Topical treatment of skin cancer using oligopeptides

Abstract

The present invention relates to the use of compounds of formula (I), pharmaceutically acceptable salts thereof and pharmaceutical compositions comprising them, in particular compounds of formula (I) and pharmaceutical compositions, for use in a method of topically treating skin cancer in a mammal wherein said method comprises topically administering said compounds to said mammal; # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a compound of formula (I), a pharmaceutically acceptable salt thereof and a pharmaceutical composition comprising the same, and in particular to a compound of formula (I), a pharmaceutically acceptable salt thereof and a pharmaceutical composition comprising the same, for use in a method for topical treatment of skin cancer (including basal cell carcinoma, cutaneous squamous cell carcinoma and melanoma). Background Art

[0002] The skin is considered an interesting route for drug delivery because the topical administration of drugs for skin application and / or transdermal application not only avoids the risk of toxicity and gastrointestinal side effects (usually associated with other classical treatments such as systemic treatments, which can be parenteral or oral), but is also more acceptable to patients and does not require assessment before, during or after treatment to check for toxicity levels in the liver, kidneys, etc. Therefore, the development of drugs suitable for topical application, especially skin application, has received a lot of attention.

[0003] Human skin is the largest and most accessible organ for drug delivery. However, it represents a natural physicochemical barrier of the human body and is therefore characterized by low permeability to limit the transport of most pathogens, toxins, and drug molecules. Therefore, drug delivery into and through the skin is a serious challenge.

[0004] The major transport barrier for drug delivery into or through the skin is the stratum corneum, which is typically 10 to 20 μm thick and consists of 10 to 15 layers of corneocytes surrounded by a lipid-rich lipid matrix composed of ceramides, cholesterol, and free fatty acids. To penetrate the stratum corneum, drugs must traverse the tortuous lipid pathways surrounding the keratin-rich cells or partition repeatedly between the keratin-rich aqueous and lipid phases. In general, small molecules are able to penetrate the stratum corneum, while in contrast, the delivery of large molecules such as peptides and proteins remains a major challenge. For pharmaceutical purposes, when it comes to topical dermatological treatments or transdermal systemic therapies, some advocate limiting the development of new compounds to molecular weights (MW) below 500 Daltons, as around 500 Daltons is the beginning of a rapid decline in skin absorption due to molecular size. This is now known as the "500 Dalton rule." Other physicochemical properties of topical drugs that favor passive transport through the stratum corneum are an octanol / water partition coefficient logP between 1 and 3, 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 localized, destructive cancerous (malignant) growths of the skin. They originate in the cells of the epidermis, the top layer of the skin. Unlike malignant melanoma of the skin, most of these skin cancers rarely spread to other parts of the body (metastasize) and become life-threatening. There are three main types of skin cancer: (1) basal cell carcinoma (BCC; most common), (2) squamous cell carcinoma of the skin (cSCC; second most common), which originates in skin cells, and (3) melanoma, which originates in pigment-producing skin cells (melanocytes) and is less common than the first two but more dangerous. Other rare forms of skin cancer include lymphoma, Merkel cell carcinoma, and cancers of other tissues in the skin, including sarcomas and 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] We unexpectedly found that the compounds of the present invention are suitable for topical application as potential drug products for the treatment of skin cancer. In particular, it has been found that the compounds of the present invention can not only penetrate into the skin (as shown in pig skin penetration experiments), but also have activity against skin squamous cell carcinoma, basal cell carcinoma and melanoma (as shown in in vitro assays). In addition, preferred compounds of the present invention show successful treatment and antiproliferative effects in an in vivo melanoma mouse model (the most commonly used metastatic melanoma model in preclinical studies) using C67BLC / 6 mice carrying B16-F10 cell tumors, and local drug treatment results in a significant reduction in the tumor volume of the treatment group compared to the control group. Histological studies further show a powerful pro-apoptotic potential. In addition, in vitro experiments using cancer tissues of several patients with basal cell carcinoma and skin squamous cell carcinoma and treated with compounds of the present invention result in a significant reduction in proliferating cells at concentrations as low as 500nM.

[0007] Therefore, in a first aspect, the present invention provides a compound of formula (I) and a pharmaceutically acceptable salt of the compound of formula (I) for use in a method for topical treatment of skin cancer in a mammal, preferably a human, wherein the method comprises topically administering the compound to the mammal, preferably the human,

[0008]

[0009] In the formula

[0010] A1 is A2 is

[0011] A3 is B1 is

[0012] B2 is B3 is

[0013] 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, 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, a1+a 12 +a2+a 22 +a3+a 32 is 0, 1, 2 or 3, b1+b 12 +b2+b 22 +b3+b 32 is 0, 1, or 2;

[0014] R1 is selected from carbocyclyl or heterocyclyl, each independently optionally and preferably substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, R4, R5 and R6 are independently H, C1-C3 alkyl at each occurrence;

[0015] R2 is selected from C4-C 12 Alkyl, C4-C 10 Alkoxy, C1-C3 alkylene-cycloalkyl, C1-C3 alkylene-aryl and C1-C3 alkylene-heteroaryl, independently in the C1-C3 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-; the alkyl, cycloalkyl, aryl and heteroaryl are each independently optionally and preferably substituted by 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 by C1-C3 alkyl or OC1-C3 alkyl;

[0016] R3 is

[0017] In the formula

[0018] R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 Two of them together with the carbon atoms to which they are attached form a carbocyclic or heterocyclic ring, preferably a carbocyclic ring;

[0019] R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxy or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form heteroaryl or heterocyclyl at each occurrence, each independently optionally and preferably substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence, and the arrow indicates the connection to the C(O)-moiety shown in formula (I);

[0020] Preferably, said method comprises topically administering to said mammal, preferably said human, an effective amount of said compound.

[0021] In another aspect, the present invention provides a compound of formula (I) for use in a method for treating skin cancer in a mammal, preferably a human, wherein the method comprises topically administering to the mammal, preferably the human, a compound of formula (I) as defined herein (in particular as defined in any one of the appended claims) or a pharmaceutically acceptable salt thereof, preferably the method comprises topically administering to the mammal, preferably the human, an effective amount of the compound or the pharmaceutically acceptable salt thereof.

[0022] In another aspect, the present invention provides a pharmaceutical composition for use in a method for topical treatment of skin cancer in a mammal, preferably a human, wherein the pharmaceutical composition comprises a compound of formula (I) as defined herein (in particular as defined in any one of the appended claims) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or adjuvant, and the method comprises topically administering the pharmaceutical composition to the mammal, preferably a human, preferably the pharmaceutical composition comprises an effective amount of the compound.

[0023] Therefore, in particular, the present invention provides compounds and pharmaceutical compositions for use in a method for topical treatment of skin cancer in a mammal, preferably a human, in particular for use in a method for topical treatment of melanoma and non-melanoma skin cancer (preferably melanoma, basal cell carcinoma and squamous cell carcinoma of the skin), wherein the method comprises topically administering to the mammal, preferably the human, an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0024] In another aspect, the present invention provides a method for topically treating skin cancer in a mammal, preferably a human, wherein the method comprises topically administering to the mammal, preferably the human, a compound of formula (I) of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof. Preferably, the method comprises topically administering to the mammal, preferably the human, an effective amount of the compound, the pharmaceutically acceptable salt thereof, or the pharmaceutical composition.

[0025] On the other hand, the present invention provides use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for topical treatment of skin cancer in a mammal, preferably a human, wherein the method comprises topically administering the compound of formula (I) or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present invention to the mammal, preferably the human; preferably, the method comprises topically administering an effective amount of the compound or a pharmaceutically acceptable salt thereof or the pharmaceutical composition to the mammal, preferably the human.

[0026] Other aspects and embodiments of the invention will become apparent as the description proceeds. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1A : Tumor sections were collected from C57Bl / 6 mice bearing B16F10 xenografts and subjected to histological study. Tumor sections were stained with the proliferation marker Ki67 (to identify tumor areas with proliferating cells) and the apoptosis marker TUNEL (to show areas with apoptotic cells within the tumor).

[0028] Figure 1B :right Figure 1A Quantification of Ki67 and TUNEL positively stained cells, Group A (placebo) and Group B (6027) as shown.

[0029] 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.

[0030] Figure 2B :right Figure 2A Quantification of Ki67 and TUNEL positively stained cells as indicated, A (DMSO), B (Vismodegib, 20 mM), C (6027, 500 nM) and D (6027, 1 mM).

[0031] Figure 3A : Immunohistochemical staining of patient-derived basal cell carcinoma (BCC) cultured for 4 days in medium containing 500 nM 6027 or DMSO as negative control and vismodegib (20 μM) as positive control.

[0032] Figure 3B :right Figure 3A Quantification of Ki67 and TUNEL positively stained cells as indicated, A (DMSO), B (Vismodegib, 20 mM) and C (6027, 500 nM).

[0033] Figure 4A : Histopathological sections of patient-derived basal cell carcinoma (BCC) incubated for 4 days with 500 nM 6027 and standard of care vismodegib (20 μM) as a positive control.

[0034] Figure 4B :right Figure 4A Quantification of Ki67-positive stained cells as shown, A (Vismodegib, 20 mM) and B (6027, 500 nM).

[0035] Figure 5A : Histopathological sections of patient-derived high-grade cutaneous squamous cell carcinoma (cSCC) treated for 4 days with 500 nM 6027 and DMSO as negative control.

[0036] Figure 5B :right Figure 5A Quantification of Ki67 positively stained cells as shown, A (DMSO) and B (6027, 500 nM). DETAILED DESCRIPTION

[0037] 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 the invention belongs. The embodiments, preferred embodiments and very preferred embodiments described and disclosed herein should apply to all aspects and other embodiments, preferred embodiments and very preferred embodiments, whether or not specifically mentioned again.

[0038] As used herein, the terms "a" or "an" mean "at least one" unless otherwise specified.

[0039] Each alkyl moiety, alone or as part of a larger group (e.g., alkoxy, aminoalkyl, or haloalkoxy or alkylene), refers to a monovalent saturated acyclic (i.e., non-cyclic) hydrocarbon radical, which may be straight-chain or branched. Thus, "alkyl" does not contain any carbon-carbon double bonds or any carbon-carbon triple bonds. "C 1-6 The term "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, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, neopentoxy and n-hexoxy. Examples of aminoalkyl include aminomethyl, aminoethyl, dimethylaminomethyl and dimethylaminoethyl. A haloalkoxy group refers to an alkoxy group further substituted with a halogen.

[0040] As used herein, the term "alkylene" refers to an alkanediyl group, i.e., a divalent saturated acyclic hydrocarbon group, which may be straight-chain or branched. 1-6 “Alkylene” means an alkylene group having 1 to 6 carbon atoms. Preferred exemplary alkylene groups are methylene (—CH2—), ethylene (e.g., —CH2-CH2— or —CH(—CH3)-), propylene (e.g., CH2-CH2-CH2-, —CH(-CH2-CH3)-, —CH2-CH(-CH3)-, or —CH(-CH3)-CH2-), or butylene (e.g., —CH2-CH2-CH2-CH2-).

[0041] Each haloalkyl moiety, alone or as part of a larger group (e.g., haloalkoxy), is an alkyl group substituted with one or more halogen atoms, which may be the same or different. The haloalkyl group contains, for example, 1 to 5 halogen substituents or 1 to 3 halogen substituents. Examples particularly include fluoromethyl, difluoromethyl, trifluoromethyl, chlorodifluoromethyl, and 2,2,2-trifluoroethyl.

[0042] Each alkenyl moiety, alone or as part of a larger group (e.g., alkenyloxy or alkenylene), is a straight or branched chain, preferably C2-C 16 Alkenyl, more preferably C2-C 14 Alkenyl. Each part can be (E)-configuration or (Z)-configuration. Examples include vinyl and allyl. Therefore, if applicable, the compounds of the present invention containing alkenyl parts can include compounds with alkenyl parts in (E)-configuration, compounds with alkenyl parts in (Z)-configuration, and mixtures of any ratio thereof.

[0043] Halogen is fluorine, chlorine, bromine or iodine.

[0044] As used herein, the term "carbocyclic group" refers to a monovalent hydrocarbon ring group, including a monocyclic ring and a bridged ring, a spirocyclic ring and / or a fused ring system (which may be composed of, for example, two or three rings), wherein the ring group may be saturated, partially unsaturated (i.e., unsaturated but non-aromatic) or aromatic. Unless otherwise defined, "carbocyclic group" preferably refers to an aryl, a cycloalkyl or a cycloalkenyl group. The number of carbon atoms in the carbocyclic group is not particularly limited, preferably 3 to 14, more preferably 4 to 12 or 5 to 10.

[0045] As used herein, the term "heterocyclyl" refers to a monovalent cyclic group, including a monocyclic ring and a bridged ring, a spirocyclic ring and / or a fused ring system (which may be composed of, for example, two or three rings), wherein the cyclic 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; one or more S ring atoms (if present) and / or one or more N ring atoms (if present) may be optionally oxidized, one or more carbon ring atoms may be optionally oxidized (i.e., forming an oxo group), and further the cyclic group may be saturated, partially unsaturated (i.e., unsaturated but non-aromatic) or aromatic. Unless otherwise defined, "heterocyclyl" preferably refers to heteroaryl, heterocycloalkyl or heterocycloalkenyl. The number of carbon atoms in the heterocyclyl is not particularly limited, preferably 5 to 14, more preferably 5 to 12 or 5 to 10.

[0046] As used herein, the term "aryl" refers to a group consisting of 6 to 14 carbon atoms (C6-C 14). The aryl group includes a bicyclic, tricyclic or tetracyclic group containing an aromatic ring, preferably a bicyclic group, on which a saturated, partially unsaturated or aromatic carbon ring or heterocycle is fused or bridged. The aryl group is optionally independently substituted by one or more substituents (usually and preferably by one or two substituents), wherein the substituents are usually and preferably independently selected from C1-C4 alkyl, halogen, oxo, CF3, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C6H5, C6H5 substituted by halogen, C1-C3 alkyl, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2. Typical aryl groups include, but are not limited to, phenyl, substituted phenyl, naphthyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, 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 (preferably one substituent), the substituents being independently selected from C1-C4 alkyl, halogen, CF3, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C6H5, C6H5 substituted with halogen, C1-C3 alkyl, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2.

[0047] As used herein, the term "heteroaryl" refers to an aromatic ring group, including a monocyclic aromatic ring and a bridged ring and / or a fused ring system containing at least one aromatic ring (for example, a ring system consisting of two or three fused rings, wherein at least one of the fused rings is an aromatic ring; or a bridged ring system consisting of two or three rings, wherein at least one of the bridged rings is an aromatic ring), wherein the aromatic ring group contains one or more (for example, one, two, three or four) ring heteroatoms independently selected from O, S and N, and the remaining ring atoms are carbon atoms; 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 is optionally substituted independently by one or more substituents, typically and preferably by one or two substituents independently selected from C1-C4 alkyl, halogen, oxo, CF3, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C6H5, C6H5 substituted by halogen, C1-C3 alkyl, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2. Unless otherwise defined, "heteroaryl" preferably has 5 to 14 ring atoms, more preferably 5 to 12 or 5 to 10 ring atoms.

[0048] In a preferred embodiment, the heteroaryl group is a monovalent monocyclic aromatic or bicyclic aromatic ring group, wherein the aromatic 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; the aromatic ring group is optionally independently substituted by one or more substituents, usually and preferably substituted by one or two substituents independently selected from C1-C4 alkyl, halogen, oxo, CF3, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C6H5, C6H5 substituted by halogen, C1-C3 alkyl, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2. Unless otherwise defined, such monocyclic or bicyclic heteroaryl groups preferably have 5 to 12, preferably 5 to 10 ring atoms.

[0049] Examples of heteroaryl groups include pyridyl, imidazolyl, imidazopyridyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, tetrahydroisoquinolyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, thiadiazolyl, furazanyl, benzofurazanyl, benzopyranyl, benzophenylthio, benzothiazolyl, benzoxazolyl, The heteroaryl groups include, but are not limited to, C1-C4 alkyl, halogen, oxo, CF3, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C6H5, C6H5 substituted with halogen, C1-C3 alkyl, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2.

[0050] Other examples of such monocyclic heteroaryl groups include, but are not limited to, 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, 5-oxazolyl, 2-furanyl, 3-furanyl, 2-thienyl, 3-thienyl, 3-triazolyl, 1 -triazolyl, 5-tetrazolyl, 1-tetrazolyl and 2-tetrazolyl, which are optionally substituted independently by one or more substituents, typically and preferably by one or two substituents, 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)2, C6H5, C6H5 substituted by halogen, C1-C3 alkyl, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2.

[0051] As used herein, the term "cycloalkyl" refers to a monovalent saturated hydrocarbon ring group, including a monocyclic ring as well as a bridged ring, a spirocyclic ring and / or a fused ring system (which may be composed of, for example, two, three or four rings; for example, a fused ring system composed of two or three fused rings). Cycloalkyl is optionally substituted independently by one or more substituents, usually and preferably substituted by one or two substituents independently selected from C1-C4 alkyl, halogen, oxo, CF3, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C6H5, C6H5 substituted by halogen, C1-C3 alkyl, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2. Unless otherwise defined, "cycloalkyl" preferably refers to C 3-14 Cycloalkyl. Examples of cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or adamantyl.

[0052] As used herein, the term "heterocycloalkyl" refers to a monovalent saturated ring group, including a monocyclic ring as well as a bridged ring, a spirocyclic ring and / or a fused ring system (which may be composed of, for example, two or three rings; for example, a fused ring system composed of two or three fused 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; one or more S ring atoms (if present) and / or one or more N ring atoms (if present) may be optionally oxidized, and further one or more carbon ring atoms may be optionally oxidized (i.e., to form an oxo group). The heterocycloalkyl group is optionally substituted independently with one or more substituents, typically and preferably with one or two substituents independently selected from C1-C4 alkyl, halogen, oxo, CF3, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C6H5, C6H5 substituted with halogen, C1-C3 alkyl, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2. Unless otherwise defined, "heterocycloalkyl" preferably refers to a 3- to 14-membered saturated ring group, which is a monocyclic ring or a fused ring system (e.g., a fused ring system consisting of two fused 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, 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 group containing one or more (e.g., one, two or three) ring heteroatoms independently selected from O, S and N, 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.

[0053] As used herein, the term "cycloalkenyl" refers to an unsaturated alicyclic (non-aromatic) hydrocarbon ring group, including a monocyclic ring as well as a bridged ring, a spirocyclic ring and / or a fused ring system (which may be composed of, for example, two or three rings; for example, a fused ring system composed of two or three fused rings), wherein the hydrocarbon ring group contains one or more (e.g., one or two) carbon-carbon double bonds and does not contain any carbon-carbon triple bonds. The cycloalkenyl group is optionally independently substituted with one or more substituents, usually and preferably substituted with one or two substituents, the substituents usually and preferably 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)2, C6H5, C6H5 substituted with halogen, C1-C3 alkyl, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2. "Cycloalkenyl" may refer to, for example, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl or cycloheptadienyl. Unless otherwise defined, "cycloalkenyl" preferably refers to C 3-14 Cycloalkenyl, more preferably C 3-7 Cycloalkenyl.

[0054] As used herein, the term "heterocycloalkenyl" refers to an unsaturated alicyclic (non-aromatic) ring group, including a monocyclic ring as well as a bridged ring, a spirocyclic and / or a fused ring system (which may be composed of, for example, two or three rings; for example, a fused ring system composed of two or three fused 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, the remaining ring atoms are carbon atoms, one or more S ring atoms (if present) and / or one or more N ring atoms (if present) may be optionally oxidized, one or more carbon ring atoms may be optionally oxidized (i.e., to form an oxo group), and further the ring group contains at least one double bond between adjacent ring atoms, and does not contain any triple bonds between adjacent ring atoms. The heterocycloalkenyl group is optionally substituted independently by one or more substituents, typically and preferably by one or two substituents, the substituents typically and preferably being 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)2, C6H5, C6H5 substituted by halogen, C1-C3 alkyl, OH, OC1-C3 alkyl, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2. Unless otherwise defined, "heterocycloalkenyl" preferably refers to a 3- to 14-membered unsaturated alicyclic ring group, which is a monocyclic ring or a fused ring system (e.g., a fused ring system consisting of two fused 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, 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, the ring group contains at least one double bond between adjacent ring atoms, and does not contain any triple bonds between adjacent ring atoms.

[0055] When a group is described as being optionally substituted, preferably there are 1 to 5 optional substituents, more preferably there are 1 to 3 optional substituents, and even more preferably there are one or two optional substituents.

[0056] As used herein, the term "amino acid" refers to an organic compound containing the functional groups amine (-NH2) and carboxylic acid (-COOH) and zwitterions thereof (usually and preferably, as well as side chains specific to each amino acid). The term "amino acid" generally and preferably includes naturally occurring amino acids, such as protein amino acids (produced by RNA translation), non-protein amino acids (produced by other metabolic mechanisms, such as post-translational modifications), standard or canonical amino acids (directly encoded by codons of the genetic code), and non-standard or 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 non-natural amino acids. In addition, the term covers amino acids such as alpha-(α-), beta-(β-), gamma-(γ-) and delta-(δ-) and mixtures thereof in any ratio, as well as any isomeric forms of amino acids, i.e., D- and L-stereoisomers (or represented by (R) and (S) nomenclature) and mixtures thereof in any ratio, preferably a 1:1 racemic ratio. The amino acids in the present invention are preferably L-configurations. The terms "D-stereoisomer," "L-stereoisomer," "D-amino acid," or "L-amino acid" refer to the chiral alpha carbon of an amino acid.

[0057] Some compounds of formula (I) of the present invention may contain one or more chiral centers, and these compounds may be provided as pure enantiomers or pure diastereomers and mixtures of any ratio thereof. The compounds of the present invention also include all tautomeric forms of compounds of formula (I). The compounds of formula (I) may also be solvated, particularly hydrated, which are also included in the compounds of formula (I). The term "chirality" refers to compounds having the non-overlapping properties of mirror image partners, while the term "achirality" refers to compounds that can overlap with mirror image partners. The term "stereoisomer" refers to compounds having the same chemical composition but different arrangements of atoms or groups in space. "Diastereomer" refers to stereoisomers with more than two chiral centers, wherein the compounds are not mirror images of each other. Diastereomers have different physical properties (e.g., melting points, boiling points, spectral properties) and chemical and biological reactivity. A mixture of diastereomers may be separated under high-resolution analytical procedures (e.g., electrophoresis and chromatography). "Enantiomers" refers to two stereoisomers of a compound that are non-superimposable mirror images of each other. The stereochemical definitions and conventions used herein generally follow: S. P. Parker, ed., McRaw-Hiff Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York; E. Eliel and S. Wilen, "Stereochemistry of Organic Compounds", John Wiley & Sons, New York, 1994. The compounds of the present invention may contain asymmetric or chiral centers and therefore exist in different stereoisomeric forms. All stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers and atropisomers, and mixtures thereof (e.g., racemic mixtures), constitute 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. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule relative to its chiral center. The prefixes d and l or (+) and (-) are used to indicate the sign of the rotation of plane polarized light by a compound, wherein (-) or l indicates that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Specific stereoisomers may also be referred to as enantiomers, and mixtures of such isomers are often referred to as enantiomeric mixtures 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 can be interconverted via a low energy barrier. For example, proton tautomers include interconversions via proton migration, such as keto-enol and imine-enamine isomerizations.

[0058] As used herein, the phrase "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable organic or inorganic salt of the compounds of the invention, particularly an acid addition salt. Exemplary salts include, but are not limited to, sulfates, citrates, acetates, oxalates, chloride salts, bromide salts, iodide salts, nitrates, bisulfates, phosphates, acid phosphates, isonicotinates, lactates, salicylates, acid citrates, tartrates, oleates, tannates, pantothenates, bitartrates, ascorbates, succinates, maleates, gentisates, fumarates, gluconates, glucuronates, sucrose salts, formates, benzoates, glutamates, methanesulfonates (mesylate), ethanesulfonates, benzenesulfonates, p-toluenesulfonates, and pamoates. A pharmaceutically acceptable salt may involve the inclusion of another molecule, such as acetate ions, succinate ions, or other counterions. Counterions may be any organic or inorganic moiety that stabilizes the charge on the parent compound. In addition, a pharmaceutically acceptable salt may have more than one charged atom in its structure. When multiple charged atoms are part of a pharmaceutically acceptable salt, multiple counterions may be present. Thus, a pharmaceutically acceptable salt may have more than one charged atom and / or more than one counterion. If the compound of the 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 (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric acid, etc.), or by treating the free base with an organic acid (e.g., acetic acid; trifluoroacetic acid; maleic acid; succinic acid; mandelic acid; fumaric acid; malonic acid; pyruvic acid; oxalic acid; glycolic acid; salicylic acid; pyranosidic acid, such as glucuronic acid or galacturonic acid; α-hydroxy acids, such as citric acid or tartaric acid; amino acids, such as aspartic acid or glutamic acid; aromatic acids, such as benzoic acid or cinnamic acid; sulfonic acids, such as p-toluenesulfonic acid or ethanesulfonic acid; etc.).

[0059] "Solvate" refers to an association or complex of one or more solvent molecules with a compound of the 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.

[0060] The terms "compounds of the present invention" and "compounds of the present invention" and "compounds of formula (I)" include stereoisomers, geometric isomers, tautomers, solvates, pharmaceutically acceptable salts and solvates of their salts.

[0061] 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.

[0062] As used herein, the term "treatment" of a condition or disease (e.g., "treatment" of skin cancer such as melanoma) is well known in the art. "Treating" a condition or disease means that the patient / subject is suspected of or has been diagnosed with the condition or disease. Patients / subjects suspected of having a condition or disease typically exhibit specific clinical and / or pathological symptoms that can be readily attributed to a specific pathological condition (i.e., diagnosed with the condition or disease) by one skilled in the art.

[0063] For example, "treating" a disorder or disease can result in cessation of the progression of the disorder or disease (e.g., without worsening of symptoms) or a delay in the progression of the disorder or disease (in the case where the cessation of progression is only temporary). "Treatment" of a disorder or disease can also result in a partial response (e.g., symptom improvement) or a complete response (e.g., disappearance of symptoms) in a subject / patient with the disorder or disease. Therefore, "treating" a disorder or disease can also refer to improving the disorder or disease, for example, which can result in cessation of the progression of the disorder or disease or a delay in the progression of the disorder or disease. This partial or complete response may relapse afterwards. It should be understood that the subject / patient can have a wide range of responses to treatment. For example, the treatment of a disorder or disease can include curative treatment (preferably resulting in a complete response and ultimately curing the disorder or disease) and palliative treatment (including symptom relief). For example, "improving" a disorder or disease can result in cessation of the progression of the disorder or disease or a delay in the progression of the disorder or disease. "Treatment" can also mean extending the survival period compared to the expected survival period without treatment.

[0064] The term "effective amount" refers to an amount of a compound of the invention or a pharmaceutical composition of the invention that (i) treats a particular disease or condition or (ii) alleviates, improves or eliminates one or more symptoms of a particular disease or condition. In the case of skin cancer, particularly where the cancer is melanoma, basal cell carcinoma or squamous cell carcinoma of the skin, an effective amount of the drug can reduce tumor volume and / or alleviate clinical symptoms.

[0065] As used herein, the term "skin cancer" refers to skin cancers, such as melanoma skin cancer and non-melanoma skin cancer (NMSC), but also refers to and includes precancerous and pre-invasive neoplastic skin proliferations, such as cutaneous squamous cell carcinoma in situ (cSCCis or Bowen's disease) or actinic keratosis (AK, also known as "solar keratosis" and "senile keratosis"). In addition, as used herein, the term "skin cancer" shall also include cutaneous lymphomas, such as cutaneous T-cell lymphoma (CTCL) or cutaneous B-cell lymphoma (CBCL), or their pre-invasive forms. Bowen's disease is a skin neoplastic disease that can be considered an early or intraepidermal form of squamous cell carcinoma. Actinic keratosis is characterized by thick, scaly or crusty precancerous plaques that usually form when the skin is damaged by ultraviolet (UV) radiation from the sun or indoor tanning beds.

[0066] The term "non-melanoma skin cancer" (abbreviated as "NMSC") is used herein to refer to a group of diseases that specifically includes 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.

[0067] The term “field cancerization” refers to a precancerous regional defect, a biological process in which a large area of ​​cells on the surface of a tissue or within an organ is affected by carcinogenic changes. This process is usually due to prolonged exposure to a harmful environment, such as ultraviolet radiation. The initial steps of field cancerization are associated with various molecular lesions, such as acquired gene mutations and epigenetic changes, occurring in extensive multifocal “fields”. This field is affected by subclinical (invisible, non-palpable) AK lesions, early AK lesions, advanced AK lesions, and even potentially invasive cSCC. The concept of field cancerization provides a theoretical basis for regional therapy, in which the entire field is treated rather than a single lesion. The goal of regional therapy is not only to eliminate clinically visible lesions, but also to eliminate subclinical lesions and prevent the development of invasive squamous cell carcinoma.

[0068] As used herein, the term "topical application" refers to application to a body surface, such as the skin. Typically and preferably, as used herein, the term "topical application" refers to epidermal application applied directly to the skin. Therefore, in a very preferred embodiment of the present invention, the topical application is application on the skin, preferably the topical application is application directly on the skin. Typically and preferably, the present invention relates to topical applications and topically acting compounds for cutaneous use to treat skin infections, as described in the EMA Quality Equivalence Guidelines for Topical Products (https: / / www.ema.europa.eu / en / documents / scientific-guideline / draft-guideline-quality-equivalence-topical-products_en.pdf).

[0069] In a first aspect, the present invention provides a compound of formula (I), and a pharmaceutically acceptable salt of the compound of formula (I), for use in a method for topical treatment of skin cancer in a mammal, wherein the method comprises topically administering the compound to the mammal,

[0070]

[0071] In the formula

[0072] A1 is A2 is

[0073] A3 is B1 is

[0074] B2 is B3 is

[0075] 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, 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, a1+a 12 +a2+a 22 +a 22 +a3+a 32 is 0, 1, 2 or 3, b1+b 12 +b2+b 22 +b3+b 32 is 0, 1, or 2;

[0076] R1 is selected from carbocyclyl or heterocyclyl, each independently optionally and preferably substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, R4, R5 and R6 are independently H, C1-C3 alkyl at each occurrence;

[0077] R2 is selected from C4-C 12 Alkyl, C4-C 10 Alkoxy, C1-C3 alkylene-cycloalkyl, C1-C3 alkylene-aryl and C1-C3 alkylene-heteroaryl, independently in the C1-C3 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-; the alkyl, cycloalkyl, aryl and heteroaryl are each independently optionally and preferably substituted by 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 by C1-C3 alkyl or OC1-C3 alkyl;

[0078] R3 is

[0079] In the formula

[0080] R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 Two of them together with the carbon atoms to which they are attached form a carbocyclic or heterocyclic ring, preferably a carbocyclic ring;

[0081] R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxy or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form heteroaryl or heterocyclyl at each occurrence, each independently optionally and preferably substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence, and the arrow indicates the connection to the C(O)-moiety shown in formula (I);

[0082] Preferably, said method comprises topically administering to said mammal, preferably a human, an effective amount of said compound.

[0083] In another aspect, the present invention provides a compound of formula (I*), and a pharmaceutically acceptable salt of the compound of formula (I*), for use in a method for topical treatment of skin cancer in a mammal, wherein the method comprises topically administering the compound to the mammal, preferably a human,

[0084]

[0085] In the formula

[0086] R1 is selected from carbocyclyl or heterocyclyl, each independently optionally and preferably substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, R4, R5 and R6 are independently H, C1-C3 alkyl at each occurrence;

[0087] R2 is selected from C5-C 12 Alkyl, C4-C 10Alkoxy, C1-C3 alkylene-cycloalkyl, C1-C3 alkylene-aryl and C1-C3 alkylene-heteroaryl, independently in the C1-C3 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-; the alkyl, cycloalkyl, aryl and heteroaryl are each independently optionally and preferably substituted by one or more (usually and preferably one or two) substituents selected from C1-C2 alkyl, C1-C2 haloalkyl, oxo, OH, halogen, C1-C2 alkoxy, C6H5 or C6H5 substituted by C1-C3 alkyl or OC1-C3 alkyl;

[0088] R3 is

[0089] In the formula

[0090] R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 Two of them together with the carbon atoms to which they are attached form a carbocyclic or heterocyclic ring, preferably a carbocyclic ring;

[0091] R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxy or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form heteroaryl or heterocyclyl at each occurrence, each independently optionally and preferably substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence, and the arrow indicates the connection to the C(O)-moiety shown in formula (I);

[0092] Preferably, said method comprises topically administering to said mammal, preferably a human, an effective amount of said compound.

[0093] In another aspect, the present invention provides a method for topically treating skin cancer in a mammal, preferably a human, wherein the method comprises topically administering to the mammal, preferably the human, a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof. Preferably, the method comprises topically administering to the mammal, preferably the human, an effective amount of the compound.

[0094] On the other hand, the present invention provides use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for topical treatment of skin cancer in a mammal, preferably a human, wherein the method comprises topically administering the compound of formula (I) or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present invention to the mammal, preferably the human; preferably, the method comprises topically administering an effective amount of the compound or a pharmaceutically acceptable salt thereof or the pharmaceutical composition to the mammal, preferably the human.

[0095] In a preferred embodiment, the compound of formula (I) is a compound of any one of formulas (II) to (IV):

[0096]

[0097] Preferably, the compound of formula (I) is a compound of formula (II).

[0098] 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).

[0099] In another preferred embodiment, R1 is selected from phenyl, naphthyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, anthracenyl, phenanthrenyl, biphenyl, indenyl, indanyl, pyridyl, imidazolyl, imidazopyridyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, tetrahydroisoquinolyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pter R4, R5 and R6 are independently H, C1-C3 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted with halogen, C1-C3 alkyl, OR4, NR5R6, R4, R5 and R6 are independently H, C1-C3 alkyl at each occurrence.

[0100] In another preferred embodiment, R1 is selected from cycloalkyl, aryl or heteroaryl, each of which is independently optionally substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5 and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, and R4, R5 and R6 are independently H or C1-C3 alkyl at each occurrence.

[0101] In another preferred embodiment, R1 is selected from cycloalkyl, monocyclic or bicyclic aryl, or monocyclic or bicyclic heteroaryl, each of which is independently optionally substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, and R4, R5 and R6 are independently H or C1-C3 alkyl at each occurrence.

[0102] In another preferred embodiment, R1 is selected from cycloalkyl, monocyclic or bicyclic aromatic aryl, or monocyclic or bicyclic heteroaryl, each of which is independently optionally substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, and R4, R5 and R6 are independently H or C1-C3 alkyl at each occurrence.

[0103] In another preferred embodiment, R1 is selected from phenyl or a monocyclic or bicyclic heteroaryl group containing one or two heteroatoms selected from N, O and S, preferably phenyl, coumarin or oxazolyl, further preferably phenyl or oxazolyl, each independently optionally substituted by C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5 and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, and R4, R5 and R6 are independently H or C1-C3 alkyl at each occurrence.

[0104] In another preferred embodiment, R1 is selected from phenyl, naphthyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, biphenyl, indenyl, indanyl, pyridyl, imidazolyl, imidazopyridyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, tetrahydroisoquinolyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl , pteridinyl, purinyl, thiadiazolyl, furazolyl, benzofurazolyl, benzopyranyl, benzophenylthio, benzothiazolyl, benzoxazolyl, coumarinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, furapyridinyl, cyclopentyl, cyclohexyl, adamantyl, each independently optionally substituted with C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted with halogen, C1-C3 alkyl, OR4, NR5R6, R4, R5 and R6 are independently H, C1-C3 alkyl at each occurrence.

[0105] In another preferred embodiment, the R1 is selected from the following formula

[0106]

[0107] In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

[0108] In another preferred embodiment, the R1 is selected from phenyl or monocyclic or bicyclic heteroaryl, each independently optionally substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, R4, R5 and R6 are independently H, C1-C3 alkyl at each occurrence. In another preferred embodiment, the R1 is selected from phenyl or monocyclic or bicyclic aromatic heteroaryl, each independently optionally substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, R4, R5 and R6 are independently H, C1-C3 alkyl at each occurrence.

[0109] In another preferred embodiment, R1 is selected from phenyl, pyridyl, imidazolyl, imidazopyridyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizine, phthalazinyl, pyridazinyl, triazinyl, isoindolyl , pteridinyl, purinyl, thiadiazolyl, benzofurazolyl, benzopyranyl, benzophenylthio, benzothiazolyl, benzoxazolyl, each independently optionally substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, R4, R5 and R6 are independently H, C1-C3 alkyl at each occurrence.

[0110] In another preferred embodiment, R1 is selected from phenyl, imidazolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzopyranyl, benzophenylthio, benzothiazolyl, benzoxazolyl, each independently optionally substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, and R4, R5 and R6 are independently H, C1-C3 alkyl when they appear each time.

[0111] In another preferred embodiment, R1 is selected from phenyl, imidazolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzopyranyl, benzophenylthio, benzothiazolyl, benzoxazolyl, coumarin, each independently optionally substituted with methyl, ethyl, chloro, fluoro, oxo, CF3, OC1-C2 alkyl, NR 11 R 12 , C6H5 and methyl, ethyl, chlorine, fluorine, OC1-C2 alkyl, NR 11 R 12 Substituted C6H5, R 11 , R 12 is independently at each occurrence H, methyl, ethyl.

[0112] In another preferred embodiment, the R1 is selected from the following formula

[0113]

[0114] In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

[0115] In another preferred embodiment, R1 is selected from phenyl or monocyclic or bicyclic heteroaryl, each of which is independently optionally substituted by C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5 and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, and R4, R5 and R6 are independently H or C1-C3 alkyl at each occurrence.

[0116] In another preferred embodiment, R1 is selected from phenyl or monocyclic or bicyclic heteroaryl, each of which is independently optionally substituted by methyl, ethyl, chlorine, fluorine, CF3, OC1-C2 alkyl, NR5R6, C6H5 and C6H5 substituted by methyl, ethyl, chlorine, fluorine, OC1-C2 alkyl, NR5R6 (preferably monosubstituted), and R5 and R6 are independently H, methyl, and ethyl at each occurrence.

[0117] In another preferred embodiment, R1 is selected from phenyl or a monocyclic or bicyclic heteroaryl group containing one or two heteroatoms selected from N, O and S, each of which is independently optionally substituted by C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5 and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, and R4, R5 and R6 are independently H or C1-C3 alkyl at each occurrence.

[0118] In another preferred embodiment, R1 is selected from phenyl or a monocyclic or bicyclic heteroaryl group containing one or two heteroatoms selected from N, O and S, each of which is independently optionally substituted by methyl, ethyl, chlorine, fluorine, CF3, OC1-C2 alkyl, NR5R6, C6H5 and C6H5 substituted by methyl, ethyl, chlorine, fluorine, OC1-C2 alkyl, NR5R6 (preferably monosubstituted), and R5 and R6 are independently H, methyl, and ethyl at each occurrence.

[0119] In another preferred embodiment, R1 is selected from phenyl, imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, coumarin, each independently optionally substituted by C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, and R4, R5 and R6 are independently H, C1-C3 alkyl at each occurrence.

[0120] In another preferred embodiment, R1 is selected from phenyl, imidazolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, coumarinyl, each independently optionally substituted by methyl, ethyl, chlorine, fluorine, CF3, OC1-C2 alkyl, NR5R6, C6H5 and C6H5 substituted by methyl, ethyl, chlorine, fluorine, OC1-C2 alkyl, NR5R6 (preferably monosubstituted), and R5 and R6 are independently H, methyl, and ethyl at each occurrence.

[0121] In another preferred embodiment, R1 is selected from phenyl, thienyl, oxazolyl, pyrrolyl, coumarinyl, each of which is independently substituted by C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, and R4, R5 and R6 are independently H or C1-C3 alkyl at each occurrence.

[0122] In another preferred embodiment, R1 is selected from phenyl, thienyl, oxazolyl, pyrrolyl, coumarinyl, each of which is independently substituted by methyl, ethyl, chlorine, fluorine, CF3, OC1-C2 alkyl, NR5R6, C6H5 and C6H5 substituted by methyl, ethyl, chlorine, fluorine, OC1-C2 alkyl, NR5R6 (preferably monosubstituted), and R5 and R6 are independently H, methyl, and ethyl at each occurrence.

[0123] In another preferred embodiment, the R1 is selected from the following formula

[0124]

[0125] In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

[0126] In another preferred embodiment, R1 is phenyl or oxazolyl, each of which is independently optionally substituted by C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, and R4, R5 and R6 are independently H or C1-C3 alkyl at each occurrence.

[0127] In another preferred embodiment, R1 is phenyl, coumarin or oxazolyl, each independently optionally substituted by methyl, ethyl, chlorine, fluorine, CF3, OC1-C2 alkyl, NR5R6, C6H5, and C6H5 substituted by methyl, ethyl, chlorine, fluorine, OC1-C2 alkyl, NR5R6 (preferably monosubstituted), R 11 , R 12 is independently at each occurrence H, methyl, ethyl.

[0128] In another preferred embodiment, R1 is phenyl, coumarin or oxazolyl, each independently optionally substituted by methyl, chlorine, fluorine, CF3, OCH3, C6H5, C6H5 substituted by methyl or fluorine (preferably monosubstituted) (preferably monosubstituted).

[0129] In another very preferred embodiment, said R1 is selected from the following formula

[0130]

[0131] In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

[0132] In another very preferred embodiment, said R1 is In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

[0133] In another very preferred embodiment, said R1 is In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

[0134] In another very preferred embodiment, said R1 is In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

[0135] In another very preferred embodiment, said R2 is selected from C5-C 12 Alkyl, C4-C 10 Alkoxy, C1-C3 alkylene-cycloalkyl, C1-C3 alkylene-aryl and C1-C3 alkylene-heteroaryl, independently in the C1-C3 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-; the alkyl, cycloalkyl, aryl and heteroaryl are each independently optionally and preferably substituted by one or more (usually and preferably one or two) substituents selected from C1-C2 alkyl, C1-C2 haloalkyl, oxo, OH, halogen, C1-C2 alkoxy, C6H5 or C6H5 substituted by C1-C3 alkyl or OC1-C3 alkyl.

[0136] In another very preferred embodiment, said R2 is selected from C5-C 12 Alkyl, C4-C 10C1-C3 alkylene-C5-C6 cycloalkyl, C1-C3 alkylene-phenyl, C1-C3 alkylene-biphenyl, C1-C3 alkylene-(monocyclic or bicyclic heteroaryl), independently in the C1-C3 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-, the phenyl, biphenyl, C5-C6-cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted by The monocyclic or bicyclic heteroaryl group is substituted by one or more (usually and preferably one or two) substituents selected from C1-C2 alkyl, C1-C2 haloalkyl, halogen, C1-C2 alkoxy, and preferably, the monocyclic or bicyclic heteroaryl group is selected from imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzopyranyl, benzophenylthio, benzothiazolyl, and benzoxazolyl.

[0137] In another preferred embodiment, said R2 is selected from C5-C 12 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), independently in the C1-C3 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-, the phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted by one or more (usually and preferably one or two) substituents selected from methyl, ethyl, fluorine, chlorine, methoxy, preferably, the monocyclic or bicyclic heteroaryl is selected from imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzopyranyl, benzophenylthio, benzothiazolyl, benzoxazolyl.

[0138] In another preferred embodiment, said R2 is selected from C5-C 12 Alkyl, C4-C 10C1-C2 alkylene-C5-C6 cycloalkyl, C1-C2 alkylene-phenyl, C1-C2 alkylene-biphenyl, C1-C2 alkylene-(monocyclic or bicyclic heteroaryl), independently in the C1-C2 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-, the phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted by one or more (usually and preferably one or two) selected from C1-C2 alkyl, C1-C 2-haloalkyl, halogen, C1-C2 alkoxy substituents, preferably, the monocyclic or bicyclic heteroaryl is selected from imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzopyranyl, benzophenylthio, benzothiazolyl, benzoxazolyl, further preferably, the monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl.

[0139] In another preferred embodiment, said R2 is selected from C5-C 12 Alkyl, C4-C 10 C1-C2 alkylene-C5-C6 cycloalkyl, C1-C2 alkylene-phenyl, C1-C2 alkylene-biphenyl, C1-C2 alkylene-(monocyclic or bicyclic heteroaryl), independently in the C1-C2 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-, the phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted by one or more (usually and preferably one or two) selected from methyl, ethyl, Preferably, the monocyclic or bicyclic heteroaryl is selected from imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzopyranyl, benzophenylthio, benzothiazolyl and benzoxazolyl. Further preferably, the monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl and benzoxazolyl.

[0140] In another preferred embodiment, said R2 is selected from 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 the C1-C2 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-, the phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted by one or more (usually and preferably one or two) substituents selected from methyl, ethyl, fluorine, chlorine, methoxy, and the monocyclic or bicyclic heteroaryl is selected from imidazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzopyranyl, benzophenylthio, benzothiazolyl, benzoxazolyl.

[0141] In another preferred embodiment, said R2 is selected from C5-C 12 Alkyl, C4-C 10 Alkoxy, C1-C2 alkylene-C5-C6 cycloalkyl, C1-C2 alkylene-phenyl, C1-C2 alkylene-biphenyl, C1-C2 alkylene-(monocyclic or bicyclic heteroaryl), independently in the C1-C2 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-, the phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted by one or more (usually and preferably one or two) substituents selected from methyl, ethyl, fluorine, chlorine, methoxy, and the monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl.

[0142] In another preferred embodiment, said R2 is selected from 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 the phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted by one or more (usually and preferably one or two) substituents selected from C1-C2 alkyl, C1-C2 haloalkyl, halogen, C1-C2 alkoxy, preferably, the monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, further preferably, the monocyclic or bicyclic heteroaryl is selected from thiazolyl, indolyl, benzothiazolyl and benzoxazolyl.

[0143] In another preferred embodiment, said R2 is selected from C5-C 12 Alkyl, C4-C 10Alkoxy, C1-C2 alkylene-C5-C6 cycloalkyl, CH2-phenyl, CH2-biphenyl, CH2-O-biphenyl, CH2-(monocyclic or bicyclic heteroaryl), wherein the phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted by one or more (usually and preferably one or two) substituents selected from methyl, ethyl, fluorine, chlorine, methoxy, preferably, the monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, further preferably, the monocyclic or bicyclic heteroaryl is selected from thiazolyl, indolyl, benzothiazolyl and benzoxazolyl.

[0144] In another preferred embodiment, said R2 is selected from 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 the phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted by one or more (usually and preferably one or two) substituents selected from methyl, ethyl, fluorine, chlorine, methoxy, and the monocyclic or bicyclic heteroaryl is selected from isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl.

[0145] In another preferred embodiment, said R2 is selected from 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 the phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted by one or more (usually and preferably one or two) substituents selected from methyl, ethyl, fluorine, chlorine, methoxy, and the monocyclic or bicyclic heteroaryl is selected from thiazolyl, indolyl, benzothiazolyl and benzoxazolyl.

[0146] In another very preferred embodiment, said R2 is selected from

[0147] In the formula, R represents a connection to the CH moiety shown in formula (I).

[0148] In another very preferred embodiment, said R2 is selected from C5-C 12alkyl, C1-C2 alkylene-C5-C6 cycloalkyl, CH2-phenyl, CH2-biphenyl and CH2-O-biphenyl, and a monocyclic or bicyclic heteroaryl selected from thiazolyl, indolyl, benzothiazolyl and benzoxazolyl, wherein the phenyl, biphenyl and monocyclic or bicyclic heteroaryl are optionally substituted with one or two substituents selected from methyl, ethyl, fluorine, chlorine and methoxy.

[0149] In another very preferred embodiment, said R2 is selected from

[0150] In the formula, R represents a connection to the CH moiety shown in formula (I).

[0151] In another very preferred embodiment, said R2 is selected from C5-C 12 alkyl, C1-C2-alkylene-C5-C6-cycloalkyl, CH2-biphenyl and CH2-O-biphenyl, and a bicyclic heteroaryl selected from benzothiazolyl and benzoxazolyl.

[0152] In another very preferred embodiment, said R2 is selected from

[0153]

[0154] In the formula, R represents a connection to the CH moiety shown in formula (I).

[0155] In another very preferred embodiment, said R2 is In the formula, R represents a connection to the CH moiety shown in formula (I).

[0156] In another very preferred embodiment, said R2 is In the formula, R represents a connection to the CH moiety shown in formula (I).

[0157] In another preferred embodiment, said R3 is

[0158] In the formula,

[0159] R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 Two of them, together with the carbon atoms to which they are attached, form a carbocyclic ring;

[0160] R 11 and R 12are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxy or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form heteroaryl or heterocyclyl at each occurrence, each independently optionally and preferably substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence, and the arrow indicates the connection to the C(O)-moiety shown in formula (I).

[0161] In another preferred embodiment, said R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 Two of them together with the carbon atoms to which they are attached form a carbocyclic or heterocyclic ring, preferably a carbocyclic ring; R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxyl or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form at each occurrence a monocyclic or bicyclic heteroaryl or a monocyclic or bicyclic heterocyclyl, the monocyclic or bicyclic heteroaryl or monocyclic or bicyclic heterocyclyl being preferably selected from pyridyl, imidazolyl, imidazopyridyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl , isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, tetrahydroisoquinolyl, indolyl, benzimidazolyl, indazolyl, indolizinyl, pyridazinyl, triazinyl, isoindolyl, purinyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, homopiperazinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, dihydrooxazolyl, pyrazolyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, each independently optionally substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence.

[0162] In another preferred embodiment, R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10The two of them together with the carbon atoms to which they are attached form a monocyclic carbocycle or a monocyclic heterocycle, preferably a monocyclic carbocycle, further preferably, the monocyclic carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and again further preferably, the monocyclic carbocycle is cyclobutyl or cyclopentyl, R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxyl or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form at each occurrence a monocyclic or bicyclic heteroaryl or a monocyclic or bicyclic heterocyclyl, the monocyclic or bicyclic heteroaryl or monocyclic or bicyclic heterocyclyl being preferably selected from pyridyl, imidazolyl, imidazopyridyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl , isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, tetrahydroisoquinolyl, indolyl, benzimidazolyl, indazolyl, indolizinyl, pyridazinyl, triazinyl, isoindolyl, purinyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, homopiperazinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, dihydrooxazolyl, pyrazolyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, each independently optionally substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence.

[0163] In another preferred embodiment, R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 The two of them together with the carbon atoms to which they are attached form a monocyclic carbocycle or a monocyclic heterocycle, preferably a monocyclic carbocycle, further preferably, the monocyclic carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and again further preferably, the monocyclic carbocycle is cyclobutyl or cyclopentyl, R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxyl or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form a monocyclic heteroaryl or a monocyclic heterocyclyl at each occurrence, each independently optionally substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence.

[0164] In another preferred embodiment, R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 The two of them together with the carbon atoms to which they are attached form a monocyclic carbocycle or a monocyclic heterocycle, preferably a monocyclic carbocycle, further preferably, the monocyclic carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and again further preferably, the monocyclic carbocycle is cyclobutyl or cyclopentyl, R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxyl or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form a monocyclic heteroaryl or a monocyclic heterocyclyl at each occurrence, the monocyclic heteroaryl or the monocyclic heterocyclyl containing one or two heteroatoms selected from nitrogen, oxygen and sulfur (including R 11 and R 12 The nitrogen atom connected thereto) are each independently optionally substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence.

[0165] In another preferred embodiment, R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 The two of them together with the carbon atoms to which they are attached form a monocyclic carbocycle or a monocyclic heterocycle, preferably a monocyclic carbocycle, further preferably, the monocyclic carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and again further preferably, the monocyclic carbocycle is cyclobutyl or cyclopentyl, R 11 and R 12are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxy or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form at each occurrence a monocyclic heteroaryl or a monocyclic heterocyclyl, the monocyclic heteroaryl or the monocyclic heterocyclyl being selected from pyridyl, 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 optionally substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence.

[0166] In another preferred embodiment, R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 The two of them together with the carbon atoms to which they are attached form a monocyclic carbocycle, further preferably, the monocyclic carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and again further preferably, the monocyclic carbocycle is cyclobutyl or cyclopentyl, R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxyl or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form a monocyclic heterocycle at each occurrence, each independently optionally substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence.

[0167] In another preferred embodiment, R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 The two of them together with the carbon atoms to which they are attached form a monocyclic carbocycle or a monocyclic heterocycle, preferably a monocyclic carbocycle, further preferably, the monocyclic carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and again further preferably, the monocyclic carbocycle is cyclobutyl or cyclopentyl, R11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxyl or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form a monocyclic heterocyclic group at each occurrence, the monocyclic heterocyclic group containing one or two heteroatoms selected from nitrogen, oxygen and sulfur (including R 11 and R 12 The nitrogen atom connected thereto) are each independently optionally substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence.

[0168] In another preferred embodiment, R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 The two of them together with the carbon atoms to which they are attached form a monocyclic carbocycle, further preferably, the monocyclic carbocycle is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and again further preferably, the monocyclic carbocycle is cyclobutyl or cyclopentyl, R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted with halogen, hydroxy or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently at each occurrence form a monocyclic heterocycle 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 optionally substituted with halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence.

[0169] In another preferred embodiment, R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10Two of the groups together with the carbon atoms to which they are attached form a monocyclic carbocyclic ring selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, preferably, the monocyclic carbocyclic ring is cyclobutyl or cyclopentyl, R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxyl or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form at each occurrence a monocyclic heterocycle selected from piperidinyl, morpholinyl, thiomorpholinyl, preferably piperidinyl or morpholinyl, further preferably morpholinyl, each independently optionally substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence.

[0170] In another very preferred embodiment, said R3 is selected from

[0171]

[0172] In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

[0173] In another very preferred embodiment, said R3 is selected from

[0174]

[0175] In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

[0176] In another very preferred embodiment, said R3 is In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

[0177] In another very preferred embodiment, the compound of formula (I) is selected from:

[0178] 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-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide;

[0179] 6027: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0180] 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]carboxamido}decanamide;

[0181] 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]carboxamido}-3-phenylpropionamido]pentanamide;

[0182] 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]carboxamido}hexanamide;

[0183] 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-{[(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]decanoamide;

[0184] 7059: (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-methylpentanamide;

[0185] 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]carboxamido}-3-({4'-methoxy-[1,1'-biphenyl]-4-yl}oxy)propionamido]-4-methylpentanamido]-2-methylpropanamido}-4-methylpentanamido]-4-methylpentanamide;

[0186] 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]carboxamido}-3-(6-methoxy-1,3-benzothiazol-2-yl)propionamido]-4-methylpentanamide;

[0187] 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]carboxamido}propionamido]-4-methylpentanamide;

[0188] 7850: (2S)-2-[(2S)-3-(1,3-benzoxazol-2-yl)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}propionamido]-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;

[0189] 8341: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0190] 8342: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0191] 8343: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0192] 21164: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0193] 21287: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(7-methoxy-2-oxo-2H-chromene-3-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-methylpentanamide;

[0194] 21314: (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0195] 21315: (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]carboxamido}butyramido]-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;

[0196] 21359: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]carboxamido}butyramido]-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;

[0197] 21362: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]carboxamido}butyramido]-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;

[0198] 21365: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]carboxamido}butyramido]-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;

[0199] 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]carboxamido}butyramido]-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;

[0200] 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]carboxamido}butanamide;

[0201] Preferably, the compound of formula (I) is:

[0202] 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-methylethyl)carbamoyl]-1-methylethyl}carbamoyl)-3-methylbutyl]carbamoyl}-3-methylbutyl]carbamoyl}-1-methylethyl)-4-methylpentanamide;

[0203] 6027: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0204] 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]carboxamido}decanamide;

[0205] 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]carboxamido}-3-({4'-methoxy-[1,1'-biphenyl]-4-yl}oxy)propionamido]-4-methylpentanamido]-2-methylpropanamido}-4-methylpentanamido]-4-methylpentanamide;

[0206] 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]carboxamido}propionamido]-4-methylpentanamide;

[0207] 8343: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0208] 21314: (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0209] 21359: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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; or

[0210] 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0211] In another very preferred embodiment, the compound of formula (I) is 6027: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0212] In another very 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]formamido}decanamide.

[0213] In another very 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}propionamido]-4-methylpentanamide.

[0214] In another very preferred embodiment, the compound of formula (I) is 8343: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0215] In another very preferred embodiment, the compound of formula (I) is 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0216] In another very preferred embodiment, the compound is selected from the formula described in 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 described in claim 11 and is selected from 6025, 6027, 6253, 7824, 7848, 8343, 21314, 21359 or 21369. In another very preferred embodiment, the compound of formula (I) is formula 6027 described in claim 11. In another very preferred embodiment, the compound of formula (I) is the formula 6253 described in claim 11. In another very preferred embodiment, the compound of formula (I) is the formula 7848 described in claim 11. In another very preferred embodiment, the compound of formula (I) is the formula 8343 described in claim 11. In another very preferred embodiment, the compound of formula (I) is the formula 21369 described in claim 11.

[0217] In another aspect, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: 6781:

[0219] 7824:

[0221] 7844:

[0223] 7850:

[0225] 21164:

[0227] 21287:

[0229] 21314:

[0231] 21315:

[0233] 21359:

[0235] 21362:

[0237] 21365:

[0239] 21369:

[0241] 21372:

[0243]

[0244] Preferably, the compound of formula (I) is selected from the following compounds: 7824:

[0245] 21314:

[0247] 21359:

[0249] 21369:

[0251]

[0252] Further preferably, the compound is 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0253] In another aspect, the present invention provides a compound of formula (I), wherein the compound is selected from:

[0254] 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]carboxamido}hexanamide;

[0255] 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]carboxamido}-3-({4'-methoxy-[1,1'-biphenyl]-4-yl}oxy)propionamido]-4-methylpentanamido]-2-methylpropanamido}-4-methylpentanamido]-4-methylpentanamide;

[0256] 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]carboxamido}-3-(6-methoxy-1,3-benzothiazol-2-yl)propionamido]-4-methylpentanamide;

[0257] 7850: (2S)-2-[(2S)-3-(1,3-benzoxazol-2-yl)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}propionamido]-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;

[0258] 21164: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0259] 21287: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(7-methoxy-2-oxo-2H-chromene-3-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-methylpentanamide;

[0260] 21314: (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0261] 21315: (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]carboxamido}butyramido]-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;

[0262] 21359: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]carboxamido}butyramido]-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;

[0263] 21362: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]carboxamido}butyramido]-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;

[0264] 21365: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]carboxamido}butyramido]-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;

[0265] 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]carboxamido}butyramido]-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;

[0266] 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]carboxamido}butanamide;

[0267] Preferably, the compound is selected from:

[0268] 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]carboxamido}-3-({4'-methoxy-[1,1'-biphenyl]-4-yl}oxy)propionamido]-4-methylpentanamido]-2-methylpropanamido}-4-methylpentanamido]-4-methylpentanamide;

[0269] 21314: (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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;

[0270] 21359: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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

[0271] 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]carboxamido}butyramido]-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; and

[0272] Further preferably, the compound is 21369: (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0273] The present invention provides a compound of formula (I) for use in a method for topical treatment of skin cancer in a mammal, wherein the method comprises topically administering to the mammal an effective amount of the compound or a pharmaceutically acceptable salt thereof. In a preferred embodiment, the mammal is a human.

[0274] In another preferred embodiment, the topical application is the application of the compound or its pharmaceutically acceptable salt to the skin of a mammal. In a preferred embodiment, the topical application is the application of the compound or its pharmaceutically acceptable salt to the skin of a human. In another preferred embodiment, the topical application is the application of the compound or its pharmaceutically acceptable salt directly to the skin of a mammal. In a preferred embodiment, the topical application is the application of the compound or its pharmaceutically acceptable salt directly to the skin of a human.

[0275] In another preferred embodiment, the skin cancer is selected from melanoma skin cancer, non-melanoma skin cancer and skin lymphoma and any pre-invasive form thereof. In another preferred embodiment, the skin cancer is selected from melanoma, non-melanoma skin cancer and any pre-invasive form thereof, preferably, the non-melanoma skin cancer is basal cell carcinoma, Merkel cell carcinoma or skin squamous cell carcinoma, further preferably, the non-melanoma skin cancer is basal cell carcinoma or skin squamous cell carcinoma or any pre-invasive form thereof. In another preferred embodiment, the skin cancer is selected from melanoma skin cancer, non-melanoma skin cancer and skin lymphoma and any pre-invasive form thereof, preferably, the non-melanoma skin cancer is basal cell carcinoma, Merkel cell carcinoma or skin squamous cell carcinoma, further preferably, the non-melanoma skin cancer is basal cell carcinoma or skin squamous cell carcinoma or any pre-invasive form thereof. In another preferred embodiment, the skin cancer is selected from melanoma, non-melanoma skin cancer and any pre-invasive form thereof, preferably, the non-melanoma skin cancer is basal cell carcinoma, Merkel cell carcinoma or cutaneous squamous cell carcinoma, further preferably, the non-melanoma skin cancer is basal cell carcinoma or cutaneous squamous cell carcinoma or any pre-invasive form thereof.

[0276] In another preferred embodiment, the skin cancer is selected from melanoma skin cancer and non-melanoma skin cancer, preferably, the non-melanoma skin cancer is basal cell carcinoma or squamous cell carcinoma of the skin. In another preferred embodiment, the skin cancer is melanoma. In another preferred embodiment, the skin cancer is non-melanoma skin cancer, preferably, the non-melanoma skin cancer is basal cell carcinoma, Merkel cell carcinoma or squamous cell carcinoma of the skin or any pre-invasive form thereof, further preferably, the non-melanoma skin cancer is basal cell carcinoma or squamous cell carcinoma of the skin or any pre-invasive form thereof. In another preferred embodiment, the skin cancer is basal cell carcinoma. In another preferred embodiment, the skin cancer is squamous cell carcinoma of the skin. 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.

[0277] In another preferred embodiment, the cutaneous squamous cell carcinoma (cSCC) is invasive cSCC. In another preferred embodiment, the cutaneous squamous cell carcinoma (cSCC) is metastatic cSCC. In another preferred embodiment, the basal cell carcinoma is selected from 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 another preferred embodiment, the basal cell carcinoma is invasive basal cell carcinoma. In another preferred embodiment, the basal cell carcinoma is nodular basal cell carcinoma. In another embodiment, the basal cell carcinoma is selected from cystic basal cell carcinoma, scar basal cell carcinoma (also known as "morphoplasmic basal cell carcinoma" or "morphological basal cell carcinoma"), micronodular basal cell tumors, pigmented basal cell carcinoma, erosive ulcers (also known as "Jacob's ulcers"), Pinkus fibroepithelioma, polypoid basal cell carcinoma, pore-like basal cell carcinoma and abnormal basal cell carcinoma.

[0278] In another preferred embodiment, the skin cancer is a pre-invasive form of non-melanoma skin cancer, the pre-invasive form is selected from cutaneous squamous cell carcinoma in situ (cSCCis, also known as "Bowen's disease") and precancerous actinic keratosis (AK). In another preferred embodiment, the skin cancer is a pre-invasive form of non-melanoma skin cancer, the pre-invasive form is cutaneous squamous cell carcinoma in situ (cSCCis, also known as "Bowen's disease"). In another preferred embodiment, the skin cancer is a pre-invasive form of non-melanoma skin cancer, the pre-invasive form is precancerous 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 a 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 a cutaneous lymphoma, and the cutaneous lymphoma is cutaneous T-cell lymphoma (CTCL). In another preferred embodiment, the skin cancer is a cutaneous lymphoma, and 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, and the pre-invasive form is actinic keratosis (AK). In another preferred embodiment, the skin cancer is a pre-invasive form of non-melanoma skin cancer, and the pre-invasive form is cSCC in situ (cSCCis).

[0279] The compounds and methods described herein are advantageously used to inhibit the proliferation of skin cancer cells. Preferably, the compounds and methods provide a topical treatment for skin cancer in mammals, preferably humans. The method provides a topical treatment for various types of skin cancer (particularly basal cell carcinoma, squamous cell carcinoma of the skin or melanoma or cutaneous lymphoma). Therefore, topical administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising them provides a treatment for skin cancer in a patient.

[0280] Example

[0281] 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 at The peptides were synthesized on Fmoc-β-Ala Wang resin (0.70 mmol g) using a Liberty Blue microwave peptide synthesizer (CEM Inc., Matthews, NC) using Fmoc solid phase technology. -1 ) on a scale of 0.25 mmol or 0.10 mmol, repeating the coupling, deprotection and washing steps. Coupling was 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) were coupled at 90°C for 4 min with microwave irradiation. For the second AIB coupling, double coupling was 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 were microwave irradiated at 90°C for 1 min. After synthesis, the peptide was cleaved from the resin by treatment with a cleavage mixture consisting of TFA / H2O (95:5) (1 mL / 0.1 g resin) at ambient temperature for 90 min. The suspended resin was removed by filtration and concentrated in vacuo. The crude peptide was dissolved in acetonitrile and purified using an ISCO chromatography system with a mobile phase of H2O / acetonitrile (95:5, v / v) and acetonitrile / H2O with a gradient of ACN from 10% to 100% to obtain a peptide acid with a purity of ≥90%. The purified peptide acid (1.0 equiv) was coupled with a suitable primary or secondary amine (2.0 equiv) in solution in the presence of DIPEA (3.0 equiv) and COMU (2.0 equiv) or HATU to obtain the final peptide. The solvent was partially removed and the crude peptide was directly purified by a Gilson PLC 2020 personal purification system as described above to obtain a final product with a purity of ≥95%.

[0282] High-resolution mass spectrometry was performed on an Agilent Technologies 6530Q-TOF. The quality 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 H2O-acetonitrile (Solvent A, 97:3 v / v, LC-MS Ultra UHPLC grade, Sigma-Aldrich, Germany) and 0.1% formic acid (LC-MS grade, Sigma-Aldrich, Germany) and acetonitrile-H2O (Solvent B, 97:3 v / v, LC-MS Ultra The elution was performed with a flow rate of 0.6 ml / min, an injection volume of 10 μL, and a gradient elution program of linear increment from 100% A to 100% B in 3 min and return to the initial state in the next 1 min. Waters TQD QCA065 MS detection of the analytes was performed on a triple quadrupole mass spectrometer in positive and negative ion modes with a mass range of 100 to 1500 m / z.

[0283] Example 1

[0284] General procedure for the synthesis of compounds of formula (I)

[0285] Peptides were synthesized starting from Fmoc-β-Ala linked to Wang resin as shown in Scheme 1 using a Liberty Blue microwave peptide synthesizer (CEM Corporation, Matthews, NC) using Fmoc solid phase technology with repeated coupling, deprotection and washing steps. Couplings were performed as follows: Fmoc-L-amino acid or blocking 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) were coupled by 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 were microwave irradiated at 90°C for 1 min. Different peptides were assembled on a solid support in typically and preferably 9 steps by sequentially using the building blocks listed in Table 1 (amino acids used in Scheme 1, steps 1 to 8) and the free acid derivatives listed in Table 2 (Scheme 1, step 9). After synthesis, the peptides were cleaved from the resin by treatment with a cleavage mixture consisting of TFA / H2O (95:5) (1 mL / 0.1 g resin) at ambient temperature for 90 min. The suspended resin was removed by filtration and concentrated in vacuo. The crude peptide was dissolved in acetonitrile and purified by an ISCO chromatography system. The purified peptide acid was coupled in solution with the appropriate primary or secondary amine reported in Table 3 in the presence of DIPEA and COMU or HATU (Scheme 1, step 10) to give the final peptide, which was then directly purified on a Gilson PLC 2020 Personal Purification System.

[0286] Solution 1 : According to the general scheme, the compound of formula (I) is synthesized

[0287]

[0288] Table 1: Amino acid building blocks used to synthesize compounds of formula (I)

[0289]

[0290]

[0291]

[0292] Table 2: Free acids used for the synthesis of compounds of formula (I) (Scheme 1, step 9)

[0293]

[0294] Table 3: Primary or secondary amines used in the synthesis of compounds of formula (I) (Scheme 1, step 10)

[0295]

[0296]

[0297] Example 2

[0298] Synthesis of amino acids as building blocks for compounds of formula (I)

[0299] Some of the amino acids required for the synthesis of compounds of formula (I) are not commercially available. In such cases, they are synthesized. Scheme 2 shows these syntheses.

[0300] Solution 2 :Synthesis of non-commercially available amino acids

[0301]

[0302] The synthesis of amino acids 8a to 8c 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 concentrated H2SO4 droplets were added. The reaction mixture was refluxed for 3 hours. It was then cooled to RT, the pH was adjusted to 8.0 with 20% w / v Na2CO3 aqueous solution, 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.

[0303] In the next step, 2 (1.0 eq.) and 4-DMAP (0.12 eq.) were dissolved in anhydrous pyridine (0.6 ml / mmol of 2) and cooled to -5°C. p-TsOH (3.3 eq.) was added to the solution and the resulting yellow reaction mixture was stirred under nitrogen for 16 hours. 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 KHSO4 solution (2×10 mL), saturated NaHCO3 solution (2×100 mL) and brine (2×100 mL). The organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to give the tosylate 3. Subsequently, an acetone solution of sodium iodide (5.0 eq.) (0.0 mL / mmol of NaI) was added dropwise to an acetone solution of tosylate 3 (1.0 eq.) (1.0 mL / mmol of 3) under nitrogen. The resulting yellow solution was stirred at RT for 48 hours. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in chloroform (100 mL), washed with water (2×50 mL), saturated Na2S2O3 solution (2×50 mL) and brine (2×30 mL). The organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to give 4 as a light yellow solid. Then, Zn(0) (powder) (4.0 eq.) was weighed into a flame-dried flask and I2 (0.22 eq.) (1.0 ml / mmol of Zn(0)) dissolved in anhydrous DMF was added. Iodine 4 (1.0 eq.) and I2 were added and the reaction mixture was stirred until complete consumption (TLC indicated).

[0304] Most of the supernatant was removed with a syringe and added directly to a pre-stirred solution of Pd2(dba)3 (0.05mmol), Xphos (0.10mmol) and the respective halide 6a-6c (1.3mmol) in anhydrous DMF (1.0ml / mmol of Zn(0)). The reaction mixture was stirred at 40°C for 2 hours. The mixture was then poured into saturated NaHCO3 solution (10mL) and extracted with EtOAc (2×10mL). The combined organic layers were washed with saturated NaCl solution, dried over MgSO4, concentrated and dried in vacuo. The crude product was purified by flash column chromatography to provide esters 7a-7c. Finally, 7a-7c (1.0 equivalent) was added to the i To a suspension in a 0.8M CaCl solution in PrOH / H2O (7 at 20 ml / mmol), NaOH (1.2 equivalents) was added and the mixture was stirred overnight at RT. The reaction was quenched by adding acetic acid and the solvent was removed under reduced pressure. Final amino acids 8a-8c were obtained by RP chromatography of the crude product.

[0305] Example 3

[0306] Synthesis of compounds of formula (I)

[0307] The synthesis of compounds of formula (I) has been described in EP 3'345'917A1, including the synthesis of compounds 6025, 6027 and 6253. Similarly, unless otherwise stated, the compounds of formula (I) described further hereafter have been synthesized according to the general procedure described in Scheme 1. Each synthesized compound is identified by its compound number (4 or 5 digits), its structural formula and the IUPAC name generated by MarvinSketch software. In addition, the synthesis of 6328, 6781, 7058, 7059, 7824, 7844, 7848, 7850, 8341, 8342, 8343, 21164, 21189, 21287, 21314, 21315, 21359, 21362, 21365, 21368, 21369 and 21372 is described by the used building blocks, acids and amines disclosed in Tables 1-3. In addition, experimentally measured molecular weights are provided. Finally, for each specifically identified compound of formula (I), the characterization of substituents R1, R2 and R3 according to formula (I) is given, and the residue R in the definition of the substituents R1, R2 and R3 corresponds to and represents the corresponding connection in formula (I). 6025:

[0309]

[0310] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(2-methyl-1,3-oxazole-4-carbonyl)pyrrolidin-2-yl]formamido}butyramido]-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

[0311] 6027:

[0313]

[0314] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0315]

[0316] 6253:

[0318]

[0319] (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]carboxamido}decanamide.

[0320] 6328:

[0322]

[0323] (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]carboxamido}-3-phenylpropionamido]pentanamide.

[0324]

[0325]

[0326] Compound Molecular mass Quality of experimental measurements 6328[M+H]+ 1147.72 1147.76

[0327] 6781:

[0329]

[0330] (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]carboxamido}hexanamide.

[0331]

[0332] Compound Molecular mass Quality of experimental measurements 6781[M+H]+ 1113.74 1113.60

[0333] 7058:

[0335]

[0336] (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-{[(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]decanoamide.

[0337]

[0338] Compound Molecular mass Quality of experimental measurements 7058[M+H]+ 1115.76 1115.70

[0339]

[0340] Solution 4 :Synthesis of compound 7058

[0341]

[0342]

[0343] Compound 7058 was prepared as shown in Scheme 4. More analytically: To compound 7057 (0.1 mmol) was added formic acid (3.7 mL) and the reaction mixture was stirred at RT for 12 hours. Upon completion, the formic acid was removed in vacuo to obtain 7058 without further purification. 7059:

[0345]

[0346] (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-methylpentanamide.

[0347]

[0348] Compound Molecular mass Quality of experimental measurements 7059[M+H]+ 1153.77 1153.50

[0349]

[0350] 7824:

[0352]

[0353] (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)propionamido]-4-methylpentanamido]-2-methylpropanamido}-4-methylpentanamido]-4-methylpentanamide.

[0354]

[0355] Compound Molecular mass Quality of experimental measurements 7824[M+H]+ 1282.76 1282.51

[0356] 7844:

[0358]

[0359] (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]carboxamido}-3-(6-methoxy-1,3-benzothiazol-2-yl)propionamido]-4-methylpentanamide.

[0360]

[0361] Compound Molecular mass Quality of experimental measurements 7844[M+H]+ 1247.58 1248.34

[0362] 7848:

[0364]

[0365] (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}propionamido]-4-methylpentanamide.

[0366]

[0367] Compound Molecular mass Quality of experimental measurements 7848[M+H]+ 1235.68 1236.38

[0368] 7850:

[0370]

[0371] (2S)-2-[(2S)-3-(1,3-benzoxazol-2-yl)-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}propionamido]-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.

[0372]

[0373]

[0374] Compound Molecular mass Quality of experimental measurements 7850[M+H]+ 1235.68 1236.38

[0375] 8341:

[0377]

[0378] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0379]

[0380] Compound Molecular mass Quality of experimental measurements 8341[M+H]+ 1182.77 1183.04

[0381] 8342:

[0383]

[0384] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0385]

[0386] Compound Molecular mass Quality of experimental measurements 8342[M+H]+ 1190.75 1191.09

[0387] 8343:

[0389]

[0390] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0391]

[0392] Compound Molecular mass Quality of experimental measurements 8343[M+H]+ 1140.75 1141.04

[0393] 21164:

[0395]

[0396] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0397]

[0398] Compound Molecular mass Quality of experimental measurements 21164[M+H]+ 1195.60 1195.13

[0399]

[0400] 21189 (reference example):

[0401]

[0402] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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-methylpentanamide.

[0403]

[0404] Compound Molecular mass Quality of experimental measurements 21189[M+H]+ 1123.65 1123.02

[0405] 21287:

[0407]

[0408] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(7-methoxy-2-oxo-2H-chromene-3-carbonyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0409]

[0410] Compound Molecular mass Quality of experimental measurements 21287[M+H]+ 1261.64 1261.22

[0411] 21314:

[0413]

[0414] (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0415]

[0416] Compound Molecular mass Quality of experimental measurements 21314[M+H]+ 1195.60 1195.13

[0417]

[0418] 21315:

[0420]

[0421] (3S)-3-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0422]

[0423] Compound Molecular mass Quality of experimental measurements 21315[M+H]+ 1223.65 1223.49

[0424] 21359:

[0426]

[0427] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0428]

[0429] Compound Molecular mass Quality of experimental measurements <![CDATA[21359[M+H-CF3]+]]> 1166.53 1166.71

[0430] 21362:

[0432]

[0433] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0434]

[0435] Compound Molecular mass Quality of experimental measurements <![CDATA[21362[M+H-CF3]+]]> 1166.53 1166.87

[0436] 21365:

[0438]

[0439] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0440]

[0441] Compound Molecular mass Quality of experimental measurements 21365[M+H]+ 1194.59 1194.92

[0442]

[0443] 21368 (reference example):

[0444]

[0445] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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-methylpentanamide.

[0446]

[0447] Compound Molecular mass Quality of experimental measurements 21368[M+H]+ 1223.65 1223.07

[0448] 21369:

[0450]

[0451] (2S)-2-[(2S)-4-cyclohexyl-2-{[(2S)-1-(4-fluorobenzoyl)pyrrolidin-2-yl]formamido}butyramido]-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.

[0452]

[0453] Compound Molecular mass Quality of experimental measurements 21369[M]+ 1206.48 1206.90

[0454] 21372:

[0456]

[0457] (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]carboxamido}butanamide.

[0458]

[0459] Compound Molecular mass Quality of experimental measurements 21372[M]+ 1258.32 1258.63

[0460]

[0461] Example 4

[0462] Skin penetration studies

[0463] Skin permeation experiments were conducted on pig skin using Franz diffusion cells (FDC), exemplarily using the highly preferred compound 6027 of the present invention.

[0464] Chemicals

[0465] Acetonitrile (ACN), formic acid (FA), ethanol (EtOH), and dimethyl sulfoxide (DMSO) were all of HPLC grade and used together with purified water, which was purchased from Satorius Stedim Biotech GmbH (Göttingen, Germany). pro ultrapure water system is produced by reverse osmosis.

[0466] Skin preparation

[0467] Skin from pig ears is widely used and accepted for skin penetration studies. Freshly slaughtered pig ears, no older than 5 months, of both sexes, were obtained from Bell Schweiz AG slaughterhouse (Basel, Switzerland). The ears were cleaned with tap water and the hair was clipped with an electric hair clipper. The skin on the back of the ear was separated from the underlying cartilage using a scalpel. After the skin was cleaned again, it was stored in aluminum foil at -20°C for no more than one month.

[0468] Penetration experiment

[0469] After thawing the frozen skin, it was cut into appropriate circles and the skin thickness was measured with a caliper. The skin was mounted into the FDC, tightly sealed with Teflon tape and fixed on a specially designed stand. The receptor chamber was then equipped with a magnetic stirrer, filled with the placebo preparation and equilibrated for one hour. To check the integrity of the barrier and ensure that there was no invisible damage to the prepared skin, TEWL (transepidermal water loss) was measured. The FDC was then placed in a water bath at +32°C ± 1, on a magnetic stirrer (500 rpm). Due to the differences in each FDC, the skin application area for drug penetration of each pool was measured and calculated in advance. Therefore, in the infinite dose experiment, the amount of 6027 in the donor chamber must be adjusted to 284.1 μL / cm 2 .

[0470] At predetermined time points, 50 μL samples were withdrawn from the receptor chamber. The missing volume was replaced by the corresponding placebo preparation.

[0471] At the end of the permeation experiment and before reassembling the FDC, the donor chamber was flushed with 10 mL ACN:H2O (55:45).

[0472] Skin Extraction

[0473] To extract 6027 from the skin, the frozen skin was cut into small pieces and ground using a cryogenic grinder from Spex SamplePrep (Metuchen, USA). Grind. Set to 10 minutes precooling, perform 4 cycles at a rate of 10 CPS, 2 minutes each. The resulting powder was resuspended in 2 mL ACN:H2O (55:45). After vortexing, the mixture was sonicated using a Branson Sonifier 250 (Model: 101-cx03-197) from BransonUltrasonics Corporation (Danbury, USA). The ultrasonic setting was an output control of 2 and a duty cycle of 30% for 60 seconds. After shaking the sample with a horizontal shaker at 37°C for 3 minutes, the skin powder was separated by centrifugation until the supernatant became clear. The extraction process was repeated twice more.

[0474] High performance liquid chromatography-mass spectrometry (HPLC-MS)

[0475] The concentration of 6027 was determined using an Agilent Technologies HPLC-MS. The system was equipped with an Infinity LABLC / MSD XT G6135B, a degasser G1379, an isocratic pump G1310A, an autosampler G1329A with a thermostat G1330B, a column oven G1316, and an Agilent C18 reversed-phase column, Zobrax SB-C18 Narrow Bore 2.1×150mM 5Micron.

[0476] The mobile phase was a mixture of ACN:H2O:FA (55:45:0.1) [v / v]. The flow rate was set to 0.5 mL / min, the temperature in the autosampler was set to 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:H2O (55:45) to within the calibration range. Other conditions and details, such as skin thickness, are listed in Table 4.

[0477] Table 4: Quantification of 6027 in full thickness porcine skin after 48 hours using a Franz cell. Three replicates were performed and analyzed by HPLC-MS after skin extraction. Data are shown as a percentage of total 6027.

[0478]

[0479] The FDC skin permeability of 6027 was shown to be 16%. This is particularly surprising because 6027 and other compounds of formula (I) not only have a molecular weight above 1000 Da, but also 6027 has an octanol / water partition coefficient log D of 4.56 and a low water solubility of 0.022 mg / ml.

[0480] Example 5

[0481] In vitro antiproliferation effect on skin squamous cell carcinoma cells

[0482] 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. Squamous cell carcinoma of the skin is not usually life-threatening, although it can be aggressive. If not treated, Squamous cell carcinoma of the skin may grow larger or spread to other parts of the body, causing serious complications. Most Squamous cell carcinomas of the skin are caused by prolonged exposure to ultraviolet (UV) radiation, either from the sun or from tanning beds or lamps. Avoiding UV rays can help reduce the risk of Squamous cell carcinoma of the skin and other forms of skin cancer. Squamous cells are found in many places in the body, and Squamous cell carcinoma can develop wherever squamous cells are found. Squamous cell carcinoma of the skin and cutaneous squamous cell carcinoma (cSCC) each refer to cancers that develop in the squamous cells of the skin.

[0483] Several representative and preferred compounds of the invention have been tested and shown to be active in vitro against the skin squamous cell carcinoma cell line SCC12.

[0484] Methods. Skin squamous cell carcinoma cell culture SCC12 was cultured in keratinocyte SFM medium (KSFM, catalog number 17005042, Gibco) supplemented with human recombinant epidermal growth factor 1-53 (EGF1-53) and bovine pituitary extract (BFE). For library screening, cells were distributed to 96-well plates at a cell density of 7500 cells (90 μl / well) and adhered overnight in a cell culture incubator at 37°C and 5% CO2. The compounds were serially diluted to 1000 times in DMSO (10mM, 5mM, 1mM, 500μM, 250μM, 100μM, 10μM, 1μM). 2μl of these dilutions were further diluted in 198μl of keratinocyte growth medium with supplements added, mixed thoroughly, and then 10μl was further distributed in triplicate to 96-well plates containing 90μl 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, 1 nM.

[0485] The plates were incubated for an additional 72 hours and the cell culture medium was replaced with a medium containing a 0.1% resazurin sodium salt stock solution (stock solution: 0.015 mg / ml in PBS, catalog number R7017, Sigma-Aldrich). The cell culture plates were incubated in a cell culture incubator at 37°C and 5% CO2 for 2-3 hours. The cell culture plates were read using a fluorescence-based plate reader (Infinite M200Pro, Tecan, Switzerland) using an excitation light between 530-560 nM and an emission light of 590 nM. Calculations and plots were performed using the GraphPad Prism program. The viability of the wells treated with the vehicle was set to 100%.

[0486] Table 5: Half maximal inhibitory concentration (IC) of the compounds of the present invention in the skin squamous cell carcinoma cell line SCC12 50 )

[0487]

[0488]

[0489] Results. The preferred invention tested showed activity in the nM range and IC against cutaneous squamous cell carcinoma SCC12 in vitro 50 The value confirms its usefulness as a drug for treating skin cancer, especially squamous cell carcinoma of the skin.

[0490] Example 6

[0491] In vivo antiproliferative assay

[0492] The in vitro antiproliferative effect of the preferred compound of the invention, 6027, has been tested against 6 cells of skin cancer origin, including cells from skin melanoma (M150672, M150507, M010817), cells of skin squamous cell carcinoma cell lines (SCC13 and A431_SCC) and cells of cutaneous T-cell lymphoma (MyLa_CTCL). Likewise, an antiproliferative effect was established against healthy skin dermal fibroblasts.

[0493] Methods. For library screening, cells were dispensed into 96-well plates at 90 μl / well (cell density (cells / well): M150672 (2000), M150507 (1500), M010817 (1500), M130107 (2000), SCC13 (11000), A431_SCC (1500)), MyLa_CTCL (10000) and allowed to adhere overnight in a cell culture incubator at 37°C and 5% CO2.

[0494] To maintain healthy human fibroblast cultures, Dulbecco's Modified Eagle's Medium (DMEM, catalog number 11966025, Gibco) supplemented with 10% heat-inactivated fetal bovine serum (catalog number S006420H01, BioWest) was used. To maintain melanoma cell cultures, RPMI-1640 (catalog number R0883, Gibco) supplemented with L-glutamine (catalog number 25030-081, Gibco), 1 mM sodium pyruvate (catalog number S8636, Sigma-Aldrich), 10% heat-inactivated fetal bovine serum (catalog number S006420H01, BioWest) and 1% penicillin-streptomycin (catalog number 15140122, Gibco) was used. MyLa_CTCL cell cultures were cultured 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). Skin squamous cell carcinoma cell cultures A431 and SCC13 were cultured in Dulbecco's modified Eagle's medium (DMEM, catalog number 11966025, Gibco) supplemented with 10% heat-inactivated fetal bovine serum (catalog number S006420H01, BioWest).

[0495] To determine the half-maximal inhibitory concentration (IC 50 ), the compounds were serially diluted to 1000-fold in DMSO (10mM, 5mM, 1mM, 500μM, 250μM, 100μM, 10μM, 1μM). 2μl of these dilutions were further diluted in 198μl of growth medium containing supplements, mixed thoroughly, and 10μl was further distributed in triplicate to 96-well plates containing 90μl of cells. The final compound concentrations of these dilutions were 10μM, 5μM, 1μM, 500nM, 250nM, 100nM, 10nM, 1nM.

[0496] For each drug condition and cell culture (for each row of triplicates), a well containing only RPMI-1640 complete / KSFM / DMEM medium and a well containing medium and DMSO were made as controls. The concentration of DMSO never exceeded 0.2% v / v. Each condition was inoculated in triplicate. On the day of the assay, the medium was replaced with a cell culture medium containing 0.1% resazurin sodium salt stock solution (stock solution: 0.015 mg / ml in PBS, catalog number R7017, Sigma-Aldrich). The cell culture plates were incubated in a cell culture incubator at 37°C and 5% CO2 for 2-3 hours. The cell culture plates were read using a fluorescence-based plate reader (Infinite M200 Pro, Tecan, Switzerland) using an excitation light between 530-560 nM and an emission light of 590 nM. Calculations and plots were performed using the GraphPad Prism program. The viability of the wells treated with the vehicle was set to 100%.

[0497] Table 6: Cytotoxic potency of 6027 shown as IC tested against healthy skin dermal fibroblasts and 5 skin cancer derived cells 50 Values ​​(in nM) for cells from cutaneous squamous cell carcinoma cell lines (SCC13 and A431_SCC) as well as cells from cutaneous melanoma (M010817, M150672, M150507) and cutaneous T-cell lymphoma (MyLa_CTCL).

[0498] Fibroblasts M010817 M130107 M150672 M150507 6027 3028 60 169 122 56

[0499] SCC13 A431 MyLa_CTCL 6027 191 70 451

[0500] Results. 6027 showed IC in the nM range against various skin cancer cell lines 50 The cytotoxicity against healthy skin dermal fibroblasts was not obvious. This indicates the selectivity between skin cancer-derived cells and healthy skin cells, which confirms the potential therapeutic window for the compounds of the present invention (especially 6027) to be developed as drugs for the treatment of skin cancer.

[0501] Example 7

[0502] In vitro antiproliferative and pro-apoptotic activities against B16-F10 mouse melanoma cells in C57BL / 6 mice bearing B16F10 xenografts

[0503] The mouse B16 melanoma model is the most commonly used metastatic melanoma model in preclinical studies. The syngeneic B16-F10 model was used to evaluate the response of tumor development after skin application of 6027. Liriostatin A, a known natural peptide, was initially tested in a similar manner.

[0504] Intradermal implantation of B16-F10 cells in C57BL / 6 mice results in dramatic tumor growth. Our growth studies demonstrate efficient growth kinetics following inoculation. Control animals remain in the study for 10-15 days before reaching euthanasia criteria for excessive tumor burden or ulceration. This produces a model that can provide up to a two-week administration window for test drugs to elicit their anti-tumor activity.

[0505] To determine whether C57BL / 6 mice bearing B16-F10 xenografts are an appropriate model to study the potential in vivo efficacy of skin-applied 6027, half-maximal inhibition (IC 50 ) in vitro. The same test was carried out on limystatin A.

[0506] In vitro cell viability assay. To maintain B16-F10 mouse 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.

[0507] The cell cultures were seeded into 96-well plates (Cat. No. 353072, Corning) and treated with increasing concentrations of the compounds (from 0.001 μM to 10 μM in RPMI-1640 complete medium) for 72 h the next day. The half-maximal inhibitory concentration (IC) of the compounds was determined. 50 ). 1000 cells were seeded per well. For each drug condition and cell culture (for each row in triplicate), a well containing only RPMI-1640 complete medium and a well containing medium and DMSO were made as controls. The concentration of DMSO never exceeded 0.2% v / v. Each condition was seeded in triplicate. On the day of the assay, the culture medium was changed to cell culture medium containing 0.1% resazurin sodium salt stock solution (stock solution: 0.015 mg / ml in PBS, catalog number R7017, Sigma-Aldrich). The cell culture plates were incubated in a cell culture incubator at 37°C and 5% CO2 for 2-3 hours. The cell culture plates were read using a fluorescence-based plate reader (Infinite M200 Pro, Tecan, Switzerland) using excitation light between 530-560 nM and emission light at 590 nM. Calculations and plots were performed using the GraphPad Prism program. The viability of wells treated with the vehicle was set to 100%. 6027 IC 50The IC of cephalostatin A is 208nM. 50 It is 161nM.

[0508] In vivo efficacy test method. C57Bl / 6 mice (Janvier Labs); female; 6-8 weeks old at the start of treatment. 1×10 5 The tumors were induced by 100 cells per day. A total of 24 mice were used (n=8 for vehicle, n=8 for 5% 6027, n=8 for 0.2% lycostatin A). The test substances were administered starting from day 1 after tumor cell inoculation. 20 μl of the test substances of 5% 6027 and 0.2% lycostatin A were applied percutaneously once a day with a pipette to about 1 cm from the site of B16-F10 cell injection. 2 The intact skin area was treated with 1% lipopolysaccharide for 21 days. If ulceration occurred, the animal was removed from the experiment and sacrificed.

[0509] The readout was the tumor volume measured three times per week and expressed as tumor volume (in cm 3 In addition, tumor samples were collected for histological analysis (tumor FFPE) (2 mice per group).

[0510] Results. Skin treatment with 20 μl of 5% 6027 in the area of ​​tumor inoculation was well tolerated. No skin irritation or other side effects were observed. After intradermal inoculation, B16F10 tumors gradually grew in the neck area of ​​mice. At the lowest tested concentration of 0.2%, limerostatin A showed strong skin irritation and the epidermis was destroyed. Therefore, treatment with 0.2% limerostatin A was not tolerated; for ethical reasons, the treatment regimen could not be continued for more than 7 days. Thereafter, the application site was not treated until the end of the experiment on day 16. There was no significant reduction in tumor volume in this group of animals during the entire test period.

[0511] Table 7: Tumor volumes in C57BL / 6 mice bearing intradermal B16F10 xenografts in skin areas (1 cm 2 ) received 20 μl of 5% 6027 (Group B: n=8) or vehicle control (Group A: n=8) daily. Tumor volume was measured 3 times a week with a caliper in cm 3 The mean tumor volume of the control group was normalized to 1 of the control group (RTV), and the ratio of the mean tumor volume of the treated group to the control group is indicated (T / C%). Tumors were implanted on day 0, and treatment with 6027 and vehicle control was started on day 1. Data are shown up to day 11.

[0512]

[0513] Histopathology. After sacrifice, non-ulcerated tumors were sampled and paraffin blocks were prepared for immunohistochemical staining. Ki-67 staining was used as an immunohistochemical cell marker of proliferation to visualize areas of proliferating cells in sections. During interphase, the Ki-67 antigen can only be detected within the nucleus, whereas in mitosis, most of the protein is relocalized to the chromosome surface. The Ki-67 protein is present in all active phases of the cell cycle (G1, S, G2, and mitosis) but is absent in resting (quiescent) cells (G0). The cellular content of the Ki-67 protein increases significantly during cell progression during the S phase of the cell cycle.

[0514] TUNEL staining was used to identify tumor areas where apoptosis was observed. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining is a method for detecting DNA breaks by labeling the 3′-hydroxyl ends of DNA double-strand breaks generated during apoptosis.

[0515] Results. Figure 1A and Figure 1B As shown, in tumor sections from C57BL / 6 mice bearing B16F10 xenografts that received 20 μl of 6027 daily, 6027 tumors had significantly fewer proliferating cells and significantly more apoptotic cells compared to tumors from vehicle-treated controls (two-way ANOVA, *p<0.01; ***p<0.0001).

[0516] Thus, topical treatment with the compound of the invention 6027 showed a significant reduction in tumor growth against B16-F10 mouse melanoma cells in C57BL / 6 mice bearing B16F10 xenografts, the most commonly used metastatic melanoma model in preclinical studies. Histological studies of the tumors showed a strong pro-apoptotic potential compared to the control group. It is noteworthy that lycostatin A is a known natural peptide that, while having similar in vitro cytotoxicity against B16-F10 cells, is not as suitable for skin application as 6027 because it has a strong skin irritation effect when the concentration is reduced to 0.2%, which is 25 times lower than 5% at which 6027 does not show skin irritation.

[0517] Example 8

[0518] Inhibitory effect of 6027 on ex vivo patient-derived basal cell carcinoma (BCC)

[0519] Tumor material was obtained from an invasive basal cell carcinoma (BCC) in the nose of a 91-year-old patient.

[0520] Fresh tumor material from the patient was cut into pieces of similar size, placed on an insert cell culture dish (0.4 μM, Millicell cell culture; Millipore, catalog number PICM03050), and incubated at 37° C. and 5% CO2 with 1 ml of co-culture medium (3 parts DMEM (Gibco, catalog number 41966052), 1 part Ham's F-12 nutrient mixture (Gibco, catalog number 11765054), 10% heat-inactivated fetal bovine serum (Gibco; catalog number 1050), 0.1 mg / ml NormocinTM (Invivogen, catalog number ant-nr-1), 21.8μg / ml adenine (Sigma-Aldrich, catalog number A2786), 5.45μg / ml apo-transferrin (Sigma-Aldrich, catalog number T1147), 2.18nM triiodothyronine (Sigma-Aldrich, catalog number T6397), 0.44μg / ml hydrocortisone (Sigma-Aldrich, catalog number H0888), 0.11nM cholera toxin (Sigma-Aldrich, catalog number C8052), 5.50μg / ml insulin (Sigma-Aldrich, catalog number I6634) and 0.01μg / ml epidermal growth factor (Sigma-Aldrich, catalog number E4127)) were incubated from below for 24 hours to allow tissue fragments to recover. After 24 hours, the cell culture medium was replaced with cell culture medium containing drug conditions and incubated for another 4 days at 37°C and 5% CO2. Make sure that the tissue pieces are covered with a drop of medium containing drug (day 0 and day 2). After 4 days of treatment, the tissues were fixed in 4% buffered formaldehyde and then subjected to immunohistochemical staining for histopathological examination and the treated conditions were compared with the untreated control group (DMSO).

[0521] Immunohistochemistry of ex vivo tumor fragments: Tumor sections were embedded in paraffin blocks. Serial sections were stained with the target antibodies. The following antibodies were used for staining: Ki67 (Dako, clone MIB-1, catalog number M7240, dilution 1:50, antigen retrieval: Leica Bond ER2: 95°C for 30 minutes). BERep4 (Agilent Dako, M0804, dilution 1:300, antigen retrieval Ph9) and HE ( Nach-Mayer, Artechemis, catalog number T.865.3; Eosin 1% in water-Morphisto, catalog number 10177). Slides were scanned on a Vectra Polaris slide scanner (Akoya Biosystems) and analyzed using Qpath software.

[0522] BERep4 staining was used as an immunohistochemical cell marker for basal cell carcinoma cell sections. BERep4 (also known as Ber-EP4) is a histological stain used primarily to aid in the diagnosis of basal cell carcinoma (BCC) and is an antibody to EpCAM (epithelial cell adhesion molecule). KI-67 staining was used as an immunohistochemical cell marker for proliferation to visualize areas of proliferating cells in sections. Vismodegib (20 μM), the standard of care, was further used as a positive control.

[0523] Results: Patient-derived BCC tumors incubated for 4 days showed a significant decrease in cell proliferation in the presence of 500 nM and 1 μM 6027 and 20 μM vismodegib compared to the negative control DMSO (two-way ANOVA; **p<0.002). TUNEL staining showed a significant increase in apoptotic cells after treatment with 500 nM and 1 μM concentrations of 6027 (two-way ANOVA; ***p<0.0001; **p<0.002), as shown in Table 1. Figure 2A and Figure 2B shown.

[0524] Example 9

[0525] Inhibitory effect of 6027 on ex vivo patient-derived basal cell carcinoma (BCC)

[0526] Tumor material was obtained from patients with invasive basal cell carcinoma (BCC).

[0527] The fresh biopsy tumor material of the patient was cut into pieces of similar size, placed on an insert cell culture dish (0.4 μM, Millicell cell culture; Millipore, catalog number PICM03050), and incubated at 37° C. and 5% CO2 with 1 ml of co-culture medium (3 parts DMEM (Gibco, catalog number 41966052), 1 part Ham's F-12 nutrient mixture (Gibco, catalog number 11765054), 10% heat-inactivated fetal bovine serum (Gibco; catalog number 1050), 0.1 mg / ml NormocinTM (Invivogen, catalog number ant-nr-1), 21.8μg / ml adenine (Sigma-Aldrich, catalog number A2786), 5.45μg / ml apo-transferrin (Sigma-Aldrich, catalog number T1147), 2.18nM triiodothyronine (Sigma-Aldrich, catalog number T6397), 0.44μg / ml hydrocortisone (Sigma-Aldrich, catalog number H0888), 0.11nM cholera toxin (Sigma-Aldrich, catalog number C8052), 5.50μg / ml insulin (Sigma-Aldrich, catalog number I6634) and 0.01μg / ml epidermal growth factor (Sigma-Aldrich, catalog number E4127)) were incubated from below for 24 hours to allow tissue fragments to recover. After 24 hours, the cell culture medium was replaced with cell culture medium containing drug conditions and incubated for another 4 days at 37°C and 5% CO2. Make sure that the tissue pieces are covered with a drop of medium containing drug (day 0 and day 2). After 4 days of treatment, the tissues were fixed in 4% buffered formaldehyde and then subjected to immunohistochemical staining for histopathological examination and the treated conditions were compared with the untreated control group (DMSO).

[0528] Immunohistochemistry of ex vivo tumor fragments: Tumor sections were embedded in paraffin blocks. Serial sections were stained with the target antibodies. The following antibodies were used for staining: Ki67 (Dako, clone MIB-1, catalog number M7240, dilution 1:50, antigen retrieval: Leica Bond ER2: 95°C for 30 minutes). BERep4 (Agilent Dako, M0804, dilution 1:300, antigen retrieval Ph9) and HE ( Nach-Mayer, Artechemis, catalog number T.865.3; Eosin 1% in water-Morphisto, catalog number 10177). Slides were scanned on a Vectra Polaris slide scanner (Akoya Biosystems) and analyzed using Qpath software.

[0529] BERep4 staining was used as an immunohistochemical cell marker for basal cell carcinoma cell sections. BERep4 (also known as Ber-EP4) is a histological stain used primarily to aid in the diagnosis of basal cell carcinoma (BCC) and is an antibody to EpCAM (epithelial cell adhesion molecule). KI-67 staining was used as an immunohistochemical cell marker for proliferation to show areas of proliferating cells in the sections. Vismodegib (20 μM) was used as the standard of care and was further used as a positive control.

[0530] Results. Incubation of patient-derived BCC tumors for 4 days showed that cell proliferation was significantly reduced in the presence of 500 nM of 6027 and vismodegib (20 μM) compared with the negative control DMSO (two-way ANOVA; *p<0.01; **p<0.002); after treatment with 6027 (500 nM) and vismodegib (20 μM), TUNEL-positive cells showed a significant increase in apoptotic cells (two-way ANOVA; ***p<0.0001; ****p<0.00002), as shown in Figure 5. Figure 3A and Figure 3B shown.

[0531] Example 10

[0532] Inhibitory effect of 6027 on ex vivo patient-derived basal cell carcinoma (BCC)

[0533] Tumor material was obtained from a nodular basal cell carcinoma (BCC) on the upper back of a 73-year-old patient.

[0534] Fresh tumor material from the patient was cut into pieces of similar size, placed on an insert cell culture dish (0.4 μM, Millicell cell culture; Millipore, catalog number PICM03050), and incubated at 37° C. and 5% CO2 with 1 ml of co-culture medium (3 parts DMEM (Gibco, catalog number 41966052), 1 part Ham's F-12 nutrient mixture (Gibco, catalog number 11765054), 10% heat-inactivated fetal bovine serum (Gibco; catalog number 1050), 0.1 mg / ml NormocinTM (Invivogen, catalog number ant-nr-1), 21.8μg / ml adenine (Sigma-Aldrich, catalog number A2786), 5.45μg / ml apo-transferrin (Sigma-Aldrich, catalog number T1147), 2.18nM triiodothyronine (Sigma-Aldrich, catalog number T6397), 0.44μg / ml hydrocortisone (Sigma-Aldrich, catalog number H0888), 0.11nM cholera toxin (Sigma-Aldrich, catalog number C8052), 5.50μg / ml insulin (Sigma-Aldrich, catalog number I6634) and 0.01μg / ml epidermal growth factor (Sigma-Aldrich, catalog number E4127)) were incubated from below for 24 hours to allow tissue fragments to recover. After 24 hours, the cell culture medium was replaced with cell culture medium containing drug conditions and incubated for another 4 days at 37°C and 5% CO2. Make sure that the tissue pieces are covered with a drop of medium containing drug (day 0 and day 2). After 4 days of treatment, the tissues were fixed in 4% buffered formaldehyde and then subjected to immunohistochemical staining for histopathological examination and the treated conditions were compared with the untreated control group (DMSO).

[0535] Immunohistochemistry of ex vivo tumor fragments: Tumor sections were embedded in paraffin blocks. Serial sections were stained with the target antibodies. The following antibodies were used for staining: Ki67 (Dako, clone MIB-1, catalog number M7240, dilution 1:50, antigen retrieval: Leica Bond ER2: 95°C for 30 minutes). BERep4 (Agilent Dako, M0804, dilution 1:300, antigen retrieval Ph9) and HE ( Nach-Mayer, Artechemis, catalog number T.865.3; Eosin 1% in water-Morphisto, catalog number 10177). Slides were scanned on a Vectra Polaris slide scanner (Akoya Biosystems) and analyzed using Qpath software.

[0536] BERep4 staining was used as an immunohistochemical cell marker for basal cell carcinoma cell sections. BERep4 (also known as Ber-EP4) is a histological stain used primarily to aid in the diagnosis of basal cell carcinoma (BCC) and is an antibody to EpCAM (epithelial cell adhesion molecule). Ki-67 staining was used as an immunohistochemical cell marker for proliferation to show areas of proliferating cells in the sections. Vismodegib (20 μM) was used as a positive control.

[0537] Results. Incubation of patient-derived BCC tumors for 4 days demonstrated that cell proliferation was significantly reduced in the presence of 500 nM of 6027 compared to standard of care treatment vismodegib (20 μM) (two-way ANOVA; **p<0.002), as shown in Figure 2. Figure 4A and Figure 4B shown.

[0538] Embodiment 11

[0539] Inhibitory effect of 6027 on ex vivo patient-derived cutaneous squamous cell carcinoma cSCC

[0540] Tumor material was obtained from a 79-year-old patient with high-grade cutaneous squamous cell carcinoma (cSCC) on his head.

[0541] Fresh tumor material from the patient was cut into pieces of similar size, placed on an insert cell culture dish (0.4 μM, Millicell cell culture; Millipore catalog number PICM03050), and incubated at 37° C. and 5% CO2 with 1 ml of co-culture medium (3 parts DMEM (Gibco, catalog number 41966052), 1 part Ham's F-12 nutrient mixture (Gibco, catalog number 11765054), 10% heat-inactivated fetal bovine serum (Gibco; catalog number 1050), 0.1 mg / ml NormocinTM (Invivogen, catalog number ant-nr-1), 21.8μg / ml adenine (Sigma-Aldrich, catalog number A2786), 5.45μg / ml apo-transferrin (Sigma-Aldrich, catalog number T1147), 2.18nM triiodothyronine (Sigma-Aldrich, catalog number T6397), 0.44μg / ml hydrocortisone (Sigma-Aldrich, catalog number H0888), 0.11nM cholera toxin (Sigma-Aldrich, catalog number C8052), 5.50μg / ml insulin (Sigma-Aldrich, catalog number I6634) and 0.01μg / ml epidermal growth factor (Sigma-Aldrich, catalog number E4127)) were incubated from below for 24 hours to allow tissue fragments to recover. After 24 hours, the cell culture medium was replaced with cell culture medium containing drug conditions and incubated for another 4 days at 37°C and 5% CO2. Make sure that the tissue pieces are covered with a drop of medium containing drug (day 0 and day 2). After 4 days of treatment, the tissues were fixed in 4% buffered formaldehyde and then subjected to immunohistochemical staining for histopathological examination and the treated conditions were compared with the untreated control group (DMSO).

[0542] Immunohistochemistry of ex vivo tumor fragments: Tumor sections were embedded in paraffin blocks. Serial sections were stained with the target antibodies. The following antibodies were used for staining: Ki67 (Dako, clone MIB-1, catalog number M7240, dilution 1:50, antigen retrieval: Leica Bond ER2: 95°C for 30 minutes), BERep4 (Dako, M0804, dilution 1:300, antigen retrieval Ph9) and HE ( Nach-Mayer, Artechemis, catalog number T.865.3; Eosin 1% in water-Morphisto, catalog number 10177). Slides were scanned on a Vectra Polaris slide scanner (Akoya Biosystems) and analyzed using Qpath software.

[0543] HE staining: Hematoxylin and eosin (HE) staining is one of the main tissue stains used in histology. HE is a combination of two histological stains: hematoxylin and eosin. Hematoxylin stains the nucleus purple-blue, eosin stains the extracellular matrix and cytoplasm pink, and other structures present different shades and combinations of these colors to easily distinguish the nuclear and cytoplasmic parts of the cell. In addition, the overall coloring pattern of the staining shows the general layout and distribution of the cells and provides a general overview of the structure of the tissue sample, which can be used as a diagnostic tool for SCC due to the typical immunohistochemical characteristics of SCC.

[0544] Ki-67 staining was used as an immunohistochemical cell marker for proliferation to visualize areas of proliferating cells in the sections.

[0545] Results: Figure 5A and Figure 5B As shown, incubation of patient-derived SCC tumors for 4 days demonstrated a significant decrease in cell proliferation in the presence of 500 nM of 6027 compared to the negative control DMSO (two-way ANOVA; **p<0.003).

Claims

1. A compound of formula (I) and a pharmaceutically acceptable salt of the compound of formula (I) for use in a method for topical treatment of skin cancer in a mammal, wherein: The method comprises topically administering the compound to the mammal, In the formula A1 is A2 is A3 is B1 is B2 is B3 is 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, 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, a1+a 12 +a2+a 22 +a3+a 32 is 0, 1, 2 or 3, b1+b 12 +b2+b 22 +b3+b 32 is 0, 1, or 2; R1 is selected from carbocyclyl or heterocyclyl, each independently optionally and preferably substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5, and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, R4, R5 and R6 are independently H, C1-C3 alkyl at each occurrence; R2 is selected from C4-C 12 Alkyl, C4-C 10 Alkoxy, C1-C3 alkylene-cycloalkyl, C1-C3 alkylene-aryl and C1-C3 alkylene-heteroaryl, independently in the C1-C3 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-; the alkyl, cycloalkyl, aryl and heteroaryl are each independently optionally and preferably substituted by 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 by C1-C3 alkyl or OC1-C3 alkyl; R3 is In the formula R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 Two of them together with the carbon atoms to which they are attached form a carbocyclic or heterocyclic ring, preferably a carbocyclic ring; R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxy or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form heteroaryl or heterocyclyl at each occurrence, each independently optionally and preferably substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence, and the arrow indicates the connection to the C(O)-moiety shown in formula (I).

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

3. The compound for use according to claim 1 or 2, wherein The R1 is selected from cycloalkyl, aryl or heteroaryl, preferably selected from cycloalkyl, monocyclic or bicyclic aryl, or monocyclic or bicyclic heteroaryl, each of which is independently optionally substituted by C1-C4 alkyl, halogen, oxo, CF3, OR4, NR5R6, C6H5 and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, and R4, R5 and R6 are independently H or C1-C3 alkyl at each occurrence.

4. The compound for use according to any one of claims 1 to 3, wherein The R1 is selected from phenyl or a monocyclic or bicyclic heteroaryl group containing one or two heteroatoms selected from N, O and S, preferably selected from phenyl or oxazolyl; each independently optionally substituted by C1-C4 alkyl, halogen, CF3, OR4, NR5R6, C6H5 and C6H5 substituted by halogen, C1-C3 alkyl, OR4, NR5R6, R4, R5 and R6 are independently H or C1-C3 alkyl at each occurrence.

5. The compound for use according to any one of claims 1 to 4, wherein The R1 is selected from the following formula: In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

6. The compound for use according to any one of claims 1 to 5, wherein The R2 is selected from C5-C 12 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), independently in the C1-C3 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-, the phenyl, biphenyl, C5-C6-cycloalkyl and monocyclic or bicyclic heteroaryl are each independently optionally substituted by one or more, preferably one or two, substituents selected from C1-C2 alkyl, C1-C2 haloalkyl, halogen, C1-C2 alkoxy.

7. The compound for use according to any one of claims 1 to 6, wherein The R2 is selected from C5-C 12 Alkyl, C1-C2 alkylene-C5-C6 cycloalkyl, C1-C2-alkylene-phenyl, C1-C2 alkylene-biphenyl, C1-C2-alkylene-(monocyclic or bicyclic heteroaryl), independently in the C1-C2 alkylene, one -CH2- part is optionally substituted by -CH(NH)- or -O-, preferably -O-, and the phenyl, biphenyl, C5-C6 cycloalkyl and monocyclic or bicyclic heteroaryl are independently optionally substituted by one or two substituents selected from methyl, ethyl, fluorine, chlorine and methoxy.

8. The compound for use according to any one of claims 1 to 7, wherein The R2 is selected from: In the formula, R represents a connection to the CH-moiety shown in formula (I).

9. The compound for use according to any one of claims 1 to 8, wherein R7, R8, R9 and R 10 Each occurrence is independently H or C1-C3 alkyl, preferably H or methyl, or each occurrence is independently, said R7, R8, R9 and R 10 The two of them together with the carbon atoms to which they are attached form a monocyclic carbocyclic ring or a monocyclic heterocyclic ring, preferably a monocyclic carbocyclic ring, R 11 and R 12 are independently H or C1-C4 alkyl optionally substituted by halogen, hydroxyl or C3-C6 cycloalkyl, or together with the nitrogen atom to which they are attached, independently form a monocyclic heteroaryl or a monocyclic heterocyclyl at each occurrence, each independently optionally substituted by halogen, C1-C4 alkyl, OR 13 NR 14 R 15 Replacement; R 13 , R 14 , R 15 is independently H, C1-C4 alkyl at each occurrence.

10. The compound for use according to any one of claims 1 to 9, wherein The R3 is selected from: In the formula, R represents the connection to the C(O)-moiety shown in formula (I).

11. The compound for use according to claim 1, wherein The compound 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： 21372： 12. The compound for use according to any one of claims 1 to 11, wherein The topical administration is application of the compound to mammalian skin, preferably human skin.

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

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

15. A pharmaceutical composition for use in a method for topical treatment of skin cancer in a mammal, wherein: The pharmaceutical composition comprises a compound of formula (I) as defined in any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or adjuvant, and the method comprises topically administering the pharmaceutical composition to the mammal.

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