). MS ES+ m/z 503 [M+H] ⁺ .

Example 4

(3R)-4-[2-tert-Butoxy-6-[2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

LiAIH ₄ (63 mg; 1.67 mmol) was added at 0°C to a solution of tert-butyl-4-[6-tert-butoxy-4-[(3R)-3-methylmorpholin-4-yl]-2-pyridyl ]-3-(trifluoromethyl)-piperazine-1-carboxylate (420 mg; 0.84 mmol) in THF (10 ml) and the resulting mixture was stirred at RT for 5 hours. The reaction mixture was quenched with ice/water and extracted with EtOAc ( 3× 10 ml). The combined organic extracts were washed with brine, treated with Na ₂ SO ₄ , filtered, concentrated and purified on a silica gel column eluting with 4-5% MeOH in DCM to give the product as a solid (200 mg; 59%). MS ES+ m/z 403 [M+H] ⁺ .

Example 5

(3R)-4-[2-tert-Butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

Methanesulfonyl chloride (85 mg; 0.75 mmol) was added dropwise to a solution of (3R)-4-[2-tert-butoxy-6-[2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3- methyl morpholine (200 mg; 0.5 mmol) and Et ₃ N (100 mg; 0.99 mmol) in DCM (15 ml) at 0°C and the resulting mixture was stirred at RT for 3 hours. The reaction mixture was quenched with the mixture ice/water and extracted with EtOAc (3×10 ml). The combined organic extracts were washed with brine, treated with Na ₂ SO ₄ , filtered, concentrated and purified on a silica gel column eluting with 0-100% EtOAc in heptane to give the product as a solid (150 mg; 63%). MS ES+ m/z 481 [M+H] ⁺ .

Example 6

(R)- and (S)-4-[(3R)-3-Methylmorpholin-4-yl]-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2- He

TFA (0.4 ml; 5.23 mmol) was added to a solution of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl ]-3-methyl-morpholine (150 mg; 0.31 mmol) in DCM (15 ml) at 0°C and the resulting mixture was stirred at RT for 6 hours. The pH was adjusted to above 7 using sat. aq. NaHCO ₃ solution and the mixture was extracted with DCM (3×10 ml). The combined organic extracts were washed with brine, treated with Na ₂ SO ₄ , filtered, concentrated and purified on a silica gel column eluting with 0-10% MeOH in DCM to give the product as a solid (20 mg; 23%).

After separation under chiral conditions by SFC, two isomers were obtained.

Compound of Example 6-1, first eluting isomer, with unknown absolute configuration.

^1H NMR (400 MHz, DMSO- _d6 ): δ 10.45 (br.s., 1H), 5.75 (br.s., 1H), 5.56 (br.s., 1H), 5.43 (br.s. , 1H), 4.09 (br.s., 1H), 3.90-3.69 (m, 3H), 3.66-3.35 (m, 2H), 3.51-3.39 (m 3H), 3.21-3.10 (m, 1H), 3.10 -3.08 (m, 1H), 3.02-2.69 (m, 1H), 2.95-2.91 (s, 3H), 2.84-2.79 (m, 1H), 1.06-1.05 (m, 3H). MS ES+ m/z 425 [M+H] ⁺ .

Compound of Example 6-1, second eluting isomer, with unknown absolute configuration.

^1H NMR (400 MHz, DMSO- _d6 ): δ 10.45 (br.s., 1H), 5.76 (br.s., 1H), 5.56 (br.s., 1H), 5.43 (br.s. , 1H), 4.09 (br.s.,1H), 3.90-3.69 (m, 3H), 3.66-3.35 (m, 2H), 3.51-3.39 (m 3H), 3.21-3.10 (m, 1H), 3.10 -3.08 (m, 1H), 3.02-2.69 (m, 1H), 2.95-2.91 (s, 3H), 2.84-2.79 (m, 1H), 1.06-1.05 (m, 3H). MS ES ⁺ m/z 425 [M+H] ⁺ .

Example 7

(3R)-4-[2-tert-Butoxy-6-[4-[(4-fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5 using (4-fluorophenyl)methanesulfonyl chloride instead of methanesulfonyl chloride to give the product as a solid (40 mg; 37%). MS ES+ m/z 575 [M+H] ⁺ .

Example 8

6-[4-[(4-Fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-[(4-fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazine-1- yl]-4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl] -3-methyl-morpholine to give the product as a solid (12 mg; 33%). ^1H NMR (400 MHz, DMSO- _d6 ): δ 7.4 (q, 2H), 7.2 (t, 2H), 5.7 (br.s., 1H), 5.43 (br.s., 1H), 4.5 ( br.s., 2H), 4.09 (t, 1H), 3.90 (t, 2H), 3.8 (d, 1H), 3.6 (q, ZN), 3.4 (m, ZN), 3.08 (m, 3H), 2.8 (t, 1H), 1.06-1.05 (t, 3H). MS ES+ m/z 519 [M+H] ⁺ .

Example 9

(3R)-4-[2-tert-Butoxy-6-[4-[(5-fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3- methyl morpholine

The title compound was prepared as described in Example 5 using 5-fluoropyridine-3-sulfonyl chloride instead of methanesulfonyl chloride to give the product as a solid (40 mg; 32%). MS ES+m/z 562 [M+H] ⁺ .

Example 10

6-[4-[(5-Fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin- 2-he

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-[(5-fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl) piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]- 4-pyridyl]-3-methyl-morpholine to give the product as a solid (26 mg; 72%). ^1H NMR (400 MHz, DMSO- _d6 ): δ 9.8 (br.s., 1H), 8.9 (d, 1H), 8.8 (br.s., 1H), 8.2 (d, 1H), 5.7 ( br.s., 1H), 5.6 (s, 1H), 5.5 (s, 1H), 5.4 (s, 1H), 4.0 (t, 2H), 3.8 (d, 2H), 3.7-3.6 (t, 2H ), 3.5 (d, 1H), 3.4 (m, 1H), 3.3 (br.s., 1H), 3.2 (d, 1H), 2.9 (m, 1H), 2.8 (d, 1H), 2.5 (m , 1H), 1.2 (t, 1H), 1.06-1.05 (q, 3H). MS ES+m/z 506 [M+H] ⁺ .

Example 11

(3R)-4-[2-tert-Butoxy-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5 using tetrahydrofuran-3-sulfonyl chloride instead of methanesulfonyl chloride to give the product as a solid (35 mg; 35%). MS ES+m/z 537 [M+H] ⁺ .

Example 12

4-[(3R)-3-Methylmorpholin-4-yl]-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]- 4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3- methyl morpholine to give the product as a solid (26 mg; 83%). ^1H NMR (400 MHz, DMSO- _d6 ): δ 9.8 (br.s., 1H), 5.7 (d, 1H), 5.5 (d, 1H), 4.0 (m, 2H), 3.9-3.7 (m , 6H), 3.6 (m, 4H), 3.5 (m, 3H), 3.0 (m, 2H), 2.4-2.1 (m, 1H), 2.0 (m, 1H), 1.9-1.1 (q, 1H), 1.0 (q, 3H). MS ES+m/z 481 [M+H] ⁺ .

Example 13

(3R)-4-[2-tert-Butoxy-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5 using pyrrolidine-1-sulfonyl chloride instead of methanesulfonyl chloride to give the product as a solid (76 mg; 71%). MS ES+m/z 536 [M+H] ⁺ .

Example 14

4-[(3R)-3-Methylmorpholin-4-yl]-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]- 4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3- methyl morpholine to give the product as a solid (48 mg; 72%). ^1H NMR (500 MHz, DMSO-d ₆ ) δ ppm ^1.05-1.12 (m, 3H), 1.82-1.89 (m, 4H), 2.85 (br t, 1H), 3.00 (td, 1H), 3.10 (br d, 1H), 3.16-3.31 (m, 5H), 3.33-3.42 (m, 1H), 3.44-3.56 (m, 2H), 3.59-3.65 (m, 1H), 3.66-3.71 (m, 1H), 3.83 (dd, 1H), 3.90 (br dd, 2H), 4.01 (br s, 1H), 5.43 (br s, 1H), 5.49 (br s, 1H), 5.70 (br s, 1H), 9.81 (br s, 1H). MS ES+m/z 480 [M+H] ⁺ .

Example 15

4-[6-tert-Butoxy-4-[(3R)-3-methylmorpholin-4-yl]-2-pyridyl]-N,N-dimethyl-3-(trifluoromethyl)piperazine-1-sulfonamide

The title compound was prepared as described in Example 5 using N,N-dimethylsulfamoyl chloride instead of methanesulfonyl chloride to give the product as a solid (60 mg; 59%). MS ES+m/z 510 [M+H] ⁺ .

Example 16

N,N-Dimethyl-4-[4-[(3R)-3-methylmorpholin-4-yl]-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)piperazine-1-sulfonamide

The title compound was prepared as described in Example 6 using 4-[6-tert-butoxy-4-[(3R)-3-methylmorpholin-4-yl]-2-pyridyl]-N,N-dimethyl- 3-(trifluoromethyl)piperazin-1-sulfonamide instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3- methyl morpholine to give the product as a solid (18 mg; 37%). ^1H NMR (500 MHz, DMSO-d ₆ ) δ ppm ^1.05-1.12 (m, 3H), 2.80 (d, 6H), 2.86-2.95 (m, 1H), 3.00 (td, 1H), 3.12- 3.24 (m, 2H), 3.34-3.41 (m, 1H), 3.44-3.56 (m, 2H), 3.59-3.64 (m, 1H), 3.66-3.74 (m, 1H), 3.81-3.95 (m, 3H ), 4.04 (br s, 1H), 5.44 (br s, 1H), 5.49 (br s, 1H), 5.70 (br s, 1H), 9.50-10.12 (m, 1H). MS ES+m/z 454 [M+H] ⁺ .

Example 17

(3R)-4-[2-tert-Butoxy-6-[4-(2-methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5 using 2-methoxyethanesulfonyl chloride instead of methanesulfonyl chloride to give the product as a solid (74 mg; 76%). MS ES+m/z 525 [M+H] ⁺ .

Example 18

6-[4-(2-Methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-(2-methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]- 4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3- methyl morpholine to give the product as a solid (48 mg; 75%). ^1H NMR (500 MHz, DMSO-d ₆ ) δ ppm ^1.05-1.16 (m, 3H), 2.90 (br t, 1H), 3.00 (td, 1H), 3.11-3.23 (m, 2H), 3.25 -3.30 (m, 3H), 3.33-3.51 (m, 4H), 3.55-3.71 (m, 5H), 3.83-3.96 (m, 3H), 4.05 (br s, 1H), 5.44 (br s, 1H) , 5.50 (br s, 1H), 5.72 (br s, 1H), 9.81 (br s, 1H). MS ES+m/z 469 [M+H] ⁺ .

Example 19

(3R)-4-[2-tert-Butoxy-6-[4-(4-fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5 using 4-fluorobenzenesulfonyl chloride instead of methanesulfonyl chloride to give the product as a solid (40 mg; 41%). MS ES+m/z 561 [M+H] ⁺ .

Example 20

6-[4-(4-Fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-(4-fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl] -4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3 -methyl-morpholine to give the product as a solid (22 mg; 61%). ^1H NMR (500 MHz, DMSO-d ₆ ) δ ppm ^1.04 (dd, 3H), 2.28-2.36 (m, 1H), 2.53-2.60 (m, 1H), 2.97 (td, 1H), 3.23 ( br t, 1H), 3.40-3.54 (m, 3H), 3.55-3.72 (m, 3H), 3.84-4.01 (m, 2H), 4.56 (t, 1H), 5.40 (br s, 1H), 5.49 ( br s, 1H), 5.67 (br s, 1H), 7.47-7.53 (m, 2H), 7.86 (dd, 2H), 9.78 (br s, 1H). MS ES+m/z 505 [M+H] ⁺ .

Example 21

(3R)-4-[2-tert-Butoxy-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl- morpholine

The title compound was prepared as described in Example 5 using 2-methylpyrazole-3-sulfonyl chloride instead of methanesulfonyl chloride to give the product as a solid (90 mg; 82%). MS ES+m/z 547 [M+H] ⁺ .

Example 22

4-[(3R)-3-Methylmorpholin-4-yl]-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2- He

The title compound was prepared as described in Example 6 using (3R)-4-[2-tert-butoxy-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazine- 1-yl]-4-pyridyl]-3-methyl-morpholine instead of (3R)-4-[2-tert-butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4- pyridyl]-3-methyl-morpholine to give the product as a solid (35 mg; 54%). ^1H NMR (500 MHz, DMSO-d ₆ ) δ ppm ^1.06 (dd, 3H), 2.64-2.74 (m, 1H), 2.92-3.03 (m, 2H), 3.20-3.30 (m, 1H), 3.33-3.52 (m, 2H), 3.57-3.64 (m, 1H), 3.65-3.74 (m, 2H), 3.81-3.91 (m, 2H), 3.93-4.02 (m, 1H), 4.03-4.06 (m , 3H), 4.06-4.14 (m, 1H), 5.44 (s, 1H), 5.55 (br s, 1H), 5.71 (s, 1H), 6.92 (dd, 1H), 7.65 (dd, 1H), 9.82 (br s, 1H). MS ES+m/z 491 [M+H] ⁺ .

Example 23

(3R)-4-[2-tert-Butoxy-6-[4-cyclopropylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5 using cyclopropanesulfonyl chloride to give the product as a solid (80 mg; 79%). MS ES+m/z 507 [M+H] ⁺ .

Example 24

6-[4-Cyclopropylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6, yielding the product as a solid (42 mg; 59%). ^1H NMR (500 MHz, DMSO-d ₆ ) δ ppm ^0.93-1.09 (m, 7H), 2.53-2.65 (m, 1H), 2.90-3.05 (m, 2H), 3.16-3.24 (m, 2H ), 3.33-3.52 (m, 2H), 3.56-3.64 (m, 2H), 3.65-3.72 (m, 1H), 3.85-3.96 (m, 3H), 4.10 (br s, 1H), 5.44 (br s , 1H), 5.56 (br d, 1H), 5.76 (s, 1H), 9.77 (br s, 1H). MS ES+m/z 451 [M+H] ⁺ .

Example 25

(3R)-4-[2-tert-Butoxy-6-[4-(1-piperidylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5 using piperidine-1-sulfonyl chloride to give the product as a solid (100 mg; 92%). MS ES+m/z 550 [M+H] ⁺ .

Example 26

4-[(3R)-3-Methylmorpholin-4-yl]-6-[4-(1-piperidylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6, yielding the product as a solid (42 mg; 47%). ^1H NMR (500 MHz, DMSO-d ₆ ) δ ppm ^1.07 (dd, 3H), 1.52 (br s, 6H), 2.85-3.03 (m, 1H), 3.08-3.12 (m, 1H) , 3.17 (br d, 5H), 3.34-3.39 (m, 1H), 3.45 (br s, 1H), 3.47-3.52 (m, 2H), 3.61 (dt, 1H), 3.69 (dd, 1H), 3.80 (d, 1H), 3.83 (d, 1H), 3.88-3.93 (m, 2H), 5.43 (s, 1H), 5.48 (br s, 1H), 5.70 (s, 1H), 9.32-9.88 (m, 1H). MS ES+m/z 494 [M+H] ⁺ .

Example 27

(3R)-4-[2-tert-Butoxy-6-[4-morpholinosulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine

The title compound was prepared as described in Example 5 using morpholine-4-sulfonyl chloride to give the product as a solid (75 mg; 77%). MS ES+m/z 483 [M+H] ⁺ .

Example 28

4-[(3R)-3-Methylmorpholin-4-yl]-6-[4-morpholinosulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6, yielding the product as a solid (16 mg; 64%). ^1H NMR (500 MHz, DMSO-d ₆ ) δ ppm ^1.07 (dd, 3H), 2.88-3.08 (m, 2H), 3.12-3.23 (m, 6H), 3.36-3.41 (m, 1H) , 3.43-3.50 (m, 1H), 3.52-3.70 (m, 7H), 3.82-3.96 (m, 3H), 4.02 (br s, 1H), 5.44 (br s, 1H), 5.49 (br s, 1H ), 5.71 (br s, 1H), 9.84 (br s, 1H). MS ES+m/z 496 [M+H] ⁺ .

Example 29

(3R)-4-[2-tert-Butoxy-6-[4-(1,2-dimethylimidazol-4-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3- methyl morpholine

The title compound was prepared as described in Example 5 using 1,2-dimethylimidazole-4-sulfonyl chloride to give the product as a solid (120 mg; 86%). MS ES+m/z 561 [M+H] ⁺ .

Example 30

6-[4-(1,2-Dimethylimidazol-4-yl)sulfonyl-2-(trifluoromethyl)-piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridine -2-he

The title compound was prepared as described in Example 6, yielding the product as a solid (24 mg; 23%). ^1H NMR (500 MHz, DMSO-d ₆ ) δ ppm ^1.04-1.09 (m, 3H), 2.25-2.32 (m, 3H), 2.41-2.49 (m, 1H), 2.65-2.82 (m, 1H ), 2.91-3.06 (m, 1H), 3.09-3.29 (m, 1H), 3.29-3.49 (m, 3H), 3.59-3.62 (m, 1H), 3.64-3.71 (m, 1H), 3.82-3.94 (m, 4H), 4.08 (br d, 1H), 4.56-4.58 (m, 1H), 5.41 (s, 1H), 5.47 (br d, 1H), 5.64-5.73 (m, 1H), 7.62-7.84 (m, 1H), 7.80 (s, 1H), 10.45 (m, 1H). MS ES+m/z 505 [M+H] ⁺ .

Example 31

6-[4-(1-Methylcyclopropyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-[(3R)-3-methylmorpholin-4-yl]-1H-pyridin-2-one

To an ice-cold solution of (3R)-4-[2-tert-butoxy-6-[2-(trifluoromethyl)piperazin-1-yl]-4-pyridyl]-3-methyl-morpholine (70 mg; 0.17 mmol in DCM (5 ml) TEA (0.12 ml; 0.87 mmol) and 1-methylcyclopropanesulfonyl chloride (40 mg; 0.26 mmol) were added. The cooling bath was removed and the resulting mixture was stirred at RT overnight. Added again 1-Methylcyclopropanesulfonyl chloride (40 mg; 0.26 mmol) and the reaction mixture was stirred at RT for 20 hours. 1-Methylcyclopropanesulfonyl chloride (40 mg; 0.26 mmol) was added again and the reaction mixture was kept at RT for 2 months. the residue was purified by preparative HPLC to give the product as a solid (21 mg; 26%) ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm ^0.80-0.90 (m, 2H), 1.06-1.07 (m, 3H) 1.12-1.28 (m, 2H), 1.41 (s, 3H), 2.94-3.10 (m, 2H), 3.12-3.24 (m, 1H), 3.28-3.31 (m, 1H), 3.35-3.41 (m , 1H), 3.42-3.51 (m, 1H), 3.58-3.73 (m, 3H), 3.87-4.00 (m, 3H), 4.03 (br d, 1H), 5.44 (br s, 1H), 5.48 (br s, 1H), 5.71 (s, 1H), 9.86 (br s, 1H). MS ES+m/z 465 [M+H] ⁺ .

Example 32

(3R)-4-[2-tert-Butoxy-6-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]-4-pyridyl]-3-methyl-morpholine

(3R)-4-(2-tert-Butoxy-6-chloro-4-pyridyl)-3-methyl-morpholine (85 mg; 0.3 mmol), K ₂ CO ₃ (83 mg; 0.6 mmol) , [4-methylsulfonyl-2-(trifluoromethyl)phenyl]boronic acid (80 mg; 0.3 mmol) and PdCl ₂ (Amphos) (11 mg; 0.02 mmol) were transferred to 1,4-dioxane (2 ml) and water (0.6 ml) and the resulting mixture was stirred at 95°C for 1 hour. The reaction mixture was left to cool, [4-methylsulfonyl-2-(trifluoromethyl)phenyl]boronic acid (161 mg; 0.6 mmol) was added. and PdCl ₂ (Amphos) (11 mg; 0.02 mmol) and the reaction mixture was again heated and stirred at 95°C for 3 hours. After cooling to RT, the mixture was extracted with EtOAc and the combined organic extracts were washed with brine, treated with Na ₂ SO ₄ , filtered, concentrated and purified on a silica gel column eluting with 0-50% EtOAc in heptane to give the product as a solid (22 mg; 16%). MS ES+m/z 473 [M+H] ⁺ .

Example 33

4-[(3R)-3-Methylmorpholin-4-yl]-6-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]-1H-pyridin-2-one

The title compound was prepared as described in Example 6, yielding the product as a solid (8 mg; 47%). ^1H NMR (500 MHz, DMSO-d ₆ ) δ ppm ^1.10 (d, 3H), 3.03 (td, 1H), 3.30-3.39 (m, 1H), 3.33-3.41 (m, 3H), 3.43- 3.54 (m, 1H), 3.58-3.70 (m, 2H), 3.86-3.95 (m, 2H), 5.48 (br s, 1H), 6.08 (br s, 1H), 7.87 (d, 1H), 8.28- 8.33 (m, 2H), 11.07 (br s, 1H). MS ES+m/z 417 [M+H] ⁺ .

Example 34

4-[6-tert-Butoxy-4-[(3R)-3-methylmorpholin-4-yl]-2-pyridyl]-N,N-dimethyl-3-(trifluoromethyl)benzenesulfonamide

A mixture of 4-bromo-N,N-dimethyl-3-(trifluoromethyl)benzenesulfonamide (199 mg; 0.6 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (183 mg; 0.72 mmol), KOAc (118 mg; 1.2 mmol) and PdCl ₂ (dppf) (44 mg; 0.06 mmol) in DMSO (2.5 ml) was stirred at 90°C overnight. After cooling to RT, (3R)-4-(2-tert-butoxy-6-chloro-4-pyridyl)-3-methyl-morpholine (171 mg; 0.6 mmol), K ₂ CO ₃ (166 mg; 0.6 mmol) were added. 1.20 mmol), PdCl ₂ (dppf) (22 mg; 0.03 mmol), 1,4-dioxane (1 ml) and water (1 ml) and the reaction mixture was stirred at 85°C for 2 hours. cooling to RT, [4-(dimethylsulfamoyl)-2-(trifluoromethyl)phenyl]boronic acid (178 mg; 0.6 mmol), K ₂ CO ₃ (83 mg; 0.6 mmol) and PdCl ₂ (dppf) were added 22 mg; 0.03 mmol) and the reaction mixture was stirred at 85°C for 5 hours. Water was added to the reaction mixture and extracted with EtOAc. The combined organic extracts were washed with brine, treated with Na ₂ SO ₄ , filtered, concentrated and purified on a silica gel column eluting with 0-80% EtOAc in heptane to give the product as a solid (200 mg; 67%). MS ES+m/z 502 [M+H] ⁺ .

Example 35

N,N-Dimethyl-4-[4-[(3R)-3-methylmorpholin-4-yl]-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)benzenesulfonamide

The title compound was prepared as described in Example 6. After completion of the reaction, the mixture was concentrated, treated with toluene and purified by preparative HPLC to give the product as a solid (85 mg; 50%). ^1H NMR (500 MHz, DMSO-d ₆ ) δ ppm 1.10 (d, 3H), ^2.66-2.78 (m, 6H), 3.03 (td, 1H), 3.38 (br s, 1H), 3.48 (td , 1H), 3.57-3.70 (m, 2H), 3.87-3.96 (m, 2H), 5.48 (br s, 1H), 6.12 (br s, 1H), 7.86 (d, 1H), 8.03 (d, 1H ), 8.12 (dd, 1H), 11.02 (br s, 1H). MS ES+m/z 446 [M+H] ⁺ .

Example 36

Biochemical analysis of Vps34

A series of dilutions of the compounds of the invention were prepared in DMSO at a concentration 100 times the final concentration in the assay (n ₁ =n ₀ /3; 10 points). The compounds were then diluted to 4 times the assay concentration in assay buffer (Buffer Q from Life Technologies, PV5125, diluted 5 times, supplemented with 2 mM dithiothreitol (DTT) and 2 mM MnCl ₂ ). 2.5 μl of diluted solutions of compounds were added to the 384-well plate used in the assay, then 2.5 μl of a 16.5 nM solution of the Vps34 enzyme (Life Technologies, PV5126) was added. Pre-incubation of the enzyme and compounds was carried out at RT for 15 min. Then, 5 μl of a substrate mixture containing 20 μM ATP (Life Technologies, PV3227) and 200 μM PI:PS (phosphatidylserine) substrate (Life Technologies, PV5122) in assay buffer was added to the wells containing the compound and enzyme. Mixing was carried out by repeated pipetting. The reaction mixture was incubated at room temperature for 1 hour. Then, 5 μl of stop and detection mixture, prepared as described in the instructions for the Adapta™ Kinase Assay Kit (Life Technologies, PV5099), containing Eu-labeled, was added to quench the reaction. anti-ADP antibody Adapta™ (2.3 nM), ADP indicator Alexa Fluor 647 (9 nM) and EDTA (ethylenediaminetetraacetic acid; 30 mM) in TR-FRET (time-resolved fluorescence resonance energy transfer) buffer. Mixing was carried out by repeated pipetting. The plate used in the assay was then incubated at room temperature for 30 min and measurements were taken using an Artemis microplate reader. The percentage of inhibition due to the compounds was calculated relative to DMSO-treated control samples. To determine the IC ₅₀ values (the concentration causing 50% inhibition), a point-by-point plot of the percentage of inhibition versus the concentration of the compounds was plotted using Dotmatics software.

The example compounds effectively inhibited Vps34 and the assay results are shown in Table 1 (mean IC ₅₀ values in nM; Adapta™).

Example 37

Simultaneous multiparameter analysis of autophagy

To determine the inhibitory effect of the proprietary compounds on autophagy, human osteosarcoma (HOS) cells stably expressing green fluorescent protein (GFP)-tagged LC3 (GFP-LC3) were used. For this purpose, autophagy was activated using the mTOR inhibitor (mammalian target of rapamycin) KU-0063794 at a concentration of 500 nM in the presence of bafilomycin A1 (Sigma-Aldrich) at a concentration of 5 nM. Briefly, cells were seeded overnight in clear-bottom 96-well plates in DMEM (Dulbecco's Modified Eagle's Medium) with high glucose (Hi-Clone, Cat. No. SH30285.01). At the start of the experiment, the medium was removed and replaced with fresh medium containing mTOR inhibitor, bafilomycin A1 and diluent or test compound as indicated. After 6 hours, the medium was removed and the cells were washed twice with ice-cold phosphate buffered saline (PBS) and fixed using 4% paraformaldehyde for 20 minutes at room temperature. Cells were then washed twice with ice-cold PBS, after which nuclear dye Hoechst 33342 was added at a concentration of 1 μg/ml in PBS. After incubation overnight at 4°C, cells were washed once with PBS to remove excess dye, and 100 μl of PBS was added to each well. Images were acquired at 20× magnification, 6 images per well, using an ImageXpress automated microscope (Molecular Devices Inc.) and analyzed using MetaXpress software to identify LC3-GFP foci. Foci area per cell was used to obtain a dose response curve, and IC ₅₀ values were calculated using nonlinear fit in GraphPad Prism software.

The tested compounds from the examples effectively inhibited autophagy in HOS cells. The results of this analysis are shown in Table 2 (mean IC ₅₀ values in nM; HOS-LC3).

Claims (110)

1. Compound of formula (I), Where R ¹ represents C ₁ -C ₃ alkyl; R ² represents hydrogen; A represents or Where R ³ is selected from R ⁶ , C ₁ -C ₆ alkyl, N,N-diC ₁ -C ₃ alkylamino and C ₁ -C ₃ alkoxyC ₁ -C ₃ alkyl, wherein said C ₁ -C ₆ alkyl is optionally substituted by one R ⁶ ; R ⁴ represents C ₁ -C ₆ haloalkyl; R ⁵ represents C ₁ -C ₆ haloalkyl; R ⁶ is selected from phenyl, monocyclic heteroaryl, C ₃ -C ₆ cycloalkyl, and heterocyclyl, each of which is optionally substituted with one or more R ⁷ ; R ⁷ is selected from halogen and C ₁ -C ₃ alkyl; wherein said heteroaryl represents an aromatic group of carbon atoms in which one or two carbon atoms are replaced by one or two nitrogen atoms; And wherein said heterocyclyl represents a cyclic group of carbon atoms in which one or two carbon atoms are replaced by one or more heteroatoms independently selected from nitrogen and oxygen; or a pharmaceutically acceptable salt thereof. 2. A compound according to claim 1, where R ¹ represents methyl, or a pharmaceutically acceptable salt thereof. 3. Connection according to any one of paragraphs. 1 or 2, where R ⁷ is selected from a fluorine atom and a methyl atom, or a pharmaceutically acceptable salt thereof. 4. Connection according to any one of paragraphs. 1-3, where R ⁴ represents trifluoromethyl; and R ⁵ represents trifluoromethyl; or a pharmaceutically acceptable salt thereof. 5. Connection according to any one of paragraphs. 1-4, where R ³ is selected from R ⁶ , C ₁ -C ₃ alkyl, N,N-diC ₁ -C ₃ alkylamino and methoxyC ₁ -C ₃ alkyl, wherein said C ₁ -C ₃ alkyl is optionally substituted by one R ⁶ , or a pharmaceutically acceptable salt thereof. 6. Connection according to any one of paragraphs. 1-5, wherein R ⁶ is selected from phenyl, pyridyl, morpholinyl, imidazolyl, pyrazolyl, cyclopropyl, cyclopentyl, pyrrolidinyl and tetrahydrofuryl, each of which is optionally substituted with one or more R ⁷ , or a pharmaceutically acceptable salt thereof. 7. Connection according to any one of paragraphs. 1-5, wherein R ⁶ is selected from phenyl, pyridyl, morpholinyl, imidazolyl, pyrazolyl, cyclopropyl, pyrrolidinyl, piperidinyl and tetrahydrofuryl, each of which is optionally substituted with one or more R ⁷ , or a pharmaceutically acceptable salt thereof. 8. Connection according to any one of paragraphs. 1-5, wherein R ⁶ is selected from phenyl, pyridyl, pyrrolidinyl, pyrazolyl, tetrahydrofuryl, each of which is optionally substituted with one or more R ⁷ , or a pharmaceutically acceptable salt thereof. 9. Connection according to any one of paragraphs. 1-8, where R ⁶ is selected from or a pharmaceutically acceptable salt thereof. 10. Connection according to any one of paragraphs. 1-8, where R ⁶ is selected from or a pharmaceutically acceptable salt thereof. 11. Connection according to any one of paragraphs. 1-8 and 10, where R ⁶ is selected from or a pharmaceutically acceptable salt thereof. 12. Connection according to any one of paragraphs. 1-8, where R ³ is selected from or a pharmaceutically acceptable salt thereof. 13. Connection according to any one of paragraphs. 1-8, where R ³ is selected from or a pharmaceutically acceptable salt thereof. 14. Connection according to any one of paragraphs. 1-8, where R ³ is selected from or a pharmaceutically acceptable salt thereof. 15. Connection according to any one of paragraphs. 1-14, where A represents or a pharmaceutically acceptable salt thereof 16. Connection according to any one of paragraphs. 1-14, where A represents or a pharmaceutically acceptable salt thereof. 17. Connection according to claim 1, where R ¹ represents methyl; A represents or and R ³ selected from or a pharmaceutically acceptable salt thereof. 18. The compound according to claim 1, where R ¹ represents methyl; R ⁴ and R ⁵ are CF ₃ ; A represents or and R ³ selected from or a pharmaceutically acceptable salt thereof. 19. The compound according to claim 1, where R ¹ represents methyl; R ⁴ represents CF ₃ ; A represents and R ³ selected from or a pharmaceutically acceptable salt thereof. 20. The compound according to claim 1, where R ¹ represents methyl; R ⁴ and R ⁵ are CF ₃ ; A represents or and R ³ selected from or a pharmaceutically acceptable salt thereof. 21. A compound according to claim 1, wherein said compound is 4-(3-methylmorpholin-4-yl)-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2- He; 6-[4-[(4-fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 6-[4-[(5-fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; N,N-dimethyl-4-[4-(3-methylmorpholin-4-yl)-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)piperazine-1-sulfonamide; 6-[4-(2-methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 6-[4-(4-fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; 6-[4-cyclopropylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-(1-piperidylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-morpholinosulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; 6-[4-(1,2-dimethylimidazol-4-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 6-[4-(1-methylcyclopropyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-methylsulfonyl-2-(trifluoromethyl)phenyl]-1H-pyridin-2-one and N,N-dimethyl-4-[4-(3-methylmorpholin-4-yl)-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)benzenesulfonamide, or a pharmaceutically acceptable salt thereof. 22. A compound according to claim 1, wherein said compound is 4-(3-methylmorpholin-4-yl)-6-[4-methylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2- He; 6-[4-[(4-fluorophenyl)methylsulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 6-[4-[(5-fluoro-3-pyridyl)sulfonyl]-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-tetrahydrofuran-3-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-pyrrolidin-1-ylsulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; N,N-dimethyl-4-[4-(3-methylmorpholin-4-yl)-6-oxo-1H-pyridin-2-yl]-3-(trifluoromethyl)piperazine-1-sulfonamide; 6-[4-(2-methoxyethylsulfonyl)-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 6-[4-(4-fluorophenyl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-4-(3-methylmorpholin-4-yl)-1H-pyridin-2-one; 4-(3-methylmorpholin-4-yl)-6-[4-(2-methylpyrazol-3-yl)sulfonyl-2-(trifluoromethyl)piperazin-1-yl]-1H-pyridin-2-one; or a pharmaceutically acceptable salt thereof. 23. Use of a compound according to any one of paragraphs. 1-22 or a pharmaceutically acceptable salt thereof for inhibiting vacuolar sorting protein 34 (Vps34) in the treatment or prevention of a disease. 24. Use of the compound according to any one of paragraphs. 1-22 or a pharmaceutically acceptable salt thereof for inhibiting Vps34 in the treatment of cancer. 25. Use according to claim 24, wherein said cancer is selected from breast cancer, such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, kidney cancer, colon cancer , glioma, prostate cancer, ovarian cancer, melanoma and lung cancer, as well as hypoxic tumors. 26. Use of a compound according to any one of paragraphs. 1-22 or a pharmaceutically acceptable salt thereof for the preparation of a medicament for inhibiting Vps34 in the treatment of cancer. 27. Use according to claim 26, wherein said cancer is selected from breast cancer, such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, kidney cancer, colon cancer , glioma, prostate cancer, ovarian cancer, melanoma and lung cancer, as well as hypoxic tumors. 28. A method of inhibiting Vps34 in the treatment of cancer, comprising administering a therapeutically effective amount of a compound according to any one of claims. 1-22 or a pharmaceutically acceptable salt thereof, to a patient in need thereof. 29. The method of claim 28, wherein said cancer is selected from breast cancer, such as triple negative breast cancer, bladder cancer, liver cancer, cervical cancer, pancreatic cancer, leukemia, lymphoma, kidney cancer, colon cancer , glioma, prostate cancer, ovarian cancer, melanoma and lung cancer, as well as hypoxic tumors. 30. Pharmaceutical composition for inhibiting Vps34, containing a therapeutically effective amount of a compound according to any one of paragraphs. 1-22 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent, carrier and/or excipient.