N-SUBSTITUTED FERROPORTIN INHIBITORS

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Description

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INTRODUCTION

The invention relates to novel compounds of the general formula (I-A)

and pharmaceutically acceptable salts thereof. The compounds of the general formula (I-A) of the present invention act as ferroportin inhibitors and are characterized by comprising an N-substituted cyclic group B. The novel compounds are particularly suitable for the use as medicaments in the prophylaxis and/or treatment of diseases caused by a lack of hepcidin or of iron metabolism disorders leading to increased iron levels or increased iron absorption. The compounds of the general formula (I-A) of the present invention are further particularly suitable for the use in the prophylaxis and/or treatment of iron overload, including thalassemia, sickle cell disease and hemochromatosis, as well as for the use in the prophylaxis and/or treatment of diseases related to or caused by increased iron levels, increased iron absorption or iron overload.

BACKGROUND AND PRIOR ART

Iron is an essential trace element for almost all organisms and is relevant in particular with respect to growth and the formation of blood. The balance of the iron metabolism is in this case primarily regulated on the level of iron recovery from haemoglobin of ageing erythrocytes and the duodenal absorption of dietary iron. The released iron is taken up via the intestine, in particular via specific transport systems (DMT-1, ferroportin), transferred into the blood circulation and thereby conveyed to the appropriate tissues and organs (transferrin, transferrin receptors).

Mammalian organisms are unable to actively discharge iron. The iron metabolism is substantially controlled by hepcidin, a peptide hormone produced in the liver, via the cellular release of iron from macrophages, hepatocytes and enterocytes. Hepcidin acts on the absorption of iron via the intestine and via the placenta and on the release of iron from the reticuloendothelial system. In the body, hepcidin is synthesized in the liver from what is known as pro-hepcidin, pro-hepcidin being coded by the gene known as the HAMP gene. The formation of hepcidin is regulated in direct correlation to the organisms iron level, i.e. if the organism is supplied with sufficient iron and oxygen, more hepcidin is formed, if iron and oxygen levels are low, or in case of increased erythropoiesis less hepcidin is formed. In the small intestinal mucosal cells and in the macrophages hepcidin binds with the transport protein ferroportin, which conventionally transports the phagocytotically recycled iron from the interior of the cell into the blood.

The transport protein ferroportin is a transmembrane protein consisting of 571 amino acids which is formed in the liver, spleen, kidneys, heart, intestine and placenta. In particular, ferroportin is localized in the basolateral membrane of intestinal epithelial cells. Ferroportin bound in this way thus acts to export the iron into the blood. In this case, it is most probable that ferroportin transports iron as Fe². If hepcidin binds to ferroportin, ferroportin is transported into the interior of the cell, where its breakdown takes place so that the release of the phagocytotically recycled iron from the cells is then almost completely blocked.

If the ferroportin is inactivated, for example by hepcidin, so that it is unable to export the iron which is stored in the mucosal cells, the stored iron is lost with the natural shedding of cells via the stools. The absorption of iron in the intestine is therefore reduced, when ferroportin is inactivated or inhibited, for example by hepcidin. In addition, ferroportin is markedly localized in the reticuloendothelial system (RES), to which the macrophages also belong. On the other hand, if the serum iron level decreases, hepcidin production in the hepatocytes of the liver is reduced so that less hepcidin is released and accordingly less ferroportin is inactivated, allowing a larger amount of stored iron to be transported into the serum.

Therefrom it becomes apparent that the hepcidin-ferroportin system directly regulates the iron metabolism and that a disorder of the hepcidin regulation mechanism therefore has a direct effect on iron metabolism in the organism. In principle the hepcidin-ferroportin regulation mechanism acts via the two following opposite principles:

On the one hand, an increase of hepcidin leads to inactivation of ferroportin, thus blocking the release of stored iron from the cells into the serum, thus decreasing the serum iron level. In pathological cases a decreased serum iron level leads to a reduced hemoglobin level, reduced erythrocyte production and thus to iron deficiency anemia.

On the other hand, a decrease of hepcidin results in an increase of active ferroportin, thus allowing an enhanced release of stored iron and an enhanced iron uptake e.g. from the food, thus increasing the serum iron level. In pathological cases an increased iron level leads to iron overload.

Iron overload states and diseases are characterized by excess iron levels. Therein, the problems arise from excess serum iron level which lead to non-transferrin bound iron (NTBI). The NTBI is rapidly taken up unspecifically by the organs, leading to an accumulation of iron in tissue and organs. Iron overload causes many diseases and undesired medical conditions, including cardiac, liver and endocrine damage. Further, iron accumulation in brain has been observed in patients suffering from neurodegenerative diseases such as for example Alzheimer's disease and Parkinson's disease. As a particular detrimental aspect of excess free iron the undesired formation of radicals must be mentioned. In particular iron(II) ions catalyze the formation (inter alia via Fenton reaction) of reactive oxygen species (ROS). These ROS cause damage to DNA, lipids, proteins and carbohydrates which has far-reaching effects in cells, tissue and organs and is well known and described in the literature to cause the so-called oxidative stress.

Besides the conventional methods for treating iron overload by removing iron from the body e.g. with chelating agents such as deferoxamine (also known as desferrioxamine B, N′-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutanoyl} amino)pentyl]-N-hydroxysuccinamide or Desferal®), deferasirox (Exjade®, 4-(3,5-bis(2-hydroxyphenyl)-1H-1,2,4-triazol-1-yl)benzoic acid) and deferiprone (Ferriprox®, 3-hydroxy-1,2-dimethylpyridin-4(1H)-one), compounds acting as hepcidin agonists or having an inhibiting or supporting effect on the biochemical regulatory pathways in the iron metabolism, such as hepcidin mimetic peptides have been described. Said therapeutic approaches are based on a direct involvement into the disturbed iron metabolism pathway by directly acting via the primary regulator hepcidin by providing a hepcidin mimetic or a hepcidin agonist, i.e. acting in the sense of a kind of hepcidin substitute or supply. The approach is based on the therapeutic rationale to treat iron overload, i.e. excess serum iron level, by inhibiting ferroportin, via the hepcidin-inactivation mechanism, thus blocking excessive iron absorption.

Ferroportin inhibitors and methods for preparing the same have been described in WO2017/068089, in WO2017/068090, in WO2021/191202, and in the unpublished international application PCT/EP2022/060546. Further, the international application WO2018/192973 describes the preparation and crystallization of various specific salts of selected ferroportin inhibitors described therein and as described in WO2017/068089 and in WO2017/068090.

WO2011/029832 relates to thiazol and oxazol compounds which act as hepcidin antagonists being described as suitable in the use for the treatment of iron deficiency diseases.

WO2021/013771 relates to the use of selected ferroportin inhibitors for treating transfusion dependant thalassemia.

WO2020/123850A1 describes further ferroportin inhibitors with a central heteroaryl bicyclic ring structure.

OBJECT OF THE INVENTION

The object of the present invention was to provide new therapeutically effective compounds that can be used for an effective therapy for the prophylaxis and treatment of iron metabolism disorders which are associated with increased iron levels, such as in particular iron overload. In a further object, the new compounds should exhibit high efficacy in the indication of the present invention, exhibit few side effects and have a low toxicity and good bioavailability and compatibility. Moreover, these new compounds, in contrast to the known iron chelating compounds, should be suitable to prevent the occurrence of increased iron levels and thus the related disorders, instead of removing excess iron from the body when the iron overload has already occurred. In a further object the new compounds should have a defined structure (stoichiometry) and should be preparable by simple synthesis processes, exhibit less sensitivity and improved long-lasting efficiency as compared to the known biomolecular compounds, such as antibodies.

This goal was achieved by the development of the novel compounds as defined herein, such as in particular according to formula (I-A) and (1-B), which have been found to act as ferroportin inhibitors. Therewith, the novel compounds are suitable for the use in the inhibition of iron transport, and thus are effective in the prophylaxis and treatment of iron metabolism disorders which are associated with increased iron levels, such as in particular iron overload, as well as in in the prophylaxis and treatment of diseases caused by a lack of hepcidin, diseases related to or caused by increased iron levels or iron overload and diseases associated with ineffective erythropoiesis.

DETAILED DESCRIPTION OF THE INVENTION

The inventors have found that specific compounds having the general structural formula (I-A) or (1-B) as defined herein, act as ferroportin inhibitors, thus effectively inhibiting iron transport and accordingly being particularly suitable for the use as medicaments, in particular for the use in the treatment and/or prophylaxis of diseases caused by a lack of hepcidin, diseases associated with ineffective erythropoiesis or iron metabolism disorders leading to increased iron levels, such as particularly iron overload states such as in particular thalassemia and hemochromatosis. Very particularly the new compounds turned out to be suitable for treating thalassemia and hemochromatosis. The new compounds are also suitable for the treatment of diseases caused by pathologically low hepcidin-levels and for the use in the inhibition of iron transport. In particular, the new compounds described herein show good metabolic stability and good bioavailability, which makes them particularly suitable as drug compounds.

Accordingly, the invention relates to novel compounds of general formula (I-A)

wherein

• • I is an integer of 1 or 2;
  • L¹ and L² each represent a linker group comprising 1 to 7 carbon atoms and which are independently selected from
    • a linear C₁-C₃-alkyl group —[CH₂]_m— or —[CH₂]_n—, respectively, wherein m and n are independently an integer of 1, 2 or 3,
    • a branched C₁-C₄-alkyl group, and
    • a C₃-C₆-cycloalkyl group, which may be a substituent to the linear C₁-C₃-alkyl group or which may form a ring together with the nitrogen atom to which it is bonded;
  • X¹ is N, S or O;
  • X² is N, S, O or CR⁵; and
  • X³ is C or N;
  • with the proviso that one of X¹ and X² is N
  • and if X³ is N, then X² is CR⁵;
  • and wherein
  • R⁵ represents
    • H,
    • halogen,
    • linear or branched C₁-C₃-alkyl, or
    • linear or branched C₁-C₃-haloalkyl;
  • A represents a group-(a-1)

• • wherein * indicates the binding position;
  • R¹ and R² independently represent 0, 1 or 2 substituents independently selected from
    • halogen,
    • linear or branched C₁-C₃-alkyl,
    • linear or branched C₁-C₃-haloalkyl, or
    • linear or branched C₁-C₃-alkoxy;
  • B represents one of the following groups (b-1), (b-2) and (b-3)

• • wherein * indicates the binding position;
  • R³ represents 0, 1, 2 or 3 substituents independently selected from
    • unsubstituted or substituted 6-membered aryl,
    • unsubstituted or substituted 5— or 6-membered heteroaryl,
    • unsubstituted or substituted bicyclic heteroaryl,
    • unsubstituted or substituted 3- to 6-membered cycloalkyl,
    • unsubstituted or substituted 5— or 6-membered heterocyclyl,
    • unsubstituted or substituted 5— or 6-membered heterocyclylalkyl,
    • unsubstituted or substituted 6-membered arylalkinyl, or
    • unsubstituted or substituted 5— or 6-membered heteroarylalkinyl,
  • wherein a substituted aryl, heteroaryl, bicyclic heteroaryl cycloalkyl, heterocyclyl, heterocyclylalkyl, arylalkinyl or heteroarylalkinyl group can carry 1, 2 or 3 substituents independently selected from
    • halogen,
    • C₁-C₃-alkyl,
    • C₁-C₃-haloalkyl, and
    • C₁-C₃-alkoxy;
  • R⁴ represents
    • unsubstituted or substituted linear or branched C₁-C₆-alkyl,
    • a dialkylether group [R⁶(CH₂)_x—O—CH₂)_y-] with R⁶ representing a substituent selected from a C₁-C₃-alkoxy group and with x and y independently representing an integer of 1, 2 or 3,
    • unsubstituted or substituted 3- to 6-membered cycloalkyl,
    • unsubstituted or substituted 5— or 6-membered heterocyclyl, or
    • unsubstituted or substituted 6-membered aryl
  • wherein alkyl, cycloalkyl, heterocyclyl and aryl can be substituted with 1 or 2 substituents, independently selected from
    • halogen,
    • C₁-C₃-alkoxy,
    • C₆-cycloalkyloxy,
    • carboxyl,
    • aminocarbonyl,
    • mono- or di-alkylaminocarbonyl,
    • an amino group comprising —NH₂, mono- and di-alkylamino,
    • unsubstituted or substituted 3- to 6-membered cycloalkyl,
    • unsubstituted or substituted 5— or 6-membered heterocyclyl,
    • unsubstituted or substituted 6-membered aryl,
    • unsubstituted or substituted 5— or 6-membered heteroaryl, and
    • unsubstituted or substituted bicyclic heteroaryl,
  • wherein a substituted cycloalkyl, heterocyclyl, aryl, heteroaryl and bicyclic heteroaryl group can carry 1, 2 or 3 substituents independently selected from
    • hydroxy,
    • cyano,
    • halogen,
    • C₁-C₃-alkyl,
    • C₁-C₃-haloalkyl,
    • C₁-C₃-alkoxy,
    • carboxyl,
    • an amino (—NH₂) or mono- or di-alkylaminogroup,
    • aminocarbonyl, and
    • mono- or di-alkylaminocarbonyl,
    • wherein a monoalkylamino group and a monoalkylaminocarbonyl group may carry a further substituent on the mono-alkyl-chain, selected from
      • C₁-C₃-alkoxy,
      • unsubstituted or substituted 6-membered aryl, and
      • unsubstituted or substituted 5— or 6-membered heteroaryl,
      • wherein a substituted aryl or heteroaryl group as a substituent of the mono-alkyl-chain can carry 1, 2 or 3 substituents independently selected from halogen, C₁-C₃-alkyl and C₁-C₃-haloalkyl;
  • and in formulae (b-2) and (b-3) one of D1, D2 and D3 is present and respresents
    • a fused 6-membered aryl ring,
    • a fused 5— or 6-membered heteroaryl ring,
    • a fused 5— or 6-membered cycloalkyl ring, or
    • a fused 5— or 6-membered heterocyclyl ring;
  • and the groups (b-2) and (b-3) carry 0, 1, 2 or 3 substituents, which are independently selected from
    • halogen,
    • linear or branched C₁-C₃-alkyl,
    • linear or branched C₁-C₃-haloalkyl,
    • linear or branched C₁-C₃-alkoxy;
  • and pharmaceutically acceptable salts thereof.
  • From the scope of the invention the following compounds (X-1), (X-2) and (X-3) are excluded:

The invention also relates to novel compounds of general formula (I-B)

• • wherein the substituents have the meaning defined above.
  • In formula (I-B), the substituents may also have the meaning as follows:
  • I is an integer of 1 or 2:
  • m and n are independently an integer of 1, 2 or 3;
  • X¹ is N, S or O;
  • X² is N, S, O or CR⁵; and
  • X³ is C or N;
  • with the proviso that one of X¹ and X² is N
  • and if X³ is N, then X² is CR⁵;
  • and wherein
  • R⁵ represents
    • H,
    • halogen,
    • linear or branched C₁-C₃-alkyl, or
    • linear or branched C₁-C₃-haloalkyl;
  • A represents a group

• • wherein * indicates the binding position;
  • R¹ and R2 independently represent 0, 1 or 2 substituents independently selected from
    • halogen,
    • linear or branched C₁-C₃-alkyl,
    • linear or branched C₁-C₃-haloalkyl, or
    • linear or branched C₁-C₃-alkoxy;
  • B represents one of the following groups (b-1), (b-2) and (b-3)

• • wherein * indicates the binding position;
  • R³ represents 0, 1, 2 or 3 substituents independently selected from
    • unsubstituted or substituted 6-membered aryl,
    • unsubstituted or substituted 5— or 6-membered heteroaryl,
    • unsubstituted or substituted bicyclic heteroaryl,
    • 3- to 6-membered cycloalkyl,
    • 5— or 6-membered heterocyclyl,
    • 5— or 6-membered heterocyclylalkyl, or
    • 6-membered arylalkinyl
  • wherein a substituted aryl, heteroaryl and bicyclic heteroaryl group can carry 1, 2 or 3 substituents independently selected from
    • halogen,
    • C₁-C₃-alkyl,
    • C₁-C₃-haloalkyl, and
    • C₁-C₃-alkoxy;
  • R⁴ represents
    • linear or branched C₁-C₆-alkyl,
    • a dialkylether group [R⁶(CH₂)_x—O—CH₂)_y-] with R⁶ representing a substituent selected from a C₁-C₃-alkoxy group and with x and y independently representing an integer of 1, 2 or 3,
    • 3- to 6-membered cycloalkyl, or
    • 5— or 6-membered heterocyclyl,
  • wherein alkyl, cycloalkyl and heterocyclyl can be substituted with 1 or 2 substituents, independently selected from
    • C₁-C₃-alkoxy,
    • carboxyl,
    • aminocarbonyl,
    • mono- or di-alkylaminocarbonyl,
    • 3- to 6-membered cycloalkyl, and
    • 5— or 6-membered heterocyclyl,
    • unsubstituted or substituted 6-membered aryl,
    • unsubstituted or substituted 5— or 6-membered heteroaryl, and
    • unsubstituted or substituted bicyclic heteroaryl,
    • wherein a substituted aryl, heteroaryl and bicyclic heteroaryl group can carry 1, 2 or 3 substituents independently selected from
      • hydroxy,
      • cyano,
      • halogen,
      • C₁-C₃-alkyl,
      • C₁-C₃-haloalkyl,
      • C₁-C₃-alkoxy,
      • carboxyl,
      • an amino (—NH₂) or mono- or di-alkylaminogroup,
      • aminocarbonyl, and
      • mono- or di-alkylaminocarbonyl,
      • wherein a monoalkylamino group and a monoalkylaminocarbonyl group may carry a further substituent on the mono-alkyl-chain, selected from
        • C₁-C₃-alkoxy,
        • unsubstituted or substituted 6-membered aryl, and
        • unsubstituted or substituted 5— or 6-membered heteroaryl,
        • wherein a substituted aryl or heteroaryl group as a substituent of the mono-alkyl-chain can carry 1, 2 or 3 substituents independently selected from halogen, C₁-C₃-alkyl and C₁-C₃-haloalkyl;
  • and in formulae (b-2) and (b-3) one of D1, D2 and D3 is present and respresents
    • a fused 6-membered aryl ring,
    • a fused 5— or 6-membered heteroaryl ring,
    • a fused 5— or 6-membered cycloalkyl ring, or
    • a fused 5— or 6-membered heterocyclyl ring;
  • and the groups (b-2) and (b-3) carry 0, 1, 2 or 3 substituents, which are independently selected from
    • halogen,
    • linear or branched C₁-C₃-alkyl,
    • linear or branched C₁-C₃-haloalkyl,
    • linear or branched C₁-C₃-alkoxy;
  • and pharmaceutically acceptable salts thereof.

Definitions

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.

The term “optionally substituted”, “optional substituent(s)” or “possible substituent(s)” 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, 4 or 5, in particular 1, 2 or 3.

If used herein, the term “one or more”, e.g. in the definition of the substituents of the compounds of general formula (I-A) and (I-B) 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”.

The term “comprising” or “containing” when used in the claims or specification includes “consisting of”.

If within the present specification any item is referred to as “as mentioned herein” or “as defined (anywhere) herein”, it means that it may be mentioned anywhere in the present specification or may have the meaning as defined anywhere in the present specification.

The terms used in the claims and specification have the following meanings: “Halogen” or “halogen atom” means a fluorine, chlorine, bromine or iodine atom, particularly a fluorine, chlorine or bromine atom, a preferred selection relates to chlorine or fluorine, a further preferred selection relates to bromine or fluorine, most preferred is fluorine.

The term “C₁-C₆-alkyl” means a linear or branched, saturated, monovalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms, e.g. a methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, pentyl or hexyl group. Methyl, ethyl, n-propyl, iso-propyl, n-butyl and iso-butyl groups are preferred. More preferred are methyl, ethyl, n-propyl and iso-propyl.

The C₁-C₆-alkyl group may optionally be substituted with 1 or 2 substituents, preferably with 1 substituent. In such a case the substituted alkyl group is preferably a substituted C₁-C₃-alkyl group, more preferably a substituted methyl or ethyl group. Such optional substituents are preferably selected from the group consisting of: halogen (forming a halogen-substituted C₁-C₃-alkyl group as defined below and herein also indicated as “C₁-C₃-haloalkyl”), such as preferably difluoroalkyl or trifluoroalkyl, C₁-C₃-alkoxy such as preferably methoxy, a cycloalkyloxy group, such as preferably a C₆-cycloalkyloxy group (cyclohexyloxy), a carboxyl group [—(C═O)OH], an aminocarbonyl group [NH₂(C═O)—], a mono-alkylaminocarbonyl group such as preferably a methylaminocarbonyl group [CH₃NH(C═O)—], an amino group comprising —NH₂, mono- and di-alkylamino, such as preferably mono- or di-methylamino, 3- to 6-membered cycloalkyl (also designated as C₃-C₆-cycloalkyl) containing 3, 4, 5 or 6 carbon atoms, such as preferably cyclopropyl and cyclohexyl, unsubstituted or substituted 5— or 6-membered heterocyclyl, unsubstituted or substituted 6-membered aryl (phenyl), unsubstituted or substituted 5— or 6-membered heteroaryl, and unsubstituted or substituted bicyclic heteroaryl such as preferably a benzimidazolyl group.

A substituted heterocyclyl, aryl, heteroaryl and bicyclic heteroaryl group as a substituent of alkyl may also carry 1, 2 or 3 substituents independently selected from hydroxy, cyano, halogen, C₁-C₃-alkyl as defined herein such as preferably methyl, C₁-C₃-haloalkyl as defined herein such as preferably difluoroethyl or trifluoromethyl (CF₃), C₁-C₃-alkoxy as defined herein such as preferably methoxy, a carboxyl group, an amino (—NH₂) or mono- or di-alkylamino group, an aminocarbonyl group as defined herein, and a mono—or di-alkylaminocarbonyl group, wherein a mono-alkylaminocarbonyl group may carry a further substituent on the mono-alkyl-chain, selected from C₁-C₃-alkoxy, unsubstituted or substituted 6-membered aryl, and unsubstituted or substituted 5— or 6-membered heteroaryl, wherein a substituted aryl or heteroaryl group as a substituent of the mono-alkyl-chain can carry 1, 2 or 3 substituents independently selected from halogen, C₁-C₃-alkyl and C₁-C₃-haloalkyl, preferably a further substituent on the mono-alkyl-chain of a mono-alkylaminocarbonyl group is selected from a halogen-substituted 5— or 6-membered heteroaryl (more preferably a fluoro-pyridinyl group).

The term “dialkylether” or “dialkylether group” as used herein means a C₃-C₇-alkyl group as defined above, wherein one CH₂-group in the alkyl-chain is replaced by —O—, resulting in a group [—(CH₂)_x—O—CH₂)_y—] with x and y independently representing an integer of 1, 2 or 3. Such a dialkylether group as a substituent R⁴ carries a further substituent R⁶, resulting in a group [R⁶(CH₂)_x—O—CH₂)_y—]. Therein, R⁶ represents a substituent selected from the group of C₁-C₃-alkoxy. If R⁶ represents a hydrogen atom the dialkylether group is unsubstituted and corresponds to a group “alkoxy” as defined herein separately.

Preferred substituents R⁶ are selected from a C₁-C₃-alkoxy group, such as in particular methoxy and ethoxy.

The term “C₁-C₃-haloalkyl” means a linear or branched, saturated, monovalent C₁-C₃-alkyl group, having the meaning as defined above, in which one or more of the hydrogen atoms are replaced, identically or differently, with a halogen atom. Particularly, said halogen atom is a chlorine or fluorine atom. More particularly, said halogen atom is a fluorine atom and even more particularly, all said halogen atoms are fluorine atoms (“C₁-C₃-fluoroalkyl”). Said C₁-C₃-haloalkyl group is, for example, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, 3,3,3-trifluoropropyl or 1,3-difluoropropan-2-yl, wherein a trifluoromethyl-group (CF₃) is particularly preferred.

The term “C₁-C₃-alkoxy” means a linear or branched, saturated, monovalent group of formula (C₁-C₃-alkyl)—O—, in which the term “C₁-C₃-alkyl” is as defined supra, e.g. a methoxy, ethoxy, n-propoxy or isopropoxy group, with a methoxy-group and an iso-propoxy group being particularly preferred.

The term cycloalkyloxy relates to a cycloalkyl-O- group, with a cycloalkyl group as defined below being bound via an oxygen (—O—). Cycloalkyloxy includes “C₃-C₆-cycloalkyloxy”, with C₆-cycloalkyloxy (cyclohexyloxy) being preferred.

A “carboxyl group” indicates a group [—(C═O)OH].

The term “mono- or di-alkylamino” indicates an amino group (—NH₂), wherein one or both hydrogens are replaced by the same of different C₁-C₃-alkyl groups. Preferred are mono- and dimethylamino groups, more preferred are dimethylamino groups.

The term “aminocarbonyl group” indicates a group [NH₂—(C═O)—].

The term “(mono-)alkylaminocarbonyl group” indicates an aminocarbonyl group [NH₂—(C═O)—], wherein one hydrogen is replaced by a C₁-C₃-alkyl group. A preferred mono-alkylaminocarbonyl group is a methylaminocarbonyl group [CH₃NH(C═O)—].

Generally, the term “aryl” includes aromatic hydrocarbon residues containing 6 to 14 carbon atoms (excluding the carbon atoms of the possible substituents), which may be monocyclic or bicyclic, including, for example: phenyl, naphthyl, phenanthrenyl and anthracenyl. Preferred is 6-membered aryl, such as phenyl.

Generally, the term “heteroaryl” includes heteroaromatic hydrocarbon residues containing 4 to 9 ring carbon atoms, which additionally contain 1 to 3 of the same or different heteroatoms selected from S, O and N in the ring, and therefore form 5- to 12-membered heteroaromatic residues which may be monocyclic or bicyclic.

Monocyclic heteroaryl groups preferably include 5- and 6-membered monocyclic heteroaryl groups, such as pyridyl (pyridinyl), pyridyl-N-oxide, pyridazinyl, pyrimidyl, pyrazinyl, thienyl (thiophenyl), furyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, oxazolyl or isoxazolyl, including from the group of 5-membered heteroaryl, for example thiazolyl such as thiazol-2-yl, 2-thiazol-2-yl, 2-thiazol-4-yl, thienyl (thiophenyl), such as thien-3-yl, pyrazolyl such as 1-pyrazol-4-yl, 3-pyrazol-5-yl, imidazolyl such as imidazole-2-yl, 2-imidazol-4-yl, 1-imidazol-4-yl, triazolyl such as 1-triazol-3-yl, 1-triazol-4-yl, such as 1,2,4-triazol-3-yl or 1,2,3-triazol-4-yl, oxazolyl such as 2-oxazol-4-yl, 2-oxazol-5-yl, iso-oxazolyl such as iso-oxazol-4-yl, oxadiazolyl such as 1,2,4-oxadiazol-3-yl, tetrazolyl and from the group of 6-membered heteroaryl, for example, pyridyl (pyridinyl) such as pyrid-1-yl, pyrid-2-yl, pyrid-3-yl, pyrid-4-yl, 2-pyrid-4-yl, 2-pyrid-6-yl, 3-pyrid-5-yl (pyridin-1-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, 2-pyridin-4-yl, 2-pyridin-6-yl, 3-pyridin-5-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl. Preferred heteroaryl groups are pyridinyl, pyrimidinyl, imidazolyl, oxazolyl, iso-oxazolyl and tetrazolyl.

Bicyclic heteroaryl groups preferably include indolizinyl, indolyl, benzo[b]thienyl, benzo[b]furyl, indazolyl, quinolyl, isoquinolyl, naphthyridinyl, quinazolinyl, quinoxalinyl, and benzimidazolyl such as benzimidazol-2-yl, benzimidazol-4-yl, benzimidazol-5-yl. A benzimidazolyl group is particularly preferred.

Generally, the term “cycloalkyl” includes aliphatic rings containing 3 to 8, more preferably 3 to 6 ring carbon atoms. Cycloalkyl includes a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group and a cyclooctyl group, with a cyclopropyl group and a cyclohexyl group being preferred.

Generally the term “heterocyclyl” includes saturated or unsaturated mono- or bicyclic 4- to 8-membered heterocyclic residues containing 1 to 3, preferably 1 to 2 same or different hetero atoms selected from N, O and S., including azetidinyl, oxetanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrofuranyl, dioxolanyl, tetrahydrothiophenyl, oxathiolanyl, piperidinyl, piperazinyl, tetrahydropyranyl, thianyl, dithianyl, trithianyl, tetrahydrothiopyranyl, morpholinyl, thiomorpholynyl, dioxanyl, etc., such as azetidin-1-yl, azetidin-2-yl, azetidin-3-yl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydro-thiophen-2-yl, tetrahydro-thiophen-3-yl, pyrrolidin-1-yl, pyrrolidin-2-yl, pyrrolidin-3-yl, morpholin-1-yl, morpholin-2-yl, morpholin-3-yl, piperidin-1-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl, piperazin-1-yl, piperazin-2-yl, tetrahydropyran-2-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl, etc.

Particularly preferred are 5— or 6-membered heterocyclyl groups, such as pyrrolidinyl, dioxolanyl, dioxanyl, piperidinyl, piperazinyl and morpholinyl residues.

The aryl, heteroaryl, bicyclic heteroaryl, cycloalkyl or heterocyclyl groups can be bound via a direct bond, a C₁-C₃-alkyl-chain, preferably a C₁-alkyl-chain or an alkinyl-chain, such as preferably an ethinyl-chain (—C—C—), or the aryl, heteroaryl, cycloalkyl or heterocyclyl groups can be condensed with aromatic rings forming fused ring systems as defined herein.

The aryl, heteroaryl, bicyclic heteroaryl, cycloalkyl or heterocyclyl group, including fused aryl, heteroaryl, bicyclic heteroaryl, cycloalkyl and heterocyclyl groups, may carry 1, 2 or 3 of the same or different substituents, independently selected from halogen as defined above such as preferably F, Br and Cl, C₁-C₃-alkyl such as preferably methyl, C₁-C₃-haloalkyl as defined above such as preferably trifluoromethyl, C₁-C₃-alkoxy as defined above such as preferably methoxy and C₆-cycloalkyloxy.

The aryl, heteroaryl, bicyclic heteroaryl, cycloalkyl or heterocyclyl group may also carry 1, 2 or 3 substituents as defined above in context with the possible substituents of alkyl.

The aryl, heteroaryl, bicyclic heteroaryl, cycloalkyl or heterocyclyl groups as defined herein may form one of the groups A and/or B as defined herein.

Therein, the group A represents a group

The group (a-1) is a pyridinyl-group, which carries 0 substituents (R¹/R² represent hydrogen) or 1 or 2 same or different substituents R¹/R², independently selected from halogen as defined above such as preferably F, Br and Cl, C₁-C₃-alkyl such as preferably methyl, C₁-C₃-haloalkyl as defined above such as preferably trifluoromethyl, and C₁-C₃-alkoxy as defined above such as preferably methoxy.

The group B represents one of the following groups (b-1), (b-2) and (b-3)

Therein, a group (b-1) represents a N-substituted benzimidazolyl group, which carries 0 substituents R³ (i.e. R³ represents hydrogen) or 1 or 2 same or different substituents R³, preferably 1 substituent R³ selected from an aryl, heteroaryl, bicyclic heteroaryl, cycloalkyl and heterocyclyl group as defined above, which is bound via a direct bond or which is bound via a C₁-C₃-alkyl-chain, preferably a C₁-alkyl-chain, or an alkinyl-chain, such as preferably an ethinyl-chain.

Preferably, R³ is selected from unsubstituted or substituted phenyl, unsubstituted or substituted 5— or 6-membered heteroaryl, unsubstituted or substituted bicyclic heteroaryl, unsubstituted or substituted 3- to 6-membered cycloalkyl, unsubstituted or substituted 5— or 6-membered heterocyclyl, unsubstituted or substituted 5— or 6-membered heterocyclylalkyl, unsubstituted or substituted 6-membered arylalkinyl or unsubstituted or substituted 5— or 6-membered heteroarylalkinyl, wherein a substituted aryl, heteroaryl and bicyclic heteroaryl group can carry 1, 2 or 3 substituents independently selected from halogen, C₁-C₃-alkyl, C₁-C₃-haloalkyl, and C₁-C₃-alkoxy.

An aryl, heteroaryl, bicyclic heteroaryl, cycloalkyl or heterocyclyl ring bound via a C₁-C₃-alkyl-chain, preferably a C₁-alkyl-chain, corresponds to R³ representing an arylalkyl, a heteroarylalkyl, a cycloalkylalkyl or a heterocyclylalkyl group, wherein “alkyl” preferably represents C₁-C₃-alkyl. A heterocyclylalkyl group, such as a piperazinylmethyl group or a morpholinylmethyl group is preferred.

An aryl, heteroaryl, bicyclic heteroaryl, cycloalkyl or heterocyclyl ring bound via an alkinyl-chain, such as preferably an ethinyl-chain, corresponds to R³ representing an arylalkinyl, a heteroarylalkinyl, a cycloalkylalkinyl or a heterocyclylalkinyl group, wherein “alkinyl” preferably represents ethinyl. An arylalkinyl group, such as a phenylethinyl group, and a heteroarylalkinyl group, such as a pyridinylethinyl group, are preferred. Therein the aryl, heteroaryl, bicyclic heteroaryl, cycloalkyl or heterocyclyl ring may carry 1, 2 or 3 of the same or different substituents selected from those defined herein for the respective groups, preferably selected from halogen as defined above such as preferably F, Br and Cl, C₁-C₃-alkyl such as preferably methyl, C₁-C₃-haloalkyl as defined above such as preferably trifluoromethyl, and C₁-C₃-alkoxy as defined above such as preferably methoxy. Therewith, such groups are also designated herein as “unsubstituted or substituted arylalkinyl”, “unsubstituted or substituted heteroarylalkinyl”, “unsubstituted or substituted cycloalkylalkinyl” and “unsubstituted or substituted heterocyclylalkinyl”.

It is also possible that an unsubstituted or substituted aryl ring is bound directly, corresponding to R³ or R⁴ representing “unsubstituted or substituted aryl”. Regarding possible substituents reference is made to the definition above. In particular for an aryl group directly bound to the group B, e.g. as the substituent R⁴, preferred substituents are selected from halogen, more preferably Cl, and C₁-C₃-alkoxy, more preferably methoxy.

The groups (b-2) and (b-3) represent fused (condensed) ring systems, wherein in formulae (b-2) and (b-3) a fused aryl, heteroaryl, cycloalkyl or heterocyclyl ring as defined above is present in one of the positions indicated by D1, D2 and D3, preferably forming a fused tricyclic ring system.

The groups (b-2) and (b-3) may optionally carry 1, 2 or 3 same or different substituents, which are independently selected from halogen, linear or branched C₁-C₃-alkyl, linear or branched C₁-C₃-haloalkyl and linear or branched C₁-C₃-alkoxy, each as defined above. Such optional substituent is hereinafter also represented by Rx.

From the group (a-1) the following groups are particularly preferred:

Therefrom the group (a-1) with the following structures are more preferred:

Therefrom the group (a-1) with the following structure is most preferred:

The compounds of the formula (I-A) and (I-B) of the present invention are characterized by comprising a substituent R⁴ in the group B, forming a “N-substituted” cyclic group B. In the embodiments of the invention, wherein R⁴ is not directly bound substituted or unsubstituted aryl, the “N-substituted” cyclic group B can be designated as “N-alkylated” (cyclic) group B. Therein, the term “N-alkylated” is understood to include a substitution with an alkyl group, a dialkylether group as well as cycloalkyl or a heterocyclyl-group as defined herein.

In such cases, particularly, the R⁴ substituent represents a linear or branched C₁-C₆-alkyl group, a dialkylether group [R⁶(CH₂)_x—O—CH₂)_y—] as defined above, a 3- to 6-membered cycloalkyl, or 5— or 6-membered heterocycly, which can be substituted or unsubstituted.

In the case of R⁴ representing a substituted alkyl group, reference is made to possible alkyl-substituents as defined above.

In the case of R⁴ representing a substituted dialkylether group, reference is made to the definition of “dialkylether” and its possible substituents R⁶ as defined above.

In the case of R⁴ representing a substituted cycloalkyl or heterocyclyl group, reference is made to the definition of possible substituents of such groups anywhere herein.

In the case of R⁴ representing a substituted aryl group, reference is made to possible aryl-(phenyl-) substituents as defined above.

Preferably, the R⁴ substituent aryl, alkyl, dialkylether, cycloalkyl and heterocyclyl can be substituted with 1 or 2 substituents, in particular such substituents as defined above for phenyl, C₁-C₃-alkyl and dialkylether.

It is preferred that the compounds (I-A) and (I-B) comprise a group (b-1) as defined herein.

If a group (b-2) is present, the following groups (b-2) are particularly preferred:

wherein R⁴ has the meaning as defined anywhere herein, such as e.g. a group (b-2):

If used herein, e.g. in the formulae (a-1), (b-1), (b-2) and (b-3), *″ indicates the binding position.

In the formula (I-A) and (I-B) “I” represents an integer of 1 or 2, preferably I=1.

In the formula (I-B) “m” and “n” independently represent an integer of 1, 2 or 3, preferably m=2 and preferably n=1 or 2.

a C₃-C₆-cycloalkyl group, which may be a substituent to the linear C₁-C₃-alkyl group or which may form a ring together with the nitrogen atom to which it is bonded

In the formula (I-A) “L¹” and “L²” each represent a linker group or a so-called spacer, i.e. an “alkyl-spacer”, which comprises 1 to 7 carbon atoms. Such an alkyl-spacer group or linker “L¹” and “L²” is independently selected from

• • a linear C₁-C₃-alkyl group —[CH₂]_m— or —[CH₂]_n—, respectively, wherein m and n are independently an integer of 1, 2 or 3; or
  • a branched C₁-C₄-alkyl group, such as preferably a 2-dimethylethyl group

or

• • a C₃-C₆-cycloalkyl group, which may be a substituent to the linear C₁-C₃-alkyl group, such as

or

• • a C₃-C₆-cycloalkyl group, which forms a ring together with the nitrogen atom to which it is bonded, such as for “L¹”

• • Preferably, the linker “L¹” has the meaning of a linear C₁-C₃-alkyl group —[CH₂]_m as defined herein.
  • Preferably, the linker “L²” has the meaning of a linker group comprising 1 to 7 carbon atoms selected from
  • a linear C₁-C₃-alkyl group —[CH₂]_n—, wherein n represents an integer of 1, 2 or 3,
  • a branched C₁-C₄-alkyl group, and
  • a C₃-C₆-cycloalkyl group, which may be a substituent to the linear C₁-C₃-alkyl group or which may form a ring together with the nitrogen atom to which it is bonded, as defined herein.
  • In the formula (I-A) and (I-B) X¹, X² and X³ are selected from:
  • X¹=N, S or 0;
  • X²=N, S, O or CR⁵;
  • X³=C or N;
  • with the proviso that one of X¹ and X² is N and if X³ is N, then X² is CR⁵, forming one of the following groups:

• • wherein * indicates the binding site to the aminocarbonyl-group and ** indicates the binding site to the —[(CH₂)]_m-amino—[(CH₂)]_n- group in the formula (I-A) and (1-B).

Therein, R⁵ represents an optional substituent in case of X²=CR⁵, and R⁵ is preferably selected from halogen, linear or branched C₁-C₃-alkyl and linear or branched C₁-C₃-haloalkyl, each as defined above. If no substituent is present, then R⁵ represents hydrogen.

Preferred are an oxazole-, an isooxazole-, a thiazole- and an isothiazole-group:

with an oxazole- and isooxazole-group being more preferred. Most preferred is a group

• • A further aspect relates to compounds of the formula (I-A) and (I-B) as defined above, wherein
  • I is an integer of 1 or 2;
  • m and n are independently an integer of 1, 2 or 3;
  • X¹ is N, S or O;
  • X² is N, S, O or CR⁵; and
  • X³ is C or N;
  • with the proviso that one of X¹ and X² is N
  • and if X³ is N, then X² is CR⁵;
  • and wherein
  • R⁵ represents H;
  • A represents a group (a-1)

• • wherein * indicates the binding position;
  • R¹ and R² independently represent 0, 1 or 2 substituents independently selected from
    • halogen,
    • linear or branched C₁-C₃-alkyl,
    • linear or branched C₁-C₃-haloalkyl, or
    • linear or branched C₁-C₃-alkoxy;
  • B represents one of the following groups (b-1), (b-2) and (b-3)

• • wherein * indicates the binding position;
  • R³ represents 0, 1, 2 or 3 substituents independently selected from
    • unsubstituted or substituted phenyl,
    • unsubstituted or substituted 5— or 6-membered heteroaryl,
    • unsubstituted or substituted bicyclic heteroaryl,
    • 6-membered heterocyclyl,
    • 6-membered heterocyclylalkyl,
    • phenylethinyl, or
    • pyridinylethinyl,
  • wherein a substituted pheny, heteroaryl and bicyclic heteroaryl group can carry 1, 2 or 3 substituents independently selected from
    • halogen,
    • C₁-C₃-alkyl,
    • C₁-C₃-haloalkyl, and
    • C₁-C₃-alkoxy;
  • R⁴ represents
    • linear or branched C₁-C₆-alkyl,
    • a dialkylether group [R⁶(CH₂)_x—O—CH₂)_y—]
    • with R⁶ representing a C₁-C₃-alkoxy group and
    • with x and y independently representing an integer of 1, 2 or 3, or
    • 5— or 6-membered unsubstituted heterocyclyl, or
    • substituted or unsubstituted phenyl,
  • wherein substituents of phenyl are selected from
    • halogen, and
    • C₁-C₃-alkoxy; and
  • wherein alkyl can be substituted with 1 or 2 substituents, independently selected from
    • halogen
    • C₁-C₃-alkoxy,
    • C₆-cycloalkyloxy,
    • carboxyl,
    • aminocarbonyl,
    • mono-alkylaminocarbonyl,
    • dialkylamino,
    • 3- to 6-membered cycloalkyl,
    • 5— or 6-membered heterocyclyl,
    • unsubstituted or substituted 6-membered aryl,
    • unsubstituted or substituted 5— or 6-membered heteroaryl, and
    • unsubstituted or substituted bicyclic heteroaryl,
    • wherein a substituted aryl, heteroaryl and bicyclic heteroaryl group can carry 1, 2 or 3 substituents independently selected from
      • halogen
      • C₁-C₃-alkyl,
      • C₁-C₃-haloalkyl,
      • C₁-C₃-alkoxy,
      • aminocarbonyl, and
      • mono-alkylaminocarbonyl,
      • wherein a mono-alkylaminocarbonyl group may carry a further substituent on the mono-alkyl-chain, selected from halogen-substituted 5— or 6-membered heteroaryl;
  • and in formulae (b-2) and (b-3) one of D1, D2 and D3 is present and represents
    • a fused phenyl ring,
    • a fused 6-membered heteroaryl ring,
    • a fused 6-membered cycloalkyl ring, or
    • a fused 5— or 6-membered heterocyclyl ring;
  • and the groups (b-2) and (b-3) carry 0 or 1 substituent selected from
    • halogen,
    • linear or branched C₁-C₃-alkyl,
    • linear or branched C₁-C₃-haloalkyl, and
    • linear or branched C₁-C₃-alkoxy;
    • and pharmaceutically acceptable salts thereof.
    • In a further aspect of the invention it is preferred that the compounds of the formula (I-A) and (I-B) as defined anywhere herein are characterized in that:
    • R³ represents H, C₁-C₃-alkoxy, pyridinylethinyl, or unsubstituted or substituted phenyl, wherein a substituted phenyl group can carry 1, 2 or 3 substituents independently selected from halogen, C₁-C₃-alkyl, C₁-C₃-haloalkyl, and C₁-C₃-alkoxy, more preferably R³ represents H or unsubstituted or substituted phenyl, wherein a substituted phenyl group can carry 1, 2 or 3 substituents independently selected from halogen, C₁-C₃-alkyl, C₁-C₃-haloalkyl, and C₁-C₃-alkoxy;
    • and/or
    • R⁴ represents linear or branched C₁-C₆-alkyl, a dialkylether group [R⁶(CH₂)_x—O—CH₂)_y—] with R⁶ representing a C₁-C₃-alkoxy group and with x and y independently representing an integer of 1, 2 or 3, or 5— or 6-membered unsubstituted heterocyclyl, or substituted or unsubstituted phenyl, wherein substituents of phenyl are selected from halogen and C₁-C₃-alkoxy.
  • Therein, alkyl can be substituted with 1 or 2 substituents, independently selected from
  • halogen,
  • C₁-C₃-alkoxy,
  • cyclohexyloxy,
  • carboxyl,
  • aminocarbonyl,
  • mono-alkylaminocarbonyl,
  • dialkylamino,
  • cyclopropyl or cyclohexyl,
  • 6-membered heterocyclyl,
  • unsubstituted or substituted phenyl,
  • unsubstituted or substituted 5— or 6-membered heteroaryl, and
  • unsubstituted or substituted bicyclic heteroaryl, wherein a substituted phenyl, heteroaryl and bicyclic heteroaryl group can carry 1, 2 or 3 substituents independently selected from
    • halogen,
    • C₁-C₃-alkyl,
    • C₁-C₃-haloalkyl,
    • C₁-C₃-alkoxy,
    • aminocarbonyl, and
    • mono-alkylaminocarbonyl, wherein a mono-alkylaminocarbonyl group may carry a further substituent on the mono-alkyl-chain, selected from halogen-substituted 5— or 6-membered heteroaryl.

More preferably R⁴ represents linear or branched C₁-C₆-alkyl, a dialkylether group [R⁶(CH₂)_x—O—CH₂)_y-] with R⁶ representing a C₁-C₃-alkoxy group and with x and y independently representing an integer of 1, 2 or 3, or 5— or 6-membered unsubstituted heterocyclyl. Therein, alkyl can be substituted with 1 or 2 substituents, independently selected from

• • C₁-C₃-alkoxy,
  • carboxyl,
  • aminocarbonyl,
  • mono-alkylaminocarbonyl,
  • cyclopropyl,
  • 6-membered heterocyclyl,
  • unsubstituted or substituted phenyl,
  • unsubstituted or substituted 5— or 6-membered heteroaryl, and
  • unsubstituted or substituted bicyclic heteroaryl,
  • wherein a substituted phenyl, heteroaryl and bicyclic heteroaryl group can carry 1, 2 or 3 substituents independently selected from
    • C₁-C₃-alkyl,
    • C₁-C₃-haloalkyl,
    • C₁-C₃-alkoxy,
    • aminocarbonyl, and
  • mono-alkylaminocarbonyl, wherein a mono-alkylaminocarbonyl group may carry a further substituent on the mono-alkyl-chain, selected from halogen-substituted 5— or 6-membered heteroaryl.

In a further aspect of the invention it is preferred that the compounds of the formula (I-A) and (I-B) as defined anywhere herein are characterized in that one or more of the substituents defined therein are particularly selected as follows:

• • halogen substituents are selected from F, Cl and Br, more preferably from F and Cl; and/or
  • linear or branched C₁-C₆-alkyl substituents are selected from methyl, ethyl propyl, iso-propyl, n-butyl and iso-butyl; and/or
  • C₁-C₃-alkoxy substituents are selected from methoxy and ethoxy; and/or
  • C₁-C₃-haloalkyl substituents are selected from difluoroethyl (—CH₂—CHF₂) and trifluoromethyl (CF₃); and/or
  • a substituted alkyl-group in the position R⁴ represents a substituted C₁-C₃-alkyl group; and/or
  • a bicyclic heteroaryl group is selected from a benzimidazolyl group.

In a particularly preferred aspect the compounds according to formula (I-A) and (I-B) as defined anywhere herein are selected from the following compounds:

No. Compound/Structure
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
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97

Pharmaceutically acceptable salts of the compounds according to the invention include, for example, salts with suitable anions, such as carboxylates, sulfonates, sulfates, chlorides, bromides, iodides, phosphates, tartrates, methane sulfonates, hydroxyethane sulfonates, glycinates, maleates, propionates, fumarates, toluene sulfonates, benzene sulfonates, trifluoroacetates, naphthalenedisulfonates-1,5, salicylates, benzoates, lactates, salts of malic acid, salts of 3-hydroxy-2-naphthoic acid-2, citrates and acetates. HCl salts are preferred.

Pharmaceutically acceptable salts of the compounds according to the invention further include, for example, salts with suitable pharmaceutically acceptable bases, such as, for example, salts with alkaline or alkaline-earth hydroxides, such as NaOH, KOH, Ca(OH)₂, Mg(OH)₂ etc., amine compounds such as ethylamine, diethylamine, triethylamine, ethyldiisopropylamine, ethanolamine, diethanolamine, triethanolamine, methylglucamine, dicyclohexylamine, dimethylaminoethanol, procaine, dibenzylamine, N-methylmorpholine, arginine, lysine, ethylenediamine, N-methylpiperidin, 2-amino-2-methyl-propanol-(1), 2-amino-2-methyl-propandiol-(1,3), 2-amino-2-hydroxyl-methyl-propandiol-(1,3) (TRIS) etc..

The novel compounds of the present invention can be present in an amorphous, crystalline or partially crystalline form or they may also be present exist as hydrates.

The novel compounds according to formula (I-A) and (I-B) as defined anywhere herein, have been found to act as ferroportin inhibitors and are thus suitable for the use as a medicament, such as in particular for the use as ferroportin inhibitors.

As already explained above, ferroportin is the iron transport protein, which is responsible for the uptake of the released iron via the intestine and its transfer into the blood circulation, thereby conveying the iron to the appropriate tissues and organs. Inactivation or inhibition of the ferroportin disables the export of the iron, thereby reducing the absorption of iron in the intestine. Ferroportin inhibition in the sense of the present invention therefore includes the inhibition of iron transport from the cells into the blood circulation and the inhibition of iron absorption in the intestine. Therein, the inhibition of iron transport and/or iron reflux may be effected by different ways of mechanism, comprising for example inhibition of iron transport activity of ferroportin and thus inhibition of iron reflux, triggering internalization, degradation and/or reduction of ferroportin, administering hepcidin agonists, i.e. compounds which compete with hepcidin or by compounds, which inhibit the binding of hepcidin to ferroportin.

Ferroportin inhibition may be determined by measuring the inhibition of ferroportin mediated iron transport activity in an iron response assay (BLAzer-Assay), as described in more detail in the Examples below. Further, ferroportin inhibition may be determined by measuring ferroportin internalization and/or degradation in the Ferroportin Internalization and Degradation Assay (FACS) or by examining the Ferroportin Ubiquitination and Degradation, each as described in more detail in the Examples below. Further, ferroportin inhibition may be determined by measuring the activity as an hepcidin agonist, for example by determining the Hepcidin binding capacity to ferroportin in the Hepcidin Internalization Assay (J774), as described in more detail in the Examples below. Further, ferroportin inhibition may be determined by confirming the inhibition of hepcidin binding to ferroportin, for example in the Biophysical Ferroportin-Hepcidin Binding Assay (Hep Bind FP), as described in more detail in the Examples below. Further, ferroportin inhibition may be determined by determining the activity of a compound regarding its ability to block iron export via ferroportin, for example with a test for measuring inhibition of iron efflux, as described in more detail in the Examples below.

Ferroportin inhibition in the sense of the present invention can thus in particular be defined by exhibiting a ferroportin inhibiting activity in at least one of the aforementioned test methods, shown in particular by: Inhibition of ferroportin mediated iron transport activity in an iron response assay (Blazer Assay): IC₅₀ value [μM] of not more than 100 (<100), preferably not more than 50 (<50), more preferably below 50 (<50).

Ferroportin Internalization and Degradation Assay (FACS): EC₅₀ value [μM] of not more than 100 (100), preferably not more than 50 (<50), more preferably below 50 (<50).

Ferroportin Ubiquitination and Degradation: visually inspected effect in Western blots of +comparable to hepcidin”, “+/−intermediate effect” and +/ +/−stronger intermediate effect”, preferred is an effect “+” or “+/ +/−”, most preferred is an effect “+”.

Hepcidin Internalization Assay (J774): IC₅₀ value [μM] of not more than 100 (<100), preferably not more than 50 (<50), more preferably below 50 (<50).

Biophysical Ferroportin-Hepcidin Binding Assay: IC₅₀ value [μM] of not more than 100 (<100), preferably not more than 50 (<50), more preferably below 50 (<50).

Inhibition of Iron Efflux: IC₅₀ value of not more than 100 (<100), preferably not more than 50 (50), more preferably below 50 (<50).

Ferroportin inhibition may further be determined in in vivo models, as described in more detail in the Examples below. Suitable in vivo models may comprise, for example, examination of hypoferremia in naive mice via measurement of serum iron reduction; examination of prevention of iron absorption in anemic rats via measurement of serum iron inhibition; examination of correction of hyperferremia in beta2-microglobulin deficient mice via measurement of serum iron reduction; examination of prevention of iron overload in beta2-microglobulin deficient mice via measurement of total iron in spleen or liver; examination of improvement of anemia, ineffective erythropoiesis and iron overload in a mouse model of p-thalassemia intermedia.

The activity of the compounds of the present invention as ferroportin inhibitors can in particular be determined by the methods as described in the Examples below.

As further already explained above, ferroportin inhibition may for example be effected by hepcidin, which is thus an essential regulating factor of iron absorption, inhibiting ferroportin and thus blocking iron transport from the cells into the blood circulation and iron absorption. It has further been found that several of the compounds as defined herein act as hepcidin mimetics or hepcidin agonists, which is also included by ferroportin inhibition in the sense of the present invention.

Accordingly, the compounds as defined in the present invention are also suitable for use in the inhibition of iron transport from the cells into the blood circulation and the inhibition of iron absorption in the intestine, as well as for the use as hepcidin mimetics or hepcidin agonists.

Due to the activity of the compounds as defined herein as ferroportin inhibitors, the compounds of the present invention are further particularly suitable for the use in the inhibition of iron transport mediated by ferroportin and thereby for the use in the prophylaxis and/or treatment of iron metabolism disorders leading to increased iron levels, of diseases related to or caused by increased iron levels, increased iron absorption or iron overload, such as in particular of tissue iron overload, of diseases associated with ineffective erythropoiesis, or of diseases caused by reduced levels of hepcidin. Further, the compounds of the present invention are suitable for the use in an adjunctive therapy by limiting the amount of iron available to pathogenic microorganisms, such as the bacterium Vibrio vulnificus, thereby preventing or treating infections caused by said pathogenic microorganisms.

Therein, diseases being associated with, being related to, being caused by or leading to increased iron levels, increased iron absorption, iron overload (e.g. tissue iron overload) or ineffective erythropoiesis comprise thalassemia, hemoglobinopathy, such as hemoglobin E disease (HbE), hemoglobin H disease (HbH), haemochromatosis, hemolytic anemia, such as sickle cell anemia (sickle cell disease) and congenital dyserythropoietic anemia.

Diseases being associated with, being related to, being caused by or leading to increased iron levels, increased iron absorption, iron overload (e.g. tissue iron overload) further comprise neurodegenerative diseases, such as for example Alzheimer's disease and Parkinson's disease, wherein the compounds are considered to be effective by limiting the deposition or increase of iron in tissue or cells.

The compounds of the present invention are further suitable for the use in the prophylaxis and/or treatment of formation of radicals, reactive oxygen species (ROS) and oxidative stress caused by excess iron or iron overload as well as in the prophylaxis and/or treatment of cardiac, liver and endocrine damage caused by excess iron or iron overload, and further in the prophylaxis and/or treatment of inflammation triggered by excess iron or iron overload.

Diseases associated with ineffective erythropoiesis comprise in particular myelodysplastic syndromes (MDS, myelodysplasia) and polycythemia vera as well as congenital dyserythropoietic anemia.

Further diseases, disorders and/or diseased conditions comprise iron overload caused by mutations in genes involved in sensing the systemic iron stores, such as hepcidin (Hamp1), hemochromatosis protein (HFE), hemojuvelin (HJV) and transferrin receptor 2 (TFR2), such as in particular diseases related to HFE and HJV gene mutations, chronic hemolysis associated diseases, sickle cell diseases, red cell membrane disorders, Glucose-6-phosphate dehydrogenase deficiency (G6PD deficiency), erythrpoietic porphyria, Friedrich's Ataxia, as well as subgroups of iron overload such as transfusional iron overload, iron intoxication, pulmonary hemosiderosis, osteopenia, insulin resistense, African iron overload, Hallervordan Spatz disease, hyperferritinemia, ceruloplasmin deficiency, neonatal hemochromatosis and red blood cell disorders comprising thalassemia, including alpha thalassemia, beta thalassemia and delta thalassemia, thalassemia intermedia, sickle cell disease and myelodyplastic syndrome.

Further diseases and/or disorders and/or diseased conditions associated with elevated iron levels include, but are not limited to, diseases with elevated iron level, comprising ataxia, Friedrich's ataxia, age-related macular degeneration, age-related cataract, age-related retinal diseases and neurodegenrative disease, such as pantothenate kinase-associated neurodegeneration, restless leg syndrome and Huntington's disease, The compounds of the present invention my further be suitable for the use in the prophylaxis and treatment of diseases caused by a lack of hepcidin.

In view thereof a further object of the present invention relates to a medicament containing one or more of the compounds as defined above, such as in particular a medicament for the prophylaxis and treatment in any of the indications, states, disorders or diseases as defined above.

A further object of the present invention relates to pharmaceutical compositions and medicaments comprising one or more of the compounds according to the invention as defined above as well as optionally one or more pharmacologically acceptable carriers and/or auxiliary substances and/or solvents.

A further object of the present invention relates to pharmaceutical compositions and medicaments comprising one or more of the compounds according to the invention as defined above as well as optionally one or more further pharmaceutically effective compounds. The said pharmaceutical compositions contain, for example up to 99 weight-% or up to 90 weight-% or up to 80 weight-% or or up to 70 weight-% of the compounds of the invention, the remainder being each formed by pharmacologically acceptable carriers and/or auxiliaries and/or solvents and/or optionally further pharmaceutically active compounds.

Therein, the pharmaceutically acceptable carriers, auxiliary substances or solvents are common pharmaceutical carriers, auxiliary substances or solvents, including various organic or inorganic carrier and/or auxiliary materials as they are customarily used for pharmaceutical purposes, in particular for solid medicament formulations. Examples include excipients, such as saccharose, starch, mannitol, sorbitol, lactose, glucose, cellulose, talcum, calcium phosphate, calcium carbonate; binding agents, such as cellulose, methylcellulose, hydroxypropylcellulose, polypropyl pyrrolidone, gelatine, gum arabic, polyethylene glycol, saccharose, starch; disintegrating agents, such as starch, hydrolyzed starch, carboxymethylcellulose, calcium salt of carboxymethylcellulose, hydroxypropyl starch, sodium glycol starch, sodium bicarbonate, calcium phosphate, calcium citrate; lubricants, such as magnesium stearate, talcum, sodium laurylsulfate; flavorants, such as citric acid, menthol, glycin, orange powder; preserving agents, such as sodium benzoate, sodium bisulfite, paraben (for example methylparaben, ethylparaben, propylparaben, butylparaben); stabilizers, such as citric acid, sodium citrate, acetic acid and multicarboxylic acids from the titriplex series, such as, for example, diethylenetriaminepentaacetic acid (DTPA); suspending agents, such as methycellulose, polyvinyl pyrrolidone, aluminum stearate; dispersing agents; diluting agents, such as water, organic solvents; waxes, fats and oils, such as beeswax, cocoa butter; polyethylene glycol; white petrolatum; etc..

Liquid medicament formulations, such as solutions, suspensions and gels usually contain liquid carrier, such as water and/or pharmaceutically acceptable organic solvents. Furthermore, such liquid formulations can also contain pH-adjusting agents, emulsifiers or dispersing agents, buffering agents, preserving agents, wetting agents, gelatinizing agents (for example methylcellulose), dyes and/or flavouring agents, for example as defined above. The compositions may be isotonic, that is, they can have the same osmotic pressure as blood. The isotonicity of the composition can be adjusted by using sodium chloride and other pharmaceutically acceptable agents, such as, for example, dextrose, maltose, boric acid, sodium tartrate, propylene glycol and other inorganic or organic soluble substances. The viscosity of the liquid compositions can be adjusted by means of a pharmaceutically acceptable thickening agent, such as methylcellulose. Other suitable thickening agents include, for example, xanthan gum, carboxymethylcellulose, hydroxypropylcellulose, carbomer and the like. The preferred concentration of the thickening agent will depend on the agent selected.

Pharmaceutically acceptable preserving agents can be used in order to increase the storage life of the liquid composition. Benzyl alcohol can be suitable, even though a plurality of preserving agents including, for example, paraben, thimerosal, chlorobutanol and benzalkonium chloride can also be used.

The above-mentioned pharmaceutical compositions are suitable, for example, for intravenous, intraperitoneal, intramuscular, intravaginal, intrabuccal, percutaneous, subcutaneous, mucocutaneous, oral, rectal, transdermal, topical, intradermal, intragasteral or intracutaneous application and are provided, for example, in the form of pills, tablets, enteric-coated tablets, film tablets, layer tablets, sustained release formulations for oral, subcutaneous or cutaneous administration (in particular as a plaster), depot formulations, dragees, suppositories, gels, salves, syrup, granulates, suppositories, emulsions, dispersions, microcapsules, microformulations, nanoformulations, liposomal formulations, capsules, enteric-coated capsules, powders, inhalation powders, microcrystalline formulations, inhalation sprays, epipastics, drops, nose drops, nose sprays, aerosols, ampoules, solutions, juices, suspensions, infusion solutions or injection solutions etc..

A further object of the present invention relates to medicaments or combined preparations containing one or more of the compounds as defined above and at least one further pharmaceutically active compound, such as in particular a compound for the prophylaxis and treatment of iron overload and the associated symptoms, preferably an iron-chelating compound, or a compound for the prophylaxis and treatment of any of the states, disorders or diseases as defined above, such as in particular a pharmaceutically active compound for the prophylaxis and treatment of thalassemia, haemochromatosis, neurodegenerative diseases (such as Alzheimer's disease or Parkinson's disease) and the associated symptoms.

A further object of the present invention relates to the use of the compounds as defined above per se, in a combination therapy (fixed dose or free dose combinations for sequential use) with one or two other active ingredients (drugs). Such combination therapy comprises co-administration of the compounds of the present invention with the at least one additional pharmaceutically active compound (drug). Combination therapy in a fixed dose combination therapy comprises co-administration of the compounds of the present invention with the at least one additional pharmaceutically active compound in a fixed-dose formulation. Combination therapy in a free dose combination therapy comprises co-administration of the compounds of the present invention and the at least one additional pharmaceutically active compound in free doses of the respective compounds, either by simultaneous administration of the individual compounds or by sequential use of the individual compounds distributed over a time period.

The at least one additional pharmaceutically active compound (drug) comprises in particular drugs for reducing iron overload (e.g. Tmprss6-ASO) or iron chelators, in particular curcumin, SSP-004184, Deferitrin, deferasirox, deferoxamine and/or deferiprone, or antioxidants such as n-acetyl cysteine, anti-diabetics such as GLP-1 receptor agonists, antibiotics such as vancomycin (Van) or tobramycin, drugs for the treatment of malaria, anticancer agents, antifungal drugs, drugs for the treatment of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease (e.g. dopamine agonists such as Levodopa), anti-viral drugs such as interferon-a or ribavirin, or immunosuppressents (cyclosporine A or cyclosporine A derivatives), iron supplements, vitamin supplements, red cell production stimulators, anti-inflammatory biologies, anti-thrombolytics, statins, vasopressors and inotropic compounds.

A further object of the present invention relates to the use of the above combinations for the prophylaxis and/or treatment of diseases caused by a lack of hepcidin or iron metabolism disorders, such as particularly iron overload states such as in particular thalassemia and hemochromatosis and other disorders as described in the present application.

A further object of the present invention relates to the use of the compounds as defined herein per se or the hereinabove described combination therapies, in combination with Blood transfusion.

The compounds, medicaments and or combined preparations according to the present invention may be administered orally, parentally, as well as intravenously.

For this purpose, the compounds according to the invention are preferably provided in medicaments or pharmaceutical compositions in the form of pills, tablets, such as enteric-coated tablets, film tablets and layer tablets, sustained release formulations for oral administration, depot formulations, dragees, granulates, emulsions, dispersions, microcapsules, microformulations, nanoformulations, liposomal formulations, capsules, such as enteric-coated capsules, powders, microcrystalline formulations, epipastics, drops, ampoules, solutions, suspensions, infusion solutions or injection solutions or in the form of a preparation suitable for inhalation.

In a preferred embodiment of the invention the compounds are administered in the form of a tablet or capsule, as defined above. These may be present, for example, as acid resistant forms or with pH dependent coatings.

The compounds of the present invention as the active substance can be administered, for example, with a unit dose of 0.001 mg/kg to 500 mg/kg body weight, for example 1 to 4 times a day. However, the dose can be increased or reduced depending on the age, weight, condition of the patient, severity of the disease or type of administration.

Accordingly, a further object of the present invention relates to compounds, medicaments, compositions and combined preparations as defined above for the preparation of a medicament, particularly for the prophylaxis and treatment of any indication, state, disorder or disease as defined above, in particular for oral or parenteral administration.

A further object of the present invention relates to a method for the prophylaxis and treatment as defined above, such as in particular for the prophylaxis and/or treatment of iron metabolism disorders being associated with or leading to increased iron levels and in particular iron overload, diseases related to or caused by increased iron levels or iron overload, iron storage diseases being associated with or leading to increased iron levels, and diseases being associated with ineffective erythropoiesis, the method comprising administering, to a patient (human or animal) in need thereof, a compound, a medicament, a composition or a combined preparation as defined above.

Therein, diseases being associated with, being related to, being caused by or leading to increased iron levels or iron overload are as defined above.

A further object of the present invention relates to the use of the compounds as defined above for the preparation of a medicament, particularly for the prophylaxis and treatment and of any indication, state, disorder or disease as defined above.

The compounds according to the invention of general structural formula (I-A) and (1-B) can basically be prepared by the processes described in the international application WO2021/191202, herein incorporated by reference.

In particular, the following general procedure describes a suitable preparation process:

Step 1:

or

or

followed by Ester Cleavage:

Step 2:

Step 3:

Step 3a - Formula (I-A):

Step 3a illustrates the general scheme for preparing compounds of the formula (I-A).

X¹, X², X³ and the groups A and B, as well as I, L¹ and L² have the meaning as defined anywhere herein.

Step 3b - Formula (I-B):

For preparing compounds of the formula (I-B), wherein the linker groups “L¹” and “L^2” independently represent a CH-group, a CH₂—CH-group or a CH₂-CH₂-CH-group, depending on the desired resulting alkylene-chain length, defined by —[CH₂]_m—, and —[CH₂]_n—, respectively, the general scheme of Step 3 is illustrated as follows

Step 3b:

X¹, X², X³ and the groups A and B, as well as I, L¹, m and n have the meaning as defined anywhere herein. Therein, the group “L¹” represents a CH-group, a CH₂-CH-group or a CH₂-CH₂-CH-group, depending on the desired resulting alkylene-chain length defined by —[CH₂]_m—.

In a further aspect the invention covers the intermediate compounds obtainable in the preparation methods described herein, such as in particular the intermediate compounds resulting from the individual steps of the general reaction scheme above and as described further in detail herein. Details of the preparation conditions provide the Examples below.

EXAMPLES

The invention is illustrated in more detail by the following examples. The examples are merely explanatory, and the person skilled in the art can extend the specific examples to further claimed compounds.

Pharmacological Assays

1. Hepcidin Internalization Assay (J774)

This cellular assay allows quantification of the binding of hepcidin to ferroportin (Fpn) through microscopic detection of internalization of a fluorescently labeled hepcidin into J774 cells. J774 is a mouse macrophage cell line which was shown to express Fpn endogenously upon incubation with iron (Knutson et al, 2005). Binding of hepcidin to Fpn triggers internalization and degradation of both hepcidin and Fpn. However, the TMR (6-carboxytetramethylrhodamine) fluorophore attached to hepcidin remains associated with the cell after degradation of the hepcidin peptide backbone. Therefore, microscopic detection of cell-associated TMR fluorescence is a measure of hepcidin binding to Fpn and internalization of hepcidin and Fpn. If TMR-hepcidin is prevented from binding to Fpn, cellular TMR fluorescence remains low (Durrenberger et al, 2013). The effect of small molecular weight Fpn inhibitor compounds in this assay was evaluated in vitro as described below.

J774 cells, harvested from ca. 80% confluent cultures, were plated at 8×10⁵ cells/ml in complete medium (DMEM, 10% FBS, 1% Penicillin-Streptomycin) containing 200 μM Fe(Ill)NTA (nitrilotriacetic acid), 100 μl per well of 96 well MicroClear plates (Greiner, Cat. 655090) and grown at 37° C. with 5% CO₂. After overnight incubation, cells were washed 3 times with pre-warmed DMEM w/o phenol red, 30 μl/well of DMEM w/o phenol red was added after the final wash and 10 μl/well of dilution series of test compounds were added in triplicates. J774 cells were pre-incubated with test compounds at 37° C. with 5% CO₂ for 15 min. before TMR-hepcidin was added at 25 nM final concentration. Cells were incubated in a total volume of 50 μl at 37° C. with 5% CO₂ for 2 hours, then Hoechst 33342 dye was added to a final concentration of 0.5 μg/ml to stain nuclei and further incubated for 10 min. at 37° C. with 5% CO₂ Cells were washed 3 times with PBS and fixed in 100 μl of 4% paraformaldehyde in PBS for 15 min. at room temperature. After removal of the paraformaldehyde solution, cells were washed 3 times with PBS leaving 100 μl per well and the plates were sealed with foil plate seal. TMR (530-550 nm excitation/575-625 nm emission/400 ms exposure time) and Hoechst 33342 (360-370 nm excitation/420-460 nm emission /10 ms exposure time) fluorescence images were acquired using a ScanR plate imager (Olympus) with a 20× high NA objective. Four pictures were acquired per well and fluorescence channel covering ca. 1500 cells per well. The acquired image data was analysed with the ScanR image analysis software. Image analysis included detection of nuclei (Hoechst 33342 fluorescence), identification of cell-associated regions, application of a virtual channel and thresholding for rolling-ball-type background reduction, followed by application of the Sum(Mean) algorithm to measure the TMR fluorescence associated with cells as a quantitative measure for internalized TMR- hepcidin. IC₅₀ values were calculated with the Sum(Mean) raw data using “log(inhibitor) vs. response” curve fitting of Prism 5 software (GraphPad Software Inc., version 5.02). For each data set the fit of the “log(inhibitor) vs. response (three parameters)” model was compared to the fit of the “log(inhibitor) vs. response - Variable slope (four parameters)” model and the IC₅₀ data of the preferred model was used. IC₅₀ data of the Fpn inhibitors that were tested in the hepcidin internalization assay are listed in Table1. The IC₅₀ of unlabeled hepcidin in this assay is 0.015±0.011 μM.

Table 1 Average (AVE) IC₅₀ data of Fpn inhibitors tested in the hepcidin internalization assay is shown for multiple measurements

TABLE 1
No. IC50 [uM]
1   0.004
2   0.001
3   0.002
4   0.001
5   0.007
6   0.0036
8   0.0012
9   0.001
10  0.0015
11  0.002
14  0.126
15  0.032
16  0.181
17  0.0008
21  0.001
23  0.0019
25  0.007
30  0.0013
31  0.0028
34  0.0074
35  0.002
37  0.0009
39  0.0017
40  0.0018
41  0.004
42  0.0024
44  0.001
45  0.0008
46  0.001
47  0.008
48  0.002
49  3.3
50  0.0008
51  0.0008
52  0.625
53  0.005
54  0.0009
55  0.0014
56  0.009
57  0.0014
58  0.0008
60  0.355
61  1.43
62  0.0085
63  0.0008
64  0.0013
65  0.0012
66  0.005
67  0.0043
68  0.007
69  0.003
70  0.0039
71  0.007
72  0.035
73  0.0026
74  0.0049
75  0.0013
76  0.013
77  0.0051
81  0.004
82  0.002
83  0.0019
84  0.15
85  0.0078
86  0.0041
87  0.017
88  0.0009
89  0.004
90  0.039
91  0.002
92  0.002
93  0.003
94  0.365
95  0.026
96  0.223
97  0.089

2. Biophysical Ferroportin-Hepcidin Binding Assay

This biophysical assay was developed to confirm inhibition of hepcidin binding to ferroportin (Fpn) more directly. Incubation of TMR-hepcidin with purified human Fpn isolated from Pichia pastoris yeast cells expressing human Fpn with a C-terminal FLAG affinity tag (Bonaccorsi di Patti, 2014) leads to increased fluorescence polarization (FP) of the TMR-hepcidin ligand. Small molecular weight Fpn inhibitors are tested for inhibition of binding of TMR-hepcidin to Fpn, as detected by dose-dependent decrease of the TMR FP signal, as described in detail below.

A mixture of 1.3 μM human Fpn and 30 nM TMR-hepcidin in FP assay buffer containing 50 mM Tris-HCl pH 7.3, 200 mM NaCl, 0.02% DDM, 0.1% BSA is plated into a 384 well black low volume round bottom plate (Corning, Cat. 3677) at 16 μl per well. 8 μl of serial dilutions of test compounds are added in duplicates to reach final Fpn and TMR-hepcidin concentrations of 1 μM and 20 nM, respectively. Plates are incubated for 90 minutes at room temperature and parallel (S) and perpendicular (P) fluorescence is measured in a Synergy H1 fluorescence reader (BioTek). FP values are calculated in mP according to the following formula.

$\mathrm{mP}=\frac{F_{\mathrm{parallel}}-F_{\mathrm{perpendicular}}}{F_{\mathrm{parallel}}+F_{\mathrm{perpendicular}}}\times 1000$

IC₅₀ values are determined with the calculated mP values as described for the hepcidin internalization assay. The IC₅₀ of unlabeled hepcidin in this assay is about 0.37±0.067 μM.

3. Inhibition of Ferroportin mediated Iron Export Activity in an Iron Response Assay

Intracellular iron levels are indirectly measured in this assay by monitoring the activity of a beta-lactamase (BLA) reporter gene fused to the human ferritin promoter and the associated iron regulatory element (IRE) contained within the 5′ untranslated region of the ferritin mRNA. Expression of ferroportin (Fpn) in such a cell line leads to iron efflux and lower iron levels as reflected by lower activity of the reporter gene. On the other hand, inhibition of Fpn-mediated iron efflux results in elevated cellular iron levels which is detected as increased reporter gene activity. Small molecular weight Fpn inhibitor compounds are tested for dose-dependent effects in this in vitro iron response assay as described below.

The HEK-293 cell line #354 is generated by stable integration of (i) a human Fpn-GFP fusion construct inserted in a derivative of the doxycycline-inducible pTRE-Tight-BI plasmid (Clontech, Cat. 631068) and (ii) a human ferritin promoter-BLA reporter gene into a derivative of the HEK-293 Tet-ON Advanced cell line (Clontech). To generate the ferritin-BLA reporter gene construct, a 1.4 kb fragment of the human ferritin H promoter is amplified by PCR from human genomic DNA (forward primer 5′-CAGGTTTGTGAGCATCCTGAA-3′ (SEQ ID NO: 1); reverse primer 5′-GGCGGCGACTAAGGAGAGG-3′ (SEQ ID NO: 2)) and inserted in front of the BLA gene present in the pcDNA™6.2/cGeneBLAzer™ DEST plasmid (Invitrogen, Cat. 12578-043) thereby replacing the original CMV promoter and placing the IRE that regulates translation of the ferritin gene ca. 170 bp upstream of the start codon of the reporter gene. #354 cells are harvested from ca. 80% confluent cultures, seeded at 1.8×10⁵ cells/ml in DMEM/F12 GlutaMAX™ medium (Invitrogen, Cat. 31331-028) containing 10% FBS (Clontech, Cat. 631106), 1% Penicillin-Streptomycin, 200 μg/ml Hygromycin B (Invitrogen, Cat. 10687-010), Blasticidin 5 μg/ml, (Invitrogen, Cat. R210-01), 4 μg/ml doxycycline (Clontech, Cat. 631311), 50 μl per well of 384 well PDL-coated plates and grown at 37° C. with 5% CO₂. After overnight incubation, 10 μl/well of dilution series of the test compounds are added in quadruplicates and plates are further incubated overnight at 37° C. with 5% CO₂. Cells are washed 3 times with HBSS leaving 25 μl per well. BLA activity is detected by adding 5 μl/well of the GeneBlazer reagent CCF4-AM (Invitrogen, Cat. K1085) to the cells. After incubation of the plates in the dark at 18° C. for 60 min., blue and green fluorescence signals are measured in a Safire2 fluorescence plate reader (Tecan) with excitation at 410 nm and emissions at 458 nm (blue) and 522 nm (green). The ratio of blue/green fluorescence as a measure for BLA activity is calculated and EC₅₀ values are determined with the calculated blue/green fluorescence ratios as described for the hepcidin internalization assay. The EC₅₀ of hepcidin in this assay is about 0.096±0.063 μM (n=37).

4. Ferroportin Internalization and Degradation Assay

HEK-293 cell line #354 (described in example 3) is used to measure the capacity of the compounds to induce internalization and degradation of ferroportin (Fpn) by fluorescence activated cell sorting (FACS). Growing HEK-293 #354 cells in doxycycline containing media induced expression of human Fpn-GFP fusion protein on the cell surface. Data from 10 independent experiments show that cultivation of HEK #354 cells for 48h in the presence of 4 μg/ml doxycycline induce in average 42.6% ±6.4% Fpn-GFP-positive cells. Small molecular weight Fpn inhibitor compounds are tested for dose-dependent effects on the Fpn-GFP mean fluorescence intensity (MFI) on HEK-293 cell line #354, as described below.

HEK #354 cells are harvested from ca. 80% confluent cultures, seeded at 0.6×10⁶ cells/ml in DMEM/F12 GlutaMAX™ medium (Invitrogen, Cat. 31331-028) containing 10% FBS (Clontech, Cat. 631106), 1% Penicillin-Streptomycin (Invitrogen, Cat. 15140-122), 200 μg/ml Hygromycin B (Invitrogen, Cat. 10687-010), Blasticidin 5 μg/ml, (Invitrogen, Cat. R210-01), 4 μg/ml doxycycline (Clontech, Cat. 631311), 50 μl per well of 384 well plates (Greiner, Cat. 781091) and grown at 37° C. with 5% CO₂. After overnight incubation, 10 μl/well of dilution series of the test compounds are added in quadruplicates and plates are further incubated overnight at 37° C. with 5% CO₂. Cells are washed once with FACS buffer (PBS containing 1% FBS, 2 mM EDTA and 0.05% NaN₃), harvested in FACS buffer with 0.5 μg/ml propidium iodide (Sigma, Cat. P4864) and analyzed in a flow cytometer (CANTO™ II, BD Biosciences) equipped with high throughput sampler. Live HEK #354 cells are gated as propidium iodide negative population and analyzed for expression of Fpn-GFP. MFI of Fpn-GFP of >2000 live cells for each compound dilution is calculated using FlowJo (Tree Star's, Oregon) and the potency of the Fpn-inhibitors to induce internalization and degradation of Fpn-GFP is calculated as described for the hepcidin internalization assay. The average EC₅₀ value of hepcidin in this assay is about 0.004±0.002 μM.

5. Ferroportin Ubiquitination and Degradation

Exposure of cells expressing ferroportin (Fpn) to hepcidin is known to trigger ubiquitination and subsequent internalization and degradation of Fpn (Qiao, 2012). The potential of Fpn inhibitors to induce Fpn ubiquitination and degradation is investigated with an immunoprecipitation assay using the J774 mouse macrophage cell line which expresses Fpn upon treatment with iron.

J774 cells (DSMZ, Cat. ACC170) are seeded at 0.8×106 cells/ml in 15 ml of medium (DMEM Gibco Cat. 11971-025, 10% heat inactivated FBS Gibco Cat. 10500-064, 1% Penicillin-Streptomycin Gibco Cat. 15140-122) containing 200 μM Fe(Ill)-NTA into 10 cm tissue culture dishes (Greiner Cat. 664160) and grown overnight at 37° C. with 5% CO₂. Cells are incubated with synthetic human hepcidin (Bachem, Cat. H-5926) or Fpn inhibitor compounds for 10 min or 120 min. Cells are washed and lysed with ice-cold lysis buffer (Pierce, Life Technoligies, Cat. 87787) including 1× HALT protease inhibitor cocktail (Life technologies, Cat. 78429) and 10 mM iodoacetamide (Sigma, Cat. 16125) to stabilize ubiquitinated proteins. Immunoprecipitation is done using the Pierce Classic IP Kit (Life Technologies, Cat. 26146) following the manufacturer's protocol. Briefly, 2 mg protein in 1.25 ml IP lysis buffer is incubated by mixing for 1 h at 4° C. with control agarose beads to pre-clear the lysate and reduce nonspecific signal. Unbound lysate is then incubated overnight with 12 μg per reaction of the affinity purified anti-Fpn antibody F308 that is raised against a GST fusion protein of mouse Fpn amino acids 224-308. Immune complexes are captured by pipetting 14 μl settled Pierce Protein A/G Plus Agarose beads (Life Technologies, Cat. 20423) per reaction and the slurry is incubated for 1.5 h at 4° C. with gentle end-over-end mixing. The beads are washed and immune complexes are eluted directly with 75 μl SDS NuPAGE LDS sample buffer (Life Technologies, Cat. NP0007) containing DTT (Life Technologies, Cat. NP0009).

After immunoprecipitation samples are analyzed by Western blotting using a rabbit anti-mouse MTP1 antiserum (Alpha Diagnostic International, Cat. MTP11-A) and a mouse anti-mono- and polyubiquitinylated conjugates monoclonal antibody (Enzo Lifesciences, Cat. BML-PW8810) for detection of ferroportin and ubiquitin, respectively. Mouse monoclonal anti-rabbit IgG light chain (Abcam, Cat. ab99697) and anti-mouse IgG H&L (Abcam, Cat. ab6789) HRP conjugates are used as secondary antibodies.

6. Inhibition of Iron Efflux by Ferroportin Inhibitors

The activity of hepcidin and ferroportin inhibitor compounds regarding their ability to block iron export via ferroportin is tested on T47D cells (ECACC, Cat. 85102201) as described below.

Cells are plated in 24-well plates (Greiner, Cat. 662160) containing 350′000 cells/well and incubated overnight with 100 μM ⁵⁸Fe (⁵⁸Fe(II)-Sulfate, Vifor Pharma Batch No. ROR 3085) in 500 μM L-Ascorbic Acid (Sigma Aldrich, Cat. 795437) containing growth medium. Cells were washed once with 500 μl iron uptake buffer (IUB; PIPES 40 mM, Cat. P1851, Glucose Monohydrate 10 mM, Cat. 49158, Sodium Chloride 260 mM, Cat. 71379, Potassium Chloride 20 mM, Cat. P9541, Magnesium Sulfate 2 mM, Cat. 63138, Sigma Aldrich), then once with removal buffer (2 min incubation, BPDS 100 μM, Cat. 11890 and Na₂S₂O₄ 500 μM, Cat. 157953, Sigma Aldrich, in IUB) and again twice with IUB. A serial dilution of hepdicin (Bachem) or ferroportin inhibitors (4 μM-0.0064 μM, 5 fold dilution) is added in a total volume of 0.6 ml per well. Cells are incubated at 37° C. with 5% CO₂ for 20 h. Supernatants are collected and ⁵⁸Fe is measured using inductively coupled plasma mass spectrometry (ICP-MS, Thermo Scientific, Element 2). Pellets are harvested for protein concentration measurements. Results are plotted as ng ⁵⁸Fe in supernatant per mg protein in cell lysates.

Preparation of Example Compounds

General Experimental Details

Commercially available reagents and solvents (HPLC grade) were used without further purification. ¹H NMR spectra were recorded on a Bruker DRX 500 MHz spectrometer, a Bruker DPX 250 MHz spectrometer or a Bruker Avance spectrometer 400 MHz in deuterated solvents. Chemical shifts (6) are in parts per million.

Compounds were purified by flash column chromatography on normal phase silica on Biotage Isolera systems using the appropriate SNAP cartridge and gradient. Alternatively, compounds were purified on reverse phase using Biotage Isolera systems with the appropriate C18 SNAP cartridge and reverse-phase eluent or by preparative HPLC (if stated otherwise).

ABBREVIATIONS

• • EtOAc Ethylacetate
  • CH₂Cl₂ Dichloromethane
  • Et₂O Diethylether
  • MeOH Methanol
  • EtOH Ethanol
  • brine Aqueous saturated sodium chloride solution
  • Chloroform-d Deuterated chloroform
  • DMSO-d₆ Deuterated dimethylsulfoxid
  • s Singlet
  • br s Bright Singlet
  • d Doublet
  • dd Double Doublets
  • dt Doublet of Triplets
  • td Triplet of Doublets
  • hept. Heptett
  • m Multiplet
  • q Quartet
  • δ Chemical shift
  • ppm Parts per million
  • M Molarity
  • mm Millimolar
  • umol Mikromolar
  • g Gram
  • mg Milligram
  • Liter
  • mL Milliliter
  • h Hours
  • min Minute
  • %-w/w Percentage by mass
  • TLC Thin layer chromatography
  • UHPLC Ultra high pressure liquid chromatography
  • MS Mass spectroscopy
  • ESI Electronic spray ionization
  • m/z mass to charge ratio
  • H⁺ Proton
  • MHz Mega Hertz
  • s.m. starting material
  • Jones Reagent CrO₃ in H₂SO₄
  • CrO₃ Chromium trioxide
  • HCl Hydrochloric acid
  • H₂SO₄ Sulfuric acid
  • NH₄Cl Ammonium chloride
  • Na₂SO₄ Sodium sulfate
  • NaOH Sodium hydroxide
  • Bn Benzyl
  • MS Mass spectra
  • ESI Electrospray ionisation
  • SNAP Biotage-column-brandname for flash column chromatography
  • R_f Retention Factor
  • TLC Thin Layer Chromatography Chemical nomenclature

The chemical names of the intermediates and the final Example Compounds were generated by using Chem Draw Professional 17.0.

All R_f values were determined using the following TLC plates: Merck, TLC Silcagel 60° F.₂₅₄.

Preparation Details

Example No. 1

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-methoxyethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-methoxyethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine (Chemspace) (100 mg, 0.46 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (113 mg, 0.46 mmol) were added to a solution of NaOH (9 mg, 0.23 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 1 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.7 (m, 2H), 8.6 (s, 1H), 8.4 (m, 1H), 7.9 (m, 1H), 7.7 (m, 2H), 7.4 (m, 3H), 4.6 (m, 4H), 4.6 (m, 2H), 3.6 (m, 2H), 3.5 (m, 8H), 3.2 (s, 3H) ppm. UHPLC/MS (ESI): [m/z]: 467 [M+H]⁺.

Example No. 2

2-(2-((2-(1-butyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-butyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine (Chemspace) (100 mg, 0.46 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (114 mg, 0.46 mmol) were added to a solution of NaOH (9 mg, 0.23 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 2 (118 mg, 0.25 mmol, 55%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.4 (m, 1H), 7.7 (m, 1H), 7.5 (m, 2H), 7.4 (m, 1H), 7.1 (m, 2H), 4.6 (d, 2H), 4.0 (m, 2H), 3.0 (m, 8H), 1.7 (m, 2H), 1.3 (m, 2H), 0.9 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 465 [M+H]⁺.

Example No. 3

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(pyridin-2-ylmethyl)-1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(pyridin-2-ylmethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine (Chemspace) (100 mg, 0.4 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (98 mg, 0.4 mmol) were added to a solution of NaOH (8 mg, 0.2 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 3 (63 mg, 0.13 mmol, 32%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 4H), 7.7 (m, 2H), 7.5 (m, 3H), 7.3 (m, 2H), 7.1 (m, 2H), 5.5 (d, 2H), 4.6 (d, 6H), 3.4 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 500 [M+H]⁺.

Example No. 4

2-(2-((2-(1-butyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide dihydrochloride

0.1 ml HCl solution (4M in dioxane) were added to a solution of 2-(2-((2-(1-butyl-1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide 2 (100 mg, 0.2 mmol) in 5 ml dichloromethane. The resulting mixture were stirred at room temperature for 30 min and concentrated under reduced pressure. The isolated material dried under HV to obtain the title compound 4 (112 mg, 0.21 mmol, 97%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.7 (m, 1H), 8.6 (s, 1H), 8.3 (m, 1H), 7.8 (m, 1H), 7.7 (m, 2H), 7.4 (m, 3H), 4.6 (m, 2H), 4.3 (m, 2H), 3.5 (m, 8H), 1.7 (m, 2H), 1.4 (m, 2H), 0.9 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 465 [M+H]⁺.

Example No. 5

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-methyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-methyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.40 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (100 mg, 0.40 mmol) were added to a solution of NaOH (40 mg, 1.00 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 5 (33.4 mg, 0.08 mmol, 20%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (m, 2H), 8.4 (m, 1H), 7.7 (m, 1H), 7.6 (m, 2H), 7.4 (m, 1H), 7.2 (m, 2H), 4.6 (d, 2H), 3.8 (s, 3H), 3.2 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 423 [M+H]⁺.

Example No. 6

2-(2-((2-(1-ethyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-ethyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.38 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (94 mg, 0.38 mmol) were added to a solution of NaOH (38 mg, 0.95 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 6 (22.2 mg, 0.05 mmol, 13%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.4 (m, 1H), 7.8-7.3 (m, 4H), 7.2 (m, 2H), 4.6 (d, 2H) 4.2 (m, 2H), 3.2 (m, 8H), 1.3 (m, 3H) ppm. UHPLC/MS (ESI): [m/z]: 437 [M+H]⁺.

Example No. 9

2-(2-((2-(1-butyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-chloropyridin-2-yl)methyl)oxazole-4-carboxamide (VIT-101131 nda112)

2-(1-butyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine (Chemspace) (75 mg, 0.35 mmol) and N-((3-chloropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (91 mg, 0.35 mmol) were added to a solution of NaOH (7 mg, 0.5 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 6 (101 mg, 0.21 mmol, 60%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 3H), 7.9 (m, 1H), 7.5 (m, 2H), 7.35 (m, 1H) 7.1 (m, 2H), 4.6 (d, 2H), 4.2 (t, 2H), 3.0 (m, 8H), 1.65 (m, 2H), 1.3 (m, 2H), 0.9 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 482 [M+H]⁺.

Example No. 10

2-(2-((2-(1-butyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((4-methylpyridin-2-yl)methyl)oxazole-4-carboxamide (VIT-101124nda110)

2-(1-butyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine (Chemspace) (100 mg, 0.46 mmol) and N-((3-methylpyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (112 mg, 0.46 mmol) were added to a solution of NaOH (10 mg, 0.23 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 2 (70 mg, 0.15 mmol, 32%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (m, 2H), 8.35 (m, 1H), 7.6 (m, 1H), 7.5 (m, 2H), 7.3 (m, 2H), 7.2 (m, 1H), 7.1 (m, 2H), 4.5 (d, 2H), 4.2 (t, 2H), 3.0 (m, 8H), 1.65 (m, 2H), 1.3 (m, 2H), 0.9 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 461 [M+H]⁺.

Example No. 11

2-(2-((2-(1-butyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-(pyridin-2-ylmethyl)oxazole-4-carboxamide

2-(1-butyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine (Chemspace) (100 mg, 0.46 mmol) and N-(pyridin-2-ylmethyl)-2-vinyloxazole-4-carboxamide (106 mg, 0.46 mmol) were added to a solution of NaOH (10 mg, 0.23 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 2 (67 mg, 0.15 mmol, 33%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.75 (m, 1H), 7.7 (m, 1H), 7.5 (m, 2H), 7.3 (m, 2H), 7.2 (m, 2H), 4.5 (d, 2H), 4.2 (t, 2H), 3.0 (m, 8H), 1.65 (m, 2H), 1.3 (m, 2H), 0.9 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 447 [M+H]⁺.

Example No. 23

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-phenethyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide (VIT-101133rxu463)

2-(1-phenethyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine hydrochloride (Chemspace) (100 mg, 0.33 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (82 mg, 0.33 mmol) were added to a solution of NaOH (20 mg, 0.5 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 6 (90.8 mg, 0.18 mmol, 54%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.5 (m, 2H), 7.4 (m, 1H), 7.3-7.0 (m, 7H), 4.6 (d, 2H) 4.4 (m, 2H), 2.9 (m, 8H), 2.65 (m, 2H) ppm. UHPLC/MS (ESI): [m/z]: 513 [M+H]⁺.

Example No. 37

2-(2-((2-(1-butyl-6,7-dihydro-1H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide (VIT-101136nda117)

2-(1-butyl-6,7-dihydro-1H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.29 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (71 mg, 0.29 mmol) were added to a solution of NaOH (29 mg, 0.72 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂CI/MeOH) to obtain the title compound 6 (77 mg, 0.15 mmol, 51%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.65 (m, 1H), 7.4 (m, 1H), 6.95 (m, 2H), 4.6 (d, 2H), 4.2 (m, 4H), 4.0 (m, 2H), 3.0 (m, 8H), 1.6 (m, 2H), 1.25 (m, 2H), 0.85 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 523 [M+H]⁺.

Example No. 44

2-(2-((2-(1-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

The preparation of Example Compound No. 43 is carried out similarly as described for Example No. 2.

¹H NMR (400 MHz, MeOH-d₄) δ 8.3 (m, 1H), 8.2 (s, 1H), 7.6 (m, 2H), 7.5 (m, 2H), 7.4 (m, 1H), 7.3 (m, 1H), 7.2 (m, 4H), 4.7 (t, 2H), 4.6 (d, 2H), 3.4 (t, 2H), 3.0 (m, 2H), 2.9 (m, 4H), 2.8 (m, 2H) ppm.

Example No. 45

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(3-methoxyphenethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(3-methoxyphenethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine (Chemspace) (100 mg, 0.34 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (84 mg, 0.34 mmol) were added to a solution of NaOH (7 mg, 0.17 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 6 (161.4 mg, 0.3 mmol, 87%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.55 (m, 2H), 7.4 (m, 1H), 7.15 (m, 3H), 6.75 (m, 1H), 6.6 (m, 2H), 4.6 (d, 2H) 4.4 (m, 2H), 3.6 (s, 3H), 2.9 (m, 8H), 2.7 (m, 2H) ppm. UHPLC/MS (ESI): [m/z]: 543 [M+H]⁺.

Example No. 46

2-(2-((2-(1-butyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-methoxypyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-butyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine (Chemspace) (75 mg, 0.35 mmol) and N-((3-methoxypyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (90 mg, 0.35 mmol) were added to a solution of NaOH (7 mg, 0.5 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 6 (63 mg, 0.13 mmol, 38%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 1H), 8.3 (m, 1H), 8.1 (m, 1H), 7.5 (m, 3H) 7.3 (m, 1H), 7.1 (m, 2H), 4.5 (d, 2H) 4.2 (m, 2H), 3.9 (s, 3H), 3.0 (m, 8H), 1.7 (m, 2H), 1.3 (m, 2H), 0.9 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 477 [M+H]- .

Example No. 47

2-(2-((2-(1-butyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-(trifluoromethyl)pyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-butyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine (Chemspace) (75 mg, 0.35 mmol) and N-((3-(trifluoromethyl)pyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (103 mg, 0.35 mmol) were added to a solution of NaOH (7 mg, 0.5 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the title compound 6 (110 mg, 0.21 mmol, 61%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.8 (m, 1H), 8.5 (m, 2H), 8.2 (m, 1H), 7.5 (m, 3H), 7.15 (m, 2H), 4.7 (d, 2H) 4.1 (t, 2H), 3.0 (m, 8H), 1.65 (m, 2H), 1.3 (m, 2H), 0.9 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 515 [M+H]⁺.

Example No. 8

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-isobutyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-isobutyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.35 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (85 mg, 0.35 mmol) were added to a solution of NaOH (34 mg, 0.86 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound (32 mg, 0.07 mmol, 20%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.6-7.4 (m, 4H), 7.1 (m, 2H), 4.6 (d, 2H), 4.0 (m, 2H), 3.0 (m, 8H), 2.1 (m, 1H), 0.8 (m, 6H) ppm. UHPLC/MS (ESI): [m/z]: 465 [M+H]⁺.

Example No. 14

2-(2-(2-((2-(4-(((3-fluoropyridin-2-yl)methyl)carbamoyl)oxazol-2-yl)ethyl)amino)ethyl)-1H-benzo[d]imidazol-1-yl)acetic acid

2-(2-(2-aminoethyl)-1H-benzo[d]imidazol-1-yl)acetic acid dihydrochloride (Chemspace) (100 mg, 0.34 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (85 mg, 0.34 mmol) were added to a solution of NaOH (48 mg, 1.2 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 14 (47 mg, 0.1 mmol, 30%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6-8.3 (m, 3H), 7.7 (m, 1H), 7.5 (m, 1H), 7.4 (m, 1H), 7.3 (m, 1H), 7.2 (m, 1H), 7.1 (m, 2H), 4.6 (d, 2H), 4.3 (m, 2H), 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 467 [M+H]⁺.

Example No. 15

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-(methylamino)-2-oxoethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(2-(2-aminoethyl)-1H-benzo[d]imidazol-1-yl)-N-methylacetamide dihydrochloride (Chemspace) (100 mg, 0.33 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (81 mg, 0.33 mmol) were added to a solution of NaOH (33 mg, 0.82 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 15 (31 mg, 0.06 mmol, 20%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.7-8.3 (m, 4H), 7.7-7.1 (m, 8H), 4.9 (d, 2H), 4.6 (d, 2H), 4.0 (m, 1H), 3.2 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 480 [M+H]*.

Example No. 16

2-(2-((2-(1-(2-amino-2-oxoethyl)-1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(2-(2-aminoethyl)-1H-benzo[d]imidazol-1-yl)acetamide dihydrochloride (Chemspace) (100 mg, 0.34 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (85 mg, 0.34 mmol) were added to a solution of NaOH (35 mg, 0.86 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 16 (22.9 mg, 0.05 mmol, 15%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.7 (m, 1H), 8.5 (m, 1H), 8.3 (m, 1H), 7.7 (m, 1H), 7.5 (m, 1H), 7.4 (m, 2H), 7.1 (m, 2H), 4.9 (d, 2H), 4.6 (d, 2H), 4.3 (t, 2H), 3.1 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 466 [M+H]⁺.

Example No. 17

2-(2-((2-(1-benzyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-benzyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.31 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (76 mg, 0.31 mmol) were added to a solution of NaOH (31 mg, 0.77 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 17 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.3 (m, 1H), 7.7 (m, 1H), 7.6 (m, 1H), 7.4 (m, 2H), 7.3 (m, 3H), 7.15 (m, 4H), 5.45 (s, 2H), 4.6 (d, 2H), 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 500 [M+H]⁺.

Example No. 21

2-(2-((2-(1-(2-cyclopropylethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(2-cyclopropylethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.33 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (82 mg, 0.33 mmol) were added to a solution of NaOH (33 mg, 0.83 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 21 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.5 (m, 2H), 7.4 (m, 1H), 7.1 (m, 2H), 4.6 (d, 2H), 4.2 (t, 2H), 3.0 (m, 8H), 1.6 (qt, 2H), 0.6 (m, 1H), 0.3 (m, 2H), 0.05 (m, 2H) ppm. UHPLC/MS (ESI): [m/z]: 477 [M+H]⁺.

Example No. 25

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(3-(methylamino)-3-oxopropyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

3-(2-(2-aminoethyl)-1H-benzo[d]imidazol-1-yl)-N-methylpropanamide dihydrochloride (Chemspace) (100 mg, 0.31 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (78 mg, 0.31 mmol) were added to a solution of NaOH (32 mg, 0.78 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 25 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (m, 2H), 8.35 (m, 1H), 7.9 (m, 1H), 7.7 (m, 1H), 7.5 (m, 2H), 7.4 (m, 1H), 7.2 (m, 2H), 4.6 (d, 2H), 4.4 (t, 2H), 3.5 (m, 8H), 2.6 (m, 2H), 1.2 (s, 3H) ppm. UHPLC/MS (ESI): [m/z]: 494 [M+H]*.

Example No. 30

2-(2-((2-(1-(2-(3,5-dimethylisoxazol-4-yl)ethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(2-(3,5-dimethylisoxazol-4-yl)ethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.28 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (70 mg, 0.28 mmol) were added to a solution of NaOH (28 mg, 0.7 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 30 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.3 (m, 1H), 7.7 (m, 1H), 7.5 (m, 1H), 7.4 (m, 2H), 7.1 (m, 2H), 4.6 (d, 2H), 4.3 (t, 2H), 3.6 (t, 2H), 3.0 (m, 5H), 2.8 (m, 3H), 1.95 (s, 3H), 1.8 (s, 3H) ppm. UHPLC/MS (ESI): [m/z]: 495 [M+H]⁺.

Example No. 31

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-(oxazol-2-yl)ethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-(oxazol-2-yl)ethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.3 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (75 mg, 0.33 mmol) were added to a solution of NaOH (30 mg, 0.76 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 31 (14 mg, 0.003 mmol, 9%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (m, 2H), 8.35 (m, 1H), 8.0 (m, 1H), 7.6-7.4 (m, 5H), 7.2 (m, 5H), 4.6 (d, 2H), 4.0 (m, 2H), 4.6 (m, 6H), 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 504 [M+H]⁺.

Example No. 34

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-(1-methyl-1 H-imidazol-2-yl)ethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2 2-(1-(2-(1-methyl-1H-imidazol-2-yl)ethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.29 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (73 mg, 0.29 mmol) were added to a solution of NaOH (29 mg, 0.73 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 34 (88 mg, 0.17 mmol, 59%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (m, 2H), 8.3 (m, 1H), 7.7-7.1 (m, 6H), 7.0 (m, 1H), 6.75 (m, 1H), 4.5 (m, 4H), 3.7 (s, 3H), 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 517 [M+H]⁺.

Example No. 35

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-(pyridin-2-yl)ethyl)-1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-(pyridin-2-yl)ethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine (Chemspace) (100 mg, 0.38 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (93 mg, 0.38 mmol) were added to a solution of NaOH (8 mg, 0.2 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 35 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (m, 3H), 8.3 (m, 1H), 7.7 (m, 2H), 7.5 (m, 1H), 7.4 (m, 2H), 7.2 (m, 1H), 4.6 (d, 2H), 4.55 (t, 2H), 3.2 (t, 2H), 3.0 (m, 6H), 2.8 (t, 2H), 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 514 [M+H]⁺.

Example No. 39

2-(2-((2-(1-butyl-5-(2-methoxyphenyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-butyl-5-(2-methoxyphenyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.25 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (63 mg, 0.25 mmol) were added to a solution of NaOH (25 mg, 0.63 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 39 (88 mg, 0.15 mmol, 62%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.6 (m, 1H), 7.5 (m, 1H), 7.3-7.5 (m, 4H), 7.1 (m, 1H), 7.0 (m, 1H), 4.6 (d, 2H), 4.2 (t, 2H), 3.75 (s, 3H), 3.0 (m, 8H), 1.7 (m, 2H), 1.3 (m, 3H), 0.9 (t, 4H) ppm. UHPLC/MS (ESI): [m/z]: 571 [M+H]⁺.

Example No. 40

2-(2-((2-(1-butyl-6-(2-methoxyphenyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-butyl-6-(2-methoxyphenyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.25 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (62 mg, 0.25 mmol) were added to a solution of NaOH (25 mg, 0.63 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 40 (96 mg, 0.17 mmol, 67%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7-7.0 (m, 9H), 4.6 (d, 2H), 4.2 (t, 2H), 3.7 (s, 3H), 3.0 (m, 8H), 1.7 (m, 2H), 1.3 (m, 4H), 0.9 (m, 4H) ppm. UHPLC/MS (ESI): [m/z]: 571 [M+H]⁺.

Example No. 41

2-(2-((2-(1-butyl-6-phenyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-butyl-6-phenyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.27 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (68 mg, 0.27 mmol) were added to a solution of NaOH (28 mg, 0.68 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 41 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.3 (m, 1H), 7.8 (m, 1H), 7.7 (m, 3H), 7.6 (m, 1H), 7.3-7.5 (m, 5H), 4.6 (d, 2H), 4.2 (t, 2H), 3.0 (m, 8H), 1.7 (m, 2H), 1.3 (m, 2H), 0.9 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 541 [M+H]⁺.

Example No. 42

2-(2-((2-(1-butyl-5-phenyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-butyl-5-phenyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.27 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (68 mg, 0.27 mmol) were added to a solution of NaOH (28 mg, 0.68 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 42 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.3 (m, 1H), 7.8 (m, 1H), 7.7 (m, 3H), 7.55 (m, 1H), 7.5-7.3 (m, 5H), 4.6 (d, 2H), 4.2 (t, 2H), 3.0 (m, 8H), 1.6 (m, 2H), 1.3 (m, 2H), 0.9 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 541 [M+H]⁺.

Example No. 48

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-(2-methoxyethoxy)ethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-(2-methoxyethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.30 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (74 mg, 0.30 mmol) were added to a solution of NaOH (30 mg, 0.74 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 48 (89 mg, 0.19 mmol, 41%) as a pale oil.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.5 (m, 2H), 7.4 (m, 1H), 7.2 (m, 2H), 4.6 (d, 2H), 4.3 (t, 2H), 3.7 (t, 2H), 3.4 (m, 4H), 3.1 (s, 3H), 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 511 [M+H]⁺.

Example No. 49

2-(2-(4-(1-butyl-1 H-benzo[d]imidazol-2-yl)piperidin-1-yl)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

1-butyl-2-(piperidin-4-yl)-1H-benzo[d]imidazole dihydrochloride (Chemspace) (100 mg, 0.3 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (75 mg, 0.3 mmol) were added to a solution of NaOH (30 mg, 0.76 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 49 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.5 (m, 2H), 7.4 (m, 1H), 7.1 (m, 2H), 4.6 (d, 2H), 4.2 (t, 2H), 2.9 (m, 8H), 2.2 (m, 2H), 1.8 (m, 4H), 1.6 (m, 2H), 1.3 (m, 2H), 0.9 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 505 [M+H]⁺.

Example No. 50

2-(2-((2-(1-(Cyclopropylmethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(Cyclopropylmethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.35 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (86 mg, 0.35 mmol) were added to a solution of NaOH (35 mg, 0.87 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 50 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.5 (m, 2H), 7.4 (m, 1H), 7.1 (m, 2H), 4.6 (d, 2H), 4.1 (t, 2H), 3.0 (m, 8H), 1.2 (m, 1H), 045 (m, 4H) ppm. UHPLC/MS (ESI): [m/z]: 463 [M+H]⁺.

Example No. 51

2-(2-((2-(1-(cyclohexylmethyl)-1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(cyclohexylmethyl)-1 H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.3 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (75 mg, 0.3 mmol) were added to a solution of NaOH (30 mg, 0.76 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 51 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.5 (m, 2H), 7.4 (m, 1H), 7.1 (m, 2H), 4.6 (d, 2H), 4.0 (d, 2H), 3.0 (m, 8H), 1.8 (m, 1H), 1.6 (m, 2H), 1.5 (m, 2H), 1.1 (m, 6H) ppm. UHPLC/MS (ESI): [m/z]: 505 [M+H]⁺.

Example No. 52

2-(2-((1-(1-butyl-1 H-benzo[d]imidazol-2-yl)-2-methylpropan-2-yl)amino)ethyl)-N-((3-fluoropyridin-Z-yl)methyl)oxazole-4-carboxamide

1-(1-butyl-1H-benzo[d]imidazol-2-yl)-2-methylpropan-2-amine dihydrochloride (Chemspace) (100 mg, 0.33 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (82 mg, 0.33 mmol) were added to a solution of NaOH (33 mg, 0.82 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 52 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.4 (m, 3H), 7.7 (m, 1H), 7.5 (m, 2H), 7.3 (m, 1H), 7.15 (m, 2H), 4.6 (d, 2H), 4.2 (t, 2H), 3.2 (s, 6H), 1.6 (m, 3H), 1.3 (m, 6H), 0.9 (m, 4H) ppm. UHPLC/MS (ESI): [m/z]: 493 [M+H]⁺.

Example No. 53

2-(2-((2-(1-(2,2-difluoroethyl)-1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(2,2-difluoroethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.34 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (83 mg, 0.34 mmol) were added to a solution of NaOH (34 mg, 0.84 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 53 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.5 (m, 2H), 7.4 (m, 1H), 7.2 (m, 2H), 6.4 (m, 1H), 4.8 (m, 2H), 4.6 (d, 2H), 4.0 (d, 2H), 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 473 [M+H]⁺.

Example No. 54

2-(2-((2-(1-(2-chlorobenzyl)-1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(2-chlorobenzyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.28 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (69 mg, 0.28 mmol) were added to a solution of NaOH (28 mg, 0.70 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 54 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.6 (m, 1H), 7.5 (m, 1H), 7.4 (m, 1H), 7.3 (m, 2H), 7.1 (m, 3H), 5.5 (s, 2H), 4.6 (d, 2H) 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 534 [M+H]⁺.

Example No. 55

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-phenyl-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-phenyl-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.32 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (80 mg, 0.32 mmol) were added to a solution of NaOH (32 mg, 0.81 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 55 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7-7.5 (m, 7H), 7.4 (m, 1H), 7.2 (m, 2H), 7.05 (m, 1H), 4.6 (d, 2H) 2.9 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 485 [M+H]⁺.

Example No. 56

2-(2-((2-(1-(2-(dimethylamino)ethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(2-(2-aminoethyl)-1H-benzo[d]imidazol-1-yl)-N,N-dimethylethan-1-amine trihydrochloride (Chemspace) (100 mg, 0.29 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (72 mg, 0.29 mmol) were added to a solution of NaOH (41 mg, 1 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 56 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.3 (m, 1H), 7.7-7.5 (m, 3H), 7.4 (m, 1H), 7.2 (m, 2H), 4.6 (d, 2H), 4.3 (t, 2H), 2.6 (m, 2H), 3.1 (m, 8H), 2.2 (s, 3H), 1.9 (s, 3H) ppm. UHPLC/MS (ESI): [m/z]: 481 [M+H]⁺.

Example No. 57

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-isopropoxyethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-isopropoxyethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.31 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (77 mg, 0.31 mmol) were added to a solution of NaOH (31 mg, 0.78 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 57 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.3 (m, 1H), 7.7 (m, 1H), 7.5 (m, 2H), 7.4 (m, 1H), 7.1 (m, 2H), 4.6 (d, 2H), 4.3 (t, 2H), 3.6 (t, 2H), 3.4 (m, 1H), 3.1 (m, 8H), 0.9 (2xs, 6H) ppm. UHPLC/MS (ESI): [m/z]: 495 [M+H]⁺.

Example No. 58

2-(2-((2-(1-(4-chlorobenzyl)-1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(4-chlorobenzyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.28 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (69 mg, 0.28 mmol) were added to a solution of NaOH (28 mg, 0.7 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 58 (89 mg, 0.19 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.3 (m, 1H), 7.7 (m, 1H), 7.5 (m, 1H), 7.4 (m, 4H), 7.1 (m, 4H), 5.5 (s, 2H), 4.6 (d, 2H), 3.0 (m, 8H), 1.95 (s, 3H), 1.8 (s, 3H) ppm. UHPLC/MS (ESI): [m/z]: 534 [M+H]⁺.

Example No. 60

2-(2-((2-(1-butyl-1 H-benzo[d]imidazol-2-yl)-2-methylpropyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-butyl-1H-benzo[d]imidazol-2-yl)-2-methylpropan-1-amine dihydrochloride (Chemspace) (100 mg, 0.3 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (78 mg, 0.3 mmol) were added to a solution of NaOH (31 mg, 0.79 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 60 (84 mg, 0.17 mmol, 57%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 7.7 (m, 1H), 7.5 (m, 1H), 7.4 (m, 2H), 7.1 (m, 2H), 4.6 (d, 2H), 4.3 (m, 2H), 2.9 (m, 4H), 1.7 (m, 2H), 1.4 (m, 6), 1.3 (m, 2H), 1.0 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 493 [M+H]⁺.

Example No. 61

2-(2-(((1-(1-butyl-1H-benzo[d]imidazol-2-yl)cyclohexyl)methyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

(1-(1-butyl-1H-benzo[d]imidazol-2-yl)cyclohexyl)methanamine dihydrochloride (Chemspace) (100 mg, 0.28 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (69 mg, 0.28 mmol) were added to a solution of NaOH (28 mg, 0.7 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 61 (63 mg, 0.12 mmol, 42%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.4 (m, 2H), 7.7 (m, 1H), 7.5 (m, 1H), 7.4 (m, 2H), 7.1 (m, 2H), 4.6 (d, 2H), 4.3 (m, 2H), 2.9 (m, 4H), 2.3 (m, 1), 1.7-1.2 (m, 12H), 1.0 (t, 3H) ppm. UHPLC/MS (ESI): [m/z]: 533 [M+H]⁺.

Example No. 62

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(pyrimidin-4-ylmethyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(pyrimidin-4-ylmethyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.31 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (76 mg, 0.31 mmol) were added to a solution of NaOH (31 mg, 0.77 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 62 (64 mg, 0.13 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 9.1 (m, 1H), 8.8 (d, 1H), 8.5 (m, 2H), 8.3 (m, 1H), 7.7-7.3 (m, 5H), 7.1 (m, 2H), 7.15 (m, 4H), 5.7 (d, 2H), 4.6 (d, 2H), 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 501 [M+H]⁺.

Example No. 63

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(3-methoxyphenyl)-1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(3-methoxyphenyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.3 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (76 mg, 0.3 mmol) were added to a solution of NaOH (30 mg, 0.73 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 63 (68 mg, 0.13 mmol, 44%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7-7.4 (m, 4H), 7.2-7.05 (m, 6H), 4.6 (d, 2H), 3.8 (s, 3H), 2.9 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 515 [M+H]⁺.

Example No. 64

2-(2-((2-(1-(3-chlorophenyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(3-chlorophenyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (103 mg, 0.27 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (67 mg, 0.27 mmol) were added to a solution of NaOH (38 mg, 0.94 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 64 (67 mg, 0.13 mmol, 48%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7-7.1 (m, 10H), 4.6 (d, 2H), 2.9 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 520 [M+H]⁺.

Example No. 65

2-(2-((2-(1-(2-(2-methoxyethoxy)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-methoxypyridin-2-yl)methyl)oxazole-4-carboxamide (VIT-101207nda152)

2-(1-(2-(2-methoxyethoxy)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.29 mmol) and N-((3-methoxypyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (72 mg, 0.29 mmol) were added to a solution of NaOH (29 mg, 0.72 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 65 (56 mg, 0.11 mmol, 37%) as a pale oil.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.4 (m, 1H), 7.7 (m, 1H), 7.4 (m, 1H), 7.3 (m, 1H), 7.0 (s, 1H), 6.9 (s, 1H), 4.6 (d, 2H), 4.2 (m, 4H), 4.0 (d, 2H), 3.0 (m, 8H), 1.2 (m, 2H), 0.4 (m, 4H) ppm. UHPLC/MS (ESI): [m/z]: 521 [M+H]⁺.

Example No. 66

2-(2-((2-(1-(2-(2-methoxyethoxy)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-methoxypyridin-2-yl)meth yl)oxazole-4-carboxamide

2-(1-(2-(2-methoxyethoxy)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.25 mmol) and N-((3-methoxypyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (66 mg, 0.25 mmol) were added to a solution of NaOH (25 mg, 0.64 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 66 (38 mg, 0.07 mmol, 26%) as a pale oil.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (s, 1H), 8.4 (m, 1H), 8.0 (m, 1H), 7.4 (m, 1H), 7.3 (m, 1H), 7.0 (s, 1H), 6.9 (s, 1H), 4.5 (m, 2H), 4.2 (m, 6H), 3.85 (s, 3H), 3.65 (m, 2H), 3.45 (s, 3H), 3.1 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 581 [M+H]⁺.

Example No. 67

2-(2-((2-(1-(2-methoxyethyl)-6,7-dihydro-1H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-methoxypyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(2-methoxyethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.3 mmol) and N-((3-methoxypyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (74 mg, 0.3 mmol) were added to a solution of NaOH (29 mg, 0.7 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 67 (58 mg, 0.11 mmol, 36%) as a pale oil.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (m, 1H), 8.4 (m, 1H), 8.0 (m, 1H), 7.4 (m, 1H), 7.2 (m, 2H), 7.0 (m, 2H), 5.95 (s, 1H), 4.5 (m, 2H), 4.2 (m, 6H), 3.85 (s, 3H), 3.6 (m, 2H), 3.2 (s, 3H), 3.1 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 537 [M+H]⁺.

Example No. 68

2-(2-((2-(1-(2,2-difluoroethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-methoxypyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(2,2-difluoroethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.28 mmol) and N-((3-methoxypyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (73 mg, 0.28 mmol) were added to a solution of NaOH (28 mg, 0.7 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 68 (20 mg, 0.04 mmol, 14%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (m, 1H), 8.3 (m, 1H), 8.1 (m, 1H), 7.4 (m, 1H), 7.3 (m, 1H), 7.05 (s, 1H), 6.95 (s, 1H), 6.35 (m, 1H), 4.6 (m, 2H), 4.5 (d, 2H), 4.2 (m, 4H), 3.8 (s, 3H), 3.1 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 543 [M+H]⁺.

Example No. 69

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(5-(2-methoxyethyl)-5H-[1,3]dioxolo[4′,5′:4,5]benzo[1,2-d]imidazol-6-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(5-(2-methoxyethyl)-5H-[1,3]dioxolo[4′,5′:4,5]benzo[1,2-d]imidazol-6-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.3 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (74 mg, 0.3 mmol) were added to a solution of NaOH (30 mg, 0.75 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 69 (84 mg, 0.17 mmol, 55%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.4 (m, 1H), 7.15 (s, 1H), 7.0 (s, 1H), 5.95 (s, 1H), 4.6 (m, 2H), 4.3 (m, 2H), 3.6 (m, 2H), 3.2 (s, 3H), 3.1 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 511 [M+H]⁺.

Example No. 70

2-(2-((2-(5-(2-methoxyethyl)-5H-[1,3]dioxolo[4′,5′:4,5]benzo[1,2-d]imidazol-6-yl)ethyl)amino)ethyl)-N-((3-methoxypyridin-2-yl)methyl)oxazole-4-carboxamide

2-(5-(2-methoxyethyl)-5H-[1,3]dioxolo[4′,5′:4,5]benzo[1,2-d]imidazol-6-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.3 mmol) and N-((3-methoxypyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (77 mg, 0.3 mmol) were added to a solution of NaOH (30 mg, 0.75 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 70 (87 mg, 0.17 mmol, 55%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.35 (m, 1H), 8.1 (m, 1H), 7.45 (m, 1H), 7.30 (m, 1H), 7.1 (s, 1H), 7.0 (s, 1H), 5.95 (s, 2H), 4.5 (m, 2H), 4.3 (m, 2H), 3.85 (s, 3H), 3.55 (m, 2H), 3.2 (s, 3H), 3.1 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 523 [M+H]⁺.

Example No. 71

2-(2-((2-(1-(2,2-difluoroethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(2,2-difluoroethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.28 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (70 mg, 0.28 mmol) were added to a solution of NaOH (28 mg, 0.7 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 71 (56 mg, 0.11 mmol, 38%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.4 (m, 1H), 7.7 (m, 1H), 7.4 (m, 1H), 7.0 (m, 2H), 6.4 (m, 1H), 4.6 (m, 4H), 4.2 (m, 4H), 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 531 [M+H]⁺.

Example No. 72

2-(2-((2-(5,6-dimethoxy-1-(3-methoxyphenyl)-1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(5,6-dimethoxy-1-(3-methoxyphenyl)-1 H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.25 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (62 mg, 0.25 mmol) were added to a solution of NaOH (25 mg, 0.63 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 72 (59 mg, 0.1 mmol, 41%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.45 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.5 (m, 1H), 7.4 (m, 2H), 7.2 (s, 1H), 7.1 (m, 3H), 6.6 (s, 1H), 4.6 (d, 2H), 3.8 (2xs, 6H), 3.7 (s, 3H), 2.9 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 575 [M+H]⁺.

Example No. 73

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-isopropoxyethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-isopropoxyethyl)-6,7-dihydro-1H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.26 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (65 mg, 0.26 mmol) were added to a solution of NaOH (26 mg, 0.66 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 73 (32 mg, 0.06 mmol, 23%) as a pale oil.

¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.4 (m, 1H), 7.0 (m, 2H), 4.6 (d, 2H), 4.2 (m, 6H), 3.6 (m, 2H), 3.2 (m, 8H), 0.9 (d, 6H) ppm. UHPLC/MS (ESI): [m/z]: 552 [M+H]⁺.

Example No. 74

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-(piperidin-1-yl)ethyl)-6,7-dihydro-1H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-(piperidin-1-yl)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine trihydrochloride (Chemspace) (100 mg, 0.23 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (56 mg, 0.23 mmol) were added to a solution of NaOH (32 mg, 0.8 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 74 (49 mg, 0.09 mmol, 37%) as a pale oil.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.3 (m, 1H), 7.7 (m, 1H), 7.4 (m, 1H), 7.0 (m, 2H), 4.6 (d, 2H), 4.2 (m, 6H), 3.2 (m, 8H), 2.4 (m, 4H), 1.9 (m, 2H), 1.5-1.2 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 578 [M+H]⁺.

Example No. 75

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(5-(3-methoxyphenyl)-5H-[1,3]dioxolo[4′,5′:4,5]benzo[1,2-d]imidazol-6-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(5-(3-methoxyphenyl)-5H-[1,3]dioxolo[4′,5′:4,5]benzo[1,2-d]imidazol-6-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.26 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (64 mg, 0.26 mmol) were added to a solution of NaOH (26 mg, 0.65 mmol) in 10 ml water.

The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 75 (53 mg, 0.1 mmol, 37%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.4 (m, 1H), 7.15-7.0 (m, 4H), 6.6 (s, 1H), 6.0 (s, 2H), 4.6 (d, 2H), 3.8 (m, 3H), 2.9 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 559 [M+H]⁺.

Example No. 76

2-(2-((2-(1-(2-(dimethylamino)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-methoxypyridin-2-yl)methyl)oxazole-4-carboxamide

2-(2-(2-aminoethyl)-6,7-dihydro-1H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-1-yl)-N,N-dimethylethan-1-amine trihydrochloride (Chemspace) (100 mg, 0.25 mmol) and N-((3-methoxypyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (65 mg, 0.25 mmol) were added to a solution of NaOH (35 mg, 0.88 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 76 (42 mg, 0.08 mmol, 31%) as a pale oil.

¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (m, 1H), 8.3 (m, 1H), 8.2 (m, 2H), 7.4 (m, 1H), 7.3 (m, 1H), 6.9 (m, 2H), 4.5 (d, 2H), 4.25-4.1 (m, 6H), 3.85 (s, 3H), 3.1 (m, 8H), 2.1 (m, 5H) ppm. UHPLC/MS (ESI): [m/z]: 550 [M+H]⁺.

Example No. 77

2-(2-((2-(1-(2-(dimethylamino)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(2-(2-aminoethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-1-yl)-N,N-dimethylethan-1-amine trihydrochloride (Chemspace) (100 mg, 0.25 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (62 mg, 0.25 mmol) were added to a solution of NaOH (35 mg, 0.88 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 77 (53 mg, 0.1 mmol, 39%) as a pale oil.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.4 (m, 1H), 8.15 (m, 2H), 7.7 (m, 1H), 7.4 (m, 1H), 6.9 (m, 2H), 4.6 (d, 2H), 4.2 (m, 6H), 3.0 (m, 8H), 2.1 (m, 4H) ppm. UHPLC/MS (ESI): [m/z]: 538 [M+H]⁺.

Example No. 81

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-morpholinoethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-morpholinoethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine trihydrochloride (Chemspace) (100 mg, 0.23 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (56 mg, 0.23 mmol) were added to a solution of NaOH (32 mg, 0.79 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 81 (61 mg, 0.11 mmol, 46%) as a pale oil.

¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.4 (m, 1H), 6.9 (m, 2H), 4.6 (d, 2H), 4.2 (m, 6H), 3.5 (m, 4H), 3.1 (m, 8H), 2.55 (m, 2H), 2.4 (m, 4H) ppm. UHPLC/MS (ESI): [m/z]: 580 [M+H]⁺.

Example No. 82

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(5-phenyl-5H-[1,3]dioxolo[4′,5′:4,5]benzo[1,2-d]imidazol-6-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(5-phenyl-5H-[1,3]dioxolo[4′,5′:4,5]benzo[1,2-d]imidazol-6-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.28 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (70 mg, 0.28 mmol) were added to a solution of NaOH (28 mg, 0.71 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 82 (40 mg, 0.08 mmol, 27%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (m, 2H), 7.8-7.4 (m, 7H), 7.2 (s, 1H), 6.6 (s, 1H), 6.0 (m, 2H), 4.5 (d, 2H), 3.4 (m, 2H), 2.9 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 529 [M+H]⁺.

Example No. 83

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-phenyl-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-phenyl-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.27 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (67 mg, 0.27 mmol) were added to a solution of NaOH (27 mg, 0.68 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 83 (43 mg, 0.08 mmol, 29%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (m, 2H), 8.4 (m, 1H), 7.8-7.4 (m, 7H), 7.0 (s, 1H), 6.5 (s, 1H), 4.5 (d, 2H), 4.1 (m, 4H), 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 543 [M+H]⁺.

Example No. 84

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-(4-methylpiperazin-1-yl)ethyl)-6,7-dihydro-1H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-(4-methylpiperazin-1-yl)ethyl)-6,7-dihydro-1H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine trihydrochloride (Chemspace) (100 mg, 0.22 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (59 mg, 0.24 mmol) were added to a solution of NaOH (24 mg, 0.6 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 84 (31 mg, 0.052 mmol, 24%) a pale oil.

¹H NMR (400 MHz, DMSO-d₆) δ 8.7-8.6 (m, 2H), 8.4 (m, 1H), 7.7 (m, 1H), 7.4 (m, 1H), 7.1-7.0 (m, 3H), 4.6 (d, 2H), 4.2 (m, 8H), 3.5 (m, 4H), 3.3 (m, 8H), 2.6 (m, 4H) ppm. UHPLC/MS (ESI): [m/z]: 593 [M+H]⁺.

Example No. 85

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-phenyl-5-(pyridin-2-ylethynyl)-1H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-phenyl-5-(pyridin-2-ylethynyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine trihydrochloride (Chemspace) (100 mg, 0.22 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (55 mg, 0.22 mmol) were added to a solution of NaOH (31 mg, 0.78 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 85 (42 mg, 0.07 mmol, 31%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6-8.3 (m, 3H), 7.9-7.8 (m, 2H), 7.6 (m, 7H), 7.4 (m, 3H), 7.1 (m, 1H), 4.6 (d, 2H), 2.9 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 585 [M+H]⁺.

Example No. 86

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-methoxyethyl)-5-(pyridin-2-ylethynyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-methoxyethyl)-5-(pyridin-2-ylethynyl)-1H-benzo[d]imidazol-2-yl)ethan-1-amine trihydrochloride (Chemspace) (100 mg, 0.23 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (58 mg, 0.23 mmol) were added to a solution of NaOH (33 mg, 0.82 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 86 (33 mg, 0.06 mmol, 25%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6-8.4 (m, 3H), 7.8-7.6 (m, 5H), 7.4 (m, 3H), 4.6 (d, 2H), 4.4 (t, 2H), 3.6 (t, 2H), 3.2 (s, 3H), 3.0 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 568 [M+H]⁺.

Example No. 87

2-(2-((2-(1-butyl-5,6-dimethoxy-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-butyl-5,6-dimethoxy-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.29 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (71 mg, 0.29 mmol) were added to a solution of NaOH (29 mg, 0.71 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 87 (29 mg, 0.006 mmol, 19%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.4 (m, 1H), 7.0 (m, 2H), 6.55 (m, 1H), 4.6 (d, 4H), 4.1 (m, 2H), 3.8 (s, 3H), 3.7 (s, 3H), 2.9 (m, 8H), 1.65 (m, 2H), 1.3 (m, 2H), 0.9 (m, 3H) ppm. UHPLC/MS (ESI): [m/z]: 525 [M+H]⁺.

Example No. 88

2-(2-((2-(1-(2-(cyclohexyloxy)ethyl)-6,7-dihydro-1H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(2-(cyclohexyloxy)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.24 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (59 mg, 0.24 mmol) were added to a solution of NaOH (24 mg, 0.6 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 88 (38 mg, 0.064 mmol, 27%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.35 (m, 1H), 7.7 (m, 1H), 7.4 (m, 1H), 6.95 (m, 2H), 4.6 (d, 2H), 4.2 (m, 6H), 3.6 (m, 2H), 3.1 (m, 1H), 1.6-1.0 (m, 10H) ppm. UHPLC/MS (ESI): [m/z]: 593 [M+H]⁺.

Example No. 89

2-(2-((2-(1-(cyclopropylmethyl)-5-(pyridin-2-ylethynyl)-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(cyclopropylmethyl)-5-(pyridin-2-ylethynyl)-1 H-benzo[d]imidazol-2-yl)ethan-1-amine trihydrochloride (Chemspace) (100 mg, 0.24 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (58 mg, 0.24 mmol) were added to a solution of NaOH (33 mg, 0.82 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 89 (45 mg, 0.08 mmol, 33%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6-8.5 (m,3H), 8.35 (m, 1H), 7.85 (m, 2H), 7.65 (m, 3H), 7.4 (m, 3H), 4.6 (d, 2H), 4.2 (d, 2H), 3.1 (m, 8H), 1.2 (m, 1H), 0.5 (m, 4H) ppm. UHPLC/MS (ESI): [m/z]: 564 [M+H]⁺.

Example No. 90

2-(2-((2-(1-(3,4-dimethoxyphenyl)-5,6-dimethoxy-1 H-benzo[d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(3,4-dimethoxyphenyl)-5,6-dimethoxy-1H-benzo[d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (200 mg, 0.47 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (115 mg, 0.47 mmol) were added to a solution of NaOH (46 mg, 1.16 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 90 (81 mg, 0.13 mmol, 29%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.4 (m, 1H), 8.2 (m, 1H), 8.0 (m, 1H), 7.6-7.2 (m, 3H), 7.0 (m, 4H), 7.0 (m, 4H), 6.55 (m, 1H), 4.5 (m, 4H), 4.2 (m, 4H), 3.9 (s, 3H), 3.8 (s, 3H), 2.9 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 605 [M+H]⁺.

Example No. 91

2-(2-((2-(1-(2-(cyclohexyloxy)ethyl)-6,7-dihydro-1H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-methoxypyridin-2-yl)methyl)oxazole-4-carboxamide

22-(1-(2-(cyclohexyloxy)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.24 mmol) and N-((3-methoxypyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (62 mg, 0.24 mmol) were added to a solution of NaOH (24 mg, 0.6 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 91 (41 mg, 0.068 mmol, 28%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (s, 1H), 8.3 (m, 1H), 8.1 (m, 1H), 7.4 (m, 1H), 7.3 (m, 1H), 6.95 (m, 2H), 4.5 (m, 2H), 4.2 (m, 8H), 3.85 (s, 3H), 3.6 (m, 1H), 3.2 (m, 8H), 1.7-1.0 (m, 10H) ppm. UHPLC/MS (ESI): [m/z]: 605 [M+H]⁺.

Example No. 92

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(3-methoxyphenyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(3-methoxyphenyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.25 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (62 mg, 0.25 mmol) were added to a solution of NaOH (25 mg, 0.63 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 92 (38 mg, 0.066 mmol, 27%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.4 (m, 1H), 8.2 (m, 1H), 8.0 (m, 1H), 7.6-7.2 (m, 3H), 7.0 (m, 4H), 7.0 (m, 4H), 6.55 (m, 1H), 4.5 (m, 4H), 4.2 (m, 4H), 3.9 (s, 3H), 3.8 (s, 3H), 2.9 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 573 [M+H]⁺.

Example No. 93

2-(2-((2-(1-(3-methoxyphenyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)-N-((3-methoxypyridin-2-yl)methyl)oxazole-4-carboxamide

2-(1-(3-methoxyphenyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.25 mmol) and N-((3-methoxypyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (65 mg, 0.25 mmol) were added to a solution of NaOH (25 mg, 0.63 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 93 (23 mg, 0.04 mmol, 16%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.5 (m, 2H), 8.4 (m, 1H), 7.7 (m, 1H), 7.5 (m, 1H), 7.4 (m, 1H), 7.0 (m, 4H), 6.5 (m, 1H), 4.6 (m, 2H), 4.2 (m, 4H), 3.8 (s, 3H), 2.9 (m, 8H) ppm. UHPLC/MS (ESI): [m/z]: 585 [M+H]⁺.

Example No. 94

N-((3-fluoropyridin-2-yl)methyl)-2-(2-((2-(1-(2-(oxazol-2-yl)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-(oxazol-2-yl)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.26 mmol) and N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (58 mg, 0.24 mmol) were added to a solution of NaOH (33 mg, 0.83 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 94 (42 mg, 0.075 mmol, 29%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (m, 2H), 8.3 (m, 1H), 8.0 (m, 1H), 7.7 (m, 1H), 7.4 (m, 1H), 7.1 (m, 1H), 7.0 (m, 2H), 4.6 (m, 2H), 4.4 (m, 2H), 4.2 (m, 4H), 3.2 (m, 15H) ppm. UHPLC/MS (ESI): [m/z]: 562 [M+H]⁺.

Example No. 95

N-((3-methoxypyridin-2-yl)methyl)-2-(2-((2-(1-(2-(oxazol-2-yl)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethyl)amino)ethyl)oxazole-4-carboxamide

2-(1-(2-(oxazol-2-yl)ethyl)-6,7-dihydro-1 H-[1,4]dioxino[2′,3′:4,5]benzo[1,2-d]imidazol-2-yl)ethan-1-amine dihydrochloride (Chemspace) (100 mg, 0.26 mmol) and N-((3-methoxypyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (67 mg, 0.26 mmol) were added to a solution of NaOH (36 mg, 0.9 mmol) in 10 ml water. The resulting mixture were heated at 80° C. under stirring for 72 hours. After cooling at room temperature, the reaction mixture was adjusted to pH 6 by using 2N HCl and extracted with dichloromethane (3×10 ml). The combined organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH) to obtain the titled compound 95 (36 mg, 0.063 mmol, 24%) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.6 (m, 1H), 8.3 (m, 1H), 8.1 (m, 1H), 8.0 (m, 1H), 7.4 (m, 1H), 7.3 (m, 1H), 7.1 (m, 1H), 6.95 (m, 2H), 4.5 (m, 4H), 4.2 (m, 4H), 3.8 (s, 3H), 3.2 (m, 10H), 2.95 (m, 2H) ppm. UHPLC/MS (ESI): [m/z]: 574 [M+H]⁺.

Intermediate Compound for the preparation of Example Compounds No. 96 and 97 2-(2-(3-(1 H-benzo[d]imidazol-2-yl)azetidin-1-yl)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

2-(Azetidin-3-yl)-1H-benzo[d]imidazole (210 mg,1.21 mmol, 1 eq.) was suspended in H₂O (12 mL) and treated with NaOH (30% Wt, 0.18 g, 1.33 mmol, 1.1 eq.). After the material dissolved completely, N-((3-fluoropyridin-2-yl)methyl)-2-vinyloxazole-4-carboxamide (300 mg, 1.21 mmol, 1 eq.) was added. The reaction mixture was heated to 80° C. until LC/MS indicated full conversion of the starting material. The reaction mixture was acidified to pH=5-6 with 1M HCl and then concentrated under reduced pressure.

The crude material was re-dissolved in methanol, concentrated on RP-silica under reduced pressure and purified by RP18 flash column chromatography yielding in the desired 2-(2-(3-(1H-benzo[d]imidazol-2-yl)azetidin-1-yl)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide (160 mg, 379 μmol, 31%) as a white solid. LCMS (ESI) m/z=421.6.

¹H NMR (400 MHz, DMSO) δ 11.14 (s, 1H), 8.62 (t, J=5.8 Hz, 1H), 8.59 (s, 1H), 8.36 (d, J=4.7 Hz, 1H), 7.69 (ddd, J=10.0, 8.3, 1.3 Hz, 1H), 7.45-7.36 (m, 2H), 7.30 (d, J=8.0 Hz, 1H), 7.01 (ddd, J=8.2, 7.0, 1.2 Hz, 1H), 6.93 (ddd, J=8.0, 7.0, 1.1 Hz, 1H), 4.62 (dd, J=5.7, 1.8 Hz, 2H), 3.34 (ddt, J=30.6, 14.0, 7.1 Hz, 7H), 3.15 (t, J=7.8 Hz, 2H) ppm.

Example No. 96

N-((3-fluoropyridin-2-yl)methyl)-2-(2-(3-(1-(2-methoxyethyl)-1H-benzo[d]imidazol-2-yl)azetidin-1-yl)ethyl)oxazole-4-carboxamide

Under an inert atmosphere 2-(2-(3-(1H-benzo[d]imidazol-2-yl)azetidin-1-yl)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide (115 mg, 274 μmol, 1 eq.) was dissolved in N,N-dimethylformamide (3 mL) and cooled to 0° C. NaH (60% Wt, 14 mg, 342 μmol, 1.25 eq.) was added. The reaction mixture was stirred 30 min at 0° C. before 1-bromo-2-methoxyethane (28.3 μL, 301 μmol, 1.1 eq.) was added. The ice-bath was removed and the reaction mixture was allowed to warm-up overnight. The reaction was quenched by the addition of aq. sat. ammonium chloride solution. The mixture was concentrated under reduced pressure. The concentrate was re-dissolved in a mixture of dichloromethane and water. Phases were separated and the aqueous phase was re-extracted with dichloromethane (3×). The combined organic phase was washed with water and brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH, 0->20% MeOH) affording the desired N-((3-fluoropyridin-2-yl)methyl)-2-(2-(3-(1-(2-methoxyethyl)-1H-benzo[d]imidazol-2-yl)azetidin-1-yl)ethyl)oxazole-4-carboxamide (25 mg, 52 μmol, 19%) as a yellowish oil. LCMS (ESI) m/z=479.6.

¹H NMR (400 MHz, MeOD) δ 8.36-8.30 (m, 2H), 7.65-7.61 (m, 1H), 7.57 (ddd, J=9.8, 8.3, 1.3 Hz, 1H), 7.49-7.44 (m, 1H), 7.35 (dt, J=8.7, 4.5 Hz, 1H), 7.29-7.22 (m, 3H), 4.74 (d, J=1.8 Hz, 2H), 4.34 (t, J=5.0 Hz, 2H), 4.17 (p, J=8.2 Hz, 1H), 3.88 (ddd, J=8.0, 6.4, 1.7 Hz, 2H), 3.66-3.56 (m, 4H), 3.19 (s, 3H), 3.11 (q, J=5.7 Hz, 2H), 2.98 (t, J=6.7 Hz, 2H) ppm.

Example No. 97

2-(2-(3-(1-(2-(dimethylamino)ethyl)-1H-benzo[d]imidazol-2-yl)azetidin-1-yl)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide

Under an inert atmosphere 2-(2-(3-(1H-benzo[d]imidazol-2-yl)azetidin-1-yl)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide (260 mg, 618 μmol, 1 eq.) was dissolved in DMF (3 mL). Potassium carbonate (256 mg, 1.86 mmol, 3 eq.) was added. The reaction mixture was stirred 10 min at 23° C., before 2-bromo-N,N-dimethylethan-1-amine hydrobromide (187 mg, 1.3 Eq, 804 μmol) was added. The reaction mixture was stirred at 23° C. for 16 hours. The reaction was concentrated under reduced pressure. The crude material was purified by flash column chromatography (CH₂Cl₂/MeOH, 0->20% MeOH) affording the desired 2-(2-(3-(1-(2-(dimethylamino)ethyl)-1H-benzo[d]imidazol-2-yl)azetidin-1-yl)ethyl)-N-((3-fluoropyridin-2-yl)methyl)oxazole-4-carboxamide (21 mg, 43 μmol, 7%) as a brownish solid. LCMS (ESI) m/z=492.5.

¹H NMR (400 MHz, DMSO) δ 8.37 (d, J=4.6 Hz, 1H), 8.24 (t, J=5.6 Hz, 1H), 8.10 (s, 1H), 7.75-7.62 (m, 1H), 7.45-7.35 (m, 2H), 7.13-7.04 (m, 2H), 6.33 (d, J=2.1 Hz, 1H), 4.54 (dd, J=5.6, 1.7 Hz, 2H), 3.51 (s, 2H), 2.91 (s, 4H), 2.70 (t, J=6.0 Hz, 2H), 2.59 (t, J=6.1 Hz, 2H), 2.32 (s, 6H) ppm. (3H covered by H₂O).

Claims

1. A compound according to formula (I-A), or a pharmaceutically acceptable salt thereof;

2. The compound or pharmaceutically acceptable salt thereof according to claim 1 which is a compound according to formula (I-B), or a pharmaceutically acceptable salt thereof;

3. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein 1 is an integer of 1 or 2;m and n are independently an integer of 1, 2 or 3;X1is N, S or O;X2 is N, S, O or CR5; andX3 is C or N;with the proviso that one of X1 and X2 is Nand if X3 is N, then X2 is CRS;and whereinR5 represents H;A represents a group (a-1)

4. The compound or pharmaceutically acceptable salt thereof according to claim 1 wherein the group B is a group (b-1) or (b-2).

5. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein X1, X2 and X3 are selected to form one of the following groups:

6. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the group A has the following structure.

7. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R3 represents H,C1-C3-alkoxy,pyridinylethinyl,unsubstituted or substituted phenyl,wherein a substituted phenyl group can carry 1, 2 or 3 substituents independently selected from halogen,C1-C3-alkyl,C1-C3-haloalkyl, andC1-C3-alkoxy;and/orR4 represents linear or branched C1-C6-alkyl,a dialkylether group [R6(CH2)x—O—CH2)y—] with R6 representing a C1-C3-alkoxy group and with x and y independently representing an integer of 1, 2 or 3, or5— or 6-membered unsubstituted heterocyclyl,substituted or unsubstituted phenyl,wherein substituents of phenyl are selected from halogen, andC1-C3-alkoxy; andunsubstituted or substituted 5— or 6-membered heterocyclyl wherein alkyl can be substituted with 1 or 2 substituents, independently selected from halogen,C1-C3-alkoxy,cyclohexyloxy,carboxyl,aminocarbonyl,mono-alkylaminocarbonyl,dialkylamino,cyclopropyl or cyclohexyl,6-membered heterocyclyl,unsubstituted or substituted phenyl,unsubstituted or substituted 5— or 6-membered heteroaryl, andunsubstituted or substituted bicyclic heteroaryl,wherein a substituted phenyl, heteroaryl and bicyclic heteroaryl group can carry 1, 2 or 3 substituents independently selected from halogen, C1-C3-alkyl,C1-C3-haloalkyl,C1-C3-alkoxyaminocarbonyl, andmono-alkylaminocarbonyl,wherein a mono-alkylaminocarbonyl group may carry a further substituent on the mono-alkyl-chain, selected from halogen-substituted 5— or 6-membered heteroaryl.

8. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein halogen substituents are selected from F, Cl and Br; and/orlinear or branched C1-C6-alkyl substituents are selected from methyl, ethyl, propyl, iso-propyl, n-butyl and iso-butyl; and/orC1-C3-alkoxy substituents are selected from methoxy and ethoxy; and/orC1-C3-haloalkyl substituents are selected from difluoroethyl (—CH2—CHF2) and trifluoromethyl (CF3); and/ora substituted alkyl-group in the position R4 represents a substituted C1-C3-alkyl group; and/ora bicyclic heteroaryl group is selected from a benzimidazolyl group.

9. The compound or pharmaceutically acceptable salt thereof according to claim 1, which is selected from

10. A medicament comprising the compound or pharmaceutically acceptable salt thereof according to claim 1.

11. A method comprising administering the compound or pharmaceutically acceptable salt thereof according to claim 1 to a patient to inhibit iron transport mediated by ferroportin.

12. A method for prophylaxis and/or treatment of an iron metabolism disorder leading to increased iron levels or increased iron absorption, and/or iron overload, comprising administering the compound or pharmaceutically acceptable salt thereof according to claim 1 to a patient in need thereof.

13. A method for prophylaxis and/or treatment of a disease related to or caused by increased iron levels, increased iron absorption or iron overload, selected from thalassemia, including alpha-thalassemia, beta-thalassemia and delta-thalassemia, hemoglobinopathy, hemoglobin E disease, hemoglobin H disease, haemochromatosis, hemolytic anemia, including sickle cell anemia or congenital dyserythropoietic anemia, comprising administering the compound or pharmaceutically acceptable salt thereof according to claim 1 to a patient in need thereof.

14. A method for prophylaxis and/or treatment of a disease or condition comprising administering the compound or pharmaceutically acceptable salt thereof according to claim 1 to a patient having the disease or condition, wherein the disease or condition is: a disease associated with ineffective erythropoiesis and/ora disease caused by reduced levels of hepcidin; and/oran infection caused by a pathogenic microorganism wherein an adjunctive therapy is administered to limit the amount of iron available to the pathogenic microorganism; and/ora neurodegenerative disease wherein the compound or pharmaceutically acceptable salt thereof is administered to limit deposition or increase of iron in tissue or cells; and/orformation of radicals, reactive oxygen species (ROS) or oxidative stress; and/orcardiac, liver or endocrine damage caused by iron overload; and/orinflammation triggered by excess iron.

15. A pharmaceutical composition comprising the compound or pharmaceutically acceptable salt thereof according to claim 1 and one or more compounds selected from pharmaceutical carriers, auxiliaries and solvents, and/orat least one additional pharmaceutically active compound.

16. The pharmaceutical composition according to claim 15, which is in the form of a formulation for oral or parenteral administration.

17. A method for combination therapy, comprising co-administration to a patient of the compound or pharmaceutically acceptable salt thereof according to claim 1 with at least one additional pharmaceutically active compound