Pyrimidopyrimidone compounds and methods of use thereof

Abstract

The present invention provides compounds having the structure of formula (I)-(VI), pharmaceutical compositions containing the same, and methods of use. In one aspect, compounds capable of inhibiting salt-inducible kinase (SIK) and methods of using the compounds to treat diseases or disorders are provided. Additionally, methods of making compounds of formula (I) or (II) are provided.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. patent application Ser. No. 63 / 342,779, filed May 17, 2022, the entire disclosure of which is incorporated herein by reference. [Background technology]

[0002] The treatment of many dermatological disorders has recognized shortcomings, including various side effects and limited effectiveness.Agents that increase skin pigmentation can have many beneficial dermatological effects, ranging from improving inflammatory skin diseases and providing sun protection to purely cosmetic applications.Therefore, it is desirable to have new treatments for dermatological disorders and defects. Summary of the Invention [Means for solving the problem]

[0003] In one aspect, we provide a compound that inhibits salt-induced kinase for the treatment or prevention of a dermatological disorder, a skin-related disorder in a subject.

[0004] In a preferred embodiment, compounds are provided having the structure of formula (I) or (II): [ka] During the ceremony: L 1 and L 2 each independently is a bond, substituted or unsubstituted alkylene, or substituted or unsubstituted alkenylene; R 2A , R 2B and R 2C are each independently hydrogen, halogen, or -CX 2 3 , -CHX 2 2 , -CH 2 X 2 , -OCX 2 3, -OCH 2 X 2 , -OCHX 2 2 , -OR 2F , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R 5 are independently halogen, -CX 5 3 , -CHX 5 2 , -CH 2 X 5 , -OCX 5 3 , -OCH 2 X 5 , -OCHX 5 2 , -OR 5F or substituted or unsubstituted C 1 -C 4 is alkyl; z is an integer from 0 to 5; X 2 and X 5 are each independently -F, -Br, -Cl, or -I; R 2F and R 5F is independently hydrogen, or substituted or unsubstituted alkyl.

[0005] In a preferred embodiment, a compound having the structure of formula (III) or (IV) is provided: [ka] During the ceremony: R 1 is independently hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted alkenyl; L 1 and L 2 each independently is a bond, substituted or unsubstituted alkylene, or substituted or unsubstituted alkenylene; R 2A , R 2B and R 2C are each independently hydrogen, halogen, or -CX 2 3 , -CHX 2 2 , -CH 2 X 2 , -OCX 2 3 , -OCH 2 X 2 , -OCHX 2 2 , -OR 2F , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R 5 are independently halogen, -CX 5 3 , -CHX 5 2 , -CH 2 X 5 , -OCX 5 3 , -OCH 2 X 5 , -OCHX 5 2 , -OR 5F , or substituted or unsubstituted C 1 -C 4 is alkyl; z is an integer from 0 to 5; R 7 -OR 7F or substituted or unsubstituted alkenyl; X 2 and X 5 are each independently -F, -Br, -Cl, or -I; R 2F , R 5F and OR 7F are each independently hydrogen, or substituted or unsubstituted alkyl.

[0006] In a preferred embodiment, a method for preparing a compound having the structure of formula (I) or (II) is provided. [ka] This method: The method includes providing a compound having a structure of formula (III) or (IV). [ka] In formulas (I) to (IV): R 1 is independently hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted alkenyl; L 1 and L 2 each independently is a bond, substituted or unsubstituted alkylene, or substituted or unsubstituted alkenylene; R 2A , R 2B and R 2C are each independently hydrogen, halogen, or -CX 2 3 , -CHX 2 2 , -CH 2 X 2 , -OCX 2 3 , -OCH 2 X 2 , -OCHX 2 2 , -OR 2F , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R 5 are independently halogen, -CX 5 3 , -CHX 5 2 , -CH 2 X 5 , -OCX 5 3 , -OCH 2 X 5 , -OCHX5 2 , -OR 5F , or substituted or unsubstituted C 1 -C 4 is alkyl; z is an integer from 0 to 5; R 7 -OR 7F or substituted or unsubstituted alkenyl; X 2 and X 5 are each independently -F, -Br, -Cl, or -I; R 2F , R 5F and R 7F are each independently hydrogen, or substituted or unsubstituted alkyl.

[0007] In a preferred embodiment, compositions or pharmaceutical compositions are provided that include one or more compounds described herein.

[0008] In a preferred embodiment, a method is provided for treating a subject suffering from or susceptible to a skin-related disorder or disease, the method comprising administering to the subject an effective amount of a compound or composition as described herein.

[0009] In a preferred aspect, a method of treating a subject suffering from or susceptible to rosacea is provided, comprising administering to the subject an effective amount of a compound or composition described herein.

[0010] In a preferred aspect, a method is provided for increasing pigmentation in a tissue of a subject, comprising administering to a subject an amount of a compound or composition described herein sufficient to increase melanin production, thereby increasing pigmentation in the tissue of the subject.

[0011] In a preferred embodiment, a method is provided for increasing cellular DNA stability or repair in skin tissue of a subject in need thereof, comprising administering to the subject a compound or composition described herein in an amount sufficient to reduce apoptosis and / or thymine dimer formation in cellular DNA of the skin tissue, thereby increasing the stability of cellular DNA in the skin tissue of the subject.

[0012] Other aspects of the invention are disclosed below. [Brief description of the drawings]

[0013] [Figure 1] 1A-1B show exemplary macrocyclic lactones.

[0014] [Diagram 2] Effect of compounds on B16 4A5 melanin content is shown. *= vs. untreated control; $= vs. DMSO 0.06%.

[0015] [Diagram 3] The results of melanin staining in the basal layer of the epidermis for all batches are shown.

[0016] [Figure 4] FIG. 1 shows the surface percentage of melanin in the basal and suprabasal layers of the epidermis quantified by image analysis.

[0017] [Diagram 5] The results of melanin staining in the basal layer of the epidermis for all batches are shown.

[0018] [Figure 6] 1 shows the percentage of the surface area occupied by melanin in the basal cell layer of the epidermis.

[0019] [Figure 7]Shown are Fontana-Masson staining images of human ex vivo explants cultured for 9 days without treatment (A), after UV irradiation (B), in vehicle (C), 0.2% SLT-048 (D), 2% SLT-048 (E), and expansion of basal and suprabasal melanin capping (F).

[0020] [Figure 8] 1 shows the results of melanin staining in the basal layer of the epidermis after treatment with SLT-045. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] definition The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulas set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts.

[0022] Where substituents are specified by their conventional chemical formula written from left to right, they equally encompass the chemically identical substituents that result from writing the structure from right to left, e.g., -CH 2 O is -OCH 2 - is equivalent to

[0023] The term "alkyl," by itself or as part of another substituent, means, unless otherwise stated, a straight (i.e., unbranched) or branched carbon chain (or carbons), or combinations thereof, which may be fully saturated, monovalent or polyunsaturated, and may include monovalent, divalent, and polyvalent groups. An alkyl group is any group having a specified number of carbons (e.g., C 1 -C 10means 1-10 carbons). Alkyl is a non-cyclized chain. Examples of saturated hydrocarbon groups include, but are not limited to, groups such as methyl ("Me"), ethyl ("Et"), n-propyl ("Pr"), isopropyl ("iPr"), n-butyl ("Bu"), t-butyl ("t-Bu"), isobutyl, sec-butyl, methyl, homologs and isomers such as n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. Unsaturated alkyl groups are those having one or more double or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and isomers. An alkoxy is an alkyl linked to the remainder of the molecule via an oxygen linker (-O-). The alkyl moiety may be an alkenyl moiety. The alkyl moiety may be an alkynyl moiety. The alkyl moiety may be fully saturated. An alkenyl may contain, in addition to one or more double bonds, more than one double bond and / or one or more triple bonds. An alkynyl may contain, in addition to one or more triple bonds, more than one triple bond and / or one or more double bonds.

[0024] The term “alkylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from alkyl, and includes, —CH 2 CH 2 CH 2 CH 2 Examples include, but are not limited to, -. Typically, alkyl (or alkylene) groups have from 1 to 24 carbon atoms, with groups having 10 or fewer carbon atoms being preferred herein. A "lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, generally having 8 or fewer carbon atoms. The term "alkenylene," by itself or as part of another substituent, means a divalent group derived from an alkene, unless otherwise stated.

[0025] The term "heteroalkyl," by itself or in combination with another term, means, unless otherwise stated, a stable linear or branched chain, or combination thereof, containing at least one carbon atom and at least one heteroatom (e.g., O, N, P, Si, and S), where the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. The heteroatom (e.g., O, N, S, Si, or P) may be located at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. Examples include, -CH 2 -CH 2 -O-CH 3 , -CH 2 -CH 2 -NH-CH 3 , -CH 2 -CH 2 -N(CH 3 )-CH 3 , -CH 2 -S-CH 2 -CH 3 , -CH 2 -S-CH 2 , -S(O)-CH 3 , -CH 2 -CH 2 -S(O) 2 -CH 3 , -CH=CH-O-CH 3 , -Si(CH 3 ) 3 , -CH 2 -CH=N-OCH 3 , -CH=CH-N(CH 3 )-CH 3 , -O-CH 3 , -O-CH 2 -CH 3 For example, -CH 2 -NH-OCH 3 and -CH 2 -O-Si(CH 3 ) 3Up to two or three heteroatoms may be consecutive, such as: A heteroalkyl moiety may contain one heteroatom (e.g., O, N, S, Si, or P). A heteroalkyl moiety may contain two optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may contain three optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may contain four optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may contain five optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may contain up to eight optionally different heteroatoms (e.g., O, N, S, Si, or P). The term "heteroalkenyl," by itself or in combination with another term, means a heteroalkyl containing at least one double bond, unless otherwise specified. Heteroalkenyl may optionally contain more than one double bond and / or one or more triple bonds in addition to one or more double bonds. The term "heteroalkynyl", by itself or in combination with another term, means, unless otherwise specified, a heteroalkyl containing at least one triple bond. Heteroalkynyl may optionally contain more than one triple bond and / or one or more double bonds in addition to one or more triple bonds.

[0026] Similarly, the term "heteroalkylene," by itself or as part of another substituent, includes, unless otherwise stated, -CH 2 -CH 2 -S-CH 2 -CH 2 - and -CH 2 -S-CH 2 -CH 2 -NH-CH 2- means a divalent group derived from heteroalkyl, exemplified but not limited to, -. For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, etc.). Additionally, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied by the direction in which the formula of the linking group is written. For example, a group of the formula -C(O) 2 R'- is -C(O) 2 R'- and -R'C(O) 2 As described above, heteroalkyl groups as used herein include both -C(O)R', -C(O)NR', ​​-NR'R'', -OR', -SR', and / or -SO. 2 It includes groups that are attached to the remainder of the molecule through a heteroatom, such as -R'. When "heteroalkyl" is recited followed by a specific heteroalkyl group, such as -NR'R'', it is understood that the terms heteroalkyl and -NR'R'' are not redundant or mutually exclusive. Rather, the specific heteroalkyl group is recited to add clarity. Thus, the term "heteroalkyl" should not be construed herein as excluding specific heteroalkyl groups, such as -NR'R''.

[0027] The terms "cycloalkyl" and "heterocycloalkyl", by themselves or in combination with other terms, mean cyclic versions of "alkyl" and "heteroalkyl", respectively, unless otherwise stated. Cycloalkyl and heterocycloalkyl are not aromatic. Additionally, for heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkyl include, but are not limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like. A "cycloalkylene" and a "heterocycloalkylene," alone or as part of another substituent, mean a divalent radical derived from a cycloalkyl and heterocycloalkyl, respectively.

[0028] In an embodiment, the heterocycloalkyl is heterocyclyl. The term "heterocyclyl" as used herein means a monocyclic, bicyclic, or polycyclic heterocycle. A heterocyclyl monocyclic heterocycle is a 3-, 4-, 5-, 6-, or 7-membered ring containing at least one heteroatom independently selected from the group consisting of O, N, and S, where the ring is saturated or unsaturated, but not aromatic. A 3- or 4-membered ring contains one heteroatom selected from the group consisting of O, N, and S. A 5-membered ring can contain zero or one double bond and one, two, or three heteroatoms selected from the group consisting of O, N, and S. A 6- or 7-membered ring contains zero, one, or two double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S. A heterocyclyl monocyclic heterocycle is attached to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the heterocyclyl monocyclic heterocycle. Representative examples of heterocyclyl monocyclic heterocycles include azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiolanyl, 1,3-dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, Heterocyclyl bicyclic heterocycles include, but are not limited to, oxazolidinyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1-dioxidethiomorpholinyl (thiomorpholine sulfone), thiopyranyl and trithianyl. Heterocyclyl bicyclic heterocycles are monocyclic heterocycles fused to either phenyl, monocyclic cycloalkyl, monocyclic cycloalkenyl, monocyclic heterocycle, or monocyclic heteroaryl. Heterocyclyl bicyclic heterocycles are attached to the parent molecular moiety via any carbon atom or any nitrogen atom contained within the monocyclic heterocycle portion of the bicyclic ring system.Representative examples of bicyclic heterocyclyls include, but are not limited to, 2,3-dihydrobenzofuran-2-yl, 2,3-dihydrobenzofuran-3-yl, indolin-1-yl, indolin-2-yl, indolin-3-yl, 2,3-dihydrobenzothien-2-yl, decahydroquinolinyl, decahydroisoquinolinyl, octahydro-1H-indolyl, and octahydrobenzofuranyl. In embodiments, the heterocyclyl group is optionally substituted with one or two groups that are independently oxo or thia. In certain embodiments, the bicyclic heterocyclyl is a 5- or 6-membered monocyclic heterocyclyl ring fused to a phenyl ring, a 5- or 6-membered monocyclic cycloalkyl, a 5- or 6-membered monocyclic cycloalkenyl, a 5- or 6-membered monocyclic heterocyclyl, or a 5- or 6-membered monocyclic heteroaryl, wherein the bicyclic heterocyclyl is optionally substituted by one or two groups that are independently oxo or thia. A polycyclic heterocyclyl ring system is a monocyclic heterocyclyl ring (base ring) fused to either (i) one ring system selected from the group consisting of bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl, and bicyclic heterocyclyl; or (ii) two other ring systems independently selected from the group consisting of phenyl, bicyclic aryl, monocyclic or bicyclic heteroaryl, monocyclic or bicyclic cycloalkyl, monocyclic or bicyclic cycloalkenyl, and monocyclic or bicyclic heterocyclyl. The polycyclic heterocyclyl is attached to the parent molecular moiety through any carbon or nitrogen atom contained within the base ring. In embodiments, the polycyclic heterocyclyl ring system is a monocyclic heterocyclyl ring (base ring) fused to either: (i) one ring system selected from the group consisting of bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl, and bicyclic heterocyclyl; or (ii) two other ring systems independently selected from the group consisting of phenyl, monocyclic heteroaryl, monocyclic cycloalkyl, monocyclic cycloalkenyl, and monocyclic heterocyclyl.Examples of polycyclic heterocyclyl groups include, but are not limited to, 10H-phenothiazin-10-yl, 9,10-dihydroacridin-9-yl, 9,10-dihydroacridin-10-yl, 10H-phenoxazin-10-yl, 10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl, 1,2,3,4-tetrahydropyrido[4,3-g]isoquinolin-2-yl, 12H-benzo[b]phenoxazin-12-yl, and dodecahydro-1H-carbazol-9-yl.

[0029] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as "haloalkyl" are meant to include monohaloalkyl and polyhaloalkyl. For example, "halo(C 1 -C 4 The term "alkyl" includes, but is not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0030] The term "aryl", unless otherwise specified, refers to a polyunsaturated aromatic hydrocarbon substituent, which may be a single ring or multiple rings (preferably 1-3 rings) fused together (i.e., fused-ring aryl) or covalently bonded to each other. Fused-ring aryl refers to multiple rings fused together, where at least one of the fused rings is an aryl ring. The term "heteroaryl" refers to an aryl group (or ring) containing at least one heteroatom, such as N, O, or S, where the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. Thus, the term "heteroaryl" includes fused-ring heteroaryl groups (i.e., multiple rings fused together, where at least one of the fused rings is a heteroaromatic ring). 5,6-fused-ring heteroarylene refers to two rings fused together, where one ring has 5 members and the other ring has 6 members, and at least one ring is a heteroaryl ring. Similarly, a 6,6-fused ring heteroarylene refers to two rings fused together, one ring having 6 members and the other ring having 6 members, and at least one ring is a heteroaryl ring. Also, a 6,5-fused ring heteroarylene refers to two rings fused together, one ring having 6 members and the other ring having 5 members, and at least one ring is a heteroaryl ring. The heteroaryl group can be attached to the remainder of the molecule through a carbon or heteroatom.Non-limiting examples of aryl and heteroaryl groups include phenyl, naphthyl, pyrrolyl, pyrazolyl, pyridazinyl, triazinyl, pyrimidinyl, imidazolyl, pyrazinyl, purinyl, oxazolyl, isoxazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrimidyl, benzothiazolyl, benzoxazolylbenzimidazolyl, benzofuran, isobenzofuranyl, indolyl, isoindolyl, benzothiophenyl, isoquinolyl, quinoxalinyl, quinolyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl,

[0033] Examples of aryl and heteroaryl ring systems include aryl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituents for each of the above noted aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below. An "arylene" and a "heteroarylene," alone or as part of another substituent, mean a divalent radical derived from an aryl and heteroaryl, respectively. The heteroaryl group substituent may be -O-bonded to a ring heteroatom nitrogen.

[0031] A fused ring heterocycloalkyl-aryl is an aryl fused to a heterocycloalkyl. A fused ring heterocycloalkyl-heteroaryl is a heteroaryl fused to a heterocycloalkyl. A fused ring heterocycloalkyl-cycloalkyl is a heterocycloalkyl fused to a cycloalkyl. A fused ring heterocycloalkyl-heterocycloalkyl is a heterocycloalkyl fused to another heterocycloalkyl. Each fused ring heterocycloalkyl-aryl, fused ring heterocycloalkyl-heteroaryl, fused ring heterocycloalkyl-cycloalkyl, or fused ring heterocycloalkyl-heterocycloalkyl may be independently unsubstituted or substituted with one or more of the substituents described herein.

[0032] A spirocyclic ring is two or more rings in which adjacent rings are linked through a single atom. The individual rings in a spirocyclic ring may be the same or different. The individual rings in a spiro ring may be substituted or unsubstituted and may have different substituents than the other individual rings in the set of spiro rings. The possible substituents for the individual rings in a spiro ring are the possible substituents for the same ring when not part of a spiro ring (e.g., the substituents of a cycloalkyl or heterocycloalkyl ring). A spiro ring may be a substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted heterocycloalkylene, and the individual rings in a spirocyclic group may be any of the preceding list, including having all rings of one type (e.g., all rings are substituted heterocycloalkylene, and each ring may be the same or different substituted heterocycloalkylene). When referring to a spiro ring system, a heterocyclic spiro ring means a spiro ring in which at least one ring is a heterocyclic ring, and each ring may be a different ring. When referring to a spiro ring system, a substituted spiro ring means that at least one ring is substituted, and each substituent can optionally be different.

[0033] symbol [ka] indicates the point of attachment of the chemical moiety to the remainder of the molecule or chemical formula.

[0034] The term "oxo" as used herein means an oxygen that is double bonded to a carbon atom.

[0035] Each of the above terms (e.g., "alkyl," "heteroalkyl," "cycloalkyl," "heterocycloalkyl," "aryl" and "heteroaryl") includes both substituted and unsubstituted forms of the indicated radical. Preferred substituents for each type of radical are provided below.

[0036] Similar to the substituents described for the alkyl group, the substituents for the aryl and heteroaryl groups vary and include, for example, -OR', -NR'R'', -SR', -halogen, -SiR'R''R''', -OC(O)R', -C(O)R', -CO 2 R', -CONR'R'', -OC(O)NR'R'', -NR''C(O)R', -NR'-C(O)NR''R''', -NR''C(O) 2 R', -NR-C(NR'R''R''')=NR''', -NR-C(NR'R'')=NR''', -S(O)R', -S(O) 2 R', -S(O) 2 NR'R'', -NRSO 2 R', -NR'NR''R''', -ONR'R'', -NR'C(O)NR''NR'''R''', -CN, -NO 2 , -R', -N 3 , -CH(Ph) 2 , Fluoro(C 1 -C 4 ) alkoxy, and fluoro (C 1 -C 4 ) alkyl, -NR'SO 2R", -NR'C(O)R", -NR'C(O)-OR", -NR'OR", which numbers range from 0 to the total number of open valences in the aromatic ring system, and R', R", R'" and R"" are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl. When the compounds described herein include more than one R group, for example, each of the R groups is independently selected such that each of the R', R", R'" and R"" groups is selected when more than one of these groups is present.

[0037] Substituents for a ring (e.g., cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene) may be shown as substituents on the ring rather than on a specific atom of the ring (commonly referred to as floating substituents). In such cases, the substituent may be attached to any of the ring atoms (subject to the rules of chemical valence), and in the case of a fused or spiro ring, a substituent shown as attached to one member of the fused or spiro ring (floating substituents on a single ring) may be a substituent on either of the fused or spiro rings (floating substituents on a polycyclic ring). When a substituent is attached to a ring but not to a specific atom (floating substituent) and the substituent subscript is an integer greater than 1, the multiple substituents may be on the same atom, the same ring, different atoms, different fused rings, different spiro rings, and each substituent may optionally be different. When the point of attachment of the ring to the rest of the molecule is not limited to a single atom (floating substituent), the point of attachment may be any atom of the ring, or, in the case of a fused or spiro ring, any atom of either the fused or spiro ring, subject to the rules of chemical valence. When the point of attachment of the ring to the rest of the molecule is not limited to a single atom (floating substituent), the point of attachment may be any atom of the ring, or, in the case of a fused or spiro ring, any atom of either the fused or spiro ring, subject to the rules of chemical valence. When a ring, fused ring, or spirocyclic ring contains one or more ring heteroatoms, and the ring, fused ring, or spirocyclic ring is depicted with one or more floating substituents (including, but not limited to, the point of attachment to the rest of the molecule), the floating substituents may be attached to the heteroatoms. When a ring heteroatom is shown bonded to one or more hydrogens in a structure or formula having a floating substituent (e.g., a ring nitrogen having two bonds to ring atoms and a third bond to a hydrogen), it is understood that when the heteroatom is bonded to the floating substituent, the substituent replaces the hydrogen, following the rules of chemical valence.

[0038] Two or more substituents may be optionally combined to form an aryl, heteroaryl, cycloalkyl, or heterocycloalkyl group. Such so-called ring-forming substituents are typically, but not necessarily, found attached to a cyclic base structure. In one embodiment, the ring-forming substituents are attached to adjacent members of the base structure. For example, two ring-forming substituents attached to adjacent members of a cyclic base structure form a fused ring structure. In another embodiment, the ring-forming substituents are attached to a single member of the base structure. For example, two ring-forming substituents attached to a single member of a cyclic base structure form a spiro ring structure. In yet another embodiment, the ring-forming substituents are attached to non-adjacent members of the base structure.

[0039] Two of the substituents on adjacent atoms of an aryl or heteroaryl ring are of the formula -TC(O)-(CRR') q Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may form a ring of the formula -A-(CH 2 ) r A and B are optionally substituted with a substituent of -B-, where A and B are independently -CRR'-, -O-, -NR-, -S-, -S(O)-, -S(O) 2 -, -S(O) 2 NR'-, or a single bond, where r is an integer from 1 to 4. One of the single bonds of the new ring so formed may optionally be replaced with a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may be of the formula -(CRR') s -X'-(C''R''R'') d wherein s and d are independently an integer from 0 to 3, and X' is -O-, -NR'-, -S-, -S(O)-, -S(O) 2 - or -S(O) 2The substituents R, R', R'', and R''' are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.

[0040] As used herein, the term "heteroatom" or "ring heteroatom" is meant to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), and silicon (Si).

[0041] As used herein, "substituent" means a group selected from the following moieties: (A) Oxo, halogen, -CCl 3 , -CBr 3 , -CF 3 , -CI 3 , -CH 2 Cl, -CH 2 Br, -CH 2 F, -CH 2 I, -CHCl 2 , -CHBr 2 , -CHF 2 , -CHI 2 , -CN, -OH, -NH 2 , -COOH, -CONH 2 , -NO 2 , -SH, -SO 3 H, -SO 4 H, -SO 2 NH 2 , -NHNH 2 , -ONH 2 , -NHC(O)NHNH 2 , -NHC(O)NH 2 , -NHSO 2 H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl 3 , -OCF 3 , -OCBr 3 , -OCI 3 , -OCHCl 2 , -OCHBr 2 , -OCH1 2 , -OCHF2 , -N 3 , unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl, or C 1 -C 4 alkyl), unsubstituted heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl, or C 5 -C 6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), unsubstituted aryl (e.g., C 6 -C 10 Aryl, C 10 aryl, or phenyl), or unsubstituted heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl), and (B) Alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl, or C 1 -C 4 alkyl), heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl, or C 5 -C 6 cycloalkyl), heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), aryl (e.g., C 6 -C 10 Aryl, C 10aryl, or phenyl), heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl): (i) Oxo, halogen, -CCl 3 , -CBr 3 , -CF 3 , -CI 3 , -CH 2 Cl, -CH 2 Br, -CH 2 F, -CH 2 I, -CHCl 2 (i) oxo, halogen, -CCl 3 , -CBr 3 , -CF 3 , -CI 3 , -CH 2 Cl, -CH 2 Br, -CH 2 F, -CH 2 I, -CHCl 2 , -CHBr 2 , -CHF 2 , -CHI 2 , -CN, -OH, -NH 2 , -COOH, -CONH 2 , -NO 2 , -SH, -SO 3 H, -SO 4 H, -SO 2 NH 2 , -NHNH 2 , -ONH 2 , -NHC(O)NHNH 2 , -NHC(O)NH 2 , -NHSO 2 H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl 3 , -OCF 3 , -OCBr 3 , -OCI 3 , -OCHCl 2 , -OCHBr 2 , -OCHI 2 , -OCHF 2 , -N 3 , unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C1 -C 6 Alkyl, or C 1 -C 4 alkyl), unsubstituted heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl, or C 5 -C 6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), unsubstituted aryl (e.g., C 6 -C 10 Aryl, C 10 aryl, or phenyl), or unsubstituted heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl), and (ii) alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl, or C 1 -C 4 alkyl), heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl, or C 5 -C 6 cycloalkyl), heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), aryl (e.g., C 6 -C 10 Aryl, C 10 aryl, or phenyl), heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl), (a) Oxo, halogen, -CCl 3 , -CBr 3 , -CF 3 , -CI 3 , -CH 2 Cl, -CH 2 Br, -CH 2 F, -CH 2 I, -CHCl 2 , -CHBr 2 , -CHF 2 , -CHI 2 , -CN, -OH, -NH 2 , -COOH, -CONH 2 , -NO 2 , -SH, -SO 3 H, -SO 4 H, -SO 2 NH 2 , -NHNH 2 , -ONH 2 , -NHC(O)NHNH 2 , -NHC(O)NH 2 , -NHSO 2 H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl 3 , -OCF 3 , -OCBr 3 , -OCI 3 , -OCHCl 2 , -OCHBr 2 , -OCHI 2 , -OCHF 2 , -N 3 , unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl, or C 1 -C 4 alkyl), unsubstituted heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl, or C 5 -C 6cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), unsubstituted aryl (e.g., C 6 -C 10 Aryl, C 10 aryl, or phenyl), or unsubstituted heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl), and (b) Oxo, halogen, -CCl 3 , -CBr 3 , -CF 3 , -CI 3 , -CH 2 Cl, -CH 2 Br, -CH 2 F, -CH 2 I, -CHCl 2 , -CHBr 2 , -CHF 2 , -CHI 2 , -CN, -OH, -NH 2 , -COOH, -CONH 2 , -NO 2 , -SH, -SO 3 H, -SO 4 H, -SO 2 NH 2 , -NHNH 2 , -ONH 2 , -NHC(O)NHNH 2 , -NHC(O)NH 2 , -NHSO 2 H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl 3 , -OCF 3 , -OCBr 3 , -OCI 3 , -OCHCl 2 , -OCHBr 2 , -OCHI 2 , -OCHF 2 , -N 3 , unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl, or C 1 -C4 alkyl), unsubstituted heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl, or C 5 -C 6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), unsubstituted aryl (e.g., C 6 -C 10 Aryl, C 10 aryl, or phenyl), or unsubstituted heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl), 1 -C 8 Alkyl, C 1 -C 6 Alkyl, or C 1 -C 4 alkyl), heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl, or C 5 -C 6 cycloalkyl), heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), aryl (e.g., C 6 -C 10 Aryl, C 10 aryl, or phenyl), heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl).

[0042] Certain compounds of the present disclosure have asymmetric carbon atoms (optical or chiral centers) or double bonds; enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisomeric forms that may be defined in terms of absolute stereochemistry as (R)- or (S)- for amino acids, or as (D)- or (L)-, and individual isomers are encompassed within the scope of the present disclosure. Compounds of the present disclosure do not include those known in the art to be too unstable to synthesize and / or isolate. The present disclosure is meant to include compounds in racemic and optically pure form. Optically active (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When compounds described herein contain olefinic bonds or other centers of geometric asymmetry, unless otherwise specified, it is intended that the compounds include both E and Z geometric isomers.

[0043] As used herein, the term "isomers" refers to compounds having the same number and kinds of atoms, and therefore the same molecular weight, but differing with respect to the structural or configuration of the atoms.

[0044] It will be apparent to one of ordinary skill in the art that certain compounds of the present disclosure may exist in tautomeric forms, all such tautomeric forms of the compounds being within the scope of the present disclosure.

[0045] Unless otherwise specified, the structures shown herein are also meant to include all stereochemical forms of the structures; that is, the R and S configurations for each asymmetric center.Thus, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the compounds are within the scope of this disclosure.Thus, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the compounds are within the scope of this disclosure.

[0046] It should be noted that throughout this application, options are described as Markush groups, e.g., at each amino acid position that contains more than one possible amino acid. It is specifically contemplated that each member of a Markush group should be considered separately, thereby including alternative embodiments, and that a Markush group should not be read as a single unit.

[0047] As used herein, the terms "a" or "an" mean one or more. Additionally, as used herein, the phrase "substituted with [n]" means that the specified group may be substituted with one or more of any or all of the specified substituents. For example, a group such as an alkyl or heteroaryl group may be "unsubstituted C 1 -C 20 When "substituted with alkyl, or unsubstituted 2- to 20-membered heteroalkyl," the group is selected from the group consisting of one or more unsubstituted C 1 -C 20 It may contain one or more unsubstituted 2-20 membered alkyls, and / or one or more unsubstituted 2-20 membered heteroalkyls.

[0048] The description of the compounds of the present disclosure is limited by the principles of chemical bonding known to those skilled in the art.Therefore, when a group can be substituted by one or more of several substituents, such substitutions are selected to give compounds that are known to those skilled in the art to be not inherently unstable and / or likely to be unstable under ambient conditions, such as aqueous, neutral, and some known physiological conditions, according to the principles of chemical bonding.For example, heterocycloalkyl or heteroaryl are bonded to the rest of the molecule through ring heteroatoms according to the principles of chemical bonding known to those skilled in the art, thereby avoiding inherently unstable compounds.

[0049] One of skill in the art will understand that when a variable (e.g., a moiety or linker) of a compound or a genus of compounds (e.g., a genus described herein) is described by a name or formula of a stand-alone compound with all valences satisfied, the unsatisfied valences of the variable will be determined by the context in which the variable is used. For example, when a variable of a compound described herein is connected (e.g., bonded) to the remainder of the compound through a single bond, the variable will be understood to represent the monovalent form of the stand-alone compound (i.e., capable of forming a single bond due to unsatisfied valences) (e.g., if a variable is named "methane" in one embodiment, but the variable is known to be attached to the remainder of the compound by a single bond, one of skill in the art will understand that the variable is actually the monovalent form of methane, i.e., methyl or -CH 3 Similarly, linker variables (e.g., L 1 , L 2 , or L 3 ), one of skill in the art will understand that the variable is a divalent form of the stand-alone compound (e.g., if a variable is assigned to "PEG" or "polyethylene glycol" in an embodiment, but the variable is connected to the remainder of the compound by two separate bonds, one of skill in the art will understand that the variable is a divalent (i.e., capable of forming two bonds through two unsatisfied valences) form of PEG, instead of the stand-alone compound PEG).

[0050] As used herein, the term "salt" refers to an acid or base salt of a compound used in the method of the present invention. Specific examples of acceptable salts are mineral acid (hydrochloric acid, hydrobromic acid, phosphoric acid, etc.) salts, organic acid (acetic acid, propionic acid, glutamic acid, citric acid, etc.) salts, and quaternary ammonium (methyl iodide, ethyl iodide, etc.) salts.

[0051] The term "pharmaceutically acceptable salt" is meant to include salts of active compounds prepared using relatively non-toxic acids or bases, depending on the specific substituents found on the compounds described herein.When a compound of the present disclosure contains a relatively acidic functional group, a base addition salt can be obtained by contacting a neutral form of such a compound, either neat or in a suitable inert solvent, with a sufficient amount of the desired base.Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or similar salts.When a compound of the present disclosure contains a relatively basic functional group, an acid addition salt can be obtained by contacting a neutral form of such a compound, either neat or in a suitable inert solvent, with a sufficient amount of the desired acid. Examples of pharma- ceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphate, dihydrogenphosphate, sulfuric, monohydrogensulfuric, hydroiodic, or phosphorous, as well as those derived from relatively non-toxic organic acids such as acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, oxalic, methanesulfonic, etc. Also included are salts of amino acids such as arginates, and salts of organic acids such as glucuronic or galacturonic acid (see, for example, Berge et al., 'Pharmaceutical Salts', Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present disclosure contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0052] Therefore, the compounds of the present disclosure may exist as salts with pharma- ceutically acceptable acids and the like. The present disclosure includes such salts. Non-limiting examples of such salts include hydrochloride, hydrobromide, phosphate, sulfate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, propionate, tartrate (e.g., (+)-tartrate, (-)-tartrate, or mixtures thereof, including racemic mixtures), succinate, benzoate, and salts with amino acids such as glutamic acid, and quaternary ammonium salts (e.g., methyl iodide, ethyl iodide, etc.). These salts can be prepared by methods known to those skilled in the art.

[0053] The neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound may differ from the various salt forms in certain physical properties, such as solubility in polar solvents.

[0054] In addition to salt forms, the present disclosure provides compounds that are in prodrug form.Prodrugs of the compounds described herein are compounds that undergo easy chemical changes under physiological conditions to provide the compounds of the present disclosure.Prodrugs of the compounds described herein can be converted in vivo after administration.Furthermore, prodrugs can be converted to the compounds of the present disclosure by chemical or biochemical methods in an ex vivo environment, for example, when contacted with suitable enzymes or chemical reagents.

[0055] Certain compounds of the present disclosure can exist in non-solvated form as well as solvated form, including hydrated form.Generally, solvated form is equivalent to non-solvated form and is included within the scope of the present disclosure.Certain compounds of the present disclosure can exist in multiple crystalline or amorphous forms.Generally, all physical forms are equivalent for the use contemplated by the present disclosure and are intended to be within the scope of the present disclosure.

[0056] "Pharmaceutically acceptable excipient" and "pharmaceutically acceptable carrier" refer to a substance that aids in the administration and absorption of an active agent to a subject and can be included in the compositions of the present disclosure without causing significant adverse toxicological effects to the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline, lactated Ringer's solution, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavorings, salt solutions (such as Ringer's solution), alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone, and colorants. Such preparations can be sterilized and mixed with auxiliary agents such as emollients, moisturizers, lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for affecting osmotic pressure, buffers, colorants, and / or aromatic substances that do not adversely react with the compounds of the present disclosure. Those of skill in the art will recognize that other pharmaceutical excipients are useful in the present disclosure, including, but not limited to, DMSO and transcutol.

[0057] The term "formulation" is intended to include formulations of active compounds that include encapsulating materials as carriers, providing capsules in which the active ingredient, with or without other carriers, is surrounded by the carrier, thus associating with it.Also included are cachets and lozenges.Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0058] As used herein, the term "about" refers to a range of values ​​that includes the specified value and that one of ordinary skill in the art would consider to be reasonably similar to the specified value. In embodiments, about refers to within a standard deviation, using measurements generally accepted in the art. In embodiments, about refers to a range that spans + / - 10% of the specified value. In embodiments, about includes the specified value.

[0059] As used herein, "EC 50The term "median effective concentration" or "EC" refers to the concentration of a molecule (e.g., a small molecule, drug, antibody, chimeric antigen receptor, or bispecific antibody) that can induce a response that is intermediate between the baseline response and the maximum response after a particular exposure time. In embodiments, the EC 50 is the concentration of a molecule (e.g., a small molecule, drug, antibody, chimeric antigen receptor, or bispecific antibody) that results in 50% of the molecule's maximum possible effect.

[0060] The term "treating" or "treatment" refers to any indication of success in treating or ameliorating an injury, disease, pathology, or condition, including any objective or subjective parameter, such as, for example, relief; remission; reduction in symptoms or making the injury, pathology, or condition more tolerable to the patient; slowing the rate of degeneration or decline; making the end point of degeneration less debilitating; improving the physical or mental health of the patient, etc. The treatment or amelioration of symptoms can be based on objective or subjective parameters, including the results of a physical exam, a neuropsychiatric exam, and / or a psychiatric evaluation. The term "treating" and its conjugations can include prevention of an injury, condition, state, or disease. In an embodiment, treatment is prevention. In an embodiment, treating does not include preventing.

[0061] "Treating" or "treatment" as used herein (and well understood in the art) also broadly includes any approach to obtain beneficial or desired results in a subject's condition, including clinical results. Beneficial or desired clinical results may include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, reduction in the extent of disease, stabilization or non-aggravation of the disease state, prevention of disease spread or spread, delay or slowing of disease progression, improvement or mitigation of the disease state, reduction in disease recurrence, and remission, whether partial or total, and detectable or undetectable. In other words, "treatment" as used herein includes any cure, amelioration, or prevention of disease. Treatment can prevent the onset of disease, inhibit the spread of disease, alleviate symptoms of disease, completely or partially eliminate the underlying cause of disease, shorten the duration of disease, or a combination of these.

[0062] The term "prevent" refers to a reduction in the occurrence of disease symptoms in a patient. As noted above, prevention may be complete (no detectable symptoms) or partial, such that fewer symptoms are observed than would occur in the absence of treatment.

[0063] "Patient" or "subject in need thereof" refers to an organism suffering from or prone to a disease or condition that can be treated by administration of the pharmaceutical compositions provided herein. Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, monkeys, goats, sheep, cows, deer, and other non-mammals. In some embodiments, the patient is a human.

[0064] An "effective amount" is an amount sufficient for the compound to achieve the stated purpose (e.g., achieve the effect for which it is administered, treat a disease, reduce enzyme activity, increase enzyme activity, reduce a signal transduction pathway, or alleviate one or more symptoms of a disease or condition) compared to the absence of the compound. An example of an "effective amount" is an amount sufficient to contribute to the treatment, prevention, or alleviation of one or more symptoms of a disease, which may also be referred to as a "therapeutically effective amount." A "reduction" of one or more symptoms (and grammatical equivalents of this phrase) means a reduction in the severity or frequency of a symptom, or the elimination of a symptom. A "prophylactically effective amount" of a drug is an amount of a drug that, when administered to a subject, has an intended prophylactic effect, e.g., an effect of preventing or delaying the onset (or recurrence) of an injury, disease, pathology, or condition, or reducing the likelihood of the onset (or recurrence) of an injury, disease, pathology, or condition, or a symptom thereof. A complete prophylactic effect does not necessarily occur by administration of one dose, but may occur only after administration of a series of doses. Thus, a prophylactically effective amount may be administered in one or more administrations. As used herein, "activity-reducing amount" refers to the amount of antagonist required to reduce the activity of enzyme compared to the absence of antagonist. As used herein, "function-destroying amount" refers to the amount of antagonist required to destroy the function of enzyme or protein compared to the absence of antagonist. The exact amount depends on the purpose of treatment and can be ascertained by those skilled in the art using known techniques (see, for example, Lieberman, Pharmaceutical Dosage Forms (vols.1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

[0065] For any compound described herein, the therapeutically effective amount can be initially determined from cell culture assays. The target concentration is the concentration of active compound that can achieve the methods described herein, as measured using methods described herein or known in the art.

[0066] As is well known in the art, the therapeutically effective amount for human use can also be determined from animal models.For example, the dose for human can be formulated to achieve the concentration found to be effective in animals.The dose in human can be adjusted by monitoring the effectiveness of the compound and adjusting the dose upward or downward as described above.It is well within the capabilities of a person skilled in the art to adjust the dose to achieve maximum effectiveness in human based on the above and other methods.

[0067] The term "therapeutically effective amount" as used herein refers to an amount of a therapeutic agent sufficient to improve the above-mentioned disorder. For example, for a given parameter, a therapeutically effective amount shows at least a 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or at least a 100% increase or decrease. The therapeutic efficacy can also be expressed as a "-fold" increase or decrease. For example, a therapeutically effective amount can have at least a 1.2-fold, 1.5-fold, 2-fold, 5-fold, or more effect over a control.

[0068] Dosage may vary depending on the requirements of the patient and the compound used. In the context of this disclosure, the dose administered to the patient should be sufficient to produce a beneficial therapeutic response in the patient over time. The size of the dose is also determined by the existence, nature, and extent of any adverse side effects. Determining the proper dosage for a particular situation is within the skill of the physician. Generally, treatment is initiated with smaller dosages that are less than the optimal dosage of the compound. Thereafter, the dosage is increased by small increments until the optimal effect is achieved under the circumstances. Dosage and administration intervals can be individually adjusted to provide a level of the administered compound that is effective for the particular clinical indication being treated. This provides a treatment regimen that is commensurate with the severity of the individual's disease state.

[0069] As used herein, the term "administer" refers to oral administration, administration as a suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal or subcutaneous administration to a subject, or implantation of a sustained release device, such as a mini-osmotic pump. Administration is by any route, including parenteral and transmucosal (e.g. buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, for example, intravenous, intramuscular, intraarteriolar, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, and the like. In embodiments, administration does not include administration of any active agent other than the listed active agents.

[0070] compound Provided herein, inter alia, are macrocyclic lactones.

[0071] In one aspect, a compound having the structure of formula (I) or (II) is provided. [ka] During the ceremony: L 1 and L 2 each independently is a bond, substituted or unsubstituted alkylene, or substituted or unsubstituted alkenylene; R 2A , R 2B and R 2C are each independently hydrogen, halogen, or -CX 2 3 , -CHX 2 2 , -CH 2 X 2 , -OCX 2 3 , -OCH 2 X 2 , -OCHX 2 2 , -OR 2F , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R 5 are independently halogen, -CX 5 3 , -CHX 5 2 , -CH 2 X 5 , -OCX 5 3 , -OCH 2 X 5 , -OCHX 5 2 , -OR 5F , or substituted or unsubstituted C 1 -C 4 is alkyl; z is an integer from 0 to 5; X 2 and X 5 are each independently -F, -Br, -Cl, or -I; R 2F and R 5F Each is hydrogen, or substituted or unsubstituted alkyl.

[0072] In embodiments, z is 1. In embodiments, z is 2. In embodiments, z is 3.

[0073] In embodiments, R 5 are independently unsubstituted C1 -C 4 In embodiments, R 5 is independently unsubstituted methyl. In embodiments, R 5 is independently unsubstituted ethyl. In embodiments, R 5 is independently unsubstituted isopropyl. In embodiments, R 5 is independently unsubstituted propyl. In embodiments, R 5 is independently unsubstituted butyl. In embodiments, R 5 is independently unsubstituted t-butyl.

[0074] In an embodiment, z is 2 and R 5 is unsubstituted methyl.

[0075] In embodiments, the compound has the structure of Formula (Ia) or (II-a): [ka] L 1 , L 2 , R 2A , R 2B , and R 2C is as described herein.

[0076] In embodiments, R 2A is a substituted or unsubstituted heterocycloalkyl, or a substituted or unsubstituted aryl. In embodiments, R 2A is a substituted or unsubstituted heterocycloalkyl. In embodiments, R 2A is substituted or unsubstituted aryl. In embodiments, R 2A is a substituted or unsubstituted piperazinyl. In embodiments, R 2A is a substituted or unsubstituted phenyl.

[0077] In embodiments, R 2A teeth [ka] wherein: R3 is hydrogen or substituted or unsubstituted alkyl; R 4A , R 4B , R 4C , and R 4D are each independently hydrogen, halogen, or -CX 4 3 , -CHX 4 2 , -CH 2 X 4 , -OCX 4 3 , -OCH 2 X 4 , -OCHX 4 2 , -OR 4F or substituted or unsubstituted alkyl; X 4 is independently -F, -Br, -Cl, or -I; R 4F is hydrogen, or substituted or unsubstituted alkyl.

[0078] In embodiments, the compound has the structure of Formula (Ib) or (II-b): [ka] In the formula, L 1 , L 2 , R 2B , R 2C , R 3 , R 4A , R 4B , R 4C , and R 4D is as described herein.

[0079] In an embodiment, L 1 is a bond, or R 6 -Substituted or unsubstituted C 1 -C 4 In embodiments, L is an alkylene. 1 is a bond. In embodiments, L 1 is a substituted or unsubstituted C 1 -C 4 In embodiments, L is an alkylene.1 is unsubstituted C 1 -C 4 In embodiments, L is an alkylene. 1 is a substitution C 1 -C 4 In embodiments, L is an alkylene. 1 is unsubstituted methylene. In embodiments, L 1 is unsubstituted ethylene. In embodiments, L 1 is unsubstituted propylene. In embodiments, L 1 is unsubstituted isopropylene. In embodiments, L 1 is unsubstituted butylene.

[0080] In an embodiment, L 1 is R 6 -Substituted or unsubstituted C 1 -C 4 In embodiments, R 6 is halogen or unsubstituted C 1 -C 4 In embodiments, L is an alkylene. 1 is a methyl-substituted C 1 -C 4 In embodiments, L is an alkylene. 1 is a methyl-substituted methylene. 1 is methyl substituted ethylene. In embodiments, L 1 is methyl-substituted propylene. In embodiments, L 1 is a methyl substituted butylene.

[0081] In embodiments, the compound has the structure of Formula (Ic): [ka] During the ceremony: L 1 is a bond, or R 6 -Substituted or unsubstituted C 1 -C 4 is alkylene; L 2 is a substituted or unsubstituted C 2 -C 5Alkylene, or substituted or unsubstituted C 2 -C 5 alkenylene; R 6 is halogen or unsubstituted C 1 -C 4 It is alkylene.

[0082] In embodiments, the compound has the structure of Formula (II-c): [ka] During the ceremony: L 1 is R 6 -Substituted or unsubstituted C 1 -C 4 is alkylene; L 2 is a substituted or unsubstituted C 2 -C 5 Alkylene, or substituted or unsubstituted C 2 -C 5 alkenylene; R 6 is halogen or unsubstituted C 1 -C 4 It is alkylene.

[0083] Exemplary compounds having the structure of formula (Ic) or (II-c) are shown in Table 1. [Table 1-1] [Table 1-2] [Table 1-3]

[0084] In embodiments, R 2A is halogen. In embodiments, R 2A is -F. In embodiments, R 2A is -Cl. In embodiments, R 2A In an embodiment, R2A is -I.

[0085] In an embodiment, the halogen R 2A Exemplary compounds of formula (Ia) or (II-a) having the formula: [Table 2]

[0086] In embodiments, R 2B and R 2C each independently represents hydrogen or -OR 2F In an embodiment, R 2F is hydrogen or unsubstituted C 1 -C 4 In embodiments, R 2B is hydrogen, -OCH 3 , or -OCH 2 CH 3 In an embodiment, R 2C is hydrogen, -OCH 3 , or -OCH 2 CH 3 It is.

[0087] In embodiments, R 2B is hydrogen and R 2C -OCH 3 In an embodiment, R 2B -OCH 3 and R 2C is hydrogen. In embodiments, R 2B and R 2C is hydrogen. In embodiments, R 2B and R 2C -OCH 3 It is.

[0088] In embodiments, R 3 is hydrogen. In embodiments, R 3 is -CH 3 It is.

[0089] In one aspect, a compound is provided having a structure of formula (III) or (IV): [ka] During the ceremony: R 1 is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted alkenyl; L 1 and L 2 each independently is a bond, substituted or unsubstituted alkylene, or substituted or unsubstituted alkenylene; R 2A , R 2B and R 2C are each independently hydrogen, halogen, or -CX 2 3 , -CHX 2 2 , -CH 2 X 2 , -OCX 2 3 , -OCH 2 X 2 , -OCHX 2 2 , -OR 2F , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R 5 are independently halogen, -CX 5 3 , -CHX 5 2 , -CH 2 X 5 , -OCX 5 3 , -OCH 2 X 5 , -OCHX 5 2 , -OR 5F , or substituted or unsubstituted C 1 -C 4 is alkyl; z is an integer from 0 to 5; R 7 -OR 7F or substituted or unsubstituted alkenyl; X 2 and X 5 are each independently -F, -Br, -Cl, or -I; R 2F , R 5F and OR 7F are each independently hydrogen, or substituted or unsubstituted alkyl.

[0090] In embodiments, R 1 is hydrogen, unsubstituted C 1 -C 4 alkyl or R 1 is hydrogen, unsubstituted C 1 -C 4 Alkyl or unsubstituted C 1 -C 4 alkenyl. In embodiments, R 1 is hydrogen. In embodiments, R 1 is a substituted or unsubstituted C 1 -C 4 In embodiments, R 1 is unsubstituted C 1 -C 4 In embodiments, R 1 is unsubstituted methyl. In embodiments, R 1 is unsubstituted ethyl. In embodiments, R 1 is unsubstituted propyl. In embodiments, R 1 is unsubstituted isopropyl. In embodiments, R 1 is unsubstituted butyl. In embodiments, R 1 is unsubstituted t-butyl. In embodiments, R 1 is a substituted or unsubstituted C 1 -C 4 alkenyl. In embodiments, R 1 is unsubstituted C 1 -C 4 alkenyl. In embodiments, R 1 is unsubstituted methenyl. In embodiments, R 1 is unsubstituted ethenyl. In embodiments, R 1 is unsubstituted propenyl. In embodiments, R 1is unsubstituted isopropenyl. In embodiments, R 1 is unsubstituted butenyl. In embodiments, R 1 is unsubstituted t-butenyl.

[0091] In embodiments, z is 1. In embodiments, z is 2. In embodiments, z is 3.

[0092] In embodiments, R 5 are independently unsubstituted C 1 -C 4 In embodiments, R 5 is independently unsubstituted methyl. In embodiments, R 5 is independently unsubstituted ethyl. In embodiments, R 5 is independently unsubstituted isopropyl. In embodiments, R 5 is independently unsubstituted propyl. In embodiments, R 5 is independently unsubstituted butyl. In embodiments, R 5 is independently unsubstituted t-butyl.

[0093] In embodiments, the compound has the structure of Formula (Ia) or (II-a): [ka] In the formula, L 1 , L 2 , R 1 , R 2A , R 2B , R 2C , and R 7 is as described herein.

[0094] In embodiments, R 2A is substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted aryl. In embodiments, R 2A is a substituted or unsubstituted heterocycloalkyl. In embodiments, R 2A is substituted or unsubstituted aryl. In embodiments, R 2Ais substituted or unsubstituted piperazinyl. In embodiments, R 2A is substituted or unsubstituted phenyl.

[0095] In embodiments, R 2A teeth [ka] wherein: R 3 is hydrogen or substituted or unsubstituted alkyl; R 4A , R 4B , R 4C , and R 4D are each independently hydrogen, halogen, or -CX 4 3 , -CHX 4 2 , -CH 2 X 4 , -OCX 4 3 , -OCH 2 X 4 , -OCHX 4 2 , -OR 4F or substituted or unsubstituted alkyl; X 4 are independently -F, -Br, -Cl, or -I; R 4F is hydrogen, or substituted or unsubstituted alkyl.

[0096] In embodiments, the compound has the structure of formula (III-b) or (IV-b): [ka] L 1 , L 2 , R 1 , R 2B , R 2C , R 3 , R 4A , R 4B , R 4C , R 4D , and R 7 is as described herein.

[0097] In an embodiment, L 1 is a bond, or R 6 -Substituted or unsubstituted C 1 -C 4 In embodiments, L is an alkylene. 1 is a bond. In embodiments, L 1 is a substituted or unsubstituted C 1 -C 4 In embodiments, L is an alkylene. 1 is unsubstituted C 1 -C 4 In embodiments, L is an alkylene. 1 is a substitution C 1 -C 4 In embodiments, L is an alkylene. 1 is unsubstituted methylene. In embodiments, L 1 is unsubstituted ethylene. In embodiments, L 1 is unsubstituted propylene. In embodiments, L 1 is unsubstituted isopropylene. In embodiments, L 1 is unsubstituted butylene.

[0098] In an embodiment, L 1 is R 6 -Substituted or unsubstituted C 1 -C 4 In embodiments, R 6 is halogen or unsubstituted C 1 -C 4 In embodiments, L is an alkylene. 1 is a methyl-substituted C 1 -C 4 In embodiments, L is an alkylene. 1 is a methyl-substituted methylene. 1 is methyl substituted ethylene. In embodiments, L 1 is methyl-substituted propylene. In embodiments, L 1 is a methyl substituted butylene.

[0099] In embodiments, the compound has the structure of Formula (III-c): [ka] During the ceremony: L 1 is a bond, or R 6 -Substituted or unsubstituted C 1 -C 4 is alkylene; L 2 is a substituted or unsubstituted C 2 -C 5 Alkylene, or substituted or unsubstituted C 2 -C 5 alkenylene; R 6 is halogen or unsubstituted C 1 -C 4 It is alkylene. R 1 , R 2B , R 2C , R 3 , and R 7 is as described herein.

[0100] In embodiments, the compound has the structure of formula (IV-c): [ka] During the ceremony: L 1 is R 6 -Substituted or unsubstituted C 1 -C 4 is alkylene; L 2 is a substituted or unsubstituted C 2 -C 5 Alkylene, or substituted or unsubstituted C 2 -C 5 alkenylene; R 6 is halogen or unsubstituted C 1 -C 4 It is alkylene. R 1 , R 2B , R 2C , R 3 , and R 7 is as described herein.

[0101] In embodiments, R 7 is -OH or unsubstituted C 1 -C 4 alkenyl. In embodiments, R 7 is -OH. In embodiments, R 7 is unsubstituted C 1 -C 4 alkenyl. In embodiments, R 7 is unsubstituted methenyl. In embodiments, R 7 is unsubstituted ethenyl. In embodiments, R 7 is unsubstituted propenyl. In embodiments, R 7 is unsubstituted isopropenyl. In embodiments, R 7 is unsubstituted butenyl. In embodiments, R 7 is unsubstituted t-butenyl.

[0102] Exemplary compounds having the structure of formula (III-c) or (IV-c) are shown in Table 3. [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4]

[0103] In embodiments, R 2A is halogen. In embodiments, R 2A is -F. In embodiments, R 2A In embodiments, R 2A In an embodiment, R 2A is -I.

[0104] In an embodiment, the halogen R 2AExemplary compounds of formula (III-a) or (IV-a) having the formula: [Table 4]

[0105] In embodiments, R 2B and R 2C each independently represents hydrogen or -OR 2F In an embodiment, R 2F is hydrogen or unsubstituted C 1 -C 4 In embodiments, R 2B is hydrogen, -OCH 3 , or -OCH 2 CH 3 In an embodiment, R 2C is hydrogen, -OCH 3 , or -OCH 2 CH 3 It is.

[0106] In embodiments, R 2B is hydrogen and R 2C -OCH 3 In an embodiment, R 2B -OCH 3 and R 2C is hydrogen. In embodiments, R 2B and R 2C is hydrogen. In embodiments, R 2B and R 2C -OCH 3 It is.

[0107] In embodiments, R 3 is hydrogen. In embodiments, R 3 is -CH 3 It is.

[0108] Pharmaceutical Compositions In one aspect, pharmaceutical compositions are also provided that include one or more of the present compounds, including one or more compounds of formula (I), (II), (III), and (IV), and a pharma- ceutically acceptable excipient. The pharmaceutical compositions can include one or more compounds of formula (I), (II), (III), and (IV) in a therapeutically effective amount (e.g., a therapeutically effective amount).

[0109] In addition, the pharmaceutical composition of the present invention is prepared by further including a pharma- ceutical acceptable carrier according to each dosage form. The type of carrier that can be used in the present invention is not particularly limited, and any carrier that is commonly used in the art and is pharma- ceutical acceptable can be used.

[0110] Physiological saline, sterile water, IV fluid, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol are non-limiting examples of carriers that can be used. These carriers can be used alone or in combination of two or more. Carriers can include non-naturally occurring carriers. Optionally, other conventionally used additives such as antioxidants and / or buffers.

[0111] The pharmaceutical composition may suitably be in the form of a spray or liquid wash, or other formulations for topical application, such as lotions, creams, ointments, pastes, gels, foams, or any other physical form as a carrier generally known for topical administration. Such thickened topical formulations are particularly advantageous, since they adhere to the area of ​​skin where the material is placed, thus allowing for the introduction of a localized high concentration of one or more of the present compounds to a specific area. For example, paraffin-based and lanolin-based creams are generally known in the art. Other thickening agents, such as polymeric thickening agents, may also be used. The formulation may also include one or more of the following: water, preservatives, active surfactants, emulsifiers, antioxidants, or solvents.

[0112] Pharmaceutical compositions can be formulated for a variety of other routes of administration, such as intranasal, oral, parenteral, intramuscular, intraarticular, intravenous, subcutaneous, or transdermal administration. Suitable pharmaceutical compositions can be formulated, for example, with diluents, dispersants, surfactants, binders, lubricants to make injectable solutions, such as aqueous solutions, or as suspensions, emulsions, and as pills, capsules, granules, or tablets, and the like.

[0113] As discussed, kits are also provided.For example, one or more compounds disclosed herein, including any one of formulas (I), (II), (III) and (IV), can be suitably packaged in a suitable container with a label for use as therapy for treating subjects suffering from, for example, pigmentation disorder, uneven skin tone, hypopigmentation, inflammatory skin disease including rosacea, vitiligo, or other skin-related disorder or disease.In addition, product or kit can further include packaging material, instructions, delivery device, for example, for treating or monitoring a specific condition.

[0114] The kit may also include a legend (e.g., a printed label or insert, or other media (e.g., an audiotape or videotape) describing the use of the product). The legend may be associated with (e.g., affixed to) the container and may describe how the composition therein should be administered (e.g., frequency and route of administration), indications therefor, and other uses. The composition may be ready for administration (e.g., present in units suitable for dosage) and may include one or more additional pharma- ceutically acceptable adjuvants, carriers, or other diluents, and / or additional therapeutic agents. Alternatively, the composition may be provided in a concentrated form, e.g., with a diluent and instructions for dilution.

[0115] As discussed, a method is provided for treating a disease or disorder associated with salt-inducible kinase (SIK), such as an inflammatory disease or disorder, by administering a compound or a pharmaceutical composition comprising the compound to a subject. Preferably, the compound can have any one of the structures of the following chemical formulas (I), (II), (III) and (IV): Inflammatory diseases associated with aberrant expression of salt-inducible kinases include, but are not limited to, ulcerative colitis, rheumatoid arthritis, psoriatic rosacea and systemic lupus erythematosus; osteoporosis; hyperproliferative diseases, prostate cancer, and ovarian cancer (Darling NJ, and Cohen P., Nuts and bolts of the salt-inducible kinases (SIKs), Biochem J. (2021) Apr 16;478(7):1377-1397; Sun Z, Jiang Q, Li J and Guo J. The potent roles of salt-inducible kinases (SIKs) in metabolic homeostasis and tumorigenesis. Signal Transduction and Targeted Therapy (2020) 5:150.The method includes administering an effective amount (e.g., a therapeutically effective amount) of a compound alone or in combination with a retinoid, (adapalene 6-[3-(1-adamantyl)-4-methoxyphenyl]-2-naphthoic acid), retinoic acid, alpha hydroxy acid, butyric acid, beta hydroxy acid, salicylic acid, corticosteroids (hydrocortisone, clobetasol propionate), vitamin D3 derivatives (calcitriol 1,25-dihydroxycholecalciferol), aryl hydrocarbon receptor modulators (Tapinarof 3,5-dihydroxy-4-isopropyl-trans-stilbene), Janus kinase inhibitors (Ruxolitinib (3R)-3-cyclopentyl-3-[4-(7H-pyrrole[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propanenitrile), phosphodiesterase-4 inhibitors (Crisaborole 4-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-5-yl)oxy]benzonitrile), cell cycle checkpoint kinase 1 and 2 inhibitors (Prexasertib 5-((5-(2-(3-aminopropoxy)-6-methoxyphenyl)-1H-pyrazol-3-yl)amino)-2-pyrazinecarbonitrile), and combinations thereof, where the two separate compositions are administered simultaneously or sequentially in any order.

[0116] In a preferred embodiment, the method is for treating or preventing pigmentation disorders, uneven skin tone, hypopigmentation, inflammatory skin diseases including rosacea, vitiligo, or other skin-related diseases or disorders, suitably in a subject suffering from or susceptible to such diseases or disorders. As discussed, the subject may suitably be a human, male or female.

[0117] The subject may be afflicted with or susceptible to vitiligo, a disorder characterized by the appearance of white spots on the skin associated with pigmentation defects.

[0118] The subject may be suffering from or susceptible to any of erythematous telangiectatic rosacea (subtype 1 rosacea), papulopustular rosacea (subtype 2 rosacea), phymatous rosacea (subtype 3 rosacea) and / or ocular rosacea (subtype 4 rosacea). In some embodiments, the treatment method may further comprise identifying and selecting a subject suffering from rosacea, including a particular subtype of rosacea, or other skin-related disease or disorder. In a subject suffering from erythematous telangiectatic rosacea (subtype 1 rosacea), the subject may exhibit redness and flushing of the skin and visible blood vessels. In a subject suffering from papulopustular rosacea (subtype 2 rosacea), the subject may exhibit redness, swelling and an acne-like rash. In a subject suffering from phymatous rosacea (subtype 3), the subject may exhibit thickening of the face or other skin, and the skin may develop a bumpy texture. In subjects suffering from ocular rosacea (subtype 4), the subject may exhibit red and watery eyes, gritty eyes, dry eyes, itchy eyes, and reduced vision. The identified and selected subjects can then be treated with the therapeutic compounds or compositions disclosed herein.

[0119] In some embodiments, the treatment method may provide a method for increasing skin pigmentation and / or reducing the risk of skin cancer in a subject in need thereof. The present disclosure provides a method for increasing skin pigmentation for cosmetic purposes. In certain embodiments, the present disclosure provides a method for increasing the appearance of skin darkening in a subject in need thereof using the compounds described (e.g., via topical administration of the compounds described).

[0120] In certain embodiments, the present disclosure provides a method of increasing the appearance of skin pigmentation in a subject, comprising topically administering to the skin of the subject a compound having the structure of any one of Formulas (I), (II), (III), and (IV), or a pharma- ceutically acceptable salt, solvate, hydrate, tautomer, or stereoisomer thereof, or a pharmaceutical composition thereof.

[0121] In certain embodiments, the present disclosure provides a method for treating polymorphous light rash (e.g., sun sensitivity). The present disclosure provides a method for inducing eumelanin synthesis. The present disclosure provides a method for inducing melanosome maturation, export, and localization.

[0122] In certain embodiments, a method of reversibly increasing skin pigmentation and / or reducing the risk of skin cancer in a subject in need thereof is provided, comprising topically administering to the skin of the subject an effective amount of a compound having the structure of any one of formulas (I), (II), (III), and (IV), or a pharmaceutical composition thereof. In certain embodiments, a method of increasing skin pigmentation and / or reducing the risk of skin cancer by topically administering to the skin of a subject on a body part a compound described herein. In certain embodiments, the body part is the face of the subject. In certain embodiments, the body part is the neck of the subject. In certain embodiments, the body part is the chest of the subject. In certain embodiments, the body part is the back of the subject. In certain embodiments, the skin of the body part is the skin of the arm of the subject. In certain embodiments, the skin of the body part is the skin of the leg of the subject. In certain embodiments, the skin is on the torso of the subject.

[0123] In certain embodiments, the present invention relates to the cosmetic and / or dermatological use of a compound having the structure of any one of formulas (I), (II), (III) and (IV) or a pharmaceutical composition thereof for coloring and / or pigmenting the skin and / or body hair and / or scalp hair. The application is suitable for improving pigmentation defects and disorders, such as the appearance of gray hair in humans (gray hair or natural whitening of hair), which may be a visible sign of the aging process (senile canities) or may be related to a genetic predisposition. The pigmentation of scalp and body hair requires the presence of melanocytes in the hair bulb of the hair follicle. It is now recognized that gray hair is associated with a decrease in the amount of melanin in the hair shaft. Since maintaining a constant coloration of scalp hair requires considerable care, it is desirable to combat the appearance of these visible signs of aging, i.e. to be able to maintain or re-establish the coloration of body hair and / or scalp hair.

[0124] In certain embodiments, a method is provided for increasing the stability or repair of cellular DNA in skin tissue of a subject in need thereof, comprising administering to the subject a compound having any one of the structures of formula (I), (II), (III) and (IV), or a pharmaceutical composition thereof, in an amount sufficient to reduce apoptosis and / or thymine dimer formation in cellular DNA of skin tissue, thereby increasing the stability or repair of cellular DNA in skin tissue of the subject.The compound described herein is administered before UV exposure to increase DNA stability, and is therefore administered after UV exposure to increase DNA repair.Methods for evaluating DNA stability and repair are known in the art, including, for example, immunostaining of thymine dimer and TUNEL assay.

[0125] Manufacturing method Provided herein are methods for making compounds having the structure of Formula (I) or (II). [ka] In the formula, L 1 , L 2 , R2A , R 2B , R 2C , R 5 and z are as described herein.

[0126] In an embodiment, the method includes providing a compound having a structure of formula (III) or (IV). [ka] In the formula, L 1 , L 2 , R 1 , R 2A , R 2B , R 2C , R 5 , R 7 and z are as described herein.

[0127] In embodiments, a compound of formula (III) or (IV) may be provided in a reaction vessel.

[0128] In embodiments, after providing a compound of formula (III) or (IV), the method may further include treating the compound of formula (III) or (IV) with a coupling reagent. In embodiments, the coupling reagent may include, but is not limited to, EDC hydrochloride.

[0129] In embodiments, after providing a compound of formula (III) or (IV), the method may further include heat treating or increasing the temperature in the reaction vessel. In embodiments, during heat treating, the temperature in the reaction vessel may range from about 40° C. to about 80° C., from about 45° C. to about 70° C., or from about 50° C. to about 60° C.

[0130] In embodiments, the method may further include purifying the reaction product obtained in the reaction vessel.

[0131] In one aspect, a method is provided for treating a subject suffering from or susceptible to a skin-related disorder or disease, comprising administering to the subject an effective amount of a compound or composition described herein.

[0132] In embodiments, a subject is identified as suffering from a skin-related disorder or disease, and the compound or composition is administered to the identified subject.

[0133] In one aspect, a method of treating a subject suffering from or susceptible to rosacea is provided, comprising administering to the subject an effective amount of a compound or composition described herein.

[0134] In embodiments, a subject is identified as suffering from rosacea and the compound or composition is administered to the identified subject.

[0135] In embodiments, the subject has erythematous telangiectatic rosacea (subtype 1), papulopustular rosacea (subtype 2), tumescent rosacea (subtype 3) and / or ocular rosacea (subtype 4).

[0136] In embodiments, the subject is identified as suffering from erythematous telangiectatic rosacea (subtype 1), papulopustular rosacea (subtype 2), cystic rosacea (subtype 3) and / or ocular rosacea (subtype 4), and the compound or composition is administered to the identified subject.

[0137] In one aspect, there is provided a method of increasing pigmentation in a tissue of a subject, comprising administering to the subject a compound or composition disclosed herein comprising Formula (Ia) and / or Formula (II-a) in an amount sufficient to increase melanin production, thereby increasing pigmentation in the tissue of the subject.

[0138] In embodiments, the tissue of interest is skin or hair.

[0139] In one embodiment, a method is provided for increasing the stability or repair of cellular DNA in skin tissue of a subject in need thereof.The method includes administering to the subject the compound or composition described herein in an amount sufficient to reduce apoptosis and / or thymine dimer formation in the cellular DNA of skin tissue, thereby increasing the stability or repair of cellular DNA in the skin tissue of the subject. EXAMPLES

[0140] Example 1: Synthesis of aniline

[0141] Scheme 1 [ka]

[0142] Methyl 2-(4-methylpiperazin-1-yl)-5-nitrobenzoate (2) [ka]

[0143] Methyl 2-fluoro-5-nitrobenzenecarboxylate (20.13 g, 119.7 mmol) was dissolved in MeCN (200.0 mL), then potassium carbonate (20.14 g, 143.6 mmol) was added, followed by N-methylpiperazine (16.0 mL, 143.6 mmol) and the reaction crude was heated at 80 °C for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (23.4 g, yield: 70%, yellow solid). LC-MS (ESI+) m / z: 279.8, RT: 0.78 min (TACC50).

[0144] Methyl 5-amino-2-(4-methylpiperazin-1-yl)benzoate (3) [ka]

[0145] Methyl 2-(4-methylpiperazin-1-yl)-5-nitrobenzoate (23.43 g, 83.89 mmol) was dissolved in AcOEt (250 mL). The reaction flask was purged (vacuum and N2 gas cycle). Pd / C (2.3 g) was then added and the reaction was subjected to H2 atmosphere at room temperature for 16 h. The reaction crude was then filtered through Celite and rinsed with AcOEt. Finally, the solvent was removed under vacuum to give the pure product (21.0 g, yield: quantitative, brown light solid). LC-MS (ESI+) m / z: 249.9, RT: 0.197 min (TACC50).

[0146] Scheme 2 [ka]

[0147] [2-(4-Methylpiperazin-1-yl)-5-nitrophenyl]acetic acid (5) [ka]

[0148] (2-Fluoro-5-nitrophenyl)acetic acid (11.0 g, 55.24 mmol) was placed in a sealed tube and dissolved in 120 mL of CH3CN. N-methylpiperazine (15.37 mL, 138.1 mmol) and potassium carbonate (9.29 g, 66.29 mmol) were then added and the reaction was stirred at 80° C. for 16 hours. The reaction was then filtered to remove solids and the solvent was removed under vacuum to give a residue which was used without further purification. LC-MS (ESI+) m / z: 279.8, RT: 0.69 min (TACC50).

[0149] Methyl 2-(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)acetate (6) [ka]

[0150] Sulfuric acid (5.0 mL) was added to a solution of [2-(4-methylpiperazin-1-yl)-5-nitrophenyl]acetic acid (9.0 g, 32.2 mmol) in MeOH (100.0 mL), which was heated at 80° C. for 2 h. The reaction was cooled to room temperature and the solvent was removed under vacuum. The reaction crude was then diluted with AcOEt and washed with water. The aqueous phase was extracted several times with a CHCl3:iPrOH 1:1 mixture. The combined organic layers were dried over MgSO4, filtered, and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (8.19 g, yield: 87%). LC-MS (ESI+) m / z: 293.9, RT: 0.77 min (TACC50).

[0151] Methyl [5-amino-2-(4-methylpiperazin-1-yl)phenyl]acetate (7) [ka]

[0152] Methyl 2-(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)acetate (8.19 g, 27.92 mmol) was dissolved in MeOH (85.0 mL). The reaction flask was purged (vacuum and N2 gas cycle). Pd / C (0.82 g) was then added and the reaction was subjected to H2 atmosphere at room temperature for 16 h. The reaction crude was then filtered through Celite and rinsed with MeOH. Finally, the solvent was removed under vacuum to give the pure product (7.40 g, yield: quantitative, brown solid). LC-MS (ESI+) m / z: 263.9, RT: 0.21 min (TACC50).

[0153] Scheme 3 [ka]

[0154] Methyl 2-(2-fluoro-5-nitrophenyl)acetate (9) [ka]

[0155] A solution of 2-fluoro-5-nitrophenylacetic acid (25.0 g, 125.54 mmol) in MeOH (500 mL) was added with sulfuric acid (15.5 mL) and heated at 80° C. for 4 h. The reaction was cooled to room temperature and the solvent was removed under vacuum. The resulting residue was diluted with AcOEt, washed with water, dried over MgSO4, filtered and concentrated under vacuum to give the pure product (25.25 g, yield: 94%, brown oil) which was used without further purification. LC-MS (ESI-) m / z: 214.0, RT: 1.26 min (TACC50).

[0156] Methyl 2-(2-fluoro-5-nitrophenyl)propanoate (10) [ka]

[0157] A solution of diisopropylamine (9.43 mL, 67.08 mmol) in anhydrous THF (170 mL) was cooled to -78 °C. Then BuLi 2.5M (24.8 mL, 61.92 mmol) was added dropwise and the reaction was stirred at -78 °C for 15 min. Then a solution of methyl 2-(2-fluoro-5-nitrophenyl)acetate (11.00 g, 51.6 mmol) in anhydrous THF (10.0 mL) was added slowly and it was stirred at -78 °C for 30 min. Finally, methyl iodide (3.2 mL, 50.66 mmol) was added and it was stirred at -78 °C for 10 min. After the addition, the reaction was stirred at room temperature for 1 h. The reaction crude was then treated with aqueous NH4Cl (sat), diluted with AcOEt and washed with aqueous NH4Cl (sat). The combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt 100:0 to 70:30) to give the pure product (5.49 g, yield: 47%, orange oil). LC-MS (ESI-) m / z: 228.0, RT: 1.36 min (TACC50).

[0158] 2-[2-(4-Methyl-piperazin-1-yl)-5-nitro-phenyl]-propionic acid methyl ester (11) [ka]

[0159] Methyl 2-(2-fluoro-5-nitrophenyl)propanoate (5.49 g, 24.17 mmol) and K2CO3 (4.07 g, 29.00 mmol) were placed in a sealed tube and dissolved in anhydrous ACN (60.0 mL). N-methylpiperazine (6.72 mL, 60.41 mmol) was then added and the reaction was stirred at 80 °C for 16 h. The solvent was then removed under vacuum and the resulting residue was diluted with AcOEt and washed with water. The combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (5.24 g, yield: 71%, orange solid). LC-MS (ESI-) m / z: 308.0, RT: 0.84 min (TACC50).

[0160] 2-[5-Amino-2-(4-methyl-piperazin-1-yl)-phenyl]-propionic acid methyl ester (12) [ka]

[0161] 2-[2-(4-Methyl-piperazin-1-yl)-5-nitro-phenyl]-propionic acid methyl ester (5.24 g, 17.05 mmol) was dissolved in MeOH (50.0 mL). The reaction flask was purged (vacuum and N2 gas cycle). Pd / C (0.54 g) was then added and the reaction was subjected to H2 atmosphere at room temperature for 16 h. The reaction crude was then filtered through Celite and rinsed with MeOH. Finally, the solvent was removed under vacuum to give the pure product (4.37 g, yield: 92%). LC-MS (ESI+) m / z: 278.0, RT: 0.20 min (TACC50).

[0162] Scheme 4 [ka]

[0163] 2-[4-(tetrahydro-pyran-2-yloxy)-but-2-enyl]-isoindole-1,3-dione (14) [ka]

[0164] 3,4-Dihydro-2H-pyran (19.07 mL, 208.6 mmol) was added to a solution of 2-(4-hydroxy-but-2-enyl)-isoindole-1,3-dione (11.90 g, 69.54 mmol) in DCM (180.0 mL). Toluene-4-sulfonic acid monohydrate (13.23 g, 69.54 mmol) was then added portionwise and the reaction was stirred at room temperature for 16 h. The reaction crude was then washed with saturated aqueous NaHCO3. The combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt 100:0 to 70:30) to give the pure product (9.609 g, yield: 54%, yellow oil). LC-MS (ESI+): RT: 1.49 min (TACC50).

[0165] 1-Methyl-4-(4-nitro-2-{[(oxan-2-yl)oxy]methyl}phenyl)piperazine (15) [ka]

[0166] N-Methylpiperazine (7.32 mL, 65.80 mmol) and potassium carbonate (6.329 g, 45.13 mmol) were added to a solution of 2-[4-(tetrahydro-pyran-2-yloxy)-but-2-enyl]-isoindole-1,3-dione (9.60 g, 37.61 mmol) in DMF (115.0 mL), which was stirred at 80 °C for 16 h. The solvent was removed under vacuum and the resulting residue was diluted with DCM and washed with water. The combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (12.04 g, yield: 95%, orange oil). LC-MS (ESI+) m / z: 336.1, RT: 0.888 min (TACC50).

[0167] 4-(4-Methyl-piperazin-1-yl)-3-(tetrahydro-pyran-2-yloxymethyl)-phenylamine (16) [ka]

[0168] 1-Methyl-4-(4-nitro-2-{[(oxan-2-yl)oxy]methyl}phenyl)piperazine (8.45 g, 25.21 mmol) was dissolved in MeOH (80.0 mL). The reaction flask was purged (vacuum and N2 gas cycle). Pd / C (0.84 g) was then added and the reaction was subjected to H2 atmosphere at room temperature for 16 h. The reaction crude was then filtered through Celite and rinsed with MeOH. Finally, the solvent was removed under vacuum to give the pure product (7.07 g, yield: 92%, brown oil). LC-MS (ESI+) m / z: 306.0, RT: 0.32 min (TACC50).

[0169] Scheme 5 [ka]

[0170] 2-Fluoro-4-methoxy-5-nitro-benzaldehyde (18) [ka]

[0171] 2-Fluoro-4-methoxy-benzaldehyde (10.0 g, 64.9 mmol) was added to a stirred solution of concentrated sulfuric acid (70.0 mL) at -10°C. Nitric acid (71.364 mmol 65%, thus 6.9 mL) was added dropwise while maintaining the reaction at -10°C. After further stirring at this temperature for 2 h, the mixture was poured onto crushed ice. The resulting precipitate was collected by filtration, dissolved in dichloromethane and washed with saturated aqueous NaHCO3. The organic layer was dried over MgSO4, filtered and the solvent was removed under vacuum to give the pure product (11.58 g, yield: 90%, yellow solid). GC-MS m / z: 199.0, RT: 9.30 min.

[0172] (2-Fluoro-4-methoxy-5-nitro-phenyl)-methanol (19) [ka]

[0173] Sodium borohydride (4.593 g, 121.4 mmol) was added portionwise to a stirred solution of 2-fluoro-4-methoxy-5-nitro-benzaldehyde (12.09 g, 60.71 mmol) in methanol at 0° C. After 2 h, methanol was removed under vacuum. The residue was treated with cold water and extracted with dichloromethane. The combined organic layers were washed with brine, dried over MgSO4, and concentrated under reduced pressure to give the product, which was used without further purification (10.75 g, yield: 88%, orange oil). LC-MS (ESI+) m / z: 201.8, RT: 0.93 min (TACC50).

[0174] tert-Butyl-(2-fluoro-4-methoxy-5-nitro-benzyloxy)-dimethyl-silane (20) [ka]

[0175] Triethylamine (33.96 mL, 249.05 mmol) was added to an ice-cold solution of (2-fluoro-4-methoxy-5-nitro-phenyl)-methanol (20.04 g, 99.62 mmol) in anhydrous DCM (300 mL), the mixture was stirred at 0° C. for 5 min, then tert-butylchlorodimethylsilane (18.0 g, 119.55 mmol) was added in portions at 0° C. The reaction mixture was stirred at room temperature for 16 h. The reaction crude was diluted with DCM and washed with water. The combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt 100:0 to 70:30) to give the pure product (9.558 g, yield: 30%). LC-MS (ESI+) m / z: 280 / 254, RT: 1.89 min (TACC50).

[0176] 1-(2-(((tert-butyldimethylsilyl)oxy)methyl)-5-methoxy-4-nitrophenyl)-4-methylpiperazine (21) [ka]

[0177] Tert-Butyl-(2-fluoro-4-methoxy-5-nitro-benzyloxy)-dimethyl-silane (9.56 g, 30.3 mmol), 1-methylpiperazine (4.05 mL, 36.4 mmol) and K2CO3 (5.03 g, 36.4 mmol) were dissolved in ACN (90 mL) and stirred at 80 °C for 16 h. The solvent was removed under vacuum and the resulting residue was diluted with AcOEt and washed with water. The combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 50% 90:10) to give the pure product (2.408 g, yield: 20%). LC-MS (ESI+) m / z: 395.8, RT: 1.31 min (TACC50).

[0178] 5-(((tert-butyldimethylsilyl)oxy)methyl)-2-methoxy-4-(4-methylpiperazin-1-yl)aniline (22) [ka]

[0179] 1-[2-(tert-Butyl-dimethyl-silanyloxymethyl)-5-methoxy-4-nitro-phenyl]-4-methyl-piperazine (2.408 g, 6.087 mmol) was dissolved in AcOEt (20.0 mL). The reaction flask was purged (vacuum and N2 gas cycle). Pd / C (0.240 g) was then added and the reaction was subjected to H2 atmosphere at room temperature for 16 h. The reaction crude was then filtered through Celite and rinsed with AcOEt. Finally, the solvent was removed under vacuum to give the pure product (2.181 g, yield: 98%, brown oil). LC-MS (ESI+) m / z: 365.9, RT: 0.929 min (TACC50).

[0180] Scheme 6 [ka]

[0181] 2-Bromo-4,5-dinitrobenzoic acid (24) [ka]

[0182] Methyl 2-bromo-4-nitrobenzoate (9.66 g, 37.147 mmol) was added to an ice-cold solution of H2SO4 (30.0 mL). HNO3 (15.0 mL) was then added dropwise. The reaction was allowed to reach room temperature and it was heated at 65° C. for 16 h. The reaction was then cooled to room temperature and poured into ice-cold water. The resulting solid was collected by filtration, washed with water and dried under vacuum to give the pure product (6.41 g, yield: 59%, yellow solid). LC-MS (ESI+) RT: 0.21 min (TACC50).

[0183] Methyl 2-bromo-4,5-dinitrobenzoate (25) [ka]

[0184] Thionyl chloride (12.1 mL, 166.31 mmol) was added dropwise to an ice-cold solution of 2-bromo-4,5-dinitrobenzoic acid (4.84 g, 16.63 mmol) in MeOH (50.0 mL). The reaction was stirred at 0° C. for 30 min and then heated to reflux for 16 h. Afterwards, the solvent was removed under vacuum and excess thionyl chloride was removed by coevaporation with DCM. The resulting residue (5.07 g, yield: 99%) was used without further purification.

[0185] Methyl 2-bromo-4-methoxy-5-nitrobenzoate (26) [ka]

[0186] A solution of KOH (1.86 g, 33.2 mmol) in MeOH (10.0 mL) was added dropwise to an ice-cold solution of methyl 2-bromo-4,5-dinitrobenzoate (5.06 g, 16.6 mmol) in MeOH (60.0 mL). The reaction was then stirred at room temperature for 4 h. Afterwards, the solvent was removed under vacuum and cold water was added to the resulting residue (100 mL). The formed precipitate was collected by filtration, washed with water and dried under vacuum to give the pure product (3.95 g, yield: 82%, white solid). LC-MS (ESI+) m / z: 290.0 RT: 1.49 min (TACC50).

[0187] (2-Bromo-4-methoxy-5-nitrophenyl)methanol (27) [ka]

[0188] Methyl 2-bromo-4-methoxy-5-nitrobenzoate (2.749 g, 9.477 mmol) was dissolved in anhydrous DCM (20.0 mL) and cooled to -78 °C. DIBAL (18.9 mmol, 1 M solution in toluene, 19.0 mL) was then added and the reaction was stirred at -78 °C for 1 h. The reaction was then quenched by adding aqueous NaOH and filtered through a plug of Celite. The filtrate was extracted with DCM and washed with brine. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure to give the product, which was used without further purification (1.00 g, yield: 40%, yellow solid). GC-MS (ESI+) m / z: 262.9 RT: 11.307 min.

[0189] (5-Amino-2-bromo-4-methoxyphenyl)methanol (28) [ka]

[0190] (2-Bromo-4-methoxy-5-nitrophenyl)methanol (3.12 g, 11.90 mmol), ammonium chloride (6.369 g, 119.1 mmol) and iron (3.325 g, 59.53 mmol) were placed in a sealed tube and dissolved in EtOH:H2O mixture (50.0 mL, 4:1). The reaction was then heated at 65° C. for 16 h. The reaction crude was then filtered through a plug of Celite and rinsed with EtOH. The solvent was removed under vacuum and the resulting residue was diluted with AcOEt and neutralized with saturated aqueous NaHCO3. The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to give the pure product (2.52 g, yield: 91%, brown solid), which was used without further purification. LC-MS (ESI+) m / z: 232.0 RT: 0.66 min (TACC50).

[0191] Scheme 7 [ka]

[0192] 2-Fluoro-4-methoxy-5-nitro-benzaldehyde (30) [ka]

[0193] 2-Fluoro-4-methoxy-benzaldehyde (5.0 g, 32.438 mmol) was added to concentrated sulfuric acid (35.0 mL) at 0° C. Nitric acid (35.682 mmol 65%, thus 3.5 mL) was then added dropwise while maintaining the solution at −10° C. After further stirring at this temperature for 2 h, the mixture was poured onto crushed ice. The resulting precipitate was collected by filtration, dissolved in dichloromethane and washed with saturated aqueous NaHCO3. The organic layer was dried over MgSO4 and concentrated under vacuum to give the pure product (5.48 g, yield: 85%, yellow solid).

[0194] (2-Fluoro-4-methoxy-5-nitro-phenyl)-methanol (31) [ka]

[0195] Sodium borohydride (2.454 g, 64.880 mmol) was added in portions to an ice-cold solution of 2-fluoro-4-methoxy-5-nitro-benzaldehyde (5.48 g, 32.44 mmol) in methanol (80.0 mL). After 2 h, the solvent was removed under vacuum and the resulting residue was treated with cold water and finally extracted with dichloromethane. The combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt 100:0 to 50:50) to give the pure product (1.71 g, yield: 26%, yellow oil). LC-MS (ESI+) m / z: 201.9 RT: 1.05 min (TACC50).

[0196] (5-Amino-2-fluoro-4-methoxyphenyl)methanol (32) [ka]

[0197] (2-Fluoro-4-methoxy-5-nitro-phenyl)-methanol (1.710 g, 8.501 mmol) was dissolved in AcOEt (30.0 mL). The reaction was subjected to vacuum-N2 atmosphere cycles. Pd on carbon (0.170 g) was then added and the reaction was subjected to H2 atmosphere for 16 h. The reaction crude was filtered through a plug of Celite and rinsed with AcOEt. Finally, the solvent was removed under vacuum to give the pure product (0.856 g, yield: 59%, brown light solid). LC-MS (ESI+) m / z: 172.0 RT: 0.25 min (TACC50).

[0198] Scheme 8 [ka]

[0199] 2,4-Dichloro-5-(chloromethyl)pyrimidine (34) [ka]

[0200] DIPEA (92.0 mL, 527.72 mmol) was slowly added via an addition funnel to an ice-cold solution of 5-hydroxymethyl-1H-pyrimidine-2,4-dione (25.00 g, 175.90 mmol) and phosphorus(V) oxychloride (82.0 mL, 879.54 mmol) in toluene (75.0 mL). After the addition was complete, the cooling bath was removed and the mixture was heated very slowly at 115° C. for 1 h and then at 125° C. for 3 h. Once the reaction was complete, the reaction mixture was cooled to room temperature and carefully added to an ice-cold biphasic mixture of water (250 mL) and AcOEt (250 mL) using an addition funnel. Finally, the mixture was stirred at 0° C. for 45 min and then extracted with toluene. The combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane / AcOEt 100:0 to 50:50) to give the pure product (24.72 g, yield: 71%, pale yellow oil). LC-MS (ESI+) m / z: 198.1, RT: 1.41 min (TACC50).

[0201] (2,4-Dichloro-pyrimidin-5-ylmethyl)-(2,6-dimethyl-phenyl)-amine (35) [ka]

[0202] Sodium iodide (1.139 g, 7.597 mmol) was dissolved in acetone (20 mL) and the mixture was stirred until a clear solution was obtained. Then, 2,4-dichloro-5-chloromethyl-pyrimidine (1.500 g, 7597 mmol) was added under N2 atmosphere and the mixture was stirred at room temperature for 15 min and at 60 °C for 45 min. The mixture was cooled and diluted with acetone (40 mL). Then, 2,6-dimethyl-phenylamine (0.939 mL, 7.597 mmol) and potassium carbonate (3.150 g, 22.791 mmol) were added and the reaction was heated at 55 °C for 12 h. The reaction mixture was then poured into cold water and extracted with AcOEt. The organic layer was washed with aqueous sodium bisulfite and brine, dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane / EtOAC 100:0 to 85:15) to give the pure product (1.677 g, yield: 87%, white solid). LC-MS (ESI+) m / z: 282.0, RT: 1.82 min (TACC50).

[0203] Example 2: Synthesis of A-type macrocycles

[0204] Scheme 9 [ka]

[0205] 3-(tert-Butyl-dimethyl-silanyloxy)-propylamine (36) [ka]

[0206] 3-Amino-1-propanol (5.00 mL, 66.57 mmol) and tert-butyldimethylsilyl chloride (12.04 g, 79.88 mmol) were dissolved in anhydrous DCM (170 mL) and cooled to 0° C. Anhydrous triethylamine (15.77 mL, 113.17 mmol) was then added and the reaction was allowed to come to room temperature and stirred for 16 h. The reaction mixture was then washed with water. The organic layer was dried over MgSO4, filtered and concentrated under vacuum to give the pure product, which was used without further purification. LC-MS (ESI+) m / z: 190.0, RT: 0.97 min (TACC50).

[0207] [3-(tert-butyl-dimethyl-silanyloxy)-propyl]-{2-chloro-5-[(2,6-dimethyl-phenylamino)-methyl]-pyrimidin-4-yl}-amine (37) [ka]

[0208] 3-(tert-Butyl-dimethyl-silanyloxy)-propylamine (12.54 g, 66.23 mmol) and triethylamine (9.48 mL, 68.04 mmol) were added to a solution of N-((2,4-dichloropyrimidin-5-yl)methyl)-2,6-dimethylaniline (12.8 g, 45.36 mmol) in anhydrous THF (130.0 mL), and the mixture was stirred at 65° C. for 16 h. The reaction crude was then diluted with AcOEt and washed with brine. The organic layer was dried over MgSO4, filtered, and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt 100:0 to 70:30) to give the pure product (13.24 g, yield: 67%, colorless oil). LC-MS (ESI+) m / z: 436.0, RT: 2.05 min (TACC50).

[0209] 7-Chloro-3-(2,6-dimethyl-phenyl)-1-(3-hydroxy-propyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (38) [ka]

[0210] Triphosgene (4.51 g, 15.20 mmol) was added to a solution of [3-(tert-butyl-dimethyl-silanyloxy)-propyl]-{2-chloro-5-[(2,6-dimethyl-phenylamino)-methyl]-pyrimidin-4-yl}-amine (8.82 g, 20.27 mmol) in DCM (60.0 mL) and it was stirred at room temperature for 1 h. Then an aqueous solution of sodium hydroxide (8.11 g, 202.72 mmol, 4.9 M) and tetrabutylammonium hydroxide (2.63 mmol 55% aqueous solution, thus 1.31 mL) was added at 0° C. The reaction was allowed to come to room temperature and stirred at room temperature for 16 h. The reaction crude was then diluted with DCM and washed with water. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt, 100:0 to 30:70) to give the pure product (6.04 g, yield: 86%, white solid). LC-MS (ESI+) m / z: 348.0, RT: 1.19 min (TACC50).

[0211] Methyl 5-((6-(2,6-dimethylphenyl)-8-(3-hydroxypropyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)benzoate (39) [ka]

[0212] 7-Chloro-3-(2,6-dimethyl-phenyl)-1-(3-hydroxy-propyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (0.480 g, 1.384 mmol) and methyl 5-amino-2-(4-methylpiperazino)benzenecarboxylate (0.459 g, 1.841 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (20.0 mL). 3A MS was then added, followed by trifluoroacetic anhydride (0.213 mL, 2.768 mmol) and the reaction was stirred at 90° C. for 16 hours. The reaction was then diluted with water and treated with aqueous NaHCO3 (2.768 mmol). The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (0.64 g, yield: 83%). LC-MS (ESI+) m / z: 561.2, RT: 0.52 min (TACC50).

[0213] 5-((6-(2,6-dimethylphenyl)-8-(3-hydroxypropyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)benzoic acid (40) [ka]

[0214] Methyl 5-((6-(2,6-dimethylphenyl)-8-(3-hydroxypropyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)benzoate was dissolved in THF (4.0 mL) and treated with HCl (5.36 mmol, 6 M aqueous solution, 0.89 mL), which was stirred at 60° C. for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (95% [0.1% aqueous HCOOH]-5% ACN to 63% [0.1% aqueous HCOOH]-37% ACN) to give the pure product (0.440 g, yield: 75%). LC-MS (ESI+) m / z: 547.3, RT: 2.094 min (VILLA).

[0215] 1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclooctaphane-1 2 ,4-dione(41) [ka]

[0216] 5-((6-(2,6-dimethylphenyl)-8-(3-hydroxypropyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)benzoic acid (0.870 g, 1.598 mmol) and DMAP (0.488 g, 3.995 mmol) were placed in a sealed tube and dissolved in anhydrous DCM (200 mL). 3A MS was then added, followed by EDC hydrochloride (0.306 g, 1.598 mmol) and 1-hydroxybenzotriazole hydrate (0.324 g, 2.397 mmol) and the reaction was heated at 50° C. for 16 hours. The reaction crude was then washed with an aqueous solution of Rochelle's salt. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:NH 3 Purification by 100:0 (deactivated with 1% DCM:MeOH 100:0 to 90:10) gave the pure product (0.250 g, yield: 30%, pale yellow solid). LC-MS (ESI+) m / z: 528.2, RT: 2.354 min (VILL_J). 1H NMR (400 MHz, CDCl3) δ 9.93 (d, J = 2.7 Hz, 1H), 7.97 (s, 1H), 7.20-7.08 (m, 4H), 7.00 (dd, J = 8.6, 2.7 Hz, 1H), 6.88 (d, J = 8.6 Hz, 1H), 4.83-4.33 (m, broad signal, 4H), 4.28-3.92 (m, broad signal 2H), 3.25 (s, 4H), 2.72 (s, 4H), 2.42 (s, 3H), 2.25 (s, 6H), 0.86 (m, 2H). [ka]

[0217] 4-(tert-Butyl-dimethyl-silanyloxy)-butylamine (42) [ka]

[0218] 4-Amino-1-butanol (5.00 mL, 56.09 mmol) and tert-butyldimethylsilyl chloride (8.88 g, 58.9 mmol) were dissolved in anhydrous DCM (100 mL) and cooled to 0° C. Anhydrous triethylamine (10.16 mL, 72.92 mmol) was then added and the reaction was allowed to reach room temperature and stirred for 16 h. The reaction mixture was then washed with water. The organic layer was dried over MgSO4, filtered and concentrated under vacuum to give the pure product (11.40 g, yield: quantitative), which was used without further purification. LC-MS (ESI+) m / z: 204.0, RT: 0.901 min (TACC50).

[0219] [4-(tert-Butyl-dimethyl-silanyloxy)-butyl]-{2-chloro-5-[(2,6-dimethyl-phenylamino)-methyl]-pyrimidin-4-yl}-amine (43) [ka]

[0220] 4-(tert-Butyl-dimethyl-silanyloxy)-butylamine (11.05 g, 54.33 mmol) and triethylamine (7.78 mL, 55.82 mmol) were added to a solution of (2,4-dichloro-pyrimidin-5-ylmethyl)-(2,6-dimethyl-phenyl)-amine (10.50 g, 37.21 mmol) in anhydrous THF (130.0 mL) and the mixture was stirred at 65° C. for 16 h. The reaction crude was then diluted with AcOEt and washed with brine. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt, 100:0 to 70:30) to give the pure product (16.70 g, yield: quantitative, colorless oil). LC-MS (ESI+) m / z: 450.0, RT: 2.14 min (TACC50).

[0221] 7-Chloro-3-(2,6-dimethyl-phenyl)-1-(4-hydroxy-butyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (44) [ka]

[0222] Triphosgene (8.28 g, 27.91 mmol) was added to a solution of [4-(tert-butyl-dimethyl-silanyloxy)-butyl]-{2-chloro-5-[(2,6-dimethyl-phenylamino)-methyl]-pyrimidin-4-yl}-amine (16.7 g, 37.21 mmol) in DCM (100 mL), which was stirred at room temperature for 1 h. Then an aqueous solution of sodium hydroxide (14.89 g, 372.12 mmol, 4.9 M) and tetrabutylammonium hydroxide (4.84 mmol 55% aqueous solution, thus 2.4 mL) was added at 0° C. The reaction was allowed to come to room temperature and stirred at room temperature for 16 h. The reaction crude was then diluted with DCM and washed with water. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was dissolved in DMF (100.0 mL) and cesium carbonate (12.25 g, 37.58 mmol) was added. The reaction mixture was heated at 60° C. for 16 h. The reaction crude was then diluted with DCM and washed with brine. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt, 100:0 to 70:30) to give the pure product (3.43 g, yield: 26%, white solid). LC-MS (ESI+) m / z: 360.0, RT: 1.21 min (TACC50).

[0223] 5-[6-(2,6-dimethyl-phenyl)-8-(4-hydroxy-butyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzoic acid methyl ester (45) [ka]

[0224] 7-Chloro-3-(2,6-dimethyl-phenyl)-1-(4-hydroxy-butyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (0.5 g, 1.386 mmol) and methyl 5-amino-2-(4-methylpiperazino)-benzenecarboxylate (0.459 g, 1.843 mmol) were placed in a sealed tube with anhydrous 2-BuOH (4.0 mL) and dissolved in anhydrous 2-BuOH (200 mL). 3A MS was then added followed by anhydrous TFA (0.316 mL, 2.772 mmol) and the reaction was stirred at 90° C. for 16 hours. The reaction was then diluted with water and treated with aqueous NaHCO3 (2.772 mmol). The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (0.371 g, yield: 47%, yellow solid). LC-MS (ESI+) m / z: 574.8 RT: 1.12 min (TACC50).

[0225] 5-((6-(2,6-dimethylphenyl)-8-(4-hydroxybutyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)benzoic acid (46) [ka]

[0226] 5-[6-(2,6-dimethyl-phenyl)-8-(4-hydroxy-butyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzoic acid methyl ester (0.516 g, 0.899 mmol) was dissolved in THF (4.0 mL) and treated with HCl (4.49 mmol, 6 M aqueous solution, 0.75 mL), which was stirred for 16 h at 60° C. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (95% [0.1% aqueous HCOOH]-5% ACN to 63% [0.1% aqueous HCOOH]-37% ACN) to give the pure product (0.338 g, yield: 67%, white solid). LC-MS (ESI+) m / z: 560.2, RT: 0.67 min (TACC50).

[0227] 1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclononaphane-1 2 ,4-dione(47) [ka]

[0228] 5-((6-(2,6-dimethylphenyl)-8-(4-hydroxybutyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)benzoic acid (0.350 g, 0.625 mmol) was dissolved in anhydrous THF (36.0 mL). Triethylamine (0.436 mL, 3.125 mmol) and 2,4,6-trichlorobenzoyl chloride (0.489 mL, 3.125 mmol) were then added and the reaction was stirred at room temperature for 16 hours. The solvent was then removed under vacuum and the resulting residue was dissolved in anhydrous DMF (5.0 mL) and diluted with anhydrous toluene (106.0 mL). It was then added dropwise via an addition funnel to a refluxing solution of DMAP (0.458 g, 3.750 mmol) in anhydrous toluene (14.0 mL, final concentration 0.005 mmol / mL, total volume 125.0 mL), which was stirred at that temperature for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (59% [25 mm aq. NH4HCO3]-41% ACN to 17% [25 mm aq. NH4HCO3]-83% ACN) to give the pure product (0.074, yield: 22%, white solid). LC-MS (ESI+) m / z: 542.2, RT: 2.459 min (VILL_J). 1H NMR (400 MHz, CDCl3) δ 9.60 (s, 1H), 7.96 (s, 1H), 7.21-7.10 (m series, 4H), 7.02-6.97 (m series, 2H), 4.50 (s, 2H), 4.33 (t, J = 5.8 Hz, 2H), 4.18-4.12 (m, 2H), 3.29 (broad signal, 4H), 2.91 (broad signal, 4H), 2.55 (s, 3H), 2.26 (s, 6H), 2.14-2.05 (m, 2H), 1.87-1.79 (m, 2H). [ka]

[0229] 5-{2-chloro-5-[(2,6-dimethyl-phenylamino)-methyl]-pyrimidin-4-ylamino}-pentan-1-ol (48) [ka]

[0230] 5-Amino-1-pentanol (1.29 g, 12.48 mmol) and triethylamine (1.74 mL, 12.48 mmol) were added to a solution of N-((2,4-dichloropyrimidin-5-yl)methyl)-2,6-dimethylaniline (3.200 g, 11.341 mmol) in anhydrous THF (32.0 mL), and the mixture was stirred at 65 °C for 16 h. The reaction crude was then diluted with AcOEt and washed with brine. The organic layer was dried over MgSO4, filtered, and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 19:1) to give the pure product (2.49 g, yield: 63%, orange oil). LC-MS (ESI+) m / z: 348.9, RT: 1.47 min (TACC50).

[0231] 7-Chloro-3-(2,6-dimethyl-phenyl)-1-(5-hydroxy-pentyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (49) [ka]

[0232] Triphosgene (B, 3.183 g, 10.727 mmol) was added to a solution of 5-{2-chloro-5-[(2,6-dimethyl-phenylamino)-methyl]-pyrimidin-4-ylamino}-pentan-1-ol (2.495 g, 7.151 mmol) in DCM (22.0 mL), which was stirred at room temperature for 1 h. Then, an aqueous solution of sodium hydroxide (5.721 g, 143.02 mmol, 4.9 M) and tetrabutylammonium hydroxide (0.930 mmol 55% aqueous solution, thus 0.441 mL) was added at 0° C. The reaction was allowed to come to room temperature and stirred at room temperature for 16 h. Afterwards, the reaction crude was diluted with DCM and washed with water. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH, 100:0 to 90:10) to give the pure product (0.953 g, yield: 36%, yellow solid). LC-MS (ESI+) m / z: 374.9, RT: 1.42 min (TACC50).

[0233] 5-[6-(2,6-dimethyl-phenyl)-8-(5-hydroxy-pentyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzoic acid methyl ester (50) [ka]

[0234] 7-Chloro-3-(2,6-dimethyl-phenyl)-1-(5-hydroxy-pentyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (0.50 g, 1.334 mmol) and methyl 5-amino-2-(4-methylpiperazino)benzenecarboxylate (0.442 g, 1.774 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (20.0 mL). 3A MS was then added, followed by trifluoroacetic anhydride (0.20 mL, 2.67 mmol) and the reaction was stirred at 90° C. for 16 hours. The reaction was then diluted with water and treated with an aqueous solution of NaHCO3 (2.67 mmol). The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (0.420 g, yield: 54%, yellow solid). LC-MS (ESI+) m / z: 587.8, RT: 2.308 min (TACC50).

[0235] 5-[6-(2,6-dimethyl-phenyl)-8-(5-hydroxy-pentyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzoic acid (51) [ka]

[0236] 5-[6-(2,6-dimethyl-phenyl)-8-(5-hydroxy-pentyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzoic acid methyl ester (0.45 g, 0.771 mmol) was dissolved in THF (3 mL) and treated with HCl (3.855 mmol, 6 M aqueous solution, 0.643 mL), which was stirred for 16 h at 60° C. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (95% [0.1% aqueous HCOOH]-5% ACN to 63% [0.1% aqueous HCOOH]-37% ACN) to give the pure product (0.296 g, yield: 67%, white solid). LC-MS (ESI+): m / z: 575.0, RT: 2.217 min (VILLA).

[0237] 1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)benzenecyclodecaphane-1 2 ,4-dione(52) [ka]

[0238] 5-[6-(2,6-dimethyl-phenyl)-8-(5-hydroxy-pentyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzoic acid (0.130 g, 0.227 mmol) was dissolved in anhydrous THF (13.0 mL). Triethylamine (0.16 mL, 1.135 mmol) and 2,4,6-trichlorobenzoyl chloride (0.18 mL, 1.135 mmol) were then added and the reaction was stirred at room temperature for 16 hours. The solvent was then removed under vacuum and the resulting residue was dissolved in anhydrous DMF (5.0 mL) and diluted with anhydrous toluene (35.0 mL). It was then added dropwise to a refluxing solution of anhydrous DMAP (0.166 g, 1.362 mmol). Toluene (5.0 mL, final concentration 0.005 mmol / mL, total volume 45.0 mL) was added via dropping funnel and it was stirred at that temperature for 16 h. Then the solvent was removed under vacuum and the resulting residue was purified by reverse phase chromatography (59% [25 mm aq. NH4HCO3]-41% ACN to 17% [25 mm aq. NH4HCO3]-83% ACN) to give the pure product (0.0066 g, yield: 6%, white solid). LC-MS (ESI+): m / z: 556.3176, RT: 3.187 min (VLIA_2T). 1 1H NMR (400MHz, CDCl 3 ) δ 9.06 (d, J = 2.5 Hz, 1H), 7.87 (s, 1H), 7.13-6.99 (m series, 5H), 6.92 (dd, J = 8.7, 2.7 Hz, 1H), 4.42 (s, 2H), 4.30-4.25 (m, 2H), 4.06-3.95 (m, 2H), 3.15 (broad signal, 4H), 2.81 (broad signal, 4H), 2.47 (s, 3H), 2.19 (s, 6H), 2.03-1.95 (m, J = 15.2, 7.2 Hz, 2H), 1.82-1.75 (m, 2H), 1.67-1.59 (m, J = 12.6, 6.3 Hz, 2H).

[0239] B-type macrocycles [ka]

[0240] The previous intermediates (35-38) have already been described in the experimental section.

[0241] [5-[6-(2,6-dimethyl-phenyl)-8-(3-hydroxy-propyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-acetic acid methyl ester (53) [ka]

[0242] 7-Chloro-3-(2,6-dimethyl-phenyl)-1-(3-hydroxy-propyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (0.277 g, 0.799 mmol) and methyl [5-amino-2-(4-methylpiperazin-1-yl)phenyl]acetate (0.280 g, 1.063 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (3.0 mL). 3A MS was then added, followed by trifluoroacetic anhydride (0.122 mL, 1.6 mmol) and the reaction was stirred at 90° C. for 16 hours. The reaction was then diluted with water and treated with aqueous NaHCO3. A solution of NaHCO3 (1.6 mmol) was added. The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (0.449 g, yield: 98%, light brownish solid). LC-MS (ESI+): m / z: 575.3, RT: 2.211 min (VILLA).

[0243] [5-[6-(2,6-dimethyl-phenyl)-8-(3-hydroxy-propyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-acetic acid (54) [ka]

[0244] [5-[6-(2,6-dimethyl-phenyl)-8-(3-hydroxy-propyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-acetic acid methyl ester (0.370 g, 0.645 mmol) was dissolved in THF (3.0 mL) and treated with HCl (3.22 mmol, 6 M solution, 0.538 mL) for 5 h at 60° C. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (95% [25 mm aq. NH4HCO3]-5% ACN to 63% [25 mm aq. NH4HCO3]-37% ACN) to give the pure product (0.099 g, yield: 27%, white solid). LC-MS (ESI+): m / z: 561.3, RT: 2.065 min (VILLA). 1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-6-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclononaphane-1 2 ,5-dione(55)

[0245] 1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-6-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclononaphane-1 2 ,5-dione(55) [ka]

[0246] [5-[6-(2,6-dimethyl-phenyl)-8-(3-hydroxy-propyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-acetic acid was dissolved in anhydrous DMF (2.0 mL) and diluted with anhydrous DCM (63.0 mL). 3A molecular sieves were added, followed by DMAP (0.060 g, 0.492 mmol) and DCC (0.081 g, 0.394 mmol) and the reaction was stirred at room temperature for 2 h. The solvent was then removed under vacuum and the resulting residue was dissolved in ACN and the solid formed was removed by filtration. The resulting filtrate was concentrated under vacuum to give a residue, which was purified by reverse phase chromatography (70% 65mm [NH4OAc+ACN (90:10)]-30% ACN to 27% [NH4OAc+ACN (90:10)]-73% ACN) to give the pure product (0.0384g, yield: 21%, white solid). LC-MS (ESI+): m / z: 542.3, RT: 2.489min (VILL-J). 1H NMR (400 MHz, CDCl3) δ 8.37 (d, J = 2.4 Hz, 1H), 7.92 (s, 1H), 7.63-7.54 (broad signal, 1H), 7.19-7.10 (m, 3H), 7.10-7.05 (m, 1H), 6.80 (dd, J = 8.5, 2.5 Hz, 1H), 4.47 (s, 2H), 4.40-4.32 (m, 2H), 4.29-4.21 (m, 2H), 3.84 (s, 2H), 2.93 (t, J = 4.7 Hz, 4H), 2.61 (broad signal, 4H), 2.37 (s, 3H), 2.24 (s, 6H), 2.13-2.04 (m, 2H). [ka]

[0247] The previous intermediates (35-38) have already been described in the experimental section.

[0248] 2-[5-[6-(2,6-dimethyl-phenyl)-8-(3-hydroxy-propyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-propionic acid methyl ester (56) (as a racemic mixture) [ka]

[0249] 7-Chloro-3-(2,6-dimethyl-phenyl)-1-(3-hydroxy-propyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (0.969 g, 2.79 mmol) and 2-[5-amino-2-(4-methyl-piperazin-1-yl)-phenyl]-propionic acid methyl ester (1.03 g, 3.72 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (8.0 mL). 3A MS was then added, followed by trifluoroacetic anhydride (0.428 mL, 5.6 mmol) and the reaction was stirred at 90° C. for 16 hours. The reaction was then diluted with water and treated with aqueous NaHCO3. A solution of NaHCO3 (5.6 mmol) was added. The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated in vacuo. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (1.33 g, yield: 81%, brown solid). LC-MS (ESI+): m / z: 588.3 RT: 1.178 min (TACC50).

[0250] 2-[5-[6-(2,6-dimethyl-phenyl)-8-(3-hydroxy-propyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-propionic acid (57) (as a racemic mixture) [ka]

[0251] 2-[5-[6-(2,6-dimethyl-phenyl)-8-(3-hydroxy-propyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-propionic acid methyl ester (0.8 g, 1.37 mmol) was dissolved in THF (4.0 mL) and treated with HCl (6.9 mmol, 6 M aqueous solution, 1.2 mL). The reaction was stirred at 60° C. for 16 h. The solvent was then removed under vacuum and the resulting residue was used without further purification. LC-MS (ESI+): m / z: 575.3, RT: 2.175 min (VILLA).

[0252] 1 3 -(2,6-dimethylphenyl)-4-methyl-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-6-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclononaphane-1 2 ,5-dione(58) [ka]

[0253] 2-[5-[6-(2,6-dimethyl-phenyl)-8-(3-hydroxy-propyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-propionic acid (0.393 g, 0.685 mmol) was dissolved in anhydrous DMF (3.0 mL) and diluted with anhydrous DCM (130.0 mL). 3A molecular sieves were added, followed by DMAP (0.125 g, 1.03 mmol) and DCC (0.170 g, 0.82 mmol) and the reaction was stirred at room temperature for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by flash column chromatography on silica (deactivated with DCM:NH3 1%) (DCM:MeOH 100:0 to 90:10) to give the pure product. SFC purification was carried out on a Jasco SFC prep system using an i-Cellulose-C column (Regis Technologies) 100 mm length x 4.6 mm ID 3 μm particle size in isocratic mode at 2.5 mL / min CO2 (60%)-ethanol (40%) + 0.1% diethylamine, 35 °C, BPR 100 Bar to give the pure enantiomers. Enantiomer F1 (0.0291 g): LC-MS (ESI+): m / z: 557.3 RT: 2.450 min (VILLA). 99% ee (RT: 8.142 min). Enantiomer F2 (0.0311 g): LC-MS (ESI+): m / z: 557.3 RT: 2.390 min (VILLA). 98% ee (RT: 8.473 min).1H NMR(400MHz,DMSO)δ9.56(s,1H),8.14(s,1H),8.10(s,1H),7.19-7.12(m,4H),7.04(dd,J=8.6 ,2.4Hz,1H),4.54(d,J=14.5Hz,1H),4.40(d,J=14.4Hz,2H),4.34(q,J=6.9Hz,1H),4.17(t,J= 10.2Hz,1H),4.11-4.01(m,2H),2.98-2.91(m,2H),2.73-2.66(m,2H),2.48-2.40(m,4H),2.23 (s,3H),2.20(s,3H),2.13(s,3H),1.99-1.88(m,1H),1.86-1.75(m,1H),1.31(d,J=6.9Hz,3H). [ka]

[0254] The previous intermediates (35-44) have already been described in the experimental section.

[0255] [5-[6-(2,6-dimethyl-phenyl)-8-(4-hydroxy-butyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-acetic acid methyl ester (59) [ka]

[0256] 7-Chloro-3-(2,6-dimethyl-phenyl)-1-(4-hydroxy-butyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (1.0 g, 2.771 mmol) and methyl [5-amino-2-(4-methylpiperazin-1-yl)phenyl]acetate (0.970 g, 3.685 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (20.0 mL). 3A MS was then added, followed by trifluoroacetic anhydride (0.63 mL, 5.54 mmol) and the reaction was stirred at 90° C. for 16 hours. The reaction was then diluted with water and treated with an aqueous solution of NaHCO3 (5.54 mmol). The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH, 100:0 to 90:10) to give the pure product (1.047 g, yield: 64%, light brown solid). LC-MS (ESI+): m / z: 588.3 RT: 2.290 min (VILLA).

[0257] 2-(5-((6-(2,6-dimethylphenyl)-8-(4-hydroxybutyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)phenyl)acetic acid (60) [ka]

[0258] [5-[6-(2,6-dimethyl-phenyl)-8-(4-hydroxy-butyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-acetic acid methyl ester (0.598 g, 1.02 mmol) was dissolved in THF (4.0 mL) and treated with HCl (5.08 mmol, 6 M aqueous solution, 0.845 mL). The reaction was stirred at 60° C. for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (95% [0.1% HCOOH aqueous solution]-5% ACN to 63% [0.1% HCOOH aqueous solution]-37% ACN) to give the pure product (0.186 g, yield: 32%). LC-MS (ESI+): m / z: 575.3, RT: 2.096 min (VILLA).

[0259] 1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-6-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclodecaphane-1 2 ,5-dione(61) [ka]

[0260] 2-(5-((6-(2,6-dimethylphenyl)-8-(4-hydroxybutyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)phenyl)acetic acid (0.160 g, 0.279 mmol) was dissolved in anhydrous THF (12.0 mL). Triethylamine (0.086 mL, 0.614 mmol) and 2,4,6-trichlorobenzoyl chloride (0.087 mL, 0.558 mmol) were then added and the reaction was stirred at room temperature for 16 hours. The solvent was then removed under vacuum and the resulting residue was dissolved in anhydrous DMF (2.0 mL) and diluted with anhydrous toluene (4.0 mL). It was then added dropwise via an addition funnel to a refluxing solution of DMAP (0.205 g, 1.674 mmol) in anhydrous toluene (50.0 mL, final concentration 0.005 mmol / mL, total volume 56.0 mL), which was stirred at that temperature for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (47% [65 mM NH4OAc+ACN (90:10)]-53% ACN to 18% [65 mM NH4OAc+ACN (90:10)]-82% ACN) to give the pure product (0.020 g, yield: 13%). LC-MS (ESI+): m / z: 556.5, RT: 2.494 min (VILLA). 1H NMR(400MHz,DMSO)δ9.47(s,1H),8.12(s,1H),7.77(s,1H),7.35(d,J=8.1Hz,1H),7.20-7.13(m,3H),7.09(d,J=8.6Hz,1H),4.49(s,2H),4. 07(t,J=5.8Hz,2H),3.99(t,J=6.6Hz,2H),3.58(s,2H),2.70-2.63(m,4H),2.37-2.27(m,4H),2.18(s,6H),2.11(s,3H),1.79-1.62(m,4H). [ka]

[0261] The previous intermediates (35-44) have already been described in the experimental section.

[0262] 2-[5-[6-(2,6-dimethyl-phenyl)-8-(4-hydroxy-butyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-propionic acid methyl ester (62) (as racemic mixture) [ka]

[0263] 7-Chloro-3-(2,6-dimethyl-phenyl)-1-(4-hydroxy-butyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (1.40 g, 3.88 mmol) and 2-[5-amino-2-(4-methyl-piperazin-1-yl)-phenyl]-propionic acid methyl ester (1.43 g, 5.16 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (20.0 mL). 3A MS was then added, followed by trifluoroacetic anhydride (0.59 mL, 7.76 mmol) and the reaction was stirred at 90° C. for 16 hours. The reaction was then diluted with water and treated with aqueous NaHCO3 (7.76 mmol). The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH, 100:0 to 90:10) to give the pure product (1.24 g, yield: 53%, yellow oil). LC-MS (ESI+): m / z: 601.7, RT: 2.148 min (VILLA).

[0264] 2-[5-[6-(2,6-dimethyl-phenyl)-8-(4-hydroxy-butyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-propionic acid (63) [ka]

[0265] 2-[5-[6-(2,6-dimethyl-phenyl)-8-(4-hydroxy-butyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-propionic acid methyl ester (0.99 g, 1.65 mmol) was dissolved in THF (10.0 mL) and treated with HCl (8.22 mmol, 6 M aqueous solution, 1.37 mL). The reaction was stirred at 60° C. for 16 h. LC-MS (ESI+): m / z: 588.0, RT: 1.57 min (TACC50).

[0266] 1 3 -(2,6-dimethylphenyl)-4-methyl-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-6-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclodecaphane-12,5-dione (64) [ka]

[0267] 2-[5-[6-(2,6-dimethyl-phenyl)-8-(4-hydroxy-butyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-propionic acid (0.965 g, 1.645 mmol) was dissolved in anhydrous DMF (2.0 mL) and diluted with anhydrous DCM (220 mL). 3A molecular sieves were added, followed by DMAP (0.30 g, 2.47 mmol) and DCC (0.41 g, 1.97 mmol) and the reaction was stirred at room temperature for 16 hours. The reaction was recharged with DMAP (C, 0.15 g, 1.24 mmol) and DCC (B, 0.41 g, 1.97 mmol) and it was stirred at room temperature for an additional 16 hours. The solvent was then removed under vacuum. The resulting residue was dissolved in ACN and the solid formed was removed by filtration. The resulting filtrate was concentrated under vacuum to give a residue which was purified by flash column chromatography on silica (deactivated with DCM:NH3 1%) (DCM:MeOH 100:0 to 90:10) to give the pure product. SFC purification was carried out on a Jasco SFC prep system using an i-Cellulose-C column (Regis Technologies) 100 mm length x 4.6 mm ID 3 μm particle size in isocratic mode at 2.5 mL / min CO2 (60%)-2-propanol (40%) + 0.1% diethylamine, 35 °C, BPR 100 Bar to give the pure enantiomers. Enantiomer F1 (0.0338 g): LC-MS (ESI+): m / z: 571.3 RT: 2.442 min (VLIA), 99% ee (RT: 7.697 min). Enantiomer F2 (0.0354 g): LC-MS (ESI+): m / z: 571.3 RT: 2.443 min (VLIA), 97% ee (RT: 7.960 min).1H NMR (400MHz, DMSO) δ 9.46 (s, 1H), 8.10 (s, 1H), 8.06 (d, J = 2.4Hz, 1H), 7.19-7.12 (m, 4H), 7.02 (dd, J = 8.6, 2.4Hz, 1H), 4.47 (m continuous, 3H), 4.34-4.27 (m, 1H), 4.11-4.02 (m, 1H), 3.97-3.90 (m, 1H), 3.85 (dd, J = 16.0, 8.4 Hz, 1H), 3.09-3.00 (m, 2H), 2.67 (broad signal, 2H), 2.44 (broad signal, 4H), 2.23 (s, 3H), 2.18 (s, 3H), 2.17 (s, 3H), 1.88-1.80 (m, 1H), 1.73-1.60 (m, 3H), 1.31 (d, J = 7.1 Hz, 3H).

[0268] C-shaped macrocycles [ka]

[0269] Methyl 3-({2-chloro-5-[(2,6-dimethylanilino)methyl]pyrimidin-4-yl}amino)propanoate (65) [ka]

[0270] Methyl 3-aminopropanoate hydrochloride (4.91 g, 35.44 mmol) and triethylamine (7.41 mL, 53.16 mmol) were added to a solution of N-((2,4-dichloropyrimidin-5-yl)methyl)-2,6-dimethylaniline (4.61 g, 16.34 mmol) in anhydrous THF (50.0 mL), and the mixture was stirred at 65 °C for 16 h. The reaction crude was then diluted with AcOEt and washed with brine. The organic layer was dried over MgSO4, filtered, and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane / EtOAC 100:0 to 70:30) to give the pure product (5.12 g, yield: 83%, yellow oil). LC-MS (ESI+): m / z: 348.9, RT: 1.616 min (TACC50).

[0271] Methyl 3-(7-chloro-3-(2,6-dimethylphenyl)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)propanoate (66) [ka]

[0272] Triphosgene (6.53 g, 22.01 mmol) was added to a solution of methyl 3-({2-chloro-5-[(2,6-dimethylanilino)methyl]pyrimidin-4-yl}amino)propanoate (5.12 g, 14.68 mmol) in DCM (45.0 mL) and it was stirred at room temperature for 1 h. Then, an aqueous solution of sodium hydroxide (11.74 g, 293.54 mmol, 4.9 M) and tetrabutylammonium hydroxide (1.908 mmol 55% aqueous solution, 0.9 mL) was added at 0° C. The reaction was allowed to reach room temperature and stirred at room temperature for 16 h. After that, the solvent was removed under vacuum and the resulting residue was dissolved in DCM and washed with water. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was dissolved in DMF (40.0 mL) and cesium carbonate (3.11 g, 16.05 mmol) was added. The reaction mixture was heated at 60° C. for 16 h. The reaction crude was then diluted with DCM and washed with brine. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt 100:0 to 80:20) to give the pure product (3.054 g, yield: 56%, yellow solid). LC-MS (ESI+) m / z: 374.8, RT: 1.58 min (TACC50).

[0273] 3-{3-(2,6-dimethyl-phenyl)-7-[4-(4-methyl-piperazin-1-yl)-3-(tetrahydro-pyran-2-yloxymethyl)-phenylamino]-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl}-propionic acid methyl ester (67) [ka]

[0274] 3-[7-Chloro-3-(2,6-dimethyl-phenyl)-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl]-propionic acid methyl ester (0.5 g, 1.33 mmol) and 4-(4-methyl-piperazin-1-yl)-3-(tetrahydro-pyran-2-yloxymethyl)-phenylamine (0.54 g, 1.77 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (10.0 mL). 3A Ms was then added followed by trifluoroacetic acid (0.2 mL, 2.66 mmol) and the reaction was stirred at 90° C. for 16 h. The reaction was then diluted with water and treated with an aqueous solution of NaHCO3 (2.66 mmol). The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (0.50 g, yield: 67%, light brownish solid). LC-MS (ESI+): m / z: 560.9 RT: 1.124 min (TACC50).

[0275] 3-{3-(2,6-dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl}-propionic acid (68) [ka]

[0276] 3-{3-(2,6-dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl}-propionic acid methyl ester (0.5 g, 0.89 mmol) was dissolved in THF (3 mL) and treated with HCl (4.46 mmol, 6 M aqueous solution, 0.743 mL), which was stirred for 3 h at 60° C. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (95% [0.1% aqueous HCOOH]-5% CAN to 63% [0.1% aqueous HCOOH]-37% ACN) to give the pure product (0.308 g, yield: 63%, white solid). LC-MS (ESI+) m / z: 546.2, RT: 2.029 min (VILLA).

[0277] 1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclooctaphane-1 2 ,6-dione(69) [ka]

[0278] 3-{3-(2,6-dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl}-propionic acid (1.00 g, 1.849 mmol) and DMAP (0.565 g, 4.622 mmol) were dissolved in anhydrous DCM (230 mL). 3A MS was then added, followed by EDC hydrochloride (0.354 g, 1.849 mmol) and 1-hydroxybenzotriazole hydrate (0.375 g, 2.773 mmol) and the reaction was stirred at room temperature for 16 hours. The reaction crude was then washed with an aqueous solution of Rochelle's salt. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:NH 3 Purification by 100:0 (deactivated with 1% DCM:MeOH 100:0 to 90:10) gave the pure product (0.320 g, yield: 33%, white solid). LC-MS (ESI+) m / z: 528.2, RT: 2.383 min (VILLJ). 1H NMR(400MHz,CDCl3)δ8.24(d,J=2.5Hz,1H),7.96(s,1H),7.47(s,1H),7.19-7.10(m,3H),7.01(d,J=8.4Hz,1H),6.82(dd,J=8 .4,2.6Hz,1H),5.37(s,2H),4.46(s,2H),4.29(t,J=6.1Hz,2H),2.95(t,J=4.3Hz,4H),2.60(s,4H),2.37(s,3H),2.26(s,6H). [ka]

[0279] The previous intermediates (35-66) have already been described in the experimental section.

[0280] Methyl 3-(7-((5-(((tert-butyldimethylsilyl)oxy)methyl)-2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-3-(2,6-dimethylphenyl)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)propanoate (70) [ka]

[0281] Methyl 3-[7-chloro-3-(2,6-dimethylphenyl)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl]propanoate (1.80 g, 4.802 mmol), 5-(tert-butyl-dimethyl-silanyloxymethyl)-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine (1.843 g, 5.04 mmol) and K2CO3 (1.00 g, 7.20 mmol) were placed in a sealed tube and dissolved in a tBuOH:DCE mixture (7:1, 32.0 mL). The reaction was purged with N2, then Pd2dba3 (0.659 g, 0.720 mmol) and XantPhos (0.391 g, 0.72 mmol) were added. The mixture was purged again with N2 and the reaction was stirred at 85°C for 16 hours. The reaction crude was then filtered through Celite and rinsed with DCM. The filtrate was washed with water, dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by reverse phase chromatography (32% [25mm NH4HCO3]-68% ACN to 4% [25mm NH4HCO3]-96% ACN) to give the pure product (0.610g, yield: 18%, brown solid). LC-MS (ESI+): m / z: 704.3, RT: 1.61 min (TACC50).

[0282] 3-(3-(2,6-dimethylphenyl)-7-((5-(hydroxymethyl)-2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)propanoic acid (71) [ka]

[0283] Methyl 3-(7-((5-(((tert-butyldimethylsilyl)oxy)methyl)-2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-3-(2,6-dimethylphenyl)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2h)-yl)propanoate (0.612 g, 0.871 mmol) was dissolved in THF (10.0 mL) and treated with HCl (4.355 mmol, 6 M aqueous solution, 0.73 mL), which was heated at 60° C. for 2 h. The solvent was then removed under vacuum and the resulting residue was used without further purification. LC-MS (ESI+): m / z: 576.30 RT: 0.68 min (TACC50).

[0284] 1 3 -(2,6-dimethylphenyl)-3 6 -Methoxy-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclooctaphane-1 2 ,6-dione(72) [ka]

[0285] 3-(3-(2,6-dimethylphenyl)-7-((5-(hydroxymethyl)-2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)propanoic acid (0.871 g, 1.513 mmol) was dissolved in anhydrous DMF (2.0 mL) and diluted with anhydrous DCM (118.0 mL). 3A molecular sieves were added, followed by DMAP (0.277 g, 2.269 mmol) and DCC (0.375 g, 1.816 mmol) and the reaction was stirred at room temperature for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (81%-19% ACN to 45% [25 mm aq. NH4HCO3]-55% ACN) to give the product. This product was found to be unstable and spontaneously decomposed to a mixture of ring-opened intermediates: macrocycles.

[0286] Scheme 10 [ka]

[0287] Methyl 4-({2-chloro-5-[(2,6-dimethylanilino)methyl]pyrimidin-4-yl}amino)butanoate (73) [ka]

[0288] Methyl 4-aminobutyrate hydrochloride (2.418 g, 15.95 mmol) and triethylamine (4.446 mL, 31.89 mmol) were added to a solution of N-((2,4-dichloropyrimidin-5-yl)methyl)-2,6-dimethylaniline (3.00 g, 10.63 mmol) in anhydrous THF (32.0 mL) and the mixture was stirred at 65 °C for 16 h. The reaction crude was then diluted with AcOEt and washed with brine. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane / EtOAC 100:0 to 70:30) to give the pure product (3.48 g, yield: 90%, yellow oil). LC-MS (ESI+): m / z: 362.8 RT: 1.616 min (TACC50).

[0289] Methyl 4-[7-chloro-3-(2,6-dimethylphenyl)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl]butanoate (74) [ka]

[0290] Triphosgene (4.26 g, 14.34 mmol) was added to a solution of methyl 4-({2-chloro-5-[(2,6-dimethylanilino)methyl]pyrimidin-4-yl}amino)butanoate (3.47 g, 9.563 mmol) in DCM (29.0 mL) and it was stirred at room temperature for 1 h. Then an aqueous solution of sodium hydroxide (7.65 g, 191.26 mmol, 4.9 M) and tetrabutylammonium hydroxide (1.243 mmol 55% aqueous solution, 0.58 mL) was added at 0° C. The reaction was allowed to reach room temperature and stirred at room temperature for 16 h. Afterwards, the reaction crude was diluted with DCM and washed with water. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was dissolved in DMF (29.0 mL) and cesium carbonate (2.031 g, 10.47 mmol) was added. The reaction mixture was heated at 60° C. for 16 h. The reaction crude was then diluted with DCM and washed with brine. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt 100:0 to 50:50) to give the pure product (2.21 g, yield: 60%, yellow solid). LC-MS (ESI+): m / z: 388.8 RT: 1.652 min (TACC50).

[0291] 4-{3-(2,6-dimethyl-phenyl)-7-[4-(4-methyl-piperazin-1-yl)-3-(tetrahydro-pyran-2-yloxymethyl)-phenylamino]-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl}-butyric acid methyl ester (75) [ka]

[0292] 4-[7-Chloro-3-(2,6-dimethyl-phenyl)-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl]-butyric acid methyl ester (1.0 g, 2.57 mmol) and 4-(4-methyl-piperazin-1-yl)-3-(tetrahydro-pyran-2-yloxymethyl)-phenylamine (1.05 g, 3.4 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (20.0 mL). Then 3A MS was added followed by anhydrous TFA (0.4 mL, 5.14 mmol) and the reaction was stirred at 90° C. for 16 h. The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (0.947 g, yield: 64%, light brown solid). LC-MS (ESI+): m / z: 574.2 RT: 2.256 min (VILL-J).

[0293] 4-{3-(2,6-dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl}-butyric acid (76) [ka]

[0294] 4-{3-(2,6-dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl}-butyric acid methyl ester (0.90 g, 1.57 mmol) was dissolved in THF (5.0 mL) and treated with HCl (7.84 mmol, 6 M aqueous solution, 1.3 mL) for 3 h at 60° C. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (95% [0.1% aqueous HCOOH]-5% ACN to 63% [0.1% aqueous HCOOH]-37% ACN) to give the pure product (0.520 g, yield: 59%, white solid). LC-MS (ESI+): m / z: 560.2 RT: 2.017 min (VILLA).

[0295] 1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclononaphane-1 2 ,6-dione(77) [ka]

[0296] 4-{3-(2,6-dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl}-butyric acid (0.114 g, 0.204 mmol) was dissolved in anhydrous DMF (3.0 mL) and diluted with anhydrous DCM (35.0 mL). The reaction mixture was cooled to 0° C. and DCC (0.051 g, 0.245 mmol), DMAP (0.050 g, 0.408 mmol) and 3A MS were added sequentially. The reaction was brought to room temperature and stirred for 16 hours. The reaction crude was concentrated under vacuum and the resulting residue was dissolved in ACN. The solid formed was removed by filtration. The filtrate was concentrated under vacuum and the resulting residue was purified by reverse phase chromatography (59% [25 mm aq. NH4HCO3]-41% ACN to 17% [25 mm aq. NH4HCO3]-83% ACN) to give the pure product (0.0101 g, yield: 9%, white solid). LC-MS (ESI+): m / z: 542.1 RT: 2.501 min (VILL-J). 1H NMR (400 MHz, CDCl3) δ 8.48 (d, J = 1.9 Hz, 1H), 7.95 (s, 1H), 7.42 (s, 1H), 7.19-7.05 (m, 4H), 6.78 (dd, J = 8.4, 2.3 Hz, 1H), 5.42 (s, 2H), 4.48 (s, 2H), 4.20-4.12 (m, 2H), 2.92 (t, J = 4.6 Hz, 4H), 2.55 (m continuous, 6H), 2.36 (s, 3H), 2.25 (s, 6H), 2.20 (broad signal, 2H).

[0297] Scheme 11 [ka]

[0298] The previous intermediates (35-74) have already been described in the experimental section.

[0299] Methyl 4-(3-(2,6-dimethylphenyl)-7-((4-fluoro-5-(hydroxymethyl)-2-methoxyphenyl)amino)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)butanoate (78) [ka]

[0300] Methyl 4-((2-chloro-5-(((chlorocarbonyl)(2,6-dimethylphenyl)amino)methyl)pyrimidin-4-yl)amino)butanoate (1.300 g, 3.343 mmol) and (5-amino-2-fluoro-4-methoxyphenyl)methanol (0.761 g, 0.761 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (50.0 mL). 3A MS was then added followed by anhydrous TFA (0.512 mL, 6.686 mmol) and the reaction was stirred at 90° C. for 16 h. The reaction was then diluted with water and treated with aqueous NaHCO3. A solution of NaHCO3 (6.686 mmol) was added. The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt, 70:30 to 0:100) to give the pure product (1.452 g, yield: 83%, pale yellow solid). LC-MS (ESI+): m / z: 523.8 RT: 1.450 min (TACC50).

[0301] 4-(3-(2,6-dimethylphenyl)-7-((4-fluoro-5-(hydroxymethyl)-2-methoxyphenyl)amino)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)butanoic acid (79) [ka]

[0302] Methyl 4-(3-(2,6-dimethylphenyl)-7-((4-fluoro-5-(hydroxymethyl)-2-methoxyphenyl)amino)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)butanoate (0.639 g, 1.220 mmol) was dissolved in THF (5.0 mL) and treated with HCl (6.10 mmol, 6 M aqueous solution, 0.22 mL), which was heated at 60° C. for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (90% [0.1% aqueous HCOOH]-10% ACN to 54% [0.1% aqueous HCOOH]-46% ACN) to give the pure product (0.386 g, yield: 62%, pale yellow solid). LC-MS (ESI+): m / z: 510.2 RT: 1.00 min (TACC50).

[0303] 1 3 -(2,6-dimethylphenyl)-3 4 -Fluoro-3 6 -Methoxy-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclononaphane-1 2 ,6-dione(80) [ka]

[0304] 4-(3-(2,6-dimethylphenyl)-7-((4-fluoro-5-(hydroxymethyl)-2-methoxyphenyl)amino)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)butanoic acid (0.316 g, 0.62 mmol) was dissolved in anhydrous DMF (10.0 mL) and diluted with anhydrous DCM (80.0 mL). 3A molecular sieves were added, followed by DMAP (0.114 g, 0.930 mmol) and DCC (0.154 g, 0.744 mmol) and the reaction was stirred at room temperature for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by flash column chromatography on silica (deactivated with DCM:NH3 1%) (DCM:MeOH 100:0 to 90:10). The resulting product was recrystallized in ACN to give the pure product (0.174 g, yield: 57%, white solid). LC-MS (ESI+): m / z: 492.2 RT: 4.047 min (VILLA). 1H NMR (400 MHz, CDCl3) δ 8.46 (d, J = 8.1 Hz, 1H), 7.95 (s, 1H), 7.71 (s, 1H), 7.20-7.10 (m, 3H), 6.65 (d, J = 11.1 Hz, 1H), 5.37 (s, 2H), 4.49 (s, 2H), 4.19-4.12 (m, 2H), 3.89 (s, 3H), 2.57-2.50 (m, 2H), 2.25 (s, 6H), 2.23-2.15 (m, 2H).

[0305] Scheme 12 [ka]

[0306] The previous intermediates (35-74) have already been described in the experimental section.

[0307] Methyl 4-(7-((4-bromo-5-(hydroxymethyl)-2-methoxyphenyl)amino)-3-(2,6-dimethylphenyl)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)butanoate (81) [ka]

[0308] Methyl 4-(7-chloro-3-(2,6-dimethylphenyl)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)butanoate (1.2 g, 3.086 mmol) and (5-amino-2-bromo-4-methoxyphenyl)methanol (1.074 g, 4.629 mmol) were placed in a sealed tube and dissolved in anhydrous 2-butanol (20.0 mL). 3A MS and anhydrous TFA (0.704 mL, 6.172 mmol) were then added and the reaction was stirred at 90° C. for 16 h. The reaction was then diluted with water and treated with aqueous NaHCO3. A solution of NaHCO3 (6.172 mmol) was added. The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt, 100:0 to 70:30) to give the pure product (1.258 g, yield: 70%, light brown solid). LC-MS (ESI+): m / z: 585.0 RT: 1.66 min (TACC50).

[0309] 4-[7-(4-bromo-5-hydroxymethyl-2-methoxy-phenylamino)-3-(2,6-dimethyl-phenyl)-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl]-butyric acid (82) [ka]

[0310] 4-[7-(4-Bromo-5-hydroxymethyl-2-methoxy-phenylamino)-3-(2,6-dimethyl-phenyl)-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl]-butyric acid methyl ester (0.7 g, 1.2 mmol) was dissolved in MeOH:HO mixture (5:1, 12.0 mL) and treated with aqueous sodium hydroxide (0.4 mL, 6.0 mmol) and the reaction was stirred at room temperature for 3 h. The solvent was then removed under vacuum and the resulting residue was used without further purification.

[0311] 3 4 -Bromo-13-(2,6-dimethylphenyl)-3 6 -Methoxy-1 1 ,1 2 ,1 3 ,14-Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzeneacyclononaphane-1 2 ,6-dione(83) [ka]

[0312] 4-[7-(4-bromo-5-hydroxymethyl-2-methoxy-phenylamino)-3-(2,6-dimethyl-phenyl)-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl]-butyric acid (0.68 g, 1.19 mmol) was dissolved in anhydrous DMF (10.0 mL) and diluted with anhydrous DCM (230.0 mL). 3A molecular sieves were added, followed by DMAP (0.220 g, 1.797 mmol) and DCC (0.297 g, 1.438 mmol) and the reaction was stirred at room temperature for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by flash column chromatography on silica (heptane:AcOEt 100:0 to 50:50) to give the pure product (0.131 g, yield: 20%, brown pale solid). LC-MS (ESI+): m / z: 554.2366 RT: 6.123 min (VILLA-2T). 1H NMR(400MHz,DMSO)δ8.41(s,1H),8.15(s,1H),8.03(s,1H),7.28(s,1H),7.19-7.14(m,3H),5.75(s, 1H),5.18(s,2H),4.52(s,2H),4.00-3.93(m,3H),3.91(s,3H),2.18(s,6H),2.05(t,J=12.0Hz,2H).

[0313] Scheme 13 [ka]

[0314] The previous intermediates (35-74) have already been described in the experimental section.

[0315] 4-[7-[5-(tert-butyl-dimethyl-silanyloxymethyl)-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamino]-3-(2,6-dimethyl-phenyl)-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl]-butyric acid methyl ester (84) [ka]

[0316] 4-[7-Chloro-3-(2,6-dimethyl-phenyl)-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl]-butyric acid methyl ester (1.30 g, 3.34 mmol), 5-(tert-butyl-dimethyl-silanyloxymethyl)-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine (1.28 g, 3.51 mmol) and K2CO3 (0.700 g, 5.02 mmol) were placed in a sealed tube and dissolved in a tBuOH:DCE mixture (7:1, 16.0 mL). The reaction was purged with N2, then Pd2dba3 (0.460 g, 0.500 mmol) and XantPhos (0.270 g, 0.500 mmol) were added. The mixture was purged again with N2 and the reaction was stirred at 85°C for 16 hours. The reaction crude was then filtered through Celite and rinsed with DCM. The filtrate was washed with water, dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by reverse phase chromatography (32% [25mm NH4HCO3]-68% ACN to 4% [25mm NH4HCO3]-96% ACN) to give the pure product (0.150g, yield: 6%, brown light solid). LC-MS (ESI+): m / z: 717.9 RT: 4.383 min (TACC50).

[0317] 4-(3-(2,6-dimethylphenyl)-7-((5-(hydroxymethyl)-2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)butanoic acid (85) [ka]

[0318] 4-[7-[5-(tert-butyl-dimethyl-silanyloxymethyl)-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamino]-3-(2,6-dimethyl-phenyl)-2-oxo-3,4-dihydro-2h-pyrimido[4,5-d]pyrimidin-1-yl]-butyric acid methyl ester (0.30 g, 0.42 mmol) was dissolved in THF (5.0 mL) and treated with HCl (2.09 mmol, 6 M aqueous solution, 0.35 mL), which was heated at 60° C. for 2 h. The solvent was then removed under vacuum and the resulting residue was used without further purification. LC-MS (ESI+): m / z: 590.0 RT: 2.76 min (TACC50).

[0319] 1 3 -(2,6-dimethylphenyl)-3 6 -Methoxy-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclononaphane-1 2 ,6-dione(86) [ka]

[0320] 4-(3-(2,6-dimethylphenyl)-7-((5-(hydroxymethyl)-2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)butanoic acid (0.210 g, 0.348 mmol) was dissolved in anhydrous DMF (2.0 mL) and diluted with anhydrous DCM (50.0 mL). 3A molecular sieves were added followed by DMAP (0.064 g, 0.522 mmol) and DCC (0.086 g, 0.418 mmol) and the reaction was stirred at room temperature for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (32% [25mm NH4HCO3]-68% ACN to 4% [25mm NH4HCO3]-96% ACN) to give the pure product (0.012g, yield: 6%, white solid). LC-MS (ESI+): m / z: 572.3 RT: 2.544min (VILLA). 1H NMR (400MHz, DMSO-d6) δ 8.16 (s, 1H), 8.04 (s, 1H), 7.82 (s, 1H), 7.02-7.16 (m, 3H), 6.77 (s, 1H), 5.19 (s, 2H), 4.43 (s, 2H), 3.85-3.90 (m, 2H), 3.82 (s, 3H), 2.75-2.82 (m, 4H), 2.37-2.42 (m, 6H)*, 2.17 (s, 3H), 2.11 (s, 6H), 1.96 (broad signal, 2H). *Solvent signals overlap.

[0321] Scheme 14 [ka]

[0322] Methyl 5-(2-chloro-5-(((2,6-dimethylphenyl)amino)methyl)pyrimidin-4-yl)pentanoate (87) [ka]

[0323] Methyl-5-aminopentanoate hydrochloride (3.12 g, 18.61 mmol) and triethylamine (5.18 mL, 37.21 mmol) were added to a solution of N-((2,4-dichloropyrimidin-5-yl)methyl)-2,6-dimethylaniline (3.50 g, 12.40 mmol) in anhydrous THF (37.0 mL) and the mixture was stirred at 65° C. for 16 h. The reaction crude was then diluted with AcOEt and washed with brine. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The obtained pure product (4.60 g, yield: 98%, yellow oil) was used without further purification. LC-MS (ESI+): m / z: 376.9 RT: 1.709 min (TACC50).

[0324] 5-[7-Chloro-3-(2,6-dimethyl-phenyl)-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl]-pentanoic acid methyl ester (88) [ka]

[0325] Triphosgene (5.433 g, 18.31 mmol) was added to a solution of methyl 5-(2-chloro-5-(((2,6-dimethylphenyl)amino)methyl)pyrimidin-4-yl)pentanoate (4.600 g, 12.205 mmol) in DCM (35.0 mL) and it was stirred at room temperature for 1 h. Then an aqueous solution of sodium hydroxide (9.76 g, 244.10 mmol, 4.9 M) and tetrabutylammonium hydroxide (1.587 mmol 55% aqueous solution, 0.75 mL) was added at 0° C. The reaction was allowed to reach room temperature and stirred at room temperature for 16 h. Afterwards, the reaction crude was diluted with DCM and washed with water. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was dissolved in DMF (35.0 mL) and cesium carbonate (4.04 g, 12.402 mmol) was added. The reaction mixture was heated at 60° C. for 16 h. The reaction crude was then diluted with DCM and washed with brine. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt 100:0 to 80:20) to give the pure product (2.626 g, yield: 53%, yellow solid). LC-MS (ESI+): m / z: 402.0 RT: 1.623 min (TACC50).

[0326] Methyl 5-(3-(2,6-dimethylphenyl)-7-((3-(hydroxymethyl)-4-(4-methylpiperazin-1-yl)phenyl)amino)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2H)-yl)pentanoate (89) [ka]

[0327] 5-[7-Chloro-3-(2,6-dimethyl-phenyl)-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl]-pentanoic acid methyl ester (0.965 g, 2.395 mmol) and 4-(4-methyl-piperazin-1-yl)-3-(tetrahydro-pyran-2-yloxymethyl)-phenylamine (B, 0.973 g, 3.185 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (20.0 mL). 3A MS was then added followed by anhydrous TFA (0.369 mL, 4.790 mmol) and the reaction was stirred at 90° C. for 16 hours. The reaction was then diluted with water and treated with aqueous NaHCO3. A solution of NaHCO3 (4.79 mmol) was added. The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated in vacuo. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (0.595 g, yield: 42%, light brown solid). LC-MS (ESI+): m / z: 587.0 RT: 1.359 min (TACC50).

[0328] 5-{3-(2,6-dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl}-pentanoic acid (90) [ka]

[0329] Methyl 5-(3-(2,6-dimethylphenyl)-7-((3-(hydroxymethyl)-4-(4-methylpiperazin-1-yl)phenyl)amino)-2-oxo-3,4-dihydropyrimido[4,5-d]pyrimidin-1(2h)-yl)pentanoate (0.595 g, 1.012 mmol) was dissolved in THF (3.0 mL) and treated with HCl (5.06 mmol, 6 M aqueous solution, 0.84 mL), which was heated at 60° C. for 2 h. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (95% [0.1% aqueous HCOOH]-5% ACN to 63% [0.1% aqueous HCOOH]-37% ACN) to give the pure product (0.395 g, yield: 68%, white solid). LC-MS (ESI+): m / z: 574.2 RT: 2.127 min (VILLA).

[0330] 1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidine-3(1,3)-benzenecyclodecaphane-1 2 ,6-dione(91) [ka]

[0331] 5-{3-(2,6-dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-2-oxo-3,4-dihydro-2H-pyrimido[4,5-d]pyrimidin-1-yl}-pentanoic acid (0.353 g, 0.615 mmol) was dissolved in anhydrous DMF (10.0 mL) and diluted with anhydrous DCM (70.0 mL). 3A molecular sieves were added followed by DMAP (0.1130 g, 0.922 mmol) and DCC (0.152 g, 0.738 mmol) and the reaction was stirred at room temperature for 16 hours. The reaction was recharged with DMAP (0.1130 g, 0.922 mmol) and DCC (0.152 g, 0.738 mmol) and the reaction was stirred at room temperature for an additional 16 hours. The solvent was then removed under vacuum and the resulting residue was dissolved in ACN. The solid formed was removed by filtration. The filtrate was concentrated under vacuum and the resulting residue was purified by reverse phase chromatography (59% [25mm NH4HCO3 aqueous solution]-41% ACN to 17% [25mm NH4HCO3 aqueous solution]-83% ACN) to give the pure product (0.0488g, yield: 14%, white solid). LC-MS (ESI+): m / z: 556.2 RT: 2.624 min (VILL_J). 1H NMR(400MHz,CDCl3)δ8.10(d,J=2.2Hz,1H), 7.92(s,1H), 7.18-7.07(m,5H), 6.84(dd,J=8.5,2.4Hz,1H), 5.37(s,2H), 4.47(s,2H), 4.0 3-3.97(m,2H), 2.93(t,J=4.6Hz,4H), 2.59(s,4H), 2.53-2.48(m,2H), 2.36(s,3H), 2.25(s,6H), 1.97-1.87(m,2H), 1.84-1.76(m,2H).

[0332] Example 3: Synthesis of macrocycles synthesized by RCM

[0333] For the ring-closing metathesis reaction, the commonly known Zhan Catalyst-1B was used (CAS: 918870-76-5), which corresponds to the structure dichloro-1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene 5-(dimethylamino)sulfonyl-2-(1-methylethoxy-O)phenylmethylene-Cruthenium(II).

[0334] Scheme 15 [ka]

[0335] Allyl-{2-chloro-5-[(2,6-dimethyl-phenylamino)-methyl]-pyrimidin-4-yl}-amine (92) [ka]

[0336] Allylamine (1.60 g, 21.26 mmol) and triethylamine (2.96 mL, 21.26 mmol) were added to a solution of N-((2,4-dichloropyrimidin-5-yl)methyl)-2,6-dimethylaniline (4.0 g, 14.18 mmol) in anhydrous THF (45.0 mL) and the mixture was stirred at 65° C. for 16 h. The reaction crude was then diluted with AcOEt and washed with brine. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The obtained pure product (4.29 g, yield: quantitative, yellow oil) was used without further purification. LC-MS (ESI+): m / z: 303.0 RT: 1.76 min (TACC50).

[0337] 1-Allyl-7-chloro-3-(2,6-dimethyl-phenyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (93) [ka]

[0338] Triphosgene (6.30 g, 21.26 mmol) was added to a solution of allyl-{2-chloro-5-[(2,6-dimethyl-phenylamino)-methyl]-pyrimidin-4-yl}-amine (4.29 g, 14.2 mmol) in DCM (45.0 mL) and it was stirred at room temperature for 1 h. Then, an aqueous solution of sodium hydroxide (11.30 g, 283.5 mmol, 4.9 M) and tetrabutylammonium hydroxide (1.84 mmol 55% aqueous solution, 0.90 mL) was added at 0° C. The reaction was allowed to reach room temperature and stirred at room temperature for 16 h. Afterwards, the reaction crude was diluted with DCM and washed with water. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was dissolved in DMF (40.0 mL) and cesium carbonate (4.66 g, 14.32 mmol) was added. The reaction mixture was heated at 60° C. for 16 h. The reaction crude was then diluted with DCM and washed with brine. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt 100:0 to 0:100) to give the pure product (2.80 g, yield: 60%, white solid). LC-MS (ESI+): m / z: 329.1 RT: 1.648 min (TACC50).

[0339] Methyl 5-((8-allyl-6-(2,6-dimethylphenyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)benzoate (94) [ka]

[0340] 1-Allyl-7-chloro-3-(2,6-dimethyl-phenyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (1.05 g, 3.193 mmol) and methyl 5-amino-2-(4-methylpiperazin-1-yl)benzoate (1.114 g, 4.47 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (20.0 mL). 3A MS was then added, followed by trifluoroacetic anhydride (0.728 mL, 6.386 mmol) and the reaction was stirred at 90° C. for 16 hours. The reaction was then diluted with water and treated with an aqueous solution of NaHCO3 (6.386 mmol). The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:deactivated with 1% NH3) (DCM:MeOH 100:0 to 90:10) to give the pure product (1.37 g, yield: 79%, yellow solid). LC-MS (ESI+): m / z: 542.0 RT: 1.15 min (TACC50).

[0341] 5-[8-Allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzoic acid (95) [ka]

[0342] Methyl 5-((8-allyl-6-(2,6-dimethylphenyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)benzoate (1.29 g, 2.38 mmol) was dissolved in THF (8.0 mL) and treated with HCl (11.92 mmol, 6 M aqueous solution, 1.98 mL), which was heated at 60° C. for 4 h. The solvent was then removed under vacuum and the resulting product was used without further purification. LC-MS (ESI+): m / z: 528.3 RT: 0.814 min (TACC50).

[0343] 5-[8-Allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzoic acid but-3-enyl ester (96) [ka]

[0344] 3-Buten-1-ol (0.3 mL, 3.57 mmol), DMAP (0.30 g, 2.38 mmol) and DCC (0.37 mg, 1.8 mmol) were added sequentially to a solution of 5-[8-allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzoic acid (0.7 g, 1.19 mmol) in a DCM:DMF mixture (5:1, 5.0 mL) and the reaction was stirred at room temperature for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (0.52 g, yield: 75%, yellow solid). LC-MS (ESI+): m / z: 582.2 RT: 1.481 min (TACC50).

[0345] (E)-1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidin-3(1,3)-benzenacyclodecaphan-8-ene-1 2 ,4-dione (97) and (Z)-1 3 -(2,6-dimethylphenyl)-34-(4-methylpiperazin-1-yl)-1 1 ,1 2 ,1 3,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidin-3(1,3)-benzenacyclodecaphan-8-ene-1 2 ,4-dione(98) [ka]

[0346] 5-[8-Allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methylpiperazin-1-yl)-benzoic acid but-3-enyl ester (0.517 g, 0.889 mmol) was placed in a sealed tube and dissolved in anhydrous toluene (100 mL). HCl (0.889 mmol, 3 M in cyclopentyl methyl ether, 0.30 mL) was then added. The reaction crude was degassed by bubbling N2 through the solution, followed by the addition of a solution of Zhan's catalyst (0.178 mmol, 0.131 g) in anhydrous toluene (18.0 mL). The reaction crude was degassed again by bubbling.

[0347] N2 was passed through the solution and heated at 110° C. for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (32% [65 mM NH4OAc+ACN (90:10)]-68% ACN to 4% [65 mM NH4OAc+ACN (90:10)]-96% ACN) to give both diastereomers.

[0348] Product A, trans diastereomer (18.2 mg, white solid). 1H NMR (400 MHz, DMSO) δ 9.64 (s, 1H), 8.54 (d, J = 2.6 Hz, 1H), 8.15 (s, 1H), 7.18 (m series, 4H), 7.06 (d, J = 8.8 Hz, 1H), 5.67 (dt, J = 15.9, 4.3 Hz, 1H), 5.59-5.49 (m, 1H), 4.50 (broad signal, 4H), 4.32 (broad signal, 2H), 2.91 (s, 4H), 2.45-2.35 (m, 6H), 2.20 (s, 3H), 2.18 (s, 6H). LC-MS (ESI+): m / z: 555.3 RT: 2.40 min (VILLA).

[0349] Product B, cis diastereomer (18.5 mg, pale yellow solid). 1H NMR(400MHz,DMSO)δ9.69(s,1H),8.79(d,J=2.7Hz,1H),8.14(s,1H),7.29( dd,J=8.8,2.7Hz,1H),7.17(m series,3H),7.07(d,J=8.9Hz,1H),5.46(dt,J=1 0.0,8.0Hz,1H),5.37(dt,J=11.0,3.5Hz,1H),4.81(s,2H),4.52(s,2H),4. 32-4.27(m,2H),2.95-2.90(m,4H),2.44(s,6H),2.22(s,3H),2.19(s,6H). LC-MS (ESI+): m / z: 555.3 RT: 2.496 min (VILLA).

[0350] Scheme 16 [ka]

[0351] The previous intermediates (35-93) have already been described in the experimental section.

[0352] Methyl 2-(5-((8-allyl-6-(2,6-dimethylphenyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)phenyl)acetate (99) [ka]

[0353] 1-Allyl-7-chloro-3-(2,6-dimethyl-phenyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (1.05 g, 3.193 mmol) and methyl 2-(5-amino-2-(4-methylpiperazin-1-yl)phenyl)acetate (1.177 g, 4.47 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (20.0 mL). 3A MS was then added followed by trifluoroacetic acid (0.728 mL, 6.386 mmol) and the reaction was stirred at 90° C. for 16 h. The reaction was then diluted with water and treated with an aqueous solution of NaHCO3 (6.386 mmol). The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM: deactivated with 1% NH3) (DCM:MeOH 100:0 to 90:10) to give the pure product (1.20 g, yield: 68%, yellow solid). LC-MS (ESI+): m / z: 556.0 RT: 1.15 min (TACC50).

[0354] [5-[8-Allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-acetic acid (100) [ka]

[0355] Methyl 2-(5-((8-allyl-6-(2,6-dimethylphenyl)-7-oxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidin-2-yl)amino)-2-(4-methylpiperazin-1-yl)phenyl)acetate (1.14 g, 2.052 mmol) was dissolved in THF (8.0 mL) and treated with HCl (10.26 mmol, 6 M aqueous solution, 1.71 mL), which was heated at 60° C. for 4 h. The solvent was then removed under vacuum and the resulting product was used without further purification. LC-MS (ESI+): m / z: 542.0 RT: 1.16 min (TACC50).

[0356] [5-[8-Allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-acetic acid allyl ester (101) [ka]

[0357] Allyl alcohol (0.420 mL, 6.15 mmol), DMAP (0.501 g, 4.10 mmol) and DCC (0.634 g, 6.15 mmol) were added sequentially to a solution of [5-[8-allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-acetic acid (1.11 g, 2.05 mmol) in a DCM:DMF mixture (4:1, 80.0 mL) and the reaction was stirred at room temperature for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (0.337 g, yield: 28%, pale yellow solid). LC-MS (ESI+): m / z: 582.2 RT: 1.25 min (TACC50).

[0358] (E)-1 3 -(2,6-dimethylphenyl)-3 4-(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-6-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidin-3(1,3)-benzenacyclodecaphan-8-ene-1 2 ,5-dione (102) and (Z)-1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-6-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidin-3(1,3)-benzenacyclodecaphan-8-ene-1 2 ,5-dione(103) [ka]

[0359] [5-[8-allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-phenyl]-acetic acid allyl ester (0.337 g, 0.579 mmol) was placed in a sealed tube, dissolved in anhydrous DMF (3.0 mL) and diluted with toluene (80.0 mL). HCl (0.579 mmol, 3 M in cyclopentyl methyl ether, 0.19 mL) was then added. The reaction crude was degassed by bubbling N2 through the solution, followed by the addition of a solution of Zhan's catalyst (0.085 g, 0.116 mmol) in anhydrous toluene (30.0 mL). The reaction crude was degassed again by bubbling N2 through the solution and heated at 110° C. for 16 h. The solvent was then removed in vacuo and the resulting residue was purified by reverse phase chromatography (59% [25 mm aq. NH4HCO3]-41% ACN to 17% [25 mm aq. NH4HCO3]-83% ACN) to give both diastereomers.

[0360] Product A trans diastereomer (9.0 mg, white solid). 1H NMR (400 MHz, CDCl3) δ 7.94 (s, 1H), 7.50 (d, J = 2.5Hz, 1H), 7.13 (m series, 4H), 6.95 (dd, J = 8.5, 2.5Hz, 1H), 6.79 (s, 1H), 6.11 (dt, J = 15.8, 6.2Hz, 1H), 5.74 (dt, J = 15.7, 4.0Hz, 1H), 4.67 (d, J = 2.5Hz, 2H), 4.46 (s, 2H), 4.36 (d, J = 6.0Hz, 2H), 3.84 (s, 2H), 3.04 (broad signal, 4H), 2.68-2.55 (m, 4H), 2.37 (s, 3H),2.25(s, 6H). LC-MS (ESI+): m / z: 554.3 RT: 2.665 min (VILLA).

[0361] Product B, cis diastereomer (10.2 mg, white solid). 1H NMR(400 MHz, CDCl3)δ8.21(d, J=2.5Hz, 1H),7.96(s, 1H),7.13(m series, 5H),6.76(dd, J=8.5, 2.6 Hz, 1H),5.84-5.76(m, 1H),5.61(dt, J=10.9, 4.3 Hz, 1H),5.11-5.07(m, 2H),4.68(d, J=7.4 Hz, 2H),4.49(s, 2H),3.87(s, 2H),2.96(t, J=4.7 Hz, 4H), 2.59 (broad signal, 4H), 2.36 (s, 3H), 2.26 (s, 6H). LC-MS (ESI+): m / z: 554.3 RT: 2.833 min (VILLA).

[0362] Scheme 17 [ka]

[0363] The previous intermediates (35-93) have already been described in the experimental section.

[0364] 3-(2,6-Dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-1-propyl-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (104) [ka]

[0365] 7-Chloro-3-(2,6-dimethyl-phenyl)-1-propyl-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (2.5 g, 7.6 mmol) and 4-(4-methyl-piperazin-1-yl)-3-(tetrahydro-pyran-2-yloxymethyl)-phenylamine (3.25 g, 10.64 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (20.0 mL). 3A MS was then added followed by trifluoroacetic acid (1.20 mL, 15.2 mmol) and the reaction was stirred at 90° C. for 16 hours. The reaction was then diluted with water and treated with aqueous NaHCO3. A solution of NaHCO3 (15.2 mmol) was added. The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH, 100:0 to 90:10) to give the pure product (2.17 g, yield: 56%, yellow solid). LC-MS (ESI+): m / z: 514.2 RT: 1.095 min TACC50.

[0366] Acrylic acid 5-[8-allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzyl ester (105) [ka]

[0367] DMAP (0.52 g, 4.22 mmol), DCC (1.31 g, 6.34 mmol) and acrylic acid (0.58 mL, 8.45 mmol) were added sequentially to a solution of 1-allyl-3-(2,6-dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (2.17 g, 4.22 mmol) in anhydrous DCM (50.0 mL) at 0 °C. The solvent was then removed under vacuum and the resulting residue was purified by flash column chromatography on silica (DCM:MeOH 100:0 to 90:10) to give the pure product (0.996 g, yield: 42% yield, yellow solid). LC-MS (ESI+): m / z: 568.2 RT: 1.343 min (TACC50).

[0368] (Z)-1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidin-3(1,3)-benzenacyclonopanal-7-ene-1 2 ,6-dione(106) [ka]

[0369] Acrylic acid 5-[8-allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzyl ester (0.450 g, 0.793 mmol) was placed in a sealed tube and dissolved in anhydrous toluene (140.0 mL). HCl (0.793 mmol, 3 M in cyclopentyl methyl ether, 0.26 mL) was then added. The reaction crude was degassed by bubbling N2 through the solution, followed by the addition of a solution of Zhan's catalyst (0.117 g, 0.159 mmol) in anhydrous toluene (20.0 mL). The reaction crude was degassed again by bubbling N2 through the solution and heated at 110° C. for 16 h. The solvent was then removed in vacuo and the resulting residue was purified by reverse phase chromatography (59% [25 mm aq. NHHCO]-41% ACN to 17% [25 mm aq. NHHCO]-83% ACN) to give the pure product as a single cis diastereomer (0.0154 g, pale yellow solid). 1H NMR(400 MHz, DMSO)δ9.55(s, 1H),8.11(s, 1H),7.74(d, J=2.1Hz, 1H),7.17(m series, 3H),7.11(d, J=8.6 Hz, 1H),7.02(dd, J=8.6, 2.3 Hz, 1H),6.31(d, J=11.6 Hz, 1H),6.19(dt, J=11.7, 6.0Hz, 1H),5.39(s, 2H),4.90(d, J=5.1Hz, 2H),4.50(s, 2H),2.81(t, J=4.5Hz, 4H),2.47(broad signal,4H),2.23(s, 3H),2.21(s, 6H). LC-MS (ESI+): m / z: 540.2 RT: 2.656 min (VILLA).

[0370] Scheme 18 [ka]

[0371] The previous intermediates (35-104) have already been described in the experimental section.

[0372] But-3-enoic acid 5-[8-allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzyl ester (107) [ka]

[0373] DMAP (0.036 g, 0.292 mmol), DCC (0.145 g, 0.701 mmol) and 3-butenoic acid (0.060 mL, 0.701 mmol) were added sequentially to an ice-cold solution of 1-allyl-3-(2,6-dimethylphenyl)-7-((3-(hydroxymethyl)-4-(4-methylpiperazin-1-yl)phenyl)amino)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one (0.30 g, 0.584 mmol) in anhydrous DCM (5.0 mL). The reaction was allowed to come to room temperature and stirred at that temperature for 16 hours. The reaction crude was then diluted with DCM and washed with brine. The combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH, 100:0 to 9:1) to give the pure product (0.285 g, yield: 84%, pale yellow solid). LC-MS (ESI+): m / z: 582.3 RT: 1.30 min (TACC50).

[0374] (E)-1 3 -(2,6-dimethylphenyl)-34-(4-methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidin-3(1,3)-benzenacyclodecaphan-8-ene-1 2 ,6-dione (108) and (Z)-1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-11 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidin-3(1,3)-benzenacyclodecaphan-8-ene-1 2 ,6-dione(109) [ka]

[0375] But-3-enoic acid 5-[8-allyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzyl ester (0.225 g, 0.387 mmol) was placed in a sealed tube and dissolved in anhydrous toluene (72.0 mL). HCl (0.387 mmol, 3 M in cyclopentyl methyl ether, 0.129 mL) was then added. The reaction crude was degassed by bubbling N2 through the solution, followed by the addition of a solution of Zhan's catalyst (0.057 g, 0.077 mmol) in anhydrous toluene (4.0 mL). The reaction crude was degassed again by bubbling N2 through the solution and heated at 110° C. for 16 h. The solvent was then removed in vacuo and the resulting residue was purified by reverse phase chromatography (50% [25 mM NH4HCO3]-50% ACN to 25% [25 mM NH4HCO3]-75% ACN) to give both diastereomers.

[0376] Product A, trans diastereomer (12.1 mg, white solid). 1H NMR (400 MHz, CDCl3) δ 7.96 (s, 1H), 7.81 (s, 1H), 7.18-7.08 (m, 4H), 6.94 (s, 1H), 6.86 (dd, J = 8.5, 2.3 Hz, 1H), 6.02 (dt, J = 15.7, 4.5 Hz, 1H), 5.76-5.64 (m, 1H), 5.36 (s, 2H), 4.56 (s, 2H), 4.50 (s, 2H), 3.10 (d, J = 6.8 Hz, 2H), 2.95-2.89 (m, 4H), 2.61 (broad signal, 4H), 2.38 (s, 3H), 2.26 (s, 6H). LC-MS (ESI+): m / z: 555.3 RT: 2.303 min (VILLA).

[0377] Product B, cis diastereomer (9.3 mg, white solid). 1H NMR(400MHz, CDCl3)δ8.52(d,J=2.1Hz,1H),7.97(s, 1H),7.15(m series, 5H),6.79(dd,J=8.5, 2.4 Hz, 1H),5.69-5.60(m, 1H),5.59-5.51(m, 1H),5.33(s, 2H),5.09-5.03(m, 2H),4.51(s, 2H),3.21(d, J=8.3 Hz, 2H),2.93-2.87(m, 4H),2.62(broad signal, 4H),2.38(s, 3H),2.26(s, 6H). LC-MS (ESI+): m / z: 555.3 RT: 2.444 min (VILLA).

[0378] Scheme 19 [ka]

[0379] The previous intermediate (35) has already been described in the experimental section.

[0380] Butyl-{2-chloro-5-[(2,6-dimethyl-phenylamino)-methyl]-pyrimidin-4-yl}-amine (110) [ka]

[0381] Anhydrous triethylamine (2.96 mL, 21.26 mmol) was added to a solution of (2,4-dichloro-pyrimidin-5-ylmethyl)-(2,6-dimethyl-phenyl)-amine (4.00 g, 14.2 mmol) and 3-butenylamine (2.0 g, 21.26 mmol) in anhydrous THF (45.0 mL). The reaction crude was stirred at 60 °C for 16 h. The reaction crude was then diluted with AcOEt and washed with brine. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt, 100:0 to 50:50) to give the pure product (4.22 g, yield: 94%, yellow oil). LC-MS (ESI+): m / z: 317.1 RT: 1.841 min (TACC50).

[0382] 1-Allyl-7-chloro-3-(2,6-dimethyl-phenyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (111) [ka]

[0383] Triphosgene (5.90 g, 19.9 mmol) was added to a solution of butyl-{2-chloro-5-[(2,6-dimethyl-phenylamino)-methyl]-pyrimidin-4-yl}-amine (4.20 g, 13.26 mmol) in DCM (45.0 mL), which was stirred at room temperature for 1 h. Then an aqueous solution of sodium hydroxide (10.60 g, 265.12 mmol, 4.9 M) and tetrabutylammonium hydroxide (1.70 mmol 55% aqueous solution, 0.85 mL) was added at 0° C. The reaction was allowed to reach room temperature and stirred at room temperature for 16 h. The reaction crude was then diluted with DCM and washed with water. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was dissolved in DMF (40.0 mL) and cesium carbonate (4.36 g, 13.39 mmol) was added. The reaction mixture was heated at 60° C. for 16 h. The reaction crude was then diluted with DCM and washed with brine. The organic layer was dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (heptane:AcOEt 100:0 to 60:40) to give the pure product (2.61 g, yield: 57%, yellow solid). LC-MS (ESI+): m / z: 343.1 RT: 1.786 min (TACC50).

[0384] 1-Butyl-3-(2,6-dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (112) [ka]

[0385] 1-Allyl-7-chloro-3-(2,6-dimethyl-phenyl)-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (1.00 g, 2.92 mmol) and 4-(4-methyl-piperazin-1-yl)-3-(tetrahydro-pyran-2-yloxymethyl)-phenylamine (1.25 g, 4.08 mmol) were placed in a sealed tube and dissolved in anhydrous 2-BuOH (10.0 mL). 3A MS was then added, followed by trifluoroacetic anhydride (0.45 mL, 5.8 mmol) and the reaction was stirred at 90° C. for 16 hours. The reaction was then diluted with water and treated with aqueous NaHCO3. A solution of NaHCO3 (5.8 mmol) was added. The aqueous phase was extracted with a CHCl3 / iPrOH mixture (1:1). Finally, the combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH, 100:0 to 90:10) to give the pure product (1.03 g, yield: 67%, yellow solid). LC-MS (ESI+): m / z: 528.2 RT: 1.009 min (TACC50).

[0386] Acrylic acid 5-[8-butyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzyl ester (113) [ka]

[0387] DMAP (0.24 g, 1.95 mmol), DCC (0.60 g, 2.93 mmol) and acrylic acid (0.27 mL, 3.90 mmol) were added sequentially to an ice-cold solution of 1-butyl-3-(2,6-dimethyl-phenyl)-7-[3-hydroxymethyl-4-(4-methyl-piperazin-1-yl)-phenylamino]-3,4-dihydro-1H-pyrimido[4,5-d]pyrimidin-2-one (1.03 g, 1.95 mmol) in anhydrous DCM (12.0 mL). The reaction was allowed to come to room temperature and stirred at that temperature for 16 hours. The reaction crude was then diluted with DCM and washed with brine. The combined organic layers were dried over MgSO4, filtered and concentrated under vacuum. The resulting residue was purified by flash column chromatography on silica (DCM:MeOH, 100:0 to 90:10) to give the pure product (0.67 g, yield: 59%, white solid). LC-MS (ESI+): m / z: 582.2 RT: 1.316 min (TACC50).

[0388] (Z)-1 3 -(2,6-dimethylphenyl)-3 4 -(4-Methylpiperazin-1-yl)-1 1 ,1 2 ,1 3 ,1 4 -Tetrahydro-5-oxa-2-aza-1(7,1)-pyrimido[4,5-d]pyrimidin-3(1,3)-benzenacyclodecaphan-7-ene-1 2 ,6-dione(114) [ka]

[0389] Acrylic acid 5-[8-butyl-6-(2,6-dimethyl-phenyl)-7-oxo-5,6,7,8-tetrahydro-pyrimido[4,5-d]pyrimidin-2-ylamino]-2-(4-methyl-piperazin-1-yl)-benzyl ester (0.20 g, 0.344 mmol) was placed in a sealed tube and dissolved in anhydrous toluene (64.0 mL). HCl (0.344 mmol, 3 M in cyclopentyl methyl ether, 0.115 mL) was then added. The reaction crude was degassed by bubbling N2 through the solution, followed by the addition of a solution of Zhan's catalyst (0.051 g, 0.069 mmol) in anhydrous toluene (5.0 mL). The reaction crude was degassed again by bubbling N2 through the solution and heated at 110° C. for 16 h. The solvent was then removed under vacuum and the resulting residue was purified by reverse phase chromatography (59% [25 mM NH4HCO3 aq.]-41% ACN to 17% [25 mM NH4HCO3 aq.]-83% ACN) to give the pure product as a single cis diastereomer (0.017 g, pale brown solid). 1H NMR (400 MHz, DMSO) δ 9.62 (s, 1H), 8.54 (d, J = 2.0 Hz, 1H), 8.14 (s, 1H), 7.20 - 7.15 (m, 3H), 7.10 (dt, J = 8.7, 5.6 Hz, 2H), 6.33 (dt, J = 11.7, 8.9 Hz, 1H), 5.89 (d, J = 11.9 Hz, 1H), 5.32 (s, 2H), 4.49 (s, 2H), 3.95 - 3.89 (m, 2H), 2.95 (dd, J = 14.0, 8.3 Hz, 2H), 2.79 (t, J = 4.5 Hz, 4H), 2.48 - 2.42 (m, 3H), 2.24 (s, 3H), 2.18 (s, 6H). LC-MS (ESI+): m / z: 555.3 RT: 2.504 min (VILLA).

[0390] Experimental procedure

[0391] HPLC_MS method description TACC50: Agilent 1260 Infinity (Quat.Pump) DAD LC / MS g6120 (G1948B) instrument. Column Thermo Scientific Accucore C18 (50 × 4.6 mm, 2.6 μm). Mobile phase: A: 0.1% HCOOH in H2O B: CH3CN. 90% A to 10% A in 1.5 min, hold for 0.9 min, 95% A in 0.1 min. Flow rate 3 mL / min, 30 °C.

[0392] VLIA: Agilent 1100HPLC DAD LC / MSg1956A instrument. Column YMC-pack ODS-AQ C18 (50 x 4.6 mm, 3 μm). Mobile phase: A: 0.1% HCOOH in H2O B: CH3CN. 95% A to 5% A in 4.8 min, hold for 1.0 min, to 95% A in 0.2 min. Flow rate 2.6 mL / min, 35 °C.

[0393] VILL-J: Agilent 1100HPLC DAD LC / MSg1956A instrument. Column Waters-XBridge C18 (50×4.6 mm, 3.5 μm). Mobile phase: A: 40 mm NH4OAc + 5% CH3CN in H2O B: CH3CN. 95% A to 0% A in 4.5 min, hold for 1.0 min, to 95% A in 0.5 min. Flow rate 2.6 mL / min, 50 °C.

[0394] VLIA-2T: Agilent 1260 Infinity DAD TOF-LC / MSg6224A instrument. Column: YMC-pack ODS-AQ C18 (50×4.6 mm, 3 μm). Mobile phase: A: 0.1% HCOOH in H2O B: CH3CN. 95% A to 5% A in 4.8 min, hold for 1.0 min, to 95% A in 0.2 min. Flow rate 2.6 mL / min, 35 °C.

[0395] Example 4: SIK1, 2 and 3 kinase activity

[0396] SIK1, 2 and 3 kinase activity was assessed using Eurofins Cerep SA ("Eurofins") KinaseProfiler. All compounds were prepared in 100% DMSO to 50x final assay concentration. Working stocks of the compounds were added to the assay wells as the first component in the reaction, followed by the remaining components as detailed in the general assay protocol below. In the standard KinaseProfiler service, there is no pre-incubation step between the compound and the kinase before the reaction begins. Positive control wells contain all components of the reaction except the compound of interest, but DMSO (at a final concentration of 2%) is included in these wells to control for solvent effects. Blank wells contain all components of the reaction with a reference inhibitor replacing the compound of interest. This nullifies kinase activity and establishes a baseline (0% kinase activity remaining).

[0397] SIK(h) was incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 100 μM AMARASAAAALARRR, 10 mM magnesium acetate and [γ-33P]-ATP (specific activity and concentration as required). Reactions were initiated by addition of the Mg / ATP mix. After 40 min incubation at room temperature, reactions were stopped by adding phosphoric acid to a concentration of 0.5%. 10 μL of the reaction was spotted onto a P30 filtermat and washed four times for 4 min in 0.425% phosphoric acid and once in methanol before drying and scintillation counting.

[0398] SIK2(h) was incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 100 μM KKKVSRSGLYRSPSMPENLNRPR, 10 mM magnesium acetate and [γ-33P-ATP] (specific activity and concentration as required). Reactions were initiated by adding the Mg / ATP mix. After 40 min incubation at room temperature, reactions were stopped by adding phosphoric acid to a concentration of 0.5%. 10 μL of the reaction was spotted onto a P30 filtermat and washed four times for 4 min in 0.425% phosphoric acid and once in methanol before drying and scintillation counting.

[0399] SIK3(h) was incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 100 μM KKKVSRSGLYRSPSMPENLNRPR, 10 mM magnesium acetate and [γ-33P-ATP] (specific activity and concentration as required). The reaction was started by adding the Mg / ATP mixture. After incubation for 40 min at room temperature, the reaction is stopped by adding phosphoric acid to a concentration of 0.5%. 10 μL of the reaction was spotted onto a P30 filtermat and washed four times for 4 min in 0.425% phosphoric acid and once in methanol before drying and scintillation counting.

[0400] The results are shown in Table 5 below. [Table 5]

[0401] The results are shown in Table 6 below. [Table 6]

[0402] Macrocyclic compounds exhibit potent inhibitory activity against salt-induced kinases. As evidenced by SLT-058 and SLT-059, the ring size and substitution of the aromatic moiety can affect potency.

[0403] Example 5: Cell-based kinase activity

[0404] Cell-based kinase activity was determined using the PathHunter detection system. PathHunter cell lines were grown from freezer stocks following standard procedures. Cells were seeded in white-walled 384-well microplates in a total volume of 20 μL and incubated for the appropriate time prior to testing. For agonist measurements, cells were incubated with samples to induce a response. Intermediate dilutions of sample stocks were performed to generate 5× samples in assay buffer. 5 μL of 5× samples were added to cells and incubated at room temperature for 3 hours. Vehicle concentration was 1%. For agonist and antagonist assays, assay signal was generated by a single addition of 12.5 μL or 15 μL (50% v / v) of PathHunter detection reagent cocktail, respectively, followed by incubation at room temperature for 1 hour. Microplates were read after signal generation using a PerkinElmer Envision™ instrument for chemiluminescent signal detection. Compound activity was analyzed using the CBIS data analysis suite (ChemInnovation, CA). For agonist mode assays, percent activity was calculated using the following formula: % activity = 100% x (mean RLU of test sample - mean RLU of vehicle control) / (mean MAX RLU of control ligand - mean RLU of vehicle control).

[0405] The results are shown in Table 7 below. [Table 7]

[0406] Example 6: Cell-Based SIK1 Activity of SLT-048 and SLT-026

[0407] SIK1 activity was determined using a NanoBRET target-binding intracellular kinase assay to screen compounds that bind to SIK1. The NanoBRET target-binding assay uses an energy transfer technique designed to measure molecular proximity in living cells. The assay measures the apparent affinity of test compounds by competitive displacement of a NanoBRET tracer reversibly bound to a NanoLuc luciferase-kinase fusion construct in cells. The intracellular binding affinity and selectivity are physiologically relevant and fundamental to the pharmacological mechanism of the compound. HEK293 cells transiently expressing the NanoLucSIK1 fusion vector were seeded into wells of a 384-well plate. The cells were pretreated with NanoBRET tracer K-4 and then treated with the reference compound dasatinib for 1 hour. The BRET signal was measured with an Envision 2104 Multilabel Reader. IC50 values ​​were calculated and IC50 curves were plotted using the GraphPad Prism 4 program based on a sigmoidal dose-response equation. The IC50(M) of the reference compound dasatinib is 3.188e -9 It was.

[0408] Next, SIK1 activity was determined for SLT-026 and SLT-048. HEK293 cells were transfected with 1 μg of SIK1-NanoLuc fusion vector and 9 μg of transfection carrier DNA. The transfected cells were treated with customer compounds (starting at 10 μM, 10 doses in 3-fold dilutions) and reference compounds (starting at 1 μM, 10 doses in 3-fold dilutions). SIK1 target association was measured by NanoBRET assay. Curve fitting was performed only if the percentage of NanoBret signal at the highest concentration of compound was less than 55%.

[0409] The IC50(M) values ​​for SLT-048 and SLT-026 were 1.63e -10 and 4.25e -11 (dasatinib was 2.77e -9 SLT-048 and SLT-026 demonstrated increased SIK1 inhibitory activity compared to the control (dasatinib).

[0410] Example 7: Evaluation of the effects of SLT-023 and SLT-048 on pigmentation of B16 4A5 cells in culture

[0411] This study aimed to evaluate the pro-pigmentation properties of the SLT-023 and SLT-048 compounds in B16 4A5 cells. First, a dose-finding study was performed by testing five concentrations of each compound on cell viability using the MTS assay. Second, the tanning properties of the test compounds applied in three concentrations in the culture medium were studied by melanin content measurements after chemical extraction. In parallel, protein dosing using the Bradford method was performed to normalize the melanin content to the total cell protein.

[0412] This study was performed on B16 4A5 mouse melanoma cells (ECACC; 94042254)-melanin producing. Cells were cultured as monolayers in DMEM medium supplemented with 20% M199, 10% FBS, and penicillin and streptomycin and grown in a cell incubator at 37°C and 5% CO2.

[0413] B16 4A5 cells were seeded in triplicate at 20,000 cells / well in 24-well plates (n=3) 24 hours before the start of treatment with five concentrations of test compounds. Cells were then incubated with five concentrations (5, 0.7, 0.1, 0.0146 and 0.002 μg / mL) of SLT-023 and SLT-048 for 72 hours, with SDS 0.05% for 72 hours, or left untreated. In addition, 0.1% DMSO, used for solubilizing the test compounds, was tested in parallel. Treatment was performed in culture medium containing only 1% FBS. At the end of treatment, cell viability was evaluated with the MTS assay (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfophenyl 2H-tetrazolium)).

[0414] Twenty-four hours before treating B16-4A5 melanocytes with SLT-023 (1, 0.3, and 0.1 μg / mL) and SLT-048 (3, 1, and 0.3 μg / mL) for 72 hours, they were seeded in 6-well plates at 100,000 cells / well. Additionally, 0.06% DMSO for 72 hours and 2 mM theophylline (Sigma; T1633) for 72 hours were used for solubilization of the test compounds and as a positive control for pigmentation, respectively. The treatments were carried out in culture medium + 1% FBS.

[0415] After 72 hours of treatment, the B16-4A5 cells were lysed with 1 M NaOH, and the total protein content was quantified by the Bradford method. The melanin content was measured after heating the samples at 80 °C for 1 hour. Absorbance readings were taken at 490 nm and compared to a synthetic melanin standard curve. These results were normalized to the amount of protein present in each sample measured using the Bradford method.

[0416] The effect of the compounds on B16 4A5 viability was evaluated. For this purpose, the cells were incubated with five concentrations of the compounds for 72 hours, left untreated, or treated with 0.1% SDS or DMSO. After 72 hours of incubation, the viability of B16 4A5 was evaluated by the MTS assay. A 0.01 < p-value < 0.05 was considered significant (* or $), a 0.001 < p-value < 0.01 was considered highly significant (** or $$), and a p-value < 0.001 was considered very highly significant (*** or $$$).

[0417] Since the solvents for SLT-023 and SLT-048 were DMSO, the viability of the cells incubated with these compounds was compared to a 0.1% DMSO control. The viability of the cells incubated with 0.05% SDS decreased strongly and significantly compared to the untreated cells, which ensured the robustness of the experiment. The viability of the cells incubated with 5 μg / mL of SLT-023 and SLT-048 decreased slightly. The cell viability of the cells incubated with 0.7 μg / mL of SLT-023 decreased slightly.

[0418] Next, the effects of the compounds on B16 4A5 melanin production were evaluated. For this purpose, cells were incubated for 72 hours with three concentrations of the compound, left untreated, treated with 2 mM theophylline (positive control for melanin production) or 0.06% DMSO. After 72 hours of incubation of B16 4A5, the intracellular melanin content was evaluated.

[0419] At the end of the 72-hour treatment, the cells were lysed with NaOH, heated at 80 °C for 1 hour, and then the melanin content was determined by spectrophotometry. The amount of protein present in each sample was determined using the Bradford method. The melanin content was then normalized against the amount of protein. Statistical analysis (Student's t-test) was performed on the normalized data. The effects of the test compounds SLT-023 and SLT-048 were compared to the 0.06% DMSO condition, with 0.01 < p-value < 0.05 considered significant (* or $), 0.001 < p-value < 0.01 considered highly significant (** or $$), and p-value < 0.001 considered very highly significant (*** or $$$) (Figure 2).

[0420] Compound SLT-023 (1, 0.3, and 0.1 μg / ml) significantly increased melanin production in a dose-dependent manner at all doses. SLT-048 at 1 and 0.3 μg / ml was also able to significantly increase intracellular melanin production after 72 hours of treatment.

[0421] In conclusion, SLT-048 at 1 and 0.3 μg / ml increased melanin production by 250% and 180% respectively compared to 0.06% DMSO used as a solvent that was able to increase melanin production. SLT-023 significantly increased melanin production in B16 4A5 cells in a dose-dependent manner at any dose.

[0422] Example 8: Evaluation of SLT-023, -026, -042, and -048 on pigmentation using ex vivo living human skin explants

[0423] The aim of this study was to evaluate the effect of different products on pigmentation (pro-pigmentation effect) using living human skin explants. This activity was evaluated by controlling cell viability after Masson's trichrome staining and melanin visualization after Fontana-Masson staining.

[0424] All tested products were diluted in the vehicle marked E (ethanol 65%, propylene glycol 25%, transcutol 10%). The diluted solutions were stored at 4 °C for the duration of the study. Thirty-six human skin explants with a mean diameter of 11 mm (± 1 mm) were prepared at an abdominoplasty from a 53-year-old Caucasian woman with III phototype (reference: P2462-AB53). The explants were kept alive in BEM culture medium (BIO-EC's Explant Medium) at 37 °C in a humidified 5% CO2 atmosphere. The study is carried out on human skin tissue obtained from surgical residues in full relation to the Declaration of Helsinki and to article L.1243-4 of the French Public Health Code. The latter does not require any prior authorization by the Ethics Committee for the sampling and use of surgical waste.

[0425] Excipient E and test product were placed in a 1 cm 2 Apply topically at a rate of 2 μL per explant, approximately 2 mg / cm 2 , on day 0 (D0), D3, D5, D7 and D10, the explants were spread and gently rubbed for 10 seconds using a small spatula. Control explants T did not receive any treatment other than medium renewal. The culture medium was half-refreshed (1 mL per well) on D3, D5, D7 and D10. The medium of the irradiated explants (UV) was replaced with HBSS (Hank's Balanced Saline; 1 mL per explant) on D0, D3, D4, D5, D6, D7 and D10. The explants of the "UV" batch were then irradiated with 2.25 J / cm2, corresponding to 0.5 MED (minimal erythema dose) on skin with III phototype, using a UV simulator Vibert Lourmat RMX 3W. 2The samples were irradiated with 100% UVA dose and 6-8% UVB. At the end of UV irradiation, explants from batch TUV were placed back in 2 mL of BEM medium. On D0, three explants from batch T0 were collected and cut into two parts. One half was fixed in a buffered formalin solution and the other half was frozen at -80 °C. On D10, three explants from the relevant batch were collected and processed in the same way as on D0. The treatment, irradiation and sampling days were adjusted to the working day schedule according to the treatments described in the study plan. After 24 h of fixation in buffered formalin, the samples were dehydrated and impregnated in paraffin using a Leica PEARL dehydration automat. The samples were embedded using a Leica EG 1160 embedding station. 5 μm thick sections were made using a Leica RM 2125 min-tome and the sections were mounted on Superfrost® histological glass slides. Frozen samples were cut into 7 μm thick sections using a Leica CM 3050 cryostat. Sections were then mounted on Superfrost® plus silanized glass slides. Microscopic observations were achieved using a Leica DMLB or Olympus BX43 microscope. Photographs were digitized with a Numeric DP72 Olympus camera equipped with CellSens saving software.

[0426] Cell viability of epidermal and dermal structures was assessed by: microscopy of formalin-fixed paraffin-embedded (FFPE) skin sections after Masson's Trichrome staining, Goldner variant. Cell viability was very slightly decreased among samples treated with 1% SLT-023, -026, -042 and -048.

[0427] Further analysis was performed to quantify melanin in the basal and suprabasal layers. Melanin was visualized after silver impregnation according to the Masson-Fontana staining method on FFPE skin sections. Staining was evaluated by microscopic observation (Figure 3) and image analysis (Figure 4). Macroscopic photographs were taken on days 0 and 10 using a Nikon D5300 camera.

[0428] UV-induced increase in melanin vs. untreated explants (UVA) was validated as a positive control experiment. Chronic UVA / B irradiation induced a significant increase of 43%** in the basal layer and a significant increase of 29% in the suprabasal layer. The vehicle "Ethanol 65%, Propylene Glycol 25%, Transcutol 10%" (E) induced no significant changes in the basal layer and a significant decrease of 22%* in the suprabasal layer. 1% product SLT-026, E (P5) induced a significant increase of 50%** in the suprabasal layer. 1% product SLT-042, F (P6) induced a significant increase of 25% in the suprabasal layer.

[0429] In conclusion, SLT-023, SLT-026, SLT-042 and SLT-048 increased melanin production in living human skin explants.

[0430] Example 9: Evaluation of SLT-048 on ex vivo pigmentation using living human skin explants

[0431] The aim of this study was to evaluate the effect of SLT-048 on pigmentation (pro-pigmentation effect) using live human skin explants in survival medium.

[0432] All tested products were diluted in vehicle (ethanol 70%, propylene glycol 30%). Twenty-four human skin explants with a mean diameter of 11 mm (±1 mm) were prepared at abdominoplasty from a 43-year-old Caucasian female with II phototype (reference: P2511-AB43). The explants were cultured in BEM culture medium (BIO-EC's Explant Medium) at 37°C in humidified 5% CO. 2 The products were grown in a 1 cm 2 2μL per unit (2mg / cm 2) and spread and gently rubbed for 10 seconds using a small spatula on day 0 (D0), D2, D5 and D7. Control explants did not receive any treatment other than medium renewal. Culture medium was half-fresh (1 mL per well) on D2, D5 and D7. After 24 h of fixation in buffered formalin, samples were dehydrated using a Leica PEARL dehydration automat and impregnated in paraffin. Samples were embedded using a Leica EG 1160 embedding station. 5 μm thick sections were made using a Leica RM 2125 min-t microtome and sections were mounted on Superfrost® histological glass slides. Frozen samples were cut into 7 μm thick sections using a Leica CM 3050 cryostat. Sections were then mounted on Superfrost® plus silanized glass slides. Microscopic observations were realized using a Leica DMLB or Olympus BX43 microscope. Photographs were digitized with a Numeric DP72 Olympus camera equipped with CellSen storing software. Melanin was visualized after silver impregnation according to the Masson-Fontana staining method on FFPE skin sections. Staining was evaluated by microscopic observation (Figure 5) and image analysis (Figure 6).

[0433] UV-induced increase in melanin versus untreated explants (UVA) was validated as a positive control experiment. 0.2% SLT-048 produced a slight increase in melanin, and 2% SLT-048 produced a moderate increase in melanin.

[0434] UV irradiation significantly increased melanin content (+78%) as a positive control. 0.2% SLT-048 produced 10.0 (+24%) and 2% SLT-048 produced 12.6 (+56%) of the surface area occupied by melanin in the basal cell layer of the epidermis, compared to vehicle E 7.9.

[0435] Melanin visualized after silver impregnation according to the Masson-Fontana staining method on FFPE skin sections is shown in Figure 7.

[0436] Melanin content was increased by 0.2% SLT-048 and highly increased by 2% SLT-048; keratinocyte nuclei capping by melanin was also detected. The increase in melanin was high in the basal layer of the epidermis and to a lesser extent in the suprabasal layers.

[0437] Following topical application, SLT-048 significantly increased epidermal melanin expression in an ex vivo skin explant model in a dose-dependent manner.

[0438] Example 10: Evaluation of SLT-045 on ex vivo pigmentation using living human skin explants

[0439] The aim of this study was to evaluate the effect of SLT-045 on pigmentation (pro-pigmentation effect) using live human skin explants in survival medium.

[0440] SLT-045 (3, 7, 15, 21 mm; 5 μL / cm2) in vehicle (44.9% DMSO / 37.5% EtOH / 17.4% propylene glycol) was applied topically using a Pasteur pipette to triplicate skin samples (8 mm punches) daily for 6 days.

[0441] As shown in FIG. 8, topical application of SLT-045 at 15 and 21 mm for 6 days ex vivo induced visible skin pigmentation.

Claims

1. A compound having the structure of formula (I) or (II). 【Chemistry 1】 During the ceremony, L １ and L ２ Each of these is independently a bonded, substituted or unsubstituted alkylene, or a substituted or unsubstituted alkenylene; Each R ２Ａ 、R ２Ｂ and R ２Ｃ are independently hydrogen, halogen, -CX ２ ３ 、-CHX ２ ２ 、-CH ２ X ２ 、-OCX ２ ３ 、-OCH ２ X ２ 、-OCHX ２ ２ 、-OR ２Ｆ 、a substituted or unsubstituted alkyl, a substituted or unsubstituted heteroalkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted aryl, or a substituted or unsubstituted heteroaryl; R ５ These are halogen and -CX, which are used independently. ５ ３ EN-CHX ５ ２ ien-CH ２ X ５ , -OCX ５ ３ , -OCH ２ X ５ , -OCHX ５ ２ , -OR ５Ｆ , or substitution or non-substitution C １ -C ４ It is alkyl; z is an integer between 0 and 5; Each X ２ and X ５ These are independently -F, -Br, -Cl, or -I; Each R ２Ｆ and R ５Ｆ (This is hydrogen, or a substituted or unsubstituted alkyl group.)

2. The compound according to claim 1, wherein z is 2.

3. R ５ However, independently, non-substituted C １ -C ４ The compound according to claim 1, wherein it is alkyl.

4. The compound according to claim 1, having the structure of formula (I-a) or (II-a). 【Chemistry 2】

5. R ２Ａ The compound according to claim 1, wherein the compound is a substituted or unsubstituted heterocycloalkyl group, or a substituted or unsubstituted aryl group.

6. R ２Ａ The compound according to claim 1, wherein the compound is a substituted or unsubstituted piperazinyl.

7. The compound according to claim 1, having the structure of formula (I-b) or (II-b). 【Transformation 3】 (In the formula, R ３ is hydrogen or a substituted or unsubstituted alkyl group; Each R ４Ａ , R ４Ｂ , R ４Ｃ , and R ４Ｄ These are, independently, hydrogen, halogen, and -CX ４ ３ EN-CHX ４ ２ ien-CH ２ X ４ , -OCX ４ ３ , -OCH ２ X ４ , -OCHX ４ ２ , -OR ４Ｆ , or substituted or unsubstituted alkyl groups; X ４ These are independently -F, -Br, -Cl, or -I; R ４Ｆ (This is hydrogen, or a substituted or unsubstituted alkyl group.)

8. The compound according to claim 7, having the structure of formula (I-c). 【Chemistry 4】 (In the formula, L １ is a combination, or R ６ - Substitute or non-substitute C １ -C ４ It is alkylene; L ２ C is either substituted or non-substituted. ２ -C ５ Alkylene, or substituted or unsubstituted C ２ -C ５ It is alkenylene; R ６ is a halogen or unsubstituted C １ -C ４ It is alkylene.

9. The compound according to claim 7, having the structure of formula (II-c). 【Transformation 5】 (In the formula, L １ R ６ - Substitutive or non-substitutive C １ -C ４ It is alkylene; L ２ C is either substituted or non-substituted. ２ -C ５ Alkylene, or substituted or unsubstituted C ２ -C ５ It is alkenylene; R ６ is a halogen or unsubstituted C １ -C ４ It is alkylene.

10. R ２Ａ The compound according to claim 1, wherein is a halogen.

11. The compound, 【Transformation 6】 【Transformation 7】 The compound according to claim 1.

12. A chemical compound having the structure of formula (III) or (IV). 【Transformation 8】 (In the formula, R １ These are independently hydrogen, a substituted or unsubstituted alkyl group, or a substituted or unsubstituted alkenyl group; L １ and L ２ are each independently a linking, substituted or unsubstituted alkylene, or a substituted or unsubstituted alkenylene; R ２Ａ 、 R ２Ｂ and R ２Ｃ are each independently hydrogen, halogen, -CX ２ ３ 、 -CHX ２ ２ 、 -CH ２ X ２ 、 -OCX ２ ３ 、 -OCH ２ X ２ 、 -OCHX ２ ２ 、 -OR ２Ｆ 、 a substituted or unsubstituted alkyl, a substituted or unsubstituted heteroalkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted aryl, or a substituted or unsubstituted heteroaryl; R ５ These are halogen and -CX, which are used independently. ５ ３ EN-CHX ５ ２ ien-CH ２ X ５ , -OCX ５ ３ , -OCH ２ X ５ , -OCHX ５ ２ , -OR ５Ｆ , or substitution or non-substitution C １ -C ４ It is alkyl; z is an integer between 0 and 5; R ７ is, -OR ７Ｆ , or substituted or unsubstituted alkenyls; X ２ and X ５ These are, independently, -F, -Br, -Cl, or -I; Each R ２Ｆ , R ５Ｆ and OR ７Ｆ (These are independently hydrogen, or a substituted or unsubstituted alkyl group.) 【Request Item 13】 【Chemistry 13】 【Chemistry 14】 【Chemistry 15】 The compound according to claim 12.

14. A method for producing a compound having the structure of formula (I) or (II), 【Chemistry 16】 A method comprising the step of providing a compound having the structure of formula (III) or (IV). 【Chemistry 17】 (In formulas (I) to (IV), R １ These are independently hydrogen, a substituted or unsubstituted alkyl group, or a substituted or unsubstituted alkenyl group; L １ and L ２ Each of these is independently a bonded, substituted or unsubstituted alkylene, or a substituted or unsubstituted alkenylene; Each R ２Ａ , R ２Ｂ and R ２Ｃ These are independently hydrogen, halogen, and -CX ２ ３ EN-CHX ２ ２ ien-CH ２ X ２ , -OCX ２ ３ , -OCH ２ X ２ , -OCHX ２ ２ , -OR ２Ｆ , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R ５ These are halogen and -CX, which are used independently. ５ ３ EN-CHX ５ ２ ien-CH ２ X ５ , -OCX ５ ３ , -OCH ２ X ５ , -OCHX ５ ２ , -OR ５Ｆ , or substitution or non-substitution C １ -C ４ It is alkyl; z is an integer between 0 and 5; R ７ is, -OR ７Ｆ , or substituted or unsubstituted alkenyls; Each X ２ and X ５ These are independently -F, -Br, -Cl, or -I; Each R ２Ｆ , R ５Ｆ and R ７Ｆ (These are independently hydrogen, or a substituted or unsubstituted alkyl group.)

15. Compounds of formula (III) or (IV) [Chemistry 18] 【Chemistry 19】 The method according to claim 14.

16. Compounds of formula (III) or (IV) 【Chemistry 20】 【Chemistry 21】 【Chemistry 22】 The method according to claim 14.

17. A pharmaceutical composition comprising the compound described in claim 1.

18. A pharmaceutical composition comprising the compound according to claim 1 for treating subjects who are suffering from or are susceptible to skin-related disorders or diseases.

19. A pharmaceutical composition comprising the compound according to claim 1 for the treatment of a person suffering from or susceptible to rosacea.

20. A pharmaceutical composition comprising the compound according to claim 1 for increasing pigmentation in a target tissue.

21. A pharmaceutical composition comprising the compound according to claim 1 for increasing the stability or repair of cellular DNA in target skin tissue where such a function is required.

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