Topical treatment of skin cancer with oligopeptides

Abstract

The present invention relates to compounds of formula (I), pharmaceutically acceptable salts thereof and pharmaceutical compositions containing them, in particular compounds of formula (I) and pharmaceutical compositions for use in a method for the topical treatment of skin cancer in a mammal, said method comprising topical administration of said compound to said mammal. [Formula 1] JPEG2025526647000170.jpg29149 [chemical 2] JPEG2025526647000171.jpg84103

Description

[Technical Field]

[0001] The present invention relates to compounds of formula (I), pharmaceutically acceptable salts thereof, and pharmaceutical compositions containing them, in particular to compounds of formula (I), pharmaceutically acceptable salts thereof, and pharmaceutical compositions containing them, for use in methods for the topical treatment of skin cancers, including basal cell carcinoma, cutaneous squamous cell carcinoma, and melanoma. [Background technology]

[0002] Skin has been recognized as an interesting route for drug delivery because local administration of drugs for dermal and / or transdermal application not only avoids the risk of toxicity and gastrointestinal side effects often associated with other classical treatments, such as parenteral or oral systemic treatments, but also has greater patient acceptance and does not require pre-, intra-, or post-treatment evaluations to check toxicity levels in the liver, kidneys, etc. Therefore, there is great interest in developing drugs suitable for topical application, especially for application to the skin.

[0003] Human skin is the largest and most accessible organ for drug delivery.However, skin is the natural physicochemical barrier of our body, and as a result, it is characterized by low permeability, which limits the transport of most pathogens, toxins and drug molecules.Therefore, drug delivery into and across the skin is extremely challenging.

[0004] The primary transport barrier for drug delivery into or across the skin is the stratum corneum, typically 10–20 μm thick and consisting of 10–15 layers of corneocytes surrounded by a lipid matrix rich in ceramides, cholesterol, and free fatty acids. To penetrate the stratum corneum, drugs must navigate the tortuous lipid pathways surrounding the keratin-rich cells, i.e., repeatedly partition between the keratin-rich aqueous phase and the lipid phase. Generally, small molecules can penetrate the stratum corneum; in contrast, delivery of larger molecules, such as peptides and proteins, remains a significant challenge. For pharmaceutical purposes, approximately 500 daltons marks the beginning of a rapid decline in dermal absorption due to molecular size, so it has been argued to limit the development of new compounds to molecular weights (MW) below 500 daltons when addressing topical dermatological or transdermal systemic therapies. This is now known as the "500 dalton rule." Additional physicochemical properties of topical drugs that are beneficial for achieving passive transport through the stratum corneum are a partition coefficient octanol / water logP of 1-3 and a water solubility greater than 1 mg / ml and the absence of polar centers (Bos and Meinardi, Exp Dermatol 2000, 9:165-169; Gorouhi and Maibach, International Journal of Cosmetic Science, 2009, 31, 327-345).

[0005] Most skin cancers are locally destructive, cancerous (malignant) growths of the skin. Most skin cancers originate in the cells of the epidermis, the surface layer of the skin. Unlike cutaneous malignant melanoma, the majority of these types of skin cancers spread (metastasize) to other parts of the body and rarely become life-threatening. There are three main types of skin cancer: (1) basal cell carcinoma (BCC; most common), (2) cutaneous squamous cell carcinoma (cSCC; second most common), which originates in skin cells, and (3) melanoma, which originates in pigment-producing skin cells (melanocytes) but is less common but more dangerous than the first two types. Other rare forms of skin cancer include lymphoma, Merkel cell carcinoma, and sarcoma, as well as cancers of other tissues of the skin, including hair and sweat gland tumors (Ascierto PA, Garbe C, Immunotherapy (2020) 12(3):167-174; U. Hillen et al., European Journal of Cancer (2018) 96:34-43, Loh TY et al., Dermatol Surg. (2017) 43(1):32-39). Summary of the Invention

[0006] The present inventors have surprisingly found that the compounds of the present invention are suitable for topical application as potential pharmaceuticals for the treatment of skin cancer. In particular, the compounds of the present invention not only penetrate the skin, as demonstrated by pig skin penetration experiments, but also have activity against cutaneous squamous cell carcinoma, basal cell carcinoma, and melanoma, as demonstrated by in vitro assays. Furthermore, preferred compounds of the present invention have shown successful treatment and antiproliferative effects in an in vivo mouse model of melanoma using C67BLC / 6 mice bearing B16-F10 cell tumors, the most commonly used metastatic melanoma model for preclinical studies, with topical drug treatment resulting in a significant reduction in tumor volume in the treated group compared with controls. Histological studies further demonstrate strong proapoptotic potential. Furthermore, ex vivo experiments using cancer tissues from several patients with basal cell carcinoma and cutaneous squamous cell carcinoma treated with the compounds of the present invention have shown a significant reduction in proliferating cells at concentrations as low as 500 nM.

[0007] Thus, in a first aspect, the present invention provides a compound of formula (I) for use in a method of topical treatment of skin cancer in a mammal, preferably a human, said method comprising topical administration of said compound to said mammal, preferably to said human, [ka] During the ceremony, A1 is [ka] A2 is [ka] A3 is [ka] B1 is [ka] B2 is [ka] B3 is [ka] During the ceremony, a1, a 12 , a2, a 22 , a3, a 32 , b1, b 12 , b2, b 22 , b3 and b 32 are independently selected from 0 and 1, and a1+a 12 , a2+a 22 , a3+a 32 , b1+b 12 , b2+b 22 and b3+b 32 are independently selected from 0 and 1, and a1+a 12 +a2+a 22 +a3+a 32 is 0, 1, 2 or 3, and b1+b 12 +b2+b 22 +b3+b 32 is 0, 1 or 2; During the ceremony, R1 is independently selected from C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, carbocyclyl or heterocyclyl optionally and preferably substituted with halogen, C1-C3 alkyl, OR4, C6H5 substituted with NR5R6, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl; R2 is C4~C 12 Alkyl, C4-C 10alkoxy, C1-C3 alkylene-cycloalkyl, C1-C3 alkylene-aryl, C1-C3 alkylene-heteroaryl, wherein independently, in said C1-C3 alkylene, one -CH2- moiety is optionally replaced by -CH(NH)- or -O-; said alkyl, cycloalkyl, aryl and heteroaryl are each independently optionally and preferably substituted with one or more, preferably one or two, substituents selected from C1-C2 alkyl, C1-C2 haloalkyl, oxo, OH, halogen, C1-C2 alkoxy, C6H5 or C6H5 substituted with C1-C3 alkyl or OC1-C3 alkyl; R3 is [ka] where: R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10 two of which, together with the carbon atoms to which they are attached, form a carbocyclic or heterocyclic ring, preferably a carbocyclic ring, wherein R 11 and R 12 are, independently of each other, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl, or C3-C6 cycloalkyl; or, together with the nitrogen atom to which they are attached, independently in each occurrence, are each independently halogen, C1-C4 alkyl, OR 13 , N.R. 14 R 15 forming a heteroaryl or heterocyclyl optionally and preferably substituted with 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl, and the arrow indicates the bond to the C(O)-moiety depicted in formula (I); as well as pharmaceutically acceptable salts of the compound of formula (I), preferably wherein the method comprises topical administration to the mammal, preferably the human, of an effective amount of the compound.

[0008] In a further aspect, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, in particular a compound of formula (I) or a pharmaceutically acceptable salt thereof as in any one of the accompanying claims, for use in a method of treating skin cancer in a mammal, preferably a human, as defined herein, said method comprising topical administration of a compound of formula (I) to said mammal, preferably to said human; preferably said method comprises topical administration to said mammal, preferably to said human, of an effective amount of said compound or said pharmaceutically acceptable salt thereof.

[0009] In a further aspect, the present invention provides a pharmaceutical composition for use in a method of topical treatment of skin cancer in a mammal, preferably a human, said pharmaceutical composition comprising a compound of formula (I) as defined herein, in particular as in any one of the appended claims, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or adjuvant, said method comprising topical administration of said pharmaceutical composition to said mammal, preferably to said human, and preferably said pharmaceutical composition comprising an effective amount of said compound.

[0010] Thus, in particular, the present invention provides compounds and pharmaceutical compositions for use in a method of topical treatment of skin cancer in a mammal, preferably a human, in particular melanoma and non-melanoma skin cancer, preferably melanoma, basal cell carcinoma and cutaneous squamous cell carcinoma, said method comprising the topical administration to said mammal, preferably said human, of an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising said compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0011] In another aspect, the present invention provides a method for topically treating skin cancer in a mammal, preferably a human, said method comprising topical administration to said mammal, preferably said human, of a compound of formula (I) of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising said compound of formula (I) or a pharmaceutically acceptable salt thereof, preferably said method comprising topical administration to said mammal, preferably said human, of an effective amount of said compound or said pharmaceutically acceptable salt thereof, or said pharmaceutical composition.

[0012] In a further aspect, the present invention provides use of a compound of formula (I) of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising said compound of formula (I) or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for topically treating skin cancer in a mammal, preferably a human, said method comprising topical administration of said compound of formula (I) or said pharmaceutically acceptable salt thereof, or said pharmaceutical composition of the present invention to said mammal, preferably said human, preferably comprising topical administration of an effective amount of said compound or said pharmaceutically acceptable salt thereof, or said pharmaceutical composition to said mammal, preferably said human.

[0013] Further aspects and embodiments of the present invention will become apparent as this description continues. [Brief explanation of the drawings]

[0014] [Figure 1] Figure 1A: Tumor slices taken from C57Bl / 6 mice bearing B16F10 xenografts were collected and studied histologically. Tumor slices were stained with the proliferation marker Ki67 to identify tumor areas with proliferating cells and with the apoptosis marker TUNEL to indicate areas within the tumor with apoptotic cells.

[0015] FIG. 1B: Quantification of cells stained positively for Ki67 and TUNEL as described in FIG. 1A for group A (placebo) and group B (6027).

[0016] [Figure 2] Figure 2A: Immunohistochemical staining of patient-derived basal cell carcinoma (BCC) cultured for 4 days in medium containing 500 nM and 1 μM 6027 or DMSO as a negative control and vismodegib (20 μM) as a positive control.

[0017] Figure 2B: Quantification of cells stained positively for Ki67 and TUNEL as described in Figure 2A for A (DMSO), B (vismodegib, 20 mM), C (6027, 500 nM) and D (6027, 1 mM).

[0018] [Figure 3] Figure 3A: Immunohistochemical staining of patient-derived basal cell carcinoma (BCC) cultured for 4 days in medium containing 500 nM 6027 or DMSO as a negative control and vismodegib (20 μM) as a positive control.

[0019] Figure 3B: Quantification of cells stained positive for Ki67 and TUNEL as described in Figure 3A for A (DMSO), B (vismodegib, 20 mM) and C (6027, 500 nM).

[0020] [Figure 4] Figure 4A: Histopathological slice of a patient-derived basal cell carcinoma (BCC) cultured for 4 days with 500 nM 6027 and standard of care vismodegib (20 μM) as a positive control.

[0021] FIG. 4B: Quantification of cells stained positively for Ki67 as described in FIG. 4A for A (vismodegib, 20 mM) and B (6027, 500 nM).

[0022] [Figure 5]FIG. 5A: Histopathological slices of patient-derived high-grade cutaneous squamous cell carcinoma (cSCC) treated with 500 nM 6027 and DMSO as a negative control for 4 days.

[0023] FIG. 5B: Quantification of cells stained positively for Ki67 as described in FIG. 5A for A (DMSO) and B (6027, 500 nM). DETAILED DESCRIPTION OF THE INVENTION

[0024] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The embodiments, preferred embodiments, and highly preferred embodiments described and disclosed herein should apply to all aspects and other embodiments, preferred embodiments, and highly preferred embodiments, whether or not specifically mentioned again.

[0025] The terms "a" or "an," as used herein, mean "at least one," unless otherwise specified.

[0026] Each alkyl moiety, alone or as part of a larger group such as alkoxy, aminoalkyl, or haloalkoxy, or alkylene, refers to a monovalent saturated acyclic (i.e., non-cyclic) hydrocarbon group which may be straight-chained or branched. Thus, an "alkyl" group does not contain any carbon-carbon double bonds or any carbon-carbon triple bonds. 1~6"Alkyl" refers to an alkyl group having 1 to 6 carbon atoms. Examples include methyl, ethyl, n-propyl, prop-2-yl, n-butyl, but-2-yl, 2-methyl-prop-1-yl, or 2-methyl-prop-2-yl. Examples of alkoxy include methoxy, ethoxy, propoxy, iso-propoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, neo-pentoxy, and n-hexoxy. Examples of aminoalkyl include aminomethyl, aminoethyl, dimethylaminomethyl, and dimethylaminoethyl. Haloalkoxy refers to an alkoxy with further substitution of a halogen.

[0027] As used herein, the term "alkylene" refers to an alkanediyl group, i.e., a divalent saturated acyclic hydrocarbon group which may be straight-chained or branched. 1~6 "Alkylene" refers to an alkylene group having 1 to 6 carbon atoms. Preferred exemplary alkylene groups are methylene (-CH-), ethylene (e.g., -CH-CH- or -CH(-CH)-), propylene (e.g., -CH-CH-CH-, -CH(-CH-CH)-, -CH-CH(-CH)-, or -CH(-CH)-CH-), or butylene (e.g., -CH-CH-CH-CH-).

[0028] Each haloalkyl moiety, alone or as part of a larger group such as haloalkoxy, is an alkyl group substituted with one or more of the same or different halogen atoms. Haloalkyl contains, for example, 1 to 5 halo substituents, or 1 to 3 halo substituents. Examples include, among others, fluoromethyl, difluoromethyl, trifluoromethyl, chlorodifluoromethyl, and 2,2,2-trifluoroethyl.

[0029] Each alkenyl moiety, either alone or as part of a larger group such as alkenyloxy or alkenylene, is straight or branched chain and preferably C2-C 16 Alkenyl, more preferably C2-C 14The alkenyl moiety is alkenyl. Each moiety can be in either the (E)- or (Z)-configuration. Examples include vinyl and allyl. Thus, compounds of the present invention containing an alkenyl moiety can include, where applicable, any of the compounds having the alkenyl moiety in its (E)-configuration, the compounds having the alkenyl moiety in its (Z)-configuration, and mixtures thereof in any ratio.

[0030] Halogen is fluorine, chlorine, bromine or iodine.

[0031] As used herein, the term "carbocyclyl" refers to a monovalent hydrocarbon ring group, including monocyclic rings as well as bridged rings, spiro rings, and / or fused ring systems (which may be composed of, for example, two or three rings), which may be saturated, partially unsaturated (i.e., unsaturated but not aromatic), or aromatic. Unless otherwise defined, "carbocyclyl" preferably refers to aryl, cycloalkyl, or cycloalkenyl. The number of carbon atoms in a carbocyclyl group is not particularly limited, but is preferably 3 to 14, more preferably 4 to 12 or 5 to 10.

[0032] As used herein, the term "heterocyclyl" refers to a monovalent ring group, including monocyclic rings and bridged, spiro, and / or fused ring systems (which may be composed of, for example, two or three rings), wherein the ring group contains one or more (e.g., one, two, three, or four) ring heteroatoms independently selected from O, S, and N, and the remaining ring atoms are carbon atoms, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) may be optionally oxidized, and one or more carbon ring atoms may be optionally oxidized (i.e., to form an oxo group), and the ring group may be saturated, partially unsaturated (i.e., unsaturated but not aromatic), or aromatic. Unless otherwise defined, "heterocyclyl" preferably refers to heteroaryl, heterocycloalkyl, or heterocycloalkenyl. The number of carbon atoms in a heterocyclyl group is not particularly limited, but is preferably 5 to 14, more preferably 5 to 12 or 5 to 10.

[0033] As used herein, the term "aryl" refers to an alkyl group having 6 to 14 carbon atoms (C6-C 14(C-C alkyl), N-C alkyl, ... Typical aryl groups include, but are not limited to, phenyl, substituted phenyl, naphthyl, 1,2-dihydronapthalenyl, 1,2,3,4-tetrahydronaphthenyl, anthracenyl, phenanthrenyl, biphenyl, indenyl, and indanyl. Unless otherwise defined, "aryl" preferably has 5 to 14 ring atoms, more preferably 5 to 10 ring atoms, and most preferably refers to phenyl or phenyl substituted with one or two substituents independently selected from C1-C4 alkyl, halogen, CF3, OH, OCl-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, CH5, halogen, C1-C3 alkyl, OH, OCl-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, CH5 substituted with N(C1-C3 alkyl)2.

[0034] As used herein, the term "heteroaryl" refers to aromatic ring groups, including monocyclic aromatic rings and bridged rings and / or fused ring systems containing at least one aromatic ring (e.g., a ring system composed of two or three fused rings, where at least one of the fused rings is aromatic; or a bridged ring system composed of two or three rings, where at least one of the bridged rings is aromatic), which contain one or more (e.g., one, two, three, or four) ring heteroatoms independently selected from O, S, and N, the remaining ring atoms are carbon atoms, and one or more S ring atoms (if present) and / or one or more N ring atoms (if present) can be optionally oxidized, and further, one or more carbon ring atoms can be optionally oxidized (i.e., to form an oxo group). Heteroaryl groups are optionally substituted with one or more substituents, typically and preferably independently with one or two substituents selected from C1-C4 alkyl, halogen, oxo, CF3, OH, O-C1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, CH5, and CH5 substituted with halogen, C1-C3 alkyl, OH, O-C1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2. Unless otherwise defined, a "heteroaryl" preferably has 5 to 14 ring atoms, more preferably 5 to 12 or 5 to 10 ring atoms.

[0035] In preferred embodiments, the heteroaryl is a monovalent monocyclic or bicyclic aromatic group containing one or more (e.g., one, two, three, or four) ring heteroatoms independently selected from O, S, and N, with the remaining ring atoms being carbon atoms, which are optionally substituted with one or more substituents, typically and preferably one or two substituents independently selected from C1-C4 alkyl, halogen, oxo, CF3, OH, OCl-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, CH5, and CH5 substituted with halogen, C1-C3 alkyl, OH, OCl-C3 alkyl, NH2, NH(C1-C3 alkyl), or N(C1-C3 alkyl)2. Unless otherwise defined, such monocyclic or bicyclic heteroaryls preferably have from 5 to 12, preferably from 5 to 10, ring atoms.

[0036] Examples of heteroaryl groups include pyridinyl, imidazolyl, imidazopyridinyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, thiadiazolyl, furazanyl, benzofurazanyl, benzopyranyl, benzothio and phenyl, benzothiazolyl, benzoxazolyl, coumarinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl, wherein said heteroaryl is independently optionally substituted with one or more, preferably one or two, substituents independently selected from C1-C4 alkyl, halogen, oxo, CF3, OH, O-C1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C6H5, and C6H5 substituted with halogen, C1-C3 alkyl, OH, O-C1-C3 alkyl, NH2, NH(C1-C3 alkyl), and N(C1-C3 alkyl)2.

[0037] Further examples of such monocyclic heteroaryl groups include 2-pyridyl, 3-pyridyl, 4-pyridyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-imidazolyl, 4-imidazolyl, 3-pyrazolyl, 4-pyrazolyl, 2-pyrrolyl, 3-pyrrolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 3-pyridazinyl, 4-pyridazinyl, 5-pyridazinyl, 2-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, 2-pyrazinyl, 2-oxazolyl, 4-oxazolyl, optionally substituted independently with one or more substituents, typically and preferably with one or two substituents. and 2-tetrazolyl, wherein the substituents are independently selected at each occurrence from C1-C4 alkyl, halogen, oxo, CF3, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl), CH5, CH5 substituted with halogen, C1-C3 alkyl, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl).

[0038] As used herein, the term "cycloalkyl" refers to a monovalent saturated hydrocarbon ring group, including monocyclic rings as well as bridged rings, spiro rings, and / or fused ring systems (which may be composed of, for example, two, three, or four rings; for example, fused ring systems composed of two or three fused rings, etc.). Cycloalkyl groups are optionally substituted with one or more substituents, typically and preferably with one or two substituents independently selected from C1-C4 alkyl, halogen, oxo, CF3, OH, OCl-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, CH5, halogen, C1-C3 alkyl, OH, OCl-C3 alkyl, NH2, NH(C1-C3 alkyl), CH5 substituted with N(C1-C3 alkyl)2. Unless otherwise defined, "cycloalkyl" preferably refers to a group selected from C1-C4 alkyl, halogen, oxo, CF3, OH, OCl-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, CH5 substituted with N(C1-C3 alkyl). 3~14 Examples of cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or adamantyl.

[0039] As used herein, the term "heterocycloalkyl" refers to a monovalent saturated ring group, including monocyclic rings and bridged rings, spiro rings, and / or fused ring systems (which may, for example, be composed of two or three rings; such as, for example, a fused ring system composed of two or three fused rings), which contains one or more (e.g., one, two, three, or four) ring heteroatoms independently selected from O, S, and N, the remaining ring atoms being carbon atoms, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) can be optionally oxidized, and further, one or more carbon ring atoms can be optionally oxidized (i.e., to form an oxo group). Heterocycloalkyl groups are optionally substituted with one or more substituents, typically and preferably with one or two substituents independently selected from C1-C4 alkyl, halogen, oxo, CF3, OH, O-C1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C6H5, halogen, C1-C3 alkyl, OH, O-C1-C3 alkyl, NH2, NH(C1-C3 alkyl), C6H5 substituted with N(C1-C3 alkyl)2. Unless otherwise defined, "heterocycloalkyl" refers to a 3- to 14-membered saturated ring group that is preferably a monocyclic ring or a fused ring system (e.g., a fused ring system composed of two fused rings), which contains one or more (e.g., one, two, three, or four) ring heteroatoms independently selected from O, S, and N, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, and one or more carbon ring atoms are optionally oxidized; more preferably, "heterocycloalkyl" refers to a 5- to 7-membered saturated monocyclic ring group that contains one or more (e.g., one, two, or three) ring heteroatoms independently selected from O, S, and N, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, and one or more carbon ring atoms are optionally oxidized.

[0040] As used herein, the term "cycloalkenyl" refers to unsaturated alicyclic (non-aromatic) hydrocarbon ring groups, including monocyclic rings as well as bridged rings, spiro rings, and / or fused ring systems (which may be composed of, e.g., two or three rings; e.g., fused ring systems composed of two or three fused rings), which contain one or more (e.g., one or two) carbon-carbon double bonds and do not contain any carbon-carbon triple bonds. Cycloalkenyl groups are optionally substituted independently with one or more substituents, typically and preferably one or two substituents, which are typically and preferably independently in each occurrence selected from C1-C4 alkyl, halogen, oxo, CF3, OH, O-C1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C6H5, C6H5 substituted with halogen, C1-C3 alkyl, OH, O-C1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2. "Cycloalkenyl" can refer to, for example, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, or cycloheptadienyl. Unless otherwise defined, "cycloalkenyl" preferably refers to C1-C4 alkyl, OH, O-C1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl). 3~14 It refers to cycloalkenyl, more preferably C 3~7 Refers to cycloalkenyl.

[0041] As used herein, the term "heterocycloalkenyl" refers to unsaturated alicyclic (non-aromatic) ring groups, including monocyclic rings and bridged rings, spiro rings, and / or fused ring systems (which may, for example, be composed of two or three rings; such as, for example, fused ring systems composed of two or three fused rings), which contain one or more (e.g., one, two, three, or four, etc.) ring heteroatoms independently selected from O, S, and N, and wherein the remaining ring atoms and carbon atoms, one or more S ring atoms (if present) and / or one or more N ring atoms (if present) can be optionally oxidized, one or more carbon ring atoms can be optionally oxidized (i.e., to form an oxo group), and further, wherein the ring group contains at least one double bond between adjacent ring atoms and does not contain any triple bonds between adjacent ring atoms. Heterocycloalkenyl groups are optionally substituted independently with one or more substituents, typically and preferably with one or two substituents, which are typically and preferably independently at each occurrence selected from C1-C4 alkyl, halogen, oxo, CF3, OH, OCl-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C6H5, and C6H5 substituted with halogen, C1-C3 alkyl, OH, OCl-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2. Unless otherwise defined, "heterocycloalkenyl" refers to a 3- to 14-membered unsaturated alicyclic ring group, preferably a monocyclic ring or a fused ring system (e.g., a fused ring system composed of two fused rings), which ring group contains one or more (e.g., one, two, three, or four) ring heteroatoms independently selected from O, S, and N, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, one or more carbon ring atoms are optionally oxidized, and wherein the ring group contains at least one double bond between adjacent ring atoms and does not contain any triple bonds between adjacent ring atoms.

[0042] Where a group is said to be optionally substituted, preferably there are optionally 1 to 5 substituents, more preferably optionally 1 to 3 substituents, and even more preferably optionally 1 or 2 substituents.

[0043] As used herein, the term "amino acid" refers to an organic compound containing the functional groups amine (-NH) and carboxylic acid (-COOH) and their zwitterions, typically and preferably with a side chain specific to each amino acid. The term "amino acid" typically and preferably includes naturally occurring amino acids, such as proteinogenic amino acids (produced by RNA translation), non-proteinogenic amino acids (produced by other metabolic mechanisms, e.g., post-translational modification), standard or canonical amino acids (directly encoded by codons in the genetic code), and non-standard or non-canonical amino acids (not directly encoded by the genetic code). Naturally occurring amino acids include non-eukaryotic and eukaryotic amino acids. As used herein, the term "amino acid" also includes chemically synthesized unnatural amino acids. Furthermore, the term encompasses alpha- (α-), beta- (β-), gamma- (γ-) and delta- (δ-) amino acids and mixtures thereof in any ratio, as well as any isomeric forms of amino acids, i.e., D- and L-stereoisomers (alternatively represented by (R) and (S) nomenclature) and mixtures thereof in any ratio, preferably in a 1:1 racemic ratio. Amino acids in the present invention are preferably in the L-configuration. The terms "D-stereoisomer," "L-stereoisomer," "D-amino acid," or "L-amino acid" refer to the chiral alpha carbon of an amino acid.

[0044] Certain compounds of formula (I) of the present invention may contain one, two, or more centers of chirality, and such compounds may be provided as pure enantiomers or pure diastereoisomers, and mixtures thereof in any ratio. The compounds of the present invention also include all tautomeric forms of the compounds of formula (I). The compounds of formula (I) may also be solvated, particularly hydrated, and these are also included in the compounds of formula (I). The term "chiral" refers to a compound that has the property of not being superimposable on its mirror-image partner, and the term "achiral" refers to a compound that is superimposable on its mirror-image partner. The term "stereoisomer" refers to compounds that have identical chemical constitution but differ in terms of the arrangement of atoms or groups in space. "Diastereomer" refers to stereoisomers with two or more centers of chirality, where the compounds are not mirror images of each other. Diastereomers have different physical properties, such as melting points, boiling points, spectral properties, and chemical and biological reactivity. Mixtures of diastereomers can be separated under high-resolution analytical procedures such as electrophoresis and chromatography. "Enantiomers" refer to two stereoisomers of a compound that are non-superimposable mirror images of one another. Stereochemical definitions and conventions used herein generally follow those in the McRaw-Hiff Dictionary of Chemical Terms (1984), edited by S.P. Parker, McGraw-Hill Book Company, New York; and "Stereochemistry of Organic Compounds," by Eliel, E. and Wilen, S., John Wiley & Sons, Inc., New York, 1994. The compounds of the present invention may contain asymmetric or chiral centers and therefore may exist in various stereoisomeric forms. All stereoisomeric forms of the compounds of the present invention, including, but not limited to, diastereomers, enantiomers, and atropisomers, as well as mixtures thereof, e.g., racemic mixtures, are intended to form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light.In describing optically active compounds, the prefixes D and L, or R and S, are used to indicate the absolute configuration of the molecule around its chiral center(s). The prefixes d and l, or (+) and (-), are used to indicate the sign of rotation of plane-polarized light by the compound; (-) or l means the compound is levorotatory. A compound with the prefix (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Specific stereoisomers are sometimes called enantiomers, and mixtures of such isomers are often called enantiomeric or scalemic mixtures. A 50:50 mixture of enantiomers is called a racemic mixture or racemate. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible via a low energy barrier. For example, proton tautomers include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerizations.

[0045] As used herein, the term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable organic or inorganic salt of the compound of the present invention, particularly an acid addition salt.Exemplary salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate (mesylate), ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate.Pharmaceutically acceptable salts may include the inclusion of another molecule, such as acetate ion, succinate ion or other counterion. Counterion can be any organic or inorganic moiety that stabilizes the charge on the parent compound.In addition, pharmaceutically acceptable salts can have more than one charged atom in their structure.If multiple charged atoms are part of a pharmaceutically acceptable salt, it can have multiple counterions.Therefore, a pharmaceutically acceptable salt can have one or more charged atoms and / or one or more counterions. When the compound of the present invention is a base, the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, by treating the free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric acid, or an organic acid such as acetic acid, trifluoroacetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid such as glucuronic acid or galacturonic acid, an alpha hydroxy acid such as citric acid or tartaric acid, an amino acid such as aspartic acid or glutamic acid, an aromatic acid such as benzoic acid or cinnamic acid, or a sulfonic acid such as p-toluenesulfonic acid or ethanesulfonic acid.

[0046] "Solvate" refers to the association or complex of one or more solvent molecules with the compound of the present invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide (DMSO), ethyl acetate, acetic acid and ethanolamine. The term "hydrate" refers to a complex in which the solvent molecule is water.

[0047] The terms "a compound of the invention" and "compounds of the invention" and "a compound of formula (I)" include stereoisomers, geometric isomers, tautomers, solvates, pharmaceutically acceptable salts and solvates of such salts.

[0048] The term "mammal" includes, but is not limited to, humans, mice, rats, guinea pigs, monkeys, dogs, cats, horses, cows, pigs, and sheep. As used herein, the term "mammal" preferably refers to humans.

[0049] As used herein, the term "treatment" of a disorder or disease (e.g., "treatment" of a skin cancer such as melanoma) is well known in the art. "Treatment" of a disorder or disease means that the disorder or disease has been suspected or diagnosed in a patient / subject. A patient / subject suspected of suffering from a disorder or disease typically exhibits certain clinical and / or pathological symptoms that one skilled in the art can readily attribute to a particular pathological condition (i.e., diagnose the disorder or disease).

[0050] "Treatment" of a disorder or disease may, for example, result in the cessation of the progression of the disorder or disease (e.g., no worsening of symptoms) or a delay in the progression of the disorder or disease (if the cessation of progression is only transient). "Treatment" of a disorder or disease may also result in a partial response (e.g., improvement of symptoms) or a complete response (e.g., disappearance of symptoms) in a subject / patient suffering from a disorder or disease. Thus, "treatment" of a disorder or disease may also refer to an improvement in the disorder or disease, which may result in, for example, the cessation of the progression of the disorder or disease or a delay in the progression of the disorder or disease. Relapse may occur after such a partial or complete response. It should be understood that a subject / patient may experience a wide range of responses to treatment. Treatment of a disorder or disease may include, inter alia, curative treatment (preferably resulting in a complete response, ultimately resulting in a cure of the disorder or disease) and symptomatic treatment (including symptomatic relief). "Amelioration" of a disorder or disease may, for example, result in the cessation of the progression of the disorder or disease or a delay in the progression of the disorder or disease. "Treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment.

[0051] The term "effective amount" refers to the amount of a compound of the present invention or a pharmaceutical composition of the present invention that (i) treats a particular disease or disorder, or (ii) reduces, ameliorates, or eliminates one or more symptoms of a particular disease or disorder. In the case of skin cancer, particularly when the cancer is melanoma, basal cell carcinoma, or cutaneous squamous cell carcinoma, an effective amount of the drug may reduce tumor volume and / or reduce clinical symptoms.

[0052] As used herein, the term "skin cancer" refers to skin cancers such as melanoma skin cancer and nonmelanoma skin cancer (NMSC), but also refers to and includes precancerous and preinvasive neoplastic skin growths such as cutaneous squamous cell carcinoma in situ (cSCCis or Bowen's disease) or actinic keratosis (AK, also known as "actinic keratosis" and "senile keratosis"). Furthermore, as used herein, the term "skin cancer" also includes cutaneous lymphomas, such as cutaneous T-cell lymphoma (CTCL) or cutaneous B-cell lymphoma (CBCL), or preinvasive forms thereof. Bowen's disease is a neoplastic skin disorder that can be considered an early stage or intraepidermal form of squamous cell carcinoma. Actinic keratosis is characterized by thick, scaly, or crusty precancerous patches of skin that typically form when the skin is damaged by ultraviolet (UV) radiation from sunlight or indoor tanning beds.

[0053] The term "non-melanoma skin cancer" (abbreviated as "NMSC") is used herein to refer to a group of diseases that includes, among others, cutaneous squamous cell carcinoma (cSCC), basal cell carcinoma (BCC), Merkel cell carcinoma (MCC), Bowen's disease (BD) and actinic keratosis (AK), as well as any precancerous and preinvasive forms thereof.

[0054] The term "field cancerization" refers to premalignant field defects, a biological process in which large areas of cells on tissue surfaces or within organs are affected by carcinogenic changes. This process often results from prolonged exposure to harmful environmental factors, such as UV radiation. The initial stage of field cancerization involves various molecular lesions, such as acquired genetic mutations and epigenetic changes, that occur over a widespread, multifocal "field." The field is affected by subclinical (non-visible, non-palpable) AK lesions, early AK lesions, late AK lesions, and occasionally invasive cSCC. The concept of field carcinogenesis provides the rationale for field therapy, in which the entire field is treated rather than individual lesions. The goal of field therapy is to eliminate not only clinically visible lesions but also subclinical lesions and to prevent the development of invasive SCC.

[0055] The term "topical administration" as used herein refers to application to a body surface such as the skin. Typically and preferably, the term "topical administration" as used herein refers to epicutaneous application, meaning direct application to the skin. Thus, in a highly preferred embodiment of the present invention, the topical administration is epicutaneous application, and preferably, the topical administration is direct application to the skin. Typically and preferably, the present invention relates to topically applied, locally acting compounds for use on the skin to treat skin infections, as referred to in the EMA's quality equivalence guidelines for topical products (https: / / www.ema.europa.eu / en / documents / scientific-guideline / draft-guideline-quality-equivalence-topical-products_en.pdf).

[0056] In a first aspect, the present invention provides a compound of formula (I) for use in a method of topical treatment of skin cancer in a mammal, said method comprising topically administering said compound to said mammal, [ka] During the ceremony, A1 is [ka] A2 is [ka] A3 is [ka] B1 is [ka] B2 is [ka] B3 is [ka] During the ceremony, a1, a 12 , a2, a 22 , a3, a 32 , b1, b 12 , b2, b 22 , b3 and b 32 are independently selected from 0 and 1, and a1+a 12 , a2+a 22 , a3+a 32 , b1+b 12 , b2+b 22 and b3+b 32 are independently selected from 0 and 1, and a1+a 12 +a2+a 22 +a3+a 32 is 0, 1, 2 or 3, and b1+b 12 +b2+b 22 +b3+b 32is 0, 1 or 2; During the ceremony, R1 is independently selected from C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, carbocyclyl or heterocyclyl optionally and preferably substituted with halogen, C1-C3 alkyl, OR4, C6H5 substituted with NR5R6, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl; R2 is C4~C 12 Alkyl, C4-C 10 alkoxy, C1-C3 alkylene-cycloalkyl, C1-C3 alkylene-aryl, C1-C3 alkylene-heteroaryl, wherein independently, in said C1-C3 alkylene, one -CH2- moiety is optionally replaced by -CH(NH)- or -O-; said alkyl, cycloalkyl, aryl and heteroaryl are each independently optionally and preferably substituted with one or more, preferably one or two, substituents selected from C1-C2 alkyl, C1-C2 haloalkyl, oxo, OH, halogen, C1-C2 alkoxy, C6H5 or C6H5 substituted with C1-C3 alkyl or OC1-C3 alkyl; R3 is [ka] where: R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10 two of which, together with the carbon atoms to which they are attached, form a carbocyclic or heterocyclic ring, preferably a carbocyclic ring, wherein R 11 and R 12are, independently of each other, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl, or C3-C6 cycloalkyl; or, together with the nitrogen atom to which they are attached, independently in each occurrence, are each independently halogen, C1-C4 alkyl, OR 13 , N.R. 14 R 15 forming a heteroaryl or heterocyclyl optionally and preferably substituted with 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl, and the arrow indicates the bond to the C(O)-moiety depicted in formula (I); as well as pharmaceutically acceptable salts of the compounds of formula (I) as described above, preferably the method comprises topical administration of an effective amount of the compound to the mammal, preferably a human.

[0057] In a further aspect, the present invention provides a compound of formula (I*) for use in a method for topically treating skin cancer in a mammal, said method comprising topically administering said compound to said mammal, preferably a human, [ka] During the ceremony, R1 is independently selected from C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, carbocyclyl or heterocyclyl optionally and preferably substituted with halogen, C1-C3 alkyl, OR4, C6H5 substituted with NR5R6, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl; R2 is C5~C 12 Alkyl, C4-C 10alkoxy, C1-C3 alkylene-cycloalkyl, C1-C3 alkylene-aryl, C1-C3 alkylene-heteroaryl, wherein independently, in said C1-C3 alkylene, one -CH2- moiety is optionally replaced by -CH(NH)- or -O-; said alkyl, cycloalkyl, aryl and heteroaryl are each independently optionally and preferably substituted with one or more, typically and preferably one or two, substituents selected from C1-C2 alkyl, C1-C2 haloalkyl, oxo, OH, halogen, C1-C2 alkoxy, C6H5 or C6H5 substituted with C1-C3 alkyl or OC1-C3 alkyl; R3 is [ka] where: R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10 two of which, together with the carbon atoms to which they are attached, form a carbocyclic or heterocyclic ring, preferably a carbocyclic ring, wherein R 11 and R 12 are, independently of each other, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl, or C3-C6 cycloalkyl; or, together with the nitrogen atom to which they are attached, independently in each occurrence, are each independently halogen, C1-C4 alkyl, OR 13 , N.R. 14 R 15 forming a heteroaryl or heterocyclyl optionally and preferably substituted with 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl, and the arrow indicates the bond to the C(O)-moiety depicted in formula (I); as well as pharmaceutically acceptable salts of said compound of formula (I*), preferably said method comprising topical administration to said mammal, preferably a human, of an effective amount of said compound.

[0058] In another aspect, the present invention provides a method for topically treating skin cancer in a mammal, preferably a human, said method comprising topical administration to said mammal, preferably said human, of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising said compound of formula (I) or a pharmaceutically acceptable salt thereof, preferably said method comprising topical administration to said mammal, preferably said human, of an effective amount of said compound.

[0059] In a further aspect, the present invention provides use of a compound of formula (I) of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising said compound of formula (I) or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for topically treating skin cancer in a mammal, preferably a human, said method comprising topical administration of said compound of formula (I) or said pharmaceutically acceptable salt thereof, or said pharmaceutical composition of the present invention to said mammal, preferably said human, preferably comprising topical administration of an effective amount of said compound or said pharmaceutically acceptable salt thereof, or said pharmaceutical composition to said mammal, preferably said human. In a preferred embodiment, the compound of formula (I) is any one of the compounds of formulas (II) to (IV): [ka] Preferably, the compound of formula (I) is a compound of formula (II).

[0060] In another preferred embodiment, the compound of formula (I) is a compound of formula (II), formula (III), or formula (IV), preferably, the compound of formula (I) is a compound of formula (II). In another preferred embodiment, the compound of formula (I) is a compound of formula (II). In another preferred embodiment, the compound of formula (I) is a compound of formula (III). In another preferred embodiment, the compound of formula (I) is a compound of formula (IV).

[0061] In a further preferred embodiment, each R1 is independently phenyl, naphthyl, 1,2-dihydronapthalenyl, 1,2,3,4-tetrahydronaphthenyl, anthracenyl, phenanthrenyl, biphenyl, indenyl, indanyl, pyridinyl, imidazolyl, imidazopyridinyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl, or oxazolyl, optionally substituted with C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, or C6H5 substituted with halogen, C1-C3 alkyl, OR4, or NR5R6. , isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, thiadiazolyl, furazanyl, benzofurazanyl, benzopyranyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, coumarinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, furopyridinyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, adamantanyl, wherein R4, R5, and R6, in each occurrence, are independently H or C1-C3 alkyl.

[0062] In another preferred embodiment, each R1 is independently selected from C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, cycloalkyl, aryl, or heteroaryl optionally substituted with halogen, C1-C3 alkyl, OR4, C6H5 substituted with NR5R6, wherein R4, R5, and R6 in each occurrence are independently H or C1-C3 alkyl.

[0063] In a further preferred embodiment, each R1 is independently selected from C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, cycloalkyl, monocyclic or bicyclic aryl, or monocyclic or bicyclic heteroaryl optionally substituted with halogen, C1-C3 alkyl, OR4, C6H5 substituted with NR5R6, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl.

[0064] In a further preferred embodiment, each R1 is independently selected from C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, cycloalkyl, monocyclic or bicyclic aromatic aryl, or monocyclic or bicyclic heteroaryl optionally substituted with halogen, C1-C3 alkyl, OR4, C6H5 substituted with NR5R6, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl.

[0065] In a further preferred embodiment, each R1 is independently selected from phenyl or monocyclic or bicyclic heteroaryl containing one or two heteroatoms selected from N, O, and S, optionally substituted with C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5, halogen, C1-C3 alkyl, OR4, C6H5 substituted with NR5R6, preferably phenyl, coumarinyl, or oxazolyl, more preferably phenyl or oxazolyl, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl.

[0066] In a further preferred embodiment, each R1 is independently phenyl, naphthyl, 1,2-dihydronapthalenyl, 1,2,3,4-tetrahydronaphthenyl, biphenyl, indenyl, indanyl, pyridinyl, imidazolyl, imidazopyridinyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl, or oxazolyl, optionally substituted with C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, or C6H5 substituted with halogen, C1-C3 alkyl, OR4, or NR5R6. , isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, thiadiazolyl, furazanyl, benzofurazanyl, benzopyranyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, coumarinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, furopyridinyl, cyclopentyl, cyclohexyl, adamantanyl, wherein R4, R5, and R6 in each occurrence are independently H or C1-C3 alkyl.

[0067] In a further preferred embodiment, R1 is of the formula [ka] wherein R represents a bond to the C(O)-moiety depicted in formula (I).

[0068] In a further preferred embodiment, each R1 is independently selected from phenyl or monocyclic or bicyclic heteroaryl optionally substituted with C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, CH5, halogen, C1-C3 alkyl, CH5 substituted with OR4, NR5R6, where R4, R5, R6 are independently in each occurrence H, C1-C3 alkyl. In a further preferred embodiment, each R1 is independently selected from phenyl or monocyclic or bicyclic aromatic heteroaryl optionally substituted with C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, CH5, halogen, C1-C3 alkyl, OR4, CH5 substituted with NR5R6, where R4, R5, R6 are independently in each occurrence H, C1-C3 alkyl.

[0069] In a further preferred embodiment, each R1 is independently selected from the group consisting of phenyl, pyridinyl, imidazolyl, imidazopyridinyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl, and oxazolyl, optionally substituted with C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted with halogen, C1-C3 alkyl, OR4, and NR5R6. , isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, thiadiazolyl, benzofurazanyl, benzopyranyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, wherein R4, R5, and R6, in each occurrence, are independently H or C1-C3 alkyl.

[0070] In a further preferred embodiment, each R1 is independently selected from phenyl, imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzopyranyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, optionally substituted with C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, CH5, halogen, C1-C3 alkyl, OR4, CH5 substituted with NR5R6, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl.

[0071] In a further preferred embodiment, each R1 is independently methyl, ethyl, chlorine, fluorine, oxo, CF3, OC1-C2 alkyl, NR 11 R 12 , C6H5, methyl, ethyl, chlorine, fluorine, OC1~C2 alkyl, NR 11 R 12 and R is selected from phenyl, imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzopyranyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, coumarinyl, optionally substituted with CH, substituted with 11 , R 12 is independently in each occurrence H, methyl, or ethyl.

[0072] In a further preferred embodiment, R1 is of the formula [ka] wherein R represents a bond to the C(O)-moiety depicted in formula (I).

[0073] In a further preferred embodiment, each R1 is independently selected from phenyl or monocyclic or bicyclic heteroaryl optionally substituted with C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5, halogen, C1-C3 alkyl, OR4, C6H5 substituted with NR5R6, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl.

[0074] In a further preferred embodiment, each R1 is independently selected from phenyl or monocyclic or bicyclic heteroaryl optionally substituted, preferably monosubstituted, with methyl, ethyl, chlorine, fluorine, CF3, O-C-C alkyl, NR5R6, C6H5, methyl, ethyl, chlorine, fluorine, O-C-C alkyl, C6H5 substituted with NR5R6, wherein R5, R6 in each occurrence are independently H, methyl, or ethyl.

[0075] In a further preferred embodiment, each R1 is independently selected from phenyl or monocyclic or bicyclic heteroaryl containing one or two heteroatoms selected from N, O, and S, optionally substituted with C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5, halogen, C1-C3 alkyl, OR4, C6H5 substituted with NR5R6, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl.

[0076] In a further preferred embodiment, each R1 is independently selected from methyl, ethyl, chlorine, fluorine, CF3, O-C-C alkyl, NR5R6, C6H5, phenyl or monocyclic or bicyclic heteroaryl containing one or two heteroatoms selected from N, O and S, optionally substituted, preferably monosubstituted, with methyl, ethyl, chlorine, fluorine, O-C-C alkyl, C6H5 substituted with NR5R6, wherein R5, R6 in each occurrence are independently H, methyl or ethyl.

[0077] In a further preferred embodiment, each R1 is independently selected from phenyl, imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, coumarinyl, optionally substituted with C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5, halogen, C1-C3 alkyl, OR4, C6H5 substituted with NR5R6, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl.

[0078] In a further preferred embodiment, each R1 is independently selected from methyl, ethyl, chlorine, fluorine, CF3, O-C-C alkyl, NR5R6, C6H5, phenyl, imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, coumarinyl, optionally substituted, preferably monosubstituted, with methyl, ethyl, chlorine, fluorine, O-C-C alkyl, C6H5 substituted with NR5R6, wherein R5 and R6 in each occurrence are independently H, methyl, or ethyl.

[0079] In a further preferred embodiment, each R1 is independently selected from C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5, phenyl, thienyl, oxazolyl, pyrrolyl, coumarinyl optionally substituted with halogen, C1-C3 alkyl, OR4, C6H5 substituted with NR5R6, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl.

[0080] In a further preferred embodiment, each R1 is independently selected from methyl, ethyl, chlorine, fluorine, CF3, O-C-C alkyl, NR5R6, C6H5, phenyl, thienyl, oxazolyl, pyrrolyl, coumarinyl, optionally substituted, preferably monosubstituted, with methyl, ethyl, chlorine, fluorine, O-C-C alkyl, C6H5 substituted with NR5R6, wherein R5 and R6 in each occurrence are independently H, methyl, or ethyl.

[0081] In a further preferred embodiment, R1 is of the formula [ka] wherein R represents a bond to the C(O)-moiety depicted in formula (I).

[0082] In a further preferred embodiment, each R1 is independently phenyl or oxazolyl optionally substituted with C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5, C6H5 substituted with halogen, C1-C3 alkyl, OR4, NR5R6, wherein R4, R5, R6 in each occurrence are independently H, C1-C3 alkyl.

[0083] In a further preferred embodiment, each R1 is independently phenyl, coumarinyl, or oxazolyl optionally substituted, preferably monosubstituted, with methyl, ethyl, chlorine, fluorine, CF3, O—C1-C2 alkyl, NR5R6, C6H5, or C6H5 substituted with methyl, ethyl, chlorine, fluorine, O—C1-C2 alkyl, NR5R6, wherein R 11 , R 12 is independently in each occurrence H, methyl, or ethyl.

[0084] In a further preferred embodiment, each R1 is independently methyl, chlorine, fluorine, CF3, OCH3C6H5, phenyl, coumarinyl or oxazolyl optionally substituted, preferably monosubstituted, with methyl or fluorine-substituted, preferably monosubstituted, C6H5.

[0085] In a further highly preferred embodiment, said R1 is of the formula [ka] wherein R represents a bond to the C(O)-moiety depicted in formula (I).

[0086] In a further highly preferred embodiment, R1 is [ka] where R represents the bond to the C(O)-moiety depicted in formula (I).

[0087] In another highly preferred embodiment, R1 is [ka] where R represents the bond to the C(O)-moiety depicted in formula (I).

[0088] In a further highly preferred embodiment, R1 is [ka] where R represents the bond to the C(O)-moiety depicted in formula (I).

[0089] In another highly preferred embodiment, R2 is C5-C 12 Alkyl, C4-C 10 and selected from alkoxy, C1-C3 alkylene-cycloalkyl, C1-C3 alkylene-aryl, C1-C3 alkylene-heteroaryl, wherein independently, in said C1-C3 alkylene, one -CH2- moiety is optionally replaced by -CH(NH)- or -O-, and said alkyl, cycloalkyl, aryl and heteroaryl are each independently optionally and preferably substituted with one or more, typically and preferably one or two, substituents selected from C1-C2 alkyl, C1-C2 haloalkyl, oxo, OH, halogen, C1-C2 alkoxy, CH5 or CH5 substituted with C1-C3 alkyl or OC1-C3 alkyl.

[0090] In another highly preferred embodiment, R2 is C5-C12 Alkyl, C4-C 10 alkoxy, C1-C3 alkylene-C5-C6 cycloalkyl, C1-C3 alkylene-phenyl, C1-C3 alkylene-biphenyl, C1-C3 alkylene-(monocyclic or bicyclic heteroaryl), wherein independently, in said C1-C3 alkylene, one -CH2- moiety is optionally replaced by -CH(NH)- or -O-, and said phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently selected from C1-C2 alkyl, Optionally substituted with one or more, typically and preferably one or two, substituents selected from C1-C2 haloalkyl, halogen, C1-C2 alkoxy, preferably said monocyclic or bicyclic heteroaryl is selected from imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzopyranyl, benzothiophenyl, benzothiazolyl, benzoxazolyl.

[0091] In a further preferred embodiment, R2 is a C5-C 12 Alkyl, C4-C 10alkoxy, C1-C3 alkylene-C5-C6 cycloalkyl, C1-C3 alkylene-phenyl, C1-C3 alkylene-biphenyl, C1-C3 alkylene-(monocyclic or bicyclic heteroaryl), wherein independently, in said C1-C3 alkylene, one -CH2- moiety is optionally replaced by -CH(NH)- or -O-, and said phenyl, biphenyl, C5-C6-cycloalkyl and monocyclic or bicyclic heteroaryl are each independently selected from methyl, methyl, methyl- ... Optionally substituted by one or more, typically and preferably one or two, substituents selected from methyl, ethyl, fluorine, chlorine, methoxy, preferably said monocyclic or bicyclic heteroaryl is selected from imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzopyranyl, benzothiophenyl, benzothiazolyl, benzoxazolyl.

[0092] In a further preferred embodiment, R2 is a C5-C 12 Alkyl, C4-C 10alkoxy, C1-C2 alkylene-C5-C6 cycloalkyl, C1-C2 alkylene-phenyl, C1-C2 alkylene-biphenyl, C1-C2 alkylene-(monocyclic or bicyclic heteroaryl), wherein independently, in said C1-C2 alkylene, one -CH2- moiety is optionally replaced by -CH(NH)- or -O-, and said phenyl, biphenyl, C5-C6-cycloalkyl and monocyclic or bicyclic heteroaryl are each independently one or more, typically and independently, selected from C1-C2 alkyl, C1-C2 haloalkyl, halogen, C1-C2 alkoxy. and preferably optionally substituted by one or two substituents, preferably said monocyclic or bicyclic heteroaryl is selected from imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzopyranyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, more preferably said monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl.

[0093] In a further preferred embodiment, R2 is a C5-C 12 Alkyl, C4-C 10alkoxy, C1-C2 alkylene-C5-C6 cycloalkyl, C1-C2 alkylene-phenyl, C1-C2 alkylene-biphenyl, C1-C2 alkylene-(monocyclic or bicyclic heteroaryl), independently, in said C1-C2 alkylene, one -CH2- moiety is optionally replaced by -CH(NH)- or -O-, and said phenyl, biphenyl, C5-C6-cycloalkyl and monocyclic or bicyclic heteroaryl are each independently one or more, typically and preferably one or more, selected from methyl, ethyl, fluorine, chlorine, methoxy. and optionally substituted with one or two substituents, preferably said monocyclic or bicyclic heteroaryl is selected from imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzopyranyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, more preferably said monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl.

[0094] In a further preferred embodiment, R2 is a C5-C 12 Alkyl, C4-C 10alkoxy, C1-C2 alkylene-C5-C6 cycloalkyl, C1-C2 alkylene-phenyl, C1-C2 alkylene-biphenyl, C1-C2 alkylene-(monocyclic or bicyclic heteroaryl), wherein independently, in said C1-C2 alkylene, one -CH2- moiety is optionally replaced by -CH(NH)- or -O-, and said phenyl, biphenyl, C5-C6-cycloalkyl and monocyclic or bicyclic heteroaryl are each independently and optionally substituted with one or more, typically and preferably one or two, substituents selected from methyl, ethyl, fluorine, chlorine, methoxy, and said monocyclic or bicyclic heteroaryl is selected from imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzopyranyl, benzothiophenyl, benzothiazolyl, benzoxazolyl.

[0095] In a further preferred embodiment, R2 is a C5-C 12 Alkyl, C4-C 10 and C-C alkylene-C-C cycloalkyl, C-C alkylene-phenyl, C-C alkylene-biphenyl, C-C alkylene-(monocyclic or bicyclic heteroaryl), wherein independently, in said C-C alkylene, one -CH- moiety is optionally replaced by -CH(NH)- or -O-, said phenyl, biphenyl, C-C cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted with one or more, typically and preferably one or two, substituents selected from methyl, ethyl, fluorine, chlorine, methoxy, and said monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl.

[0096] In a further preferred embodiment, R2 is a C5-C 12Alkyl, C4-C 10 alkoxy, C1-C2 alkylene-C5-C6 cycloalkyl, CH2-phenyl, CH2-biphenyl, CH2-O-biphenyl, CH2-(monocyclic or bicyclic heteroaryl), wherein said phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted with one or more, typically and preferably one or two, substituents selected from C1-C2 alkyl, C1-C2 haloalkyl, halogen, C1-C2 alkoxy; preferably, said monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl; more preferably, said monocyclic or bicyclic heteroaryl is selected from thiazolyl, indolyl, benzothiazolyl and benzoxazolyl.

[0097] In a further preferred embodiment, R2 is a C5-C 12 Alkyl, C4-C 10 and selected from alkoxy, C1-C2 alkylene-C5-C6 cycloalkyl, CH2-phenyl, CH2-biphenyl, CH2-O-biphenyl, CH2-(monocyclic or bicyclic heteroaryl), wherein said phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted with one or more, typically and preferably one or two, substituents selected from methyl, ethyl, fluorine, chlorine, methoxy; preferably, said monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl; more preferably, said monocyclic or bicyclic heteroaryl is selected from thiazolyl, indolyl, benzothiazolyl and benzoxazolyl.

[0098] In a further preferred embodiment, R2 is a C5-C 12 Alkyl, C4-C10 alkoxy, C1-C2 alkylene-C5-C6 cycloalkyl, CH2-phenyl, CH2-biphenyl, CH2-O-biphenyl, CH2-(monocyclic or bicyclic heteroaryl), wherein said phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted with one or more, typically and preferably one or two, substituents selected from methyl, ethyl, fluorine, chlorine, methoxy, and said monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl.

[0099] In a further preferred embodiment, R2 is a C5-C 12 Alkyl, C4-C 10 alkoxy, C1-C2 alkylene-C5-C6 cycloalkyl, CH2-phenyl, CH2-biphenyl, CH2-O-biphenyl, CH2-(monocyclic or bicyclic heteroaryl), wherein said phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted with one or more, typically and preferably one or two, substituents selected from methyl, ethyl, fluorine, chlorine, methoxy, and said monocyclic or bicyclic heteroaryl is selected from thiazolyl, indolyl, benzothiazolyl and benzoxazolyl.

[0100] In a further highly preferred embodiment, R2 is [ka] wherein R represents a bond to the CH-moiety depicted in formula (I).

[0101] In a further highly preferred embodiment, R2 is a C5-C 12and selected from alkyl, C1-C2 alkylene-C5-C6 cycloalkyl, CH2-phenyl, CH2-biphenyl and CH2-O-biphenyl, and monocyclic or bicyclic heteroaryl selected from thiazolyl, indolyl, benzothiazolyl and benzoxazolyl, wherein said phenyl, biphenyl and monocyclic or bicyclic heteroaryl are optionally substituted with one or two substituents selected from methyl, ethyl, fluorine, chlorine and methoxy.

[0102] In a further highly preferred embodiment, R2 is [ka] wherein R represents a bond to the CH-moiety depicted in formula (I).

[0103] In a further highly preferred embodiment, R2 is a C5-C 12 alkyl, C1-C2 alkylene-C5-C6 cycloalkyl, CH2-biphenyl and CH2-O-biphenyl, and bicyclic heteroaryl selected from benzothiazolyl and benzoxazolyl.

[0104] In a further highly preferred embodiment, R2 is [ka] wherein R represents a bond to the CH-moiety depicted in formula (I).

[0105] In a further highly preferred embodiment, R2 is [ka] where R represents the bond to the CH-moiety depicted in formula (I).

[0106] In a further highly preferred embodiment, R2 is [ka] where R represents the bond to the CH-moiety depicted in formula (I).

[0107] In a further preferred embodiment, R3 is [ka] and in the formula R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10 two of them, together with the carbon atoms to which they are attached, form a carbocyclic ring, R 11 and R 12 are, independently of each other, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl, or C3-C6 cycloalkyl; or, together with the nitrogen atom to which they are attached, independently in each occurrence, are each independently halogen, C1-C4 alkyl, OR 13 , N.R. 14 R 15 forming heteroaryl or heterocyclyl, optionally and preferably substituted with; R 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl, where the arrow indicates the bond to the C(O)-moiety depicted in formula (I).

[0108] In a further preferred embodiment, R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10two of these together with the carbon atoms to which they are attached form a carbocyclic or heterocyclic ring, preferably a carbocyclic ring, and R 11 and R 12 are, independently of one another, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl or C3-C6 cycloalkyl, or, together with the nitrogen atom to which they are attached, independently in each occurrence, preferably pyridinyl, imidazolyl, imidazopyridinyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoxyl each independently selected from halogen, C1-C4 alkyl, OR, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, indazolyl, indolizinyl, pyridazinyl, triazinyl, isoindolyl, purinyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, homopiperazinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, dihydrooxazolyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl; 13 , N.R. 14 R 15 forming a monocyclic or bicyclic heteroaryl or monocyclic or bicyclic heterocyclyl optionally substituted by R 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl.

[0109] In a further preferred embodiment, R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10two of which together with the carbon atoms to which they are attached form a monocyclic carbocyclic or heterocyclic ring, preferably a monocyclic carbocyclic ring, more preferably said monocyclic carbocyclic ring is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, again more preferably said monocyclic carbocyclic ring is cyclobutyl or cyclopentyl, and R 11 and R 12 are, independently of one another, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl or C3-C6 cycloalkyl, or, together with the nitrogen atom to which they are attached, independently in each occurrence, preferably pyridinyl, imidazolyl, imidazopyridinyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoxyl each independently selected from halogen, C1-C4 alkyl, OR, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, indazolyl, indolizinyl, pyridazinyl, triazinyl, isoindolyl, purinyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, homopiperazinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, dihydrooxazolyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl; 13 , N.R. 14 R 15 forming a monocyclic or bicyclic heteroaryl or monocyclic or bicyclic heterocyclyl optionally substituted by R 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl.

[0110] In a further preferred embodiment, R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10two of which together with the carbon atoms to which they are attached form a monocyclic carbocyclic or heterocyclic ring, preferably a monocyclic carbocyclic ring, more preferably said monocyclic carbocyclic ring is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, again more preferably said monocyclic carbocyclic ring is cyclobutyl or cyclopentyl, and R 11 and R 12 are, independently of each other, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl, or C3-C6 cycloalkyl; or, together with the nitrogen atom to which they are attached, independently in each occurrence, are each independently halogen, C1-C4 alkyl, OR 13 , N.R. 14 R 15 forming a monocyclic heteroaryl or monocyclic heterocyclyl optionally substituted with R 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl.

[0111] In a further preferred embodiment, R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10 two of which together with the carbon atoms to which they are attached form a monocyclic carbocyclic or heterocyclic ring, preferably a monocyclic carbocyclic ring, more preferably said monocyclic carbocyclic ring is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, again more preferably said monocyclic carbocyclic ring is cyclobutyl or cyclopentyl, and R 11 and R 12 are, independently of each other, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl, or C3-C6 cycloalkyl; or, together with the nitrogen atom to which they are attached, independently in each occurrence, are each independently halogen, C1-C4 alkyl, OR 13 , N.R.14 R 15 and forming a monocyclic heteroaryl or a monocyclic heterocyclyl optionally substituted with one or two heteroatoms (R 11 and R 12 and R 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl.

[0112] In a further preferred embodiment, R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10 two of which together with the carbon atoms to which they are attached form a monocyclic carbocyclic or heterocyclic ring, preferably a monocyclic carbocyclic ring, more preferably said monocyclic carbocyclic ring is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, again more preferably said monocyclic carbocyclic ring is cyclobutyl or cyclopentyl, and R 11 and R 12 are, independently of one another, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl, or C3-C6 cycloalkyl, or, together with the nitrogen atom to which they are attached, are independently in each occurrence selected from pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, pyridazinyl, triazinyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, 2-pyrrolinyl, 3-pyrrolinyl, dihydrooxazolyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, each independently halogen, C1-C4 alkyl, OR 13 , N.R. 14 R 15forming a monocyclic heteroaryl or monocyclic heterocyclyl optionally substituted with R 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl.

[0113] In a further preferred embodiment, R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10 two of which together with the carbon atoms to which they are attached form a monocyclic carbocycle, more preferably said monocyclic carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, again more preferably said monocyclic carbocycle is cyclobutyl or cyclopentyl, and R 11 and R 12 are, independently of each other, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl, or C3-C6 cycloalkyl; or, together with the nitrogen atom to which they are attached, independently in each occurrence, are each independently halogen, C1-C4 alkyl, OR 13 , N.R. 14 R 15 forming a monocyclic heterocycle optionally substituted with R 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl.

[0114] In a further preferred embodiment, R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10two of which together with the carbon atoms to which they are attached form a monocyclic carbocyclic or heterocyclic ring, preferably a monocyclic carbocyclic ring, more preferably said monocyclic carbocyclic ring is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, again more preferably said monocyclic carbocyclic ring is cyclobutyl or cyclopentyl, and R 11 and R 12 are, independently of each other, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl, or C3-C6 cycloalkyl; or, together with the nitrogen atom to which they are attached, independently in each occurrence, are each independently halogen, C1-C4 alkyl, OR 13 , N.R. 14 R 15 and forming a monocyclic heterocyclyl optionally substituted by one or two heteroatoms (R 11 and R 12 and R 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl.

[0115] In a further preferred embodiment, R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10 two of which together with the carbon atoms to which they are attached form a monocyclic carbocycle, more preferably said monocyclic carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, again more preferably said monocyclic carbocycle is cyclobutyl or cyclopentyl, and R 11 and R 12are, independently of one another, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl, or C3-C6 cycloalkyl, or, together with the nitrogen atom to which they are attached, are independently in each occurrence selected from imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, pyridazinyl, triazinyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, 2-pyrrolinyl, 3-pyrrolinyl, dihydrooxazolyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, each independently halogen, C1-C4 alkyl, OR 13 , N.R. 14 R 15 forming a monocyclic heterocycle optionally substituted with R 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl.

[0116] In a further preferred embodiment, R7, R8, R9 and R 10 is independently at each occurrence H or C1-C3 alkyl, preferably H or methyl, or is independently at each occurrence any of R7, R8, R9 and R 10 two of which, together with the carbon atoms to which they are attached, form a monocyclic carbocyclic ring selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, preferably said monocyclic carbocyclic ring is cyclobutyl or cyclopentyl, and R 11 and R 12 are, independently of each other, H or C1-C4 alkyl optionally substituted with halogen, hydroxyl, or C3-C6 cycloalkyl; or, together with the nitrogen atom to which they are attached, independently in each occurrence, are each independently halogen, C1-C4 alkyl, OR 13 , N.R. 14 R 15forming a monocyclic heterocycle selected from piperidinyl, morpholinyl, thiomorpholinyl, preferably from piperidinyl or morpholinyl, more preferably from morpholinyl, optionally substituted by R 13 , R 14 , R 15 is independently in each occurrence H, C1-C4 alkyl.

[0117] In a further highly preferred embodiment, R3 is [ka] wherein R represents a bond to the C(O)-moiety depicted in formula (I).

[0118] In a further highly preferred embodiment, R3 is [ka] wherein R represents a bond to the C(O)-moiety depicted in formula (I).

[0119] In a further highly preferred embodiment, R3 is [ka] where R represents the bond to the C(O)-moiety depicted in formula (I).

[0120] In a further highly preferred embodiment, the compound of formula (I) is 6025: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 6027: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 6253: (2S)-N-[(1S)-1-[(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamido}decanamide; 6328: (2S)-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methyl-2-[(2S)-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamido}-3-phenylpropanamido]pentanamide; 6781: (2S)-N-[(1S)-1-[(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamido}hexanamide; 7058: (2S)-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamido}-N-[(1S)-3-methyl-1-[(1-methyl-1-{[(1S)-3-methyl-1-{[1-methyl-1-({1-methyl-1-[(2-{[2-(methylamino)ethyl]carbamoyl}ethyl)carbamoyl]ethyl}carbamoyl)ethyl]carbamoyl}butyl]carbamoyl}ethyl)carbamoyl]butyl]decanamide; 7059: (2S)-2-[(2S)-3-cyclohexyl-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamido}propanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 7824: (2S)-N-{1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}-2-[(2S)-2-{2-[(2S)-2-[(2S)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}-3-({4'-methoxy-[1,1'-biphenyl]-4-yl}oxy)propanamido]-4-methylpentanamido]-2-methylpropanamido}-4-methylpentanamido]-4-methylpentanamide; 7844: (2S)-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-2-[(2S)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}-3-(6-methoxy-1,3-benzothiazol-2-yl)propanamido]-4-methylpentanamide; 7848: (2S)-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-2-[(2S)-3-(6-fluoro-1,3-benzothiazol-2-yl)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}propanamido]-4-methylpentanamide; 7850: (2S)-2-[(2S)-3-(1,3-benzoxazol-2-yl)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}propanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 8341: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-4-methyl-N-(1-methyl-1-{[(1S)-3-methyl-1-{[(1S)-3-methyl-1-{[1-methyl-1-({1-methyl-1-[(2-{[2-(morpholin-4-yl)ethyl]carbamoyl}ethyl)carbamoyl]ethyl}carbamoyl)ethyl]carbamoyl}butyl]carbamoyl}butyl]carbamoyl}ethyl)pentanamide; 8342: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({2-[(2,2-difluoroethyl)(methyl)amino]ethyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 8343: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-{[1-({1-[(2-{[2-(dimethylamino)ethyl]carbamoyl}ethyl)carbamoyl]-1-methylethyl}carbamoyl)-1-methylethyl]carbamoyl}-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21164: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(2-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2,2-dimethylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21287: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(7-methoxy-2-oxo-2H-chromene-3-carbonyl)pyrrolidin-2-yl]formamide}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21314: (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butan-amido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-5-methylhexanamide; 21315: (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(2S)-1-{[(2S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-4-methylpentan-2-yl]carbamoyl}-4-methylpentan-2-yl]carbamoyl}-1-methylethyl)-5-methylhexanamide; 21359: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-[(1-{[(1S)-1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2,2,2-trifluoro-1-methylethyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21362: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-[(1S)-1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-2,2,2-trifluoro-1-methylethyl]-4-methylpentanamide; 21365: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2-methylpropan-2-yl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3,3,3-trifluoropropyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21372: (2S)-4-Cyclohexyl-N-[(1S)-1-[(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3,3,3-trifluoropropyl]carbamoyl}-3,3,3-trifluoropropyl]carbamoyl}-1-methylethyl)carbamoyl]-3,3,3-trifluoropropyl]-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide; and preferably, said compound of formula (I) is selected from 6025: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamido}butanamido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1- 6027:(2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamide}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1 -{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide;6253:(2S)-N-[(1S)-1-[(1-{[(1S)-1-{[(1-{[2-({1 -[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamide}decanamide;7824: (2S)-N-{1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}-2-[(2S)-2-{2-[(2S)-2-[(2S)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}-3-({4'-methoxy-[1,1'-biphenyl]-4-yl] 7848:(2S)-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)- 8343: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamide]-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-2-[(2S)-3-(6-fluoro-1,3-benzothiazol-2-yl)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamide}propanamide]-4-methylpentanamide; (2-(2-(dimethylamino)ethyl)pyrrolidin-2-yl]formamide}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-{[1-({1-[(2-{[2-(dimethylamino)ethyl]carbamoyl}ethyl)carbamoyl]-1-methylethyl}carbamoyl)-1-methylethyl]carbamoyl}-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide;21314:(3S)-3-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butan-amido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3- 21359:(2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-N-(1-{[(1S)-1-{[(1S)-1-{[2-({1-[(dimethylamino)methyl]cyclo butyl}carbamoyl)ethyl]carbamoyl}-2,2,2-trifluoro-1-methylethyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; or 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]phosphatidyl] cyclobutyl]carbamoyl}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3,3,3-trifluoropropyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide;

[0121] In a further highly preferred embodiment, the compound of formula (I) is 6027: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide.

[0122] In a further highly preferred embodiment, the compound of formula (I) is 6253: (2S)—N—[(1S)-1-[(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamide}decanamide.

[0123] In a further highly preferred embodiment, the compound of formula (I) is 7848: (2S)—N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-2-[(2S)-3-(6-fluoro-1,3-benzothiazol-2-yl)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}propanamido]-4-methylpentanamide.

[0124] In a further highly preferred embodiment, the compound of formula (I) is 8343: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-{[1-({1-[(2-{[2-(dimethylamino)ethyl]carbamoyl}ethyl)carbamoyl]-1-methylethyl}carbamoyl)-1-methylethyl]carbamoyl}-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide.

[0125] In a further highly preferred embodiment, the compound of formula (I) is 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3,3,3-trifluoropropyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide.

[0126] In a further highly preferred embodiment, the compound is selected from the formula of claim 11 and is selected from 6025, 6027, 6253, 6328, 6781, 7058, 7059, 7824, 7844, 7848, 7850, 8341, 8342, 8343, 21164, 21287, 21314, 21315, 21359, 21362, 21365, 21369 or 21372, preferably the compound of formula (I) is selected from the formula of claim 11 and is selected from 6025, 6027, 6253, 7824, 7848, 8343, 21314, 21359 or 21369. In a further highly preferred embodiment, the compound of formula (I) is formula 6027 of claim 11. In a further highly preferred embodiment, the compound of formula (I) is formula 6253 as defined in claim 11. In a further highly preferred embodiment, the compound of formula (I) is formula 7848 as defined in claim 11. In a further highly preferred embodiment, the compound of formula (I) is formula 8343 as defined in claim 11. In a further highly preferred embodiment, the compound of formula (I) is formula 21369 as defined in claim 11.

[0127] In a further aspect, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, said compound comprising: [ka] [ka] [ka] [ka] is selected from Preferably, the compound of formula (I) is [ka] [ka] is a compound selected from More preferably, the compound is 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butan-amido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3,3,3-trifluoropropyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide.

[0128] In a further aspect, the present invention provides a compound of formula (I), said compound comprising: 6781: (2S)-N-[(1S)-1-[(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamido}hexanamide; 7824: (2S)-N-{1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}-2-[(2S)-2-{2-[(2S)-2-[(2S)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}-3-({4'-methoxy-[1,1'-biphenyl]-4-yl}oxy)propanamido]-4-methylpentanamido]-2-methylpropanamido}-4-methylpentanamido]-4-methylpentanamide; 7844: (2S)-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-2-[(2S)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}-3-(6-methoxy-1,3-benzothiazol-2-yl)propanamido]-4-methylpentanamide; 7850: (2S)-2-[(2S)-3-(1,3-benzoxazol-2-yl)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}propanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21164: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(2-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2,2-dimethylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21287: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(7-methoxy-2-oxo-2H-chromene-3-carbonyl)pyrrolidin-2-yl]formamide}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21314: (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butan-amido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-5-methylhexanamide; 21315: (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(2S)-1-{[(2S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-4-methylpentan-2-yl]carbamoyl}-4-methylpentan-2-yl]carbamoyl}-1-methylethyl)-5-methylhexanamide; 21359: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-[(1-{[(1S)-1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2,2,2-trifluoro-1-methylethyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21362: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-[(1S)-1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-2,2,2-trifluoro-1-methylethyl]-4-methylpentanamide; 21365: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2-methylpropan-2-yl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3,3,3-trifluoropropyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; 21372: (2S)-4-Cyclohexyl-N-[(1S)-1-[(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3,3,3-trifluoropropyl]carbamoyl}-3,3,3-trifluoropropyl]carbamoyl}-1-methylethyl)carbamoyl]-3,3,3-trifluoropropyl]-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide; is selected from Preferably, the compound is 7824: (2S)-N-{1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}-2-[(2S)-2-{2-[(2S)-2-[(2S)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}-3-({4'-methoxy-[1,1'-biphenyl]-4-yl}oxy)propanamido]-4-methylpentanamido]-2-methylpropanamido}-4-methylpentanamido]-4-methylpentanamide; 21314: (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butan-amido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-5-methylhexanamide; 21359: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-[(1-{[(1S)-1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2,2,2-trifluoro-1-methylethyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; and 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3,3,3-trifluoropropyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide; is selected from More preferably, the compound is 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3,3,3-trifluoropropyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide.

[0129] The present invention provides a compound of formula (I) for use in a method for the topical treatment of skin cancer in a mammal, said method comprising topically administering to said mammal an effective amount of said compound or a pharmaceutically acceptable salt thereof. In a preferred embodiment, said mammal is a human.

[0130] In a further preferred embodiment, the topical administration is application of the compound or a pharmaceutically acceptable salt thereof to the skin of a mammal. In a preferred embodiment, the topical administration is application of the compound or a pharmaceutically acceptable salt thereof to the skin of a human. In a further preferred embodiment, the topical administration is application of the compound or a pharmaceutically acceptable salt thereof directly to the skin of a mammal. In a preferred embodiment, the topical administration is application of the compound or a pharmaceutically acceptable salt thereof directly to the skin of a human.

[0131] In further preferred embodiments, the skin cancer is selected from melanoma skin cancer, non-melanoma skin cancer, and cutaneous lymphoma, and any pre-invasive forms thereof. In further preferred embodiments, the skin cancer is selected from melanoma, non-melanoma skin cancer, and any pre-invasive forms thereof, preferably the non-melanoma skin cancer is basal cell carcinoma, Merkel cell carcinoma, or cutaneous squamous cell carcinoma, and more preferably the non-melanoma skin cancer is basal cell carcinoma or cutaneous squamous cell carcinoma, or any pre-cancerous forms thereof. In further preferred embodiments, the skin cancer is selected from melanoma skin cancer, non-melanoma skin cancer, and cutaneous lymphoma, and any pre-invasive forms thereof, preferably the non-melanoma skin cancer is basal cell carcinoma, Merkel cell carcinoma, or cutaneous squamous cell carcinoma, and more preferably the non-melanoma skin cancer is basal cell carcinoma or cutaneous squamous cell carcinoma, or any pre-invasive forms thereof. In further preferred embodiments, the skin cancer is selected from melanoma, non-melanoma skin cancer and any pre-invasive forms thereof, preferably the non-melanoma skin cancer is basal cell carcinoma, Merkel cell carcinoma or cutaneous squamous cell carcinoma, even more preferably the non-melanoma skin cancer is basal cell carcinoma or cutaneous squamous cell carcinoma, or any pre-invasive forms thereof.

[0132] In a further preferred embodiment, the skin cancer is selected from melanoma and non-melanoma skin cancer, preferably, the non-melanoma skin cancer is basal cell carcinoma or cutaneous squamous cell carcinoma. In a further preferred embodiment, the skin cancer is melanoma. In a further preferred embodiment, the skin cancer is non-melanoma skin cancer, preferably, the non-melanoma skin cancer is basal cell carcinoma, Merkel cell carcinoma, or cutaneous squamous cell carcinoma, or any pre-invasive form thereof, and more preferably, the non-melanoma skin cancer is basal cell carcinoma or cutaneous squamous cell carcinoma, or any pre-invasive form thereof. In a further preferred embodiment, the skin cancer is basal cell carcinoma. In a further preferred embodiment, the skin cancer is cutaneous squamous cell carcinoma. In a preferred embodiment, the skin cancer is a pre-invasive form of non-melanoma skin cancer. In another preferred embodiment, the skin cancer is non-melanoma skin cancer. In another preferred embodiment, the skin cancer is cutaneous lymphoma.

[0133] In a further preferred embodiment, the cutaneous squamous cell carcinoma (cSCC) is an invasive cSCC. In a further preferred embodiment, the cutaneous squamous cell carcinoma (cSCC) is a metastatic cSCC. In a further preferred embodiment, the basal cell carcinoma is selected from the group consisting of superficial basal cell carcinoma (also known as "basal cell carcinoma in situ" or "superficial multicentric basal cell carcinoma"), invasive basal cell carcinoma, and nodular basal cell carcinoma. In a preferred embodiment, the basal cell carcinoma is superficial basal cell carcinoma (also known as "basal cell carcinoma in situ" or "superficial multicentric basal cell carcinoma"). In a further preferred embodiment, the basal cell carcinoma is an invasive basal cell carcinoma. In a further preferred embodiment, the basal cell carcinoma is a nodular basal cell carcinoma. In another embodiment, the basal cell carcinoma is selected from the group consisting of cystic basal cell carcinoma, cicatricial basal cell carcinoma (also known as "morphea basal cell carcinoma" or "morphologic basal cell carcinoma"), micronodular basal cell carcinoma, pigmented basal cell carcinoma, ulcerative colitis (also known as Jacob's ulcer), fibroepithelioma of Pinkus, polypoid basal cell carcinoma, pore-like basal cell carcinoma, and aberrant basal cell carcinoma.

[0134] In another preferred embodiment, the skin cancer is a pre-invasive form of non-melanoma skin cancer, wherein the pre-invasive form is selected from the group consisting of cutaneous squamous cell carcinoma in situ (cSCCis, also known as "Bowen's disease") and pre-cancerous actinic keratosis (AK). In another preferred embodiment, the skin cancer is a pre-invasive form of non-melanoma skin cancer, wherein the pre-invasive form is cutaneous squamous cell carcinoma in situ (cSCCis, also known as "Bowen's disease"). In a further preferred embodiment, the skin cancer is a pre-invasive form of non-melanoma skin cancer, wherein the pre-invasive form is pre-cancerous actinic keratosis (AK). In another preferred embodiment, the skin cancer is actinic keratosis (AK). In another preferred embodiment, the skin cancer is cutaneous squamous cell carcinoma in situ (cSCCis). In another preferred embodiment, the skin cancer is cutaneous lymphoma, preferably, the cutaneous lymphoma is cutaneous T-cell lymphoma (CTCL) or cutaneous B-cell lymphoma (CBCL). In another preferred embodiment, the skin cancer is cutaneous lymphoma, preferably, the cutaneous lymphoma is cutaneous T-cell lymphoma (CTCL). In another preferred embodiment, the skin cancer is cutaneous lymphoma, preferably, the cutaneous lymphoma is cutaneous B-cell lymphoma (CBCL). In another preferred embodiment, the skin cancer is a pre-invasive form of non-melanoma skin cancer, preferably, actinic keratosis (AK). In another preferred embodiment, the skin cancer is a pre-invasive form of non-melanoma skin cancer, preferably, the pre-invasive form is cSCC in situ (cSCCis).

[0135] The compounds and methods described herein are advantageously used to inhibit the growth of skin cancer cells.Preferably, these compounds and methods provide localized treatment for mammals, preferably humans, against skin cancer.These methods provide localized treatment for various types of skin cancer, particularly basal cell carcinoma, cutaneous squamous cell carcinoma, melanoma, or cutaneous lymphoma.Therefore, topical administration of the compound of formula (I) or its pharmaceutically acceptable salt or pharmaceutical composition containing them provides treatment for patients against skin cancer. [Example]

[0136] All reagents and solvents used in the synthesis were purchased from Sigma Aldrich, Bachem, Iris Biotech, Fluorochem, Enamine, and Combi-Blocks and used as received. Solvents were stored over 4 Å molecular sieves. Fmoc-β-Ala Wang resin (0.70 mmol g) was synthesized using Fmoc solid-phase technology with repeated coupling, deprotection, and washing steps on a Liberty Blue microwave peptide synthesizer (CEM Corp., Matthews, NC). -1Peptides were synthesized on a 0.25 mmol or 0.10 mmol scale on a 1000 ketamine column. Couplings were performed as follows: Fmoc-L-amino acid or capping group (5.0 equiv., 0.2 M in DMF), DIC (5.0 equiv., 0.5 M in DMF), and Oxyma (5.0 equiv., 1.0 M in DMF) with microwave irradiation at 90 °C for 4 min. For the second AIB coupling, double couplings were performed with microwave irradiation at 90 °C for 15 min each. Fmoc deprotection was performed as follows: 10% piperazine in NMP / ethanol (9:1) or 20% piperidine in DMF with microwave irradiation at 90 °C for 1 min. After synthesis, the peptides were cleaved from the resin by treatment with a cleavage mixture (1 mL / 0.1 g resin) consisting of TFA / HO (95:5) for 90 min at ambient temperature. The suspended resin was removed by filtration and concentrated in vacuo. The crude peptides were dissolved in acetonitrile and purified by an ISCO chromatography system using HO / acetonitrile (95:5, v / v) and acetonitrile / HO with a gradient of 10% to 100% ACN as the mobile phase to obtain peptide acids with purity of >90%. The purified peptide acids (1.0 equiv.) were coupled in solution with the appropriate primary or secondary amine (2.0 equiv.) in the presence of DIPEA (3.0 equiv.) and COMU (2.0 equiv.) or HATU to obtain the final peptides. The solvent was partially removed, and the crude peptides were directly purified by a Gilson PLC 2020 Personal Purification System as described above to obtain final products with purity of >95%.

[0137] High-resolution mass spectrometry was performed on an Agilent Technologies 6530 Q-TOF. Mass and purity of the final compounds were determined by UPLC-MS using a Waters Acquity system equipped with a Waters BEH C18 column and a diode array detector (254 nm). The mobile phase consisted of HO-acetonitrile (solvent A, 97:3 v / v, LC-MS UltraChromasolv®, UHPLC grade, Sigma-Aldrich, Germany) and 0.1% formic acid (LC-MS grade, Sigma-Aldrich, Germany) and acetonitrile-HO (solvent B, 97:3 v / v, LC-MS UltraChromasolv®, UHPLC grade, Sigma-Aldrich, Germany) and 0.1% formic acid (LC-MS grade, Sigma-Aldrich, Germany) with a run time of 4 min, a flow rate of 0.6 ml / min, an injection volume of 10 μL, and gradient elution with the following program: starting at 100% A, increasing linearly to 100% B in 3 min, and returning to initial conditions within the next 1 min. MS detection of analytes was performed on a Waters Xevo® triple quadrupole mass spectrometer equipped with an electrospray ionization (ESI) interface (Waters, Xevo® TQDQCA065) in positive and negative ion mode in the mass range of 100–1500 m / z. Example 1 General Procedure for the Synthesis of Compounds of Formula (I)

[0138] Peptides were synthesized using Fmoc solid-phase technology with repeated coupling, deprotection, and washing steps, starting with Fmoc-beta-Ala linked to Wang resin, as shown in Scheme 1, using a Liberty Blue microwave peptide synthesizer (CEM Corp., Matthews, NC). Couplings were performed as follows: Fmoc-L-amino acid or capping group (5.0 equiv., 0.2 M in DMF), DIC (5.0 equiv., 0.5 M in DMF), and Oxyma (5.0 equiv., 1.0 M in DMF) at 90°C for 4 minutes in a microwave oven. For the second Aib coupling, a double coupling was performed with microwave irradiation at 90°C for 15 minutes each. Fmoc deprotection was performed as follows: 10% piperazine in NMP / ethanol (9:1) or 20% piperidine in DMF at 90°C for 1 minute in a microwave oven. Different peptides were constructed on solid supports, typically and preferably in nine steps, by sequentially using the building blocks listed in Table 1 (amino acids used in steps 1–8, Scheme 1) and the free acid derivatives listed in Table 2 (step 9, Scheme 1). After synthesis, the peptides were cleaved from the resin by treatment with a cleavage mixture (1 mL / 0.1 g resin) consisting of TFA / HO (95:5) for 90 min at ambient temperature. The suspended resin was removed by filtration and concentrated in vacuo. The crude peptides were dissolved in acetonitrile and purified using an ISCO chromatography system. The purified peptide acids were coupled in solution with the appropriate primary or secondary amines reported in Table 3 in the presence of DIPEA and COMU or HATU (step 10, Scheme 1) to give the final peptides, which were then directly purified using a Gilson PLC 2020 Personal Purification System. Scheme 1: Compounds of formula (I) were synthesized according to the general scheme. [ka] [Table 1-1] [Table 1-2] [Table 1-3] [Table 2] [Table 3] Example 2 Synthesis of Amino Acids as Building Blocks of Compounds of Formula (I)

[0139] Some of the amino acids required for the synthesis of compounds of formula (I) were not commercially available. In this case, they were synthesized. Scheme 2 shows their synthesis. Scheme 2: Synthesis of commercially unavailable amino acids [ka]

[0140] The synthesis of amino acids 8a–c is shown in Scheme 2. First, FMOC-Ser-OH (1, 30.6 mmol, 1.0 equiv.) was suspended in MeOH (6 mL / mmol of 1) and a few drops of concentrated H2SO4 were added. The reaction mixture was set under reflux for 3 h. It was then cooled to room temperature, the pH was adjusted to 8.0 with 20% w / v aqueous Na2CO3, and extracted with EtOAc (3 × 150 mL). The organic layer was washed with brine, dried over MgSO4, and evaporated to give 2 as a white solid.

[0141] In the next step, 2 (1.0 equiv.) and 4-DMAP (0.12 equiv.) were dissolved in anhydrous pyridine (0.6 mL / mmol of 2) and cooled to -5 °C. p-TsOH (3.3 equiv.) was added to the solution, and the resulting yellow reaction mixture was stirred under nitrogen for 16 h. The reaction was then poured into ice-water, and the aqueous mixture was extracted into ethyl acetate (3 × 100 mL). The combined organic layers were washed with water (2 × 100 mL), 10% w / v KHSO solution (2 × 100 mL), saturated NaHCO solution (2 × 100 mL), and brine (2 × 100 mL). The organic layer was dried over NaSO, filtered, and concentrated under reduced pressure to give tosylate 3. Subsequently, under nitrogen, a solution of sodium iodide (5.0 equiv.) in acetone (0.0 mL / mmol NaI) was added dropwise to a solution of tosylate 3 (1.0 equiv.) in acetone (1.0 mL / mmol 3). The resulting yellow solution was stirred at room temperature for 48 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in chloroform (100 mL) and washed with water (2 × 50 mL), saturated NaSO solution (2 × 50 mL), and brine (2 × 30 mL). The organic layer was dried over NaSO, filtered, and concentrated under reduced pressure to give a pale yellow solid, 4. Zn(0) (dust) (4.0 equiv.) was then weighed into a flame-dried flask, and I (0.22 equiv.) dissolved in anhydrous DMF (1.0 mL / mmol Zn(0)) was added. Iodine 4 (1.0 equiv.) along with I 2 was added and the reaction mixture was stirred until complete consumption (as indicated by TLC).

[0142] The majority of the supernatant was removed into a syringe and added directly to a pre-stirred solution of Pd2(dba)3 (0.05 mml), Xphos (0.10 mmol), and the respective halides 6a-c (1.3 mmol) in anhydrous DMF (1.0 ml / mmol Zn(0)). The reaction mixture was stirred at 40 °C for 2 h. The mixture was then poured into saturated NaHCO3 solution (10 ml) and extracted with EtOAc (2 × 10 ml). The combined organic layers were washed with saturated NaCl solution, dried over MgSO4, concentrated, and dried under vacuum. The crude products were purified by flash column chromatography to give esters 7a-c. Finally,i To a suspension of 7a-c (1.0 equiv.) in 0.8 M CaCl solution in PrOH / HO (20 ml / mmol of 7), NaOH (1.2 equiv.) was added and the mixture was stirred at room temperature overnight. The reaction was quenched by the addition of acetic acid, and the solvent was removed under reduced pressure. RP chromatography of the crude product afforded the final amino acids 8a-c. Example 3 Synthesis of Compounds of Formula (I)

[0143] The synthesis of compounds of formula (I) has been previously described in European Patent No. 3,345,917, including the synthesis of compounds 6025, 6027, and 6253. Similarly, unless otherwise stated, the compounds of formula (I) further described thereafter have been synthesized according to the general procedure shown in Scheme 1. Each of the synthesized compounds is identified by its compound number (a four or five digit number), its structural formula, and its IUPAC name, as generated by MarvinSketch software. Additionally, the syntheses of 6328, 6781, 7058, 7059, 7824, 7844, 7848, 7850, 8341, 8342, 8343, 21164, 21189, 21287, 21314, 21315, 21359, 21362, 21365, 21368, 21369, and 21372 are described with the building blocks, acids, and amines used as disclosed in Tables 1-3. Additionally, experimentally determined molecular weights are provided. Finally, for each specifically identified compound of Formula (I), the characterization of the substituents R1, R2, and R3 according to Formula (I) is provided, with the residue R in the definitions of the substituents R1, R2, and R3 corresponding to and indicating the respective bond in Formula (I). [ka]

[0144] (2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamido}butanamido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamido [Table 4] [ka]

[0145] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamide}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide. [Table 5] [ka]

[0146] (2S)—N-[(1S)-1-[(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamide}decanamide. [Table 6] [ka]

[0147] (2S)—N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methyl-2-[(2S)-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamide}-3-phenylpropanamide]pentanamide. [Table 7] [Table 8] [Table 9] [ka]

[0148] (2S)—N-[(1S)-1-[(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamide}hexanamide. [Table 10] [Table 11] [Table 12] [ka]

[0149] (2S)-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamide}-N-[(1S)-3-methyl-1-[(1-methyl-1-{[(1S)-3-methyl-1-{[1-methyl-1-({1-methyl-1-[(2-{[2-(methylamino)ethyl]carbamoyl}ethyl)carbamoyl]ethyl}carbamoyl)ethyl]carbamoyl}butyl]carbamoyl}butyl]carbamoyl}ethyl)carbamoyl]butyl]decanamide. [Table 13] [Table 14] [Table 15] Scheme 4 - Synthesis of Compound 7058 [ka]

[0150] Compound 7058 was prepared as shown in Scheme 4. More analytically, formic acid (3.7 mL) was added to compound 7057 (0.1 mmol) and the reaction mixture was allowed to stir at room temperature for 12 hours. Upon completion, the formic acid was removed under vacuum to give 7058 without further purification. [ka]

[0151] (2S)-2-[(2S)-3-Cyclohexyl-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamido}propanamido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamido [Table 16] [Table 17] [Table 18] [ka]

[0152] (2S)-N-{1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}-2-[(2S)-2-{2-[(2S)-2-[(2S)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamide}-3-({4'-methoxy-[1,1'-biphenyl]-4-yl}oxy)propanamide]-4-methylpentanamide]-2-methylpropanamide}-4-methylpentanamide]-4-methylpentanamide. [Table 19] [Table 20] [Table 21] [ka]

[0153] (2S)—N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-2-[(2S)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamide}-3-(6-methoxy-1,3-benzothiazol-2-yl)propanamide]-4-methylpentanamide. [Table 22] [Table 23] [Table 24] [ka]

[0154] (2S)—N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-2-[(2S)-3-(6-fluoro-1,3-benzothiazol-2-yl)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamide}propanamide]-4-methylpentanamide. [Table 25] [Table 26] [Table 27] [ka]

[0155] (2S)-2-[(2S)-3-(1,3-benzoxazol-2-yl)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamide}propanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide. [Table 28] [Table 29] [Table 30] [ka]

[0156] (2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-4-methyl-N-(1-methyl-1-{[(1S)-3-methyl-1-{[1-methyl-1-({1-methyl-1-[(2-{[2-(morpholin-4-yl)ethyl]carbamoyl}ethyl)carbamoyl]ethyl}carbamoyl)ethyl]carbamoyl}butyl]carbamoyl}butyl]carbamoyl}ethyl)pentanamido [Table 31] [Table 32] [Table 33] [ka]

[0157] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamide}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({2-[(2,2-difluoroethyl)(methyl)amino]ethyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide. [Table 34] [Table 35] [Table 36] [ka]

[0158] (2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-N-(1-{[(1S)-1-{[(1S)-1-{[1-({1-[(2-{[2-(dimethylamino)ethyl]carbamoyl}ethyl)carbamoyl]-1-methylethyl}carbamoyl)-1-methylethyl]carbamoyl}-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamido [Table 37] [Table 38] [Table 39] [ka]

[0159] (2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(2-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2,2-dimethylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamido [Table 40] [Table 41] [Table 42] [ka]

[0160] (2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-N-(2-{[(1S)-1-{[(1S)-1-({2-[(2-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2,2-dimethylethyl)carbamoyl]-2,2-dimethylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-2,2-dimethylethyl)-4-methylpentanamido [Table 43] [Table 44] [Table 45] [ka] (2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(7-methoxy-2-oxo-2H-chromene-3-carbonyl)pyrrolidin-2-yl]formamide}butanamide]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide [Table 46] [Table 47] [Table 48] [ka]

[0161] (3S)-3-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-5-methylhexanamide [Table 49] [Table 50] [Table 51] [ka]

[0162] (3S)-3-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-N-(1-{[(2S)-1-{[(2S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-4-methylpentan-2-yl]carbamoyl}-4-methylpentan-2-yl]carbamoyl}-1-methylethyl)-5-methylhexanamido [Table 52] [Table 53] [Table 54] [ka]

[0163] (2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-N-(1-{[(1S)-1-{[(1S)-1-[(1-{[(1S)-1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2,2,2-trifluoro-1-methylethyl]carbamoyl}-1-methylethyl)carbamoyl]-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamido [Table 55] [Table 56] [Table 57] [ka]

[0164] (2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-N-[(1S)-1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-2,2,2-trifluoro-1-methylethyl]-4-methylpentanamido [Table 58] [Table 59] [Table 60] [ka]

[0165] (2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2-methylpropan-2-yl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamido [Table 61] [Table 62] [Table 63] [ka]

[0166] (2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-2-methylpropan-2-yl)carbamoyl]-2-methylpropan-2-yl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-2-methylpropan-2-yl)-4-methylpentanamido [Table 64] [Table 65] [Table 66] [ka]

[0167] (2S)-2-[(2S)-4-Cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butanamido]-N-(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3,3,3-trifluoropropyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamido [Table 67] [Table 68] [Table 69] [ka]

[0168] (2S)-4-Cyclohexyl-N-[(1S)-1-[(1-{[(1S)-1-{[(1S)-1-({1-[(1-{[2-({1-[(dimethylamino)methyl]cyclobutyl}carbamoyl)ethyl]carbamoyl}-1-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3,3,3-trifluoropropyl]carbamoyl}-3,3,3-trifluoropropyl]carbamoyl}-1-methylethyl)carbamoyl]-3,3,3-trifluoropropyl]-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamide}butanamide [Table 70] [Table 71] [Table 72] Example 4 Skin permeation test

[0169] Skin permeation experiments were carried out using Franz Diffusion Cells (FDC) using pig skin, as an example, using highly preferred compound 6027 of the present invention. chemicals

[0170] Acetonitrile (ACN), formic acid (FA), ethanol (EtOH) and dimethyl sulfoxide (DMSO) were all of HPLC grade, and purified water was produced by reverse osmosis with an arium® pro ultrapure water system from Satorius Stedim Biotech GmbH (Göttingen, Germany). 1. Preparation of Skin

[0171] Skin from pig ears is widely used and accepted for skin permeation studies. Pig ears were obtained from freshly slaughtered pigs of both sexes, aged 5 months or less, from the slaughterhouse Bell Schweiz AG (Basel, Switzerland). Ears were washed with tap water and the hair was removed with electric clippers. Using a scalpel, the skin was separated from the underlying cartilage from the back of the ear. The skin was washed again and then stored in aluminum foil at -20°C for up to one month. Permeation experiment

[0172] After thawing, the frozen skin was cut into appropriate circular shapes and the skin thickness was measured with a caliper. The skin was placed in the FDC, tightly sealed with a Teflon band, and fixed in a specially designed holder. The acceptor compartment was then equipped with a magnetic stirrer, filled with a placebo formulation, and allowed to equilibrate for 1 hour. TEWL (transepidermal water loss) was measured to confirm the integrity of the barrier and the absence of any visible damage from the skin preparation. The FDC was then placed in a water bath at +32°C ± 1°C with the magnetic stirrer rotating (500 rpm). Due to differences between each FDC, the skin application area for drug permeation was measured and calculated in advance for each cell. As a result, the amount of 6027 for the donor compartment was 284.1 μL / cm for the infinite dose experiment. 2 I had to adjust to.

[0173] At predetermined time points, 50 μL samples were taken from the acceptor compartment and the lost volume was replaced with the corresponding placebo formulation.

[0174] At the end of the permeation experiment and before reassembling the FDC, the donor compartment was flushed with 10 mL of ACN:H2O (55:45). skin extraction

[0175] To extract 6027 from the skin, the frozen skin was cut into small pieces and pulverized using a cryogenic grinder, Freezer / Mill®, from Spex SamplePrep (Metuchen, USA). The settings were a 10-minute pre-cooling period and a speed of 10 CPS for four 2-minute cycles. The resulting powder was resuspended in 2 mL of ACN:H2O (55:45). After vortexing, the mixture was sonicated using a Branson Sonifier 250 (Model: 101-cx03-197) from Branson Ultrasonics Corporation (Danbury, USA). The sonication settings were output control 2 and a duty cycle of 30% for 60 seconds. After shaking the sample on a horizontal shaker at 37°C for 3 minutes, the skin powder was separated by centrifugation until the supernatant appeared clear. The extraction procedure was repeated two more times. High-performance liquid chromatography-mass spectrometry (HPLC-MS)

[0176] The concentration of 6027 was determined by using HPLC-MS from Agilent Technologies. The system was equipped with an Infinity LAB LC / MSD XT G6135B from Agilent, a degasser G1379, an isocratic pump G1310A, an autosampler G1329A with a thermostat G1330B, a column oven G1316 and a C18 reversed-phase column Zobrax SB-C18 Narrow Bore 2.1x150mm 5 Micron.

[0177] The mobile phase was a mixture of ACN:HO:FA (55:45:0.1) [v / v]. The flow rate was set at 0.5 mL / min, the temperature in the autosampler was set at 25 °C, and the column temperature was 50 °C. The injection volume was 30 μL, and the total run time was 30 min. All samples were diluted with a mixture of ACN:HO (55:45) to be within the calibration range. Further conditions and details such as skin thickness are listed in Table 4.

[0178] [Table 73] A substantial skin penetration of 16% in the FDC was demonstrated for 6027. This was particularly surprising since not only do 6027 and other compounds of formula (I) have molecular weights greater than 1000 Da, but 6027 also has a partition coefficient octanol / water log D of 4.56 and a low water solubility of 0.022 mg / ml. Example 5 In vitro antiproliferative effect on cutaneous squamous cell carcinoma cells

[0179] Cutaneous squamous cell carcinoma of the skin is a common form of skin cancer that develops in the squamous cells that make up the middle and outer layers of the skin. Cutaneous squamous cell carcinoma of the skin is usually not life-threatening, but it can be invasive. Untreated, cutaneous squamous cell carcinoma can grow large or spread to other parts of the body, causing serious complications. Most squamous cell carcinoma of the skin results from long-term exposure to ultraviolet (UV) radiation, either from sunlight or tanning beds or lamps. Avoiding UV light can help reduce the risk of cutaneous squamous cell carcinoma and other forms of skin cancer. Squamous cells are found in many places in the body, and squamous cell carcinoma can occur anywhere squamous cells are found. Cutaneous squamous cell carcinoma and cutaneous squamous cell carcinoma (cSCC) refer to cancers that form in the squamous cells found in the skin, respectively.

[0180] Several representative and preferred compounds of the present invention have been tested and demonstrated activity in vitro against the cutaneous squamous cell carcinoma cell line SCC12.

[0181] Methods. Skin squamous cell carcinoma cell culture SCC12 was cultured in keratinocyte-SFM medium (KSFM, catalog no. 17005042, Gibco) supplemented with human recombinant epidermal growth factor 1-53 (EGF1-53) and bovine pituitary extract (BFE). For library screening, cells were distributed into 96-well plates at a cell density of 7,500 cells per well in 90 μl and allowed to adhere overnight in a cell culture incubator at 37°C and 5% CO2. A 1000-fold serial dilution of compounds in DMSO was performed (10 mM, 5 mM, 1 mM, 500 μM, 250 μM, 100 μM, 10 μM, 1 μM). 2 μl of these dilutions were further diluted with 198 μl of keratinocyte growth medium containing supplements, mixed well, and 10 μl was further dispensed in triplicate into 96-well plates containing 90 μl of SCC cells. These dilutions resulted in final compound concentrations on the cells of 10 μM, 5 μM, 1 μM, 500 nM, 250 nM, 100 nM, 10 nM, and 1 nM.

[0182] The plates were further incubated for 72 hours, after which the cell culture medium was replaced with medium containing 0.1% resazurin sodium salt stock solution (stock: 0.015 mg / ml in PBS, catalog number R7017, Sigma-Aldrich). The cell culture plates were incubated for 2-3 hours at 37°C and 5% CO2 in a cell culture incubator. The cell culture plates were read using a fluorescence-based plate reader (Infinite M200 Pro, Tecan, Switzerland) using an excitation of 530-560 nM and an emission of 590 nM. Calculations and graphs were performed using the GraphPad Prism program. Vehicle-treated wells were set to 100% viability.

[0183] [Table 74]

[0184] result The preferred compounds tested exhibited in vitro activity and IC50 values in the nM range against cutaneous squamous cell carcinoma SCC12, confirming their utility as drugs for the treatment of skin cancer, particularly cutaneous squamous cell skin cancer. Example 6 In vivo antiproliferation assay

[0185] The preferred compound of the present invention, 6027, was tested for its in vitro antiproliferative effect on six skin cancer-derived cells, including those derived from cutaneous melanoma (M150672, M150507, M010817), cutaneous squamous cell carcinoma cell lines (SCC13 and A431_SCC), and cutaneous T-cell lymphoma (MyLa_CTCL). Similarly, an antiproliferative effect on healthy skin dermal fibroblasts was established.

[0186] Methods: For library screening, cells were dispensed at 90 μl / well into 96-well plates (cell densities (cells / well): M150672 (2,000), M150507 (1,500), M010817 (1,500), M130107 (2,000), SCC13 (11,000), A431_SCC (1,500), MyLa_CTCL (10,000)) and allowed to adhere overnight in a cell culture incubator at 37°C and 5% CO2.

[0187] For the maintenance of healthy human fibroblast cultures, Dulbecco's Modified Eagle's Medium (DMEM, Catalog No. 11966025, Gibco) supplemented with 10% heat-inactivated fetal bovine serum (Cat. No. S006420H01, BioWest) was used. For the maintenance of melanoma cell cultures, RPMI-1640 medium (Cat. No. R0883, Gibco) supplemented with L-glutamine (Cat. No. 25030-081, Gibco), 1 mM sodium pyruvate (Cat. No. S8636, Sigma-Aldrich), 10% heat-inactivated fetal bovine serum (Cat. No. S006420H01, BioWest), and 1% penicillin-streptomycin (Cat. No. 15140122, Gibco) was used. MyLa_CTCL cell cultures were grown in RPMI 1640 medium (R0883) supplemented with 2 mM L-glutamine (G7513), 10 U / ml IL-2 (H7041), 10 U / ml IL-4 (H7291), and 10% human AB serum (H4522). Cutaneous squamous cell carcinoma cell cultures A431 and SCC13 were grown in Dulbecco's modified Eagle's medium (DMEM, catalog number 11966025, Gibco) supplemented with 10% heat-inactivated fetal bovine serum (catalog number S006420H01, BioWest).

[0188] To determine the half maximal inhibitory concentration (IC50), serial dilutions of compounds in DMSO up to 1000-fold were performed (10 mM, 5 mM, 1 mM, 500 μM, 250 μM, 100 μM, 10 μM, 1 μM). 2 μl of these dilutions were further diluted with 198 μl of growth medium containing supplements, mixed well, and 10 μl were further dispensed in triplicate into 96-well plates containing 90 μl of cells. These dilutions resulted in final compound concentrations of 10 μM, 5 μM, 1 μM, 500 nM, 250 nM, 100 nM, 10 nM, and 1 nM.

[0189] For each drug condition and cell culture (for each row in triplicate), wells containing RPMI-1640 complete / KSFM / DMEM medium alone and wells containing medium and DMSO were prepared as controls. DMSO was never at a concentration higher than 0.2% v / v. Triplicates were seeded for each condition. On the day of the assay, the medium was replaced with cell culture medium containing 0.1% resazurin sodium salt stock solution (stock: 0.015 mg / ml in PBS, catalog no. R7017, Sigma-Aldrich). The cell culture plates were incubated in a cell culture incubator at 37°C and 5% CO for 2-3 hours. The cell culture plates were read using a fluorescence-based plate reader (Infinite M200 Pro, Tecan, Switzerland) using an excitation of 530-560 nM and an emission of 590 nM. Calculations and graphs were performed using the GraphPad Prism program. Vehicle-treated wells were set at 100% viability.

[0190] [Table 75] result 6027 exhibited IC50 values in the nM range against various skin cancer cell lines. Cytotoxicity was less pronounced against healthy skin dermal fibroblasts. This suggests selectivity between skin cancer-derived cells and healthy skin cells and confirms the potential therapeutic window of the compounds of the present invention, particularly 6027, for development as drugs to treat skin cancer. Example 7 In vitro antiproliferative and proapoptotic activity against B16-F10 murine melanoma cells in C57BL / 6 mice bearing B16F10 xenografts

[0191] The murine B16 melanoma model is the most commonly used metastatic melanoma model for preclinical studies. The syngeneic B16-F10 model was used to evaluate the response of 6027 to tumor development after cutaneous application. Leucinostatin A, a known naturally occurring peptide, was first tested in the same study.

[0192] Intradermal implantation of B16-F10 cells in C57BL / 6 mice results in aggressively growing tumors. Our growth studies demonstrate efficient growth kinetics after inoculation. Control animals remain on study for 10-15 days before reaching euthanasia criteria for excessive tumor burden or ulceration. This provides a model that allows test agents up to two weeks of treatment to elicit their antitumor activity.

[0193] To determine whether C57BL / 6 mice bearing B16-F10 xenografts are an appropriate model for studying the potential in vivo efficacy of transcutaneously applied 6027, in vitro tests for half-maximal inhibition (IC50) were performed on B16F10 melanoma cells. Leucinostatin A was also tested.

[0194] In vitro cell viability assay method. For the maintenance of B16-F10 melanoma cell cultures, RPMI-1640 (catalog no. R0883, Gibco) supplemented with L-glutamine (catalog no. 25030-081, Gibco), 1 mM sodium pyruvate (catalog no. S8636, Sigma-Aldrich), 10% heat-inactivated fetal bovine serum (catalog no. S006420H01, BioWest), and 1% penicillin-streptomycin (catalog no. 15140122, Gibco) was used.

[0195] The half-maximal inhibitory concentration (IC50) of compounds was determined by seeding cell cultures in 96-well plates (Corning, Catalog No. 353072) and treating them the next day with increasing concentrations of compound (0.001 μM to 10 μM in RPMI-1640 complete) for 72 hours. 1,000 cells were seeded per well. For each drug condition and cell culture (for each row of triplicates), wells containing RPMI-1640 complete medium alone and wells containing medium and DMSO were prepared as controls. DMSO was never at a concentration higher than 0.2% v / v. Triplicates were seeded for each condition. On the day of the assay, the medium was replaced with cell culture medium containing 0.1% resazurin sodium salt stock solution (stock: 0.015 mg / ml in PBS, Catalog No. R7017, Sigma-Aldrich). The cell culture plates were incubated in a cell culture incubator at 37°C and 5% CO for 2-3 hours. The cell culture plates were read using a fluorescence-based plate reader (Infinite M200 Pro, Tecan, Switzerland) using an excitation of 530-560 nM and an emission of 590 nM. Calculations and graphs were performed using the GraphPad Prism program. Vehicle-treated wells were set to 100% viability. 6027 had an IC50 of 208 nM, and leucinostatin A had an IC50 of 161 nM.

[0196] In Vivo Efficacy Testing Method. C57Bl / 6 mice (Janvier Labs); female; 6-8 weeks old at the start of treatment. Tumors were induced in the neck skin by intradermal inoculation of 1 x 10 cells per mouse. A total of 24 mice were used (n = 8 for vehicle, n = 8 for 5% 6027, n = 8 for 0.2% leucinostatin A). Starting on day 1 after tumor cell inoculation, test articles were applied with a pipette approximately 1 cm from the site of B16-F10 cell injection once daily for up to 21 days. 2 20 μl of 5% 602 Test articles were applied transdermally at 7 and 0.2% leucinostatin A. If ulceration occurred, the animals were removed from the study and sacrificed.

[0197] The reading is (cm 3 Tumor volumes were measured three times a week and expressed as tumor volume (at 0.05°C). Additionally, tumor samples were collected (2 mice per group) for histological analysis (tumor FFPE).

[0198] result Skin treatment with 20 μl of 5% 6027 on the tumor-inoculated area was well tolerated. No skin irritation or other side effects were observed. B16F10 tumors gradually grew in the neck region of mice after intradermal inoculation. At the lowest tested concentration of 0.2%, leucinostatin A showed strong skin irritation accompanied by epidermal destruction. Therefore, treatment with 0.2% leucinostatin A was not tolerated, and for ethical reasons, the treatment regimen could not be continued for longer than 7 days. Thereafter, the application site was left untreated until the end of the experiment on day 16. Animals in this group did not show a significant decrease in tumor volume throughout the study period.

[0199] [Table 76]

[0200] Histopathology. After sacrifice, nonulcerated tumors were harvested and paraffin blocks were prepared for immunohistochemical staining. Ki-67 staining was used as an immunohistochemical cellular marker for proliferation to visualize areas of the slices with proliferating cells. During interphase, Ki-67 antigen can be detected exclusively within the cell nucleus, whereas during mitosis, the majority of this protein is relocated to the surface of chromosomes. Ki-67 protein is present during all active phases of the cell cycle (G1, S, G2, and mitosis) but is absent in resting (quiescent) cells (G0). The cellular content of Ki-67 protein increases significantly during cell progression through the S phase of the cell cycle.

[0201] TUNEL staining was used to determine the areas of the tumor where apoptosis was observed. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining is a method for detecting DNA fragmentation by labeling the 3'-hydroxyl ends of DNA double-strand breaks generated during apoptosis.

[0202] result In tumor slices from C57BL / 6 mice bearing B16F10 xenografts that received daily treatments of 20 μl of 6027, tumors showed a significant decrease in proliferative cells and a significant increase in apoptotic cells with 6027 compared with tumors from vehicle-treated controls (two-way ANOVA, *p<0.01; ***p<0.0001), as shown in Figures 1A and 1B.

[0203] Thus, topical treatment with compound 6027 of the present invention significantly reduced tumor growth in B16-F10 mouse melanoma cells in C57BL / 6 mice bearing B16F10 xenografts, the most commonly used metastatic melanoma model for preclinical studies. Histological studies of tumors demonstrated a strong pro-apoptotic potential compared to controls. Leucinostatin A, a known natural peptide, has similar in vitro cytotoxicity to B16-F10 cells, but is not as suitable for dermal application as 6027, as it exhibits strong skin irritation up to a concentration of 0.2%, 25-fold lower than the 5% concentration at which 6027 did not exhibit skin irritation. Example 8 Inhibitory effects of 6027 on patient-derived basal cell carcinoma (BCC) ex vivo

[0204] Tumor material was obtained from the nose of a 91-year-old patient with invasive basal cell carcinoma (BCC).

[0205] Fresh tumor material from the patient was cut into similarly sized pieces and placed on cell culture inserts (0.4 μM, Millicell cell culture; Millipore, catalog no. PICM03050) and mixed with 1 ml of co-culture medium (3 parts DMEM (Gibco, catalog no. 41966052), 1 part Ham's F-12 Nutrient Mix (Gibco, catalog no. 11765054), 10% heat-inactivated fetal bovine serum (Gibco, catalog no. 1050), 0.1 mg / ml Normocin™ (Invivogen, catalog no. ant-nr-1), 21.8 μg / mL Adenin (Sigma-Aldrich, catalog no. A2786), 5.45 μg / mL apotransferrin (Sigma-Aldrich, catalog no. T1147), 2.18 nM triiodothyronine (Sigma-Aldrich, catalog no. T1147), 0.1 mg / mL erythritol stearate ... The tissue fragments were incubated from underneath with 0.44 μg / mL hydrocortisone (Sigma-Aldrich, Catalog No. H0888), 0.11 nM cholera toxin (Sigma-Aldrich, Catalog No. C8052), 5.50 μg / mL insulin (Sigma-Aldrich, Catalog No. I6634), and 0.01 μg / mL epidermal growth factor (Sigma-Aldrich, Catalog No. E4127) at 37°C and 5% CO2 for 24 hours to recover. After 24 hours, the cell culture medium was replaced with cell culture medium containing the drug conditions and incubated at 37°C and 5% CO2 for an additional 4 days. It was confirmed that the tissue fragments were covered with a drop of medium containing the drug (days 0 and 2). After 4 days of treatment, tissues were fixed in 4% buffered formaldehyde followed by histopathological examination and immunohistochemical staining for comparison of treatment conditions with untreated controls (DMSO).

[0206] Immunohistochemistry of ex vivo tumor sections: Tumor slices were embedded in paraffin blocks. Serial sections were stained with the desired antibodies. The following antibodies were used for staining: Ki67 (Dako, clone MIB-1, catalog number M7240, dilution 1:50, antigen retrieval: Leica Bond ER2; 30 min at 95°C), BERep4 (Dako Agilent, M0804, dilution 1:300, antigen retrieval Ph9), and HE (Hamalaun nach Mayer, Artechemis, catalog number T.865.3; eosin 1% wassrig-Morphisto, catalog number 10177). Slides were scanned using a Vectra Polaris Slide Scanner (Akoya Biosystems) and analyzed with Qpath software.

[0207] BERep4 staining was used as an immunohistochemical cellular marker for areas of the slices with basal cell carcinoma cells. BERep4 (also called Ber-EP4) is a histological stain primarily used to aid in the diagnosis of basal cell carcinoma (BCC) and is an antibody against EpCAM (epithelial cell adhesion molecule). KI-67 staining was used as an immunohistochemical cellular marker for proliferation to visualize areas of the slices with proliferating cells. Vismodegib (20 μM), the standard of care, was also used as a positive control.

[0208] Results: Four-day incubation of patient-derived BCC tumors demonstrated a significant decrease in cell proliferation in the presence of 500 nM and 1 μM 6027 and vismodegib (20 μM) compared with the negative control DMSO (two-way ANOVA; **p<0.002). As shown in Figure 2A and Figure 2B, there was a significant increase in apoptotic cells after treatment with 500 nM and 1 μM concentrations of 6027, as indicated by TUNEL staining (two-way ANOVA; ***p<0.0001; **p<0.002). Example 9 Inhibitory effect of 6027 on patient-derived basal cell carcinoma (BCC) ex vivo

[0209] Tumor material was obtained from the patient's invasive basal cell carcinoma (BCC).

[0210] Fresh punch biopsy tumor material from the patient was cut into similarly sized pieces and placed on cell culture inserts (0.4 μM, Millicell cell culture; Millipore, catalog no. PICM03050) and co-cultured with 1 ml of co-culture medium (3 parts DMEM (Gibco, catalog no. 41966052), 1 part Ham's F-12 Nutrient Mix (Gibco, catalog no. 11765054), 10% heat-inactivated fetal bovine serum (Gibco, catalog no. 1050), 0.1 mg / ml Normocin™ (Invivogen, catalog no. ant-nr-1), 21.8 μg / mL Adenin (Sigma-Aldrich, catalog no. A2786), 5.45 μg / mL apotransferrin (Sigma-Aldrich, catalog no. T1147), 2.18 nM triiodothyronine (Sigma-Aldrich, catalog no. T1147), 0.1 mg / mL erythritol stearate ... The tissue fragments were incubated from underneath with 0.44 μg / mL hydrocortisone (Sigma-Aldrich, Catalog No. H0888), 0.11 nM cholera toxin (Sigma-Aldrich, Catalog No. C8052), 5.50 μg / mL insulin (Sigma-Aldrich, Catalog No. I6634), and 0.01 μg / mL epidermal growth factor (Sigma-Aldrich, Catalog No. E4127) at 37°C and 5% CO2 for 24 hours to recover. After 24 hours, the cell culture medium was replaced with cell culture medium containing the drug conditions and incubated at 37°C and 5% CO2 for an additional 4 days. It was confirmed that the tissue fragments were covered with a drop of medium containing the drug (days 0 and 2). After 4 days of treatment, tissues were fixed in 4% buffered formaldehyde followed by histopathological examination and immunohistochemical staining for comparison of treatment conditions with untreated controls (DMSO).

[0211] Immunohistochemistry of ex vivo tumor sections: Tumor slices were embedded in paraffin blocks. Serial sections were stained with the desired antibodies. The following antibodies were used for staining: Ki67 (Dako, clone MIB-1, catalog number M7240, dilution 1:50, antigen retrieval: Leica Bond ER2; 30 min at 95°C), BERep4 (Dako Agilent, M0804, dilution 1:300, antigen retrieval Ph9), and HE (Hamalaun nach Mayer, Artechemis, catalog number T.865.3; eosin 1% wassrig-Morphisto, catalog number 10177). Slides were scanned using a Vectra Polaris Slide Scanner (Akoya Biosystems) and analyzed with Qpath software.

[0212] BERep4 staining was used as an immunohistochemical cellular marker for areas of the slices with basal cell carcinoma cells. BERep4 (also called Ber-EP4) is a histological stain primarily used to aid in the diagnosis of basal cell carcinoma (BCC) and is an antibody against EpCAM (epithelial cell adhesion molecule). KI-67 staining was used as an immunohistochemical cellular marker for proliferation to visualize areas of the slices with proliferating cells. Vismodegib (20 μM), the standard of care, was also used as a positive control.

[0213] result Four-day incubation of patient-derived BCC tumors demonstrated a significant decrease in cell proliferation in the presence of 500 nM 6027 and 20 μM vismodegib compared with the negative control DMSO (two-way ANOVA; *p<0.01; **p<0.002). As shown in Figures 3A and 3B, there was a significant increase in apoptotic cells after treatment with 500 nM 6027 and 20 μM vismodegib, as indicated by TUNEL-positive cells (two-way ANOVA; ***p<0.0001; ****p<0.00002). Example 10 Inhibitory effect of 6027 on patient-derived basal cell carcinoma (BCC) ex vivo

[0214] Tumor material was obtained from a nodular basal cell carcinoma (BCC) from the upper back of a 73-year-old patient.

[0215] Fresh tumor material from the patient was cut into similarly sized pieces and placed on cell culture inserts (0.4 μM, Millicell cell culture; Millipore, catalog no. PICM03050) and mixed with 1 ml of co-culture medium (3 parts DMEM (Gibco, catalog no. 41966052), 1 part Ham's F-12 Nutrient Mix (Gibco, catalog no. 11765054), 10% heat-inactivated fetal bovine serum (Gibco, catalog no. 1050), 0.1 mg / ml Normocin™ (Invivogen, catalog no. ant-nr-1), 21.8 μg / mL Adenin (Sigma-Aldrich, catalog no. A2786), 5.45 μg / mL apotransferrin (Sigma-Aldrich, catalog no. T1147), 2.18 nM triiodothyronine (Sigma-Aldrich, catalog no. T1147), 0.1 mg / mL erythritol stearate ... The tissue fragments were incubated from underneath with 0.44 μg / mL hydrocortisone (Sigma-Aldrich, Catalog No. H0888), 0.11 nM cholera toxin (Sigma-Aldrich, Catalog No. C8052), 5.50 μg / mL insulin (Sigma-Aldrich, Catalog No. I6634), and 0.01 μg / mL epidermal growth factor (Sigma-Aldrich, Catalog No. E4127) at 37°C and 5% CO2 for 24 hours to recover. After 24 hours, the cell culture medium was replaced with cell culture medium containing the drug conditions and incubated at 37°C and 5% CO2 for an additional 4 days. It was confirmed that the tissue fragments were covered with a drop of medium containing the drug (days 0 and 2). After 4 days of treatment, tissues were fixed in 4% buffered formaldehyde followed by histopathological examination and immunohistochemical staining for comparison of treatment conditions with untreated controls (DMSO).

[0216] Immunohistochemistry of ex vivo tumor sections: Tumor slices were embedded in paraffin blocks. Serial sections were stained with the desired antibodies. The following antibodies were used for staining: Ki67 (Dako, clone MIB-1, catalog number M7240, dilution 1:50, antigen retrieval: Leica Bond ER2; 30 min at 95°C), BERep4 (Dako Agilent, M0804, dilution 1:300, antigen retrieval Ph9), and HE (Hamalaun nach Mayer, Artechemis, catalog number T.865.3; eosin 1% wassrig-Morphisto, catalog number 10177). Slides were scanned using a Vectra Polaris Slide Scanner (Akoya Biosystems) and analyzed with Qpath software.

[0217] BERep4 staining was used as an immunohistochemical cellular marker for areas of the slices with basal cell carcinoma cells. BERep4 (also called Ber-EP4) is a histological stain primarily used to aid in the diagnosis of basal cell carcinoma (BCC) and is an antibody against EpCAM (epithelial cell adhesion molecule). Ki-67 staining was used as an immunohistochemical cellular marker for proliferation to visualize areas of the slices with proliferating cells. Vismodegib (20 μM) was used as a positive control.

[0218] result As shown in Figure 4A and Figure 4B, 4-day incubation of patient-derived BCC tumors showed a significant decrease in cell proliferation in the presence of 500 nM 6027 compared to the standard of care treatment, vismodegib (20 μM) (two-way ANOVA; **p<0.002). Example 11 Inhibitory effect of 6027 on patient-derived cutaneous squamous cell carcinoma (cSCC) ex vivo

[0219] Tumor material was obtained from a high-grade cutaneous squamous cell carcinoma (cSCC) from the head of a 79-year-old patient.

[0220] Fresh tumor material from the patient was cut into similarly sized pieces and placed on cell culture inserts (0.4 μM, Millicell cell culture; Millipore catalog number PICM03050) and co-cultured with 1 ml of co-culture medium (3 parts DMEM (Gibco, catalog number 41966052), 1 part Ham's F-12 Nutrient Mix (Gibco, catalog number 11765054), 10% heat-inactivated fetal bovine serum (Gibco, catalog number 1050), 0.1 mg / ml Normocin™ (Invivogen, catalog number ant-nr-1), 21.8 μg / ml Adenine (Sigma-Aldrich, catalog number A2786), 5.45 μg / ml Apotransferrin (Sigma-Aldrich, catalog number T1147), 2.18 nM Triiodothyronine (Sigma-Aldrich, catalog number T1147), 0.1 mg / ml Calcium Carbonate ... The tissue fragments were incubated from underneath with 0.44 μg / mL hydrocortisone (Sigma-Aldrich, Catalog No. H0888), 0.11 nM cholera toxin (Sigma-Aldrich, Catalog No. C8052), 5.50 μg / mL insulin (Sigma-Aldrich, Catalog No. I6634), and 0.01 μg / mL epidermal growth factor (Sigma-Aldrich, Catalog No. E4127) at 37°C and 5% CO2 for 24 hours to recover. After 24 hours, the cell culture medium was replaced with cell culture medium containing the drug conditions and incubated at 37°C and 5% CO2 for an additional 4 days. It was confirmed that the tissue fragments were covered with a drop of medium containing the drug (days 0 and 2). After 4 days of treatment, tissues were fixed in 4% buffered formaldehyde followed by histopathological examination and immunohistochemical staining for comparison of treatment conditions with untreated controls (DMSO).

[0221] Immunohistochemistry of ex vivo tumor sections: Tumor slices were embedded in paraffin blocks. Serial sections were stained with the desired antibodies. The following antibodies were used for staining: Ki67 (Dako, clone MIB-1, catalog number M7240, dilution 1:50, antigen retrieval: Leica Bond ER2: 30 min at 95°C), BERep4 (Dako, M0804, dilution 1:300, antigen retrieval: Ph9), and HE (Hamalaun nach Mayer, Artechemis, catalog number T.865.3; eosin 1% wassrig-Morphisto, catalog number 10177). Slides were scanned using a Vectra Polaris Slide Scanner (Akoya Biosystems) and analyzed with Qpath software.

[0222] Hematoxylin and eosin (HE) staining is one of the primary tissue stains used in histology. HE is a combination of two histological stains: hematoxylin and eosin. Hematoxylin stains cell nuclei as a purplish blue, while eosin stains the extracellular matrix and cytoplasm pink to easily distinguish between the nuclear and cytoplasmic portions of cells. Other structures take on different hues and combinations of these colors. Furthermore, the overall pattern of coloration from this stain shows the general arrangement and distribution of cells, providing a general overview of the structure of a tissue sample. Because of the typical immunohistochemical characteristics of SCC, HE can be used as an SCC diagnostic tool.

[0223] Ki-67 staining as an immunohistochemical cell marker for proliferation was used to visualize areas of the slice with proliferating cells.

[0224] Results: As shown in Figure 5A and Figure 5B, 4-day incubation of patient-derived SCC tumors showed a significant decrease in cell proliferation in the presence of 500 nM 6027 compared to the negative control DMSO (two-way ANOVA; **p<0.003).

Claims

1. 1. A compound of formula (I) for use in a method of topical treatment of skin cancer in a mammal, said method comprising topically administering said compound to said mammal; 【Chemical 1】 During the ceremony, A １ teeth, 【Chemistry 2】 A ２ teeth, 【Chemistry 3】 A ３ teeth, 【Chemistry 4】 B １ teeth, 【Chemistry 5】 B ２ teeth, 【Chemistry 6】 B ３ teeth, 【Chemistry 7】 During the ceremony, a １ , a １２ , a ２ , a ２２ , a ３ , a ３２ , b １ , b １２ , b ２ , b ２２ , b ３ and b ３２ are independently selected from 0 and 1, １ +a １２ , a ２ +a ２２ , a ３ +a ３２ , b １ +b １２ , b ２ +b ２２ and b ３ +b ３２ are independently selected from 0 and 1, １ +a １２ +a ２ +a ２２ +a ３ +a ３２ is 0, 1, 2 or 3, and b １ +b １２ +b ２ +b ２２ +b ３ +b ３２ is 0, 1 or 2; During the ceremony, R １ are each independently C １ ~C ４ Alkyl, halogen, oxo, CF ３ , OR ４ , N.R. ５ R ６ , C ６ H ５ , halogen, C １ ~C ３ Alkyl, OR ４ , N.R. ５ R ６ C substituted with ６ H ５ is selected from carbocyclyl or heterocyclyl, optionally and preferably substituted with ４ , R ５ , R ６ independently in each occurrence: H, C １ ~C ３ is alkyl; R ２ is C ４ ~C １２ Alkyl, C ４ ~C １０ Alkoxy, C １ ~C ３ Alkylene-cycloalkyl, C １ ~C ３ Alkylene-aryl, C １ ~C ３ alkylene-heteroaryl, and independently １ ~C ３ In alkylene, one —CH ２ - moiety is optionally replaced by -CH(NH)- or -O-; and said alkyl, cycloalkyl, aryl and heteroaryl are each independently selected from the group consisting of C １ ~C ２ Alkyl, C １ ~C ２ Haloalkyl, oxo, OH, halogen, C １ ~C ２ Alkoxy, C ６ H ５ or C １ ~C ３ Alkyl or OC １ ~C ３ Alkyl-substituted C ６ H ５ Optionally and preferably substituted with one or more, preferably one or two, substituents selected from: R ３ teeth 【Chemistry 8】 where: R ７ , R ８ , R ９ and R １０ is independently in each occurrence H or C １ ~C ３ alkyl, preferably H or methyl, or independently in each occurrence, ７ , R ８ , R ９ and R １０ two of which, together with the carbon atoms to which they are attached, form a carbocyclic or heterocyclic ring, preferably a carbocyclic ring, wherein R １１ and R １２ are, independently of one another, H or halogen, hydroxyl or C ３ ~C ６ C optionally substituted with cycloalkyl １ ~C ４ alkyl; or together with the nitrogen atom to which they are attached, independently in each occurrence, each independently halogen, C １ ~C ４ Alkyl, OR １３ , N.R. １４ R １５ forming a heteroaryl or heterocyclyl optionally and preferably substituted with １３ , R １４ , R １５ independently in each occurrence: H, C １ ~C ４ alkyl, where the arrow indicates the bond to the C(O)-moiety depicted in formula (I); and pharmaceutically acceptable salts of the compounds of formula (I).

2. The compound of formula (I) is a compound of formula (I*) 【Chemistry 9】 and Preferably, the compound of formula (I) is a compound of any one of formulas (II) to (IV): 【Chemistry 10】 and More preferably, the compound for use according to claim 1, wherein said compound of formula (I) is a compound of formula (II).

3. The R １ However, each independently, C １ ~C ４ Alkyl, halogen, oxo, CF ３ , OR ４ , N.R. ５ R ６ , C ６ H ５ , halogen, C １ ~C ３ Alkyl, OR ４ , N.R. ５ R ６ C substituted with ６ H ５ is selected from cycloalkyl, aryl or heteroaryl, preferably selected from cycloalkyl, monocyclic or bicyclic aryl or monocyclic or bicyclic heteroaryl, optionally substituted with ４ , R ５ , R ６ independently in each occurrence: H, C １ ~C ３ 3. The compound for use according to claim 1 or claim 2, which is alkyl.

4. The R １ However, each independently, C １ ~C ４ Alkyl, halogen, CF ３ , OR ４ , N.R. ５ R ６ , C ６ H ５ , halogen, C １ ~C ３ Alkyl, OR ４ , N.R. ５ R ６ C substituted with ６ H ５ and optionally substituted with phenyl or monocyclic or bicyclic heteroaryl containing one or two heteroatoms selected from N, O and S, preferably phenyl or oxazolyl, wherein R ４ , R ５ , R ６ independently in each occurrence: H, C １ ~C ３ The compound for use according to any one of claims 1 to 3, which is alkyl.

5. The R １ But, the formula 【Chemistry 11】 5. The compound for use according to any one of claims 1 to 4, selected from: wherein R represents a bond to the C(O)-moiety depicted in formula (I).

6. The R ２ But C ５ ~C １２ Alkyl, C ４ ~C １０ Alkoxy, C １ ~C ３ Alkylene-C ５ ~C ６ Cycloalkyl, C １ ~C ３ Alkylene-phenyl, C １ ~C ３ Alkylene-biphenyl, C １ ~C ３ alkylene-(monocyclic or bicyclic heteroaryl), and independently １ ~C ３ In alkylene, one —CH ２ - moiety is optionally replaced by -CH(NH)- or -O-, and the phenyl, biphenyl, C ５ ~C ６ -cycloalkyl and monocyclic or bicyclic heteroaryl are each independently C １ ~C ２ Alkyl, C １ ~C ２ Haloalkyl, halogen, C １ ~C ２ 6. The compound for use according to any one of claims 1 to 5, optionally substituted with one or more substituents, preferably one or two substituents selected from alkoxy.

7. The R ２ But C ５ ~C １２ Alkyl, C １ ~C ２ Alkylene-C ５ ~C ６ Cycloalkyl, C １ ~C ２ Alkylene-phenyl, C １ ~C ２ Alkylene-biphenyl, C １ ~C ２ alkylene-(monocyclic or bicyclic heteroaryl), and independently １ ~C ２ In alkylene, one —CH ２ - moiety is optionally replaced by -CH(NH)- or -O-, preferably by -O-, and said phenyl, biphenyl, C ５ ~C ６ - Compounds for use according to any one of claims 1 to 6, wherein cycloalkyl and monocyclic or bicyclic heteroaryl are optionally substituted with one or two substituents independently selected from methyl, ethyl, fluorine, chlorine and methoxy.

8. The R ２ but, 【Chemistry 12】 8. The compound for use according to any one of claims 1 to 7, selected from: wherein R represents a bond to the CH-moiety depicted in formula (I).

9. The R ７ , R ８ , R ９ and R １０ is independently in each occurrence H or C １ ~C ３ alkyl, preferably H or methyl, or independently in each occurrence, ７ , R ８ , R ９ and R １０ two of these together with the carbon atoms to which they are attached form a monocyclic carbocyclic or monocyclic heterocyclic ring, preferably a monocyclic carbocyclic ring, and R １１ and R １２ are, independently of one another, H or halogen, hydroxyl or C ３ ~C ６ C optionally substituted with cycloalkyl １ ~C ４ alkyl; or together with the nitrogen atom to which they are attached, independently in each occurrence, each independently halogen, C １ ~C ４ Alkyl, OR １３ , N.R. １４ R １５ forming a monocyclic heteroaryl or monocyclic heterocyclyl optionally substituted with R １３ , R １４ , R １５ independently in each occurrence, H, C １ ~C ４ The compound for use according to any one of claims 1 to 8, which is alkyl.

10. The R ３ but, 【Chemistry 13】 10. The compound for use according to any one of claims 1 to 9, selected from: wherein R represents a bond to the C(O)-moiety depicted in formula (I).

11. The compound is 【Chemistry 14-1】 【Chemistry 14-2】 【Chemistry 14-3】 【Chemistry 14-4】 【Chemistry 14-5】 【Chemistry 14-6】 【Chemistry 14-7】 【Chemistry 14-8】 2. The compound for use according to claim 1, selected from:

12. The compound for use according to any one of claims 1 to 11, wherein said topical administration is the application of said compound to mammalian skin, preferably human skin.

13. The compound for use according to any one of claims 1 to 12, wherein said skin cancer is selected from melanoma and non-melanoma skin cancer, preferably said skin cancer is melanoma.

14. 13. The compound for use according to any one of claims 1 to 12, wherein the skin cancer is a non-melanoma skin cancer, preferably the non-melanoma skin cancer is selected from basal cell carcinoma or cutaneous squamous cell carcinoma.

15. 15. A pharmaceutical composition for use in a method for the topical treatment of skin cancer in a mammal, said pharmaceutical composition comprising a compound of formula (I) according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or adjuvant, said method comprising topical administration of said pharmaceutical composition to said mammal.

Images