RU2575349C2 - Ingenol-3-acylates i - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: in formula

(I) R represents (C₁-C₇)alkyl or (C₂-C₇)alkenyl, wherein R is substituted by one or more substitutes optionally specified in R1; wherein R1 represents cyano or (C₁-C₄)alkyl, (C₆)cycloalkyl or (C₆)aryl each of which may be optionally substituted by one or more substitutes optionally specified in R2, wherein R2 represents (C₁-C₄)alkyl or -COORc or =N-ORa, -NRaRb or =O; Ra and Rb independently represent (C₁-C₄)alkyl or (C₆)aryl; Rc represents (C₁-C₄)alkyl. The invention also refers to a pharmaceutical composition containing the above compound.

EFFECT: invention refers to the compound of structural formula I, which can be used to prevent, treat or reduce an intensity of symptoms of a disease or condition susceptible to IL-8 keratinocyte release stimulation, susceptible to neutrophil oxidative burst stimulation or susceptible to necrosis induction.

24 cl, 121 ex

Description

FIELD OF THE INVENTION

The present invention relates to new derivatives of 3-acylingenol and their derivatives and their use as medicines in therapy. The invention also relates to pharmaceutical compositions containing said compounds and methods for treating diseases with said compounds.

BACKGROUND OF THE INVENTION

Ingenol-3-angelate (PEP005, ingenol mebutate) is an diterpenic ester of the ingenol family, which is isolated from various species of Euphorbia, in particular, from Euphorbia peplus. This compound is currently under clinical development for use in the treatment of senile keratosis and non-melanoma skin cancer.

A number of other ingenol-3-acylates, mainly long chain saturated and unsaturated aliphatic fatty acids, have been isolated from various species of Euphorbia [H. Gotta, Z. Naturforschung, (1984), 39b, 683-94; K. Abo, Fitoterapia, (1988), 244-46, S. Zayed, J. Cancer Res. Clin. Oncol. (2001), 127, 40-47]. In addition, a small number of ingenol-3-acylates was obtained by a semi-synthetic method (B. Sorg et al., Z. Naturforsch., (1982), 37b, 748-56). Some of these ingenol derivatives have been described and tested as potent irritants and potent tumor growth agents [B. Sorg et al., Naturforsch., (1982), 37b, 748-56; B. Sorg et al., Carcinogenesis, (1987), 8, 1-4].

In addition to aliphatic esters of ingenol, aromatic esters of ingenol are also known. Described milliamin C, a derivative of ingenol-3-anthranilate (Marston, A. Planta Medica, (1983), 47, 141-47). Ingenol-3-benzoate is also described (Sorg, B .; Z Naturforschung, (1982), 37b, 748-56).

Angelic acid and angelic acid esters, when present in ingenol-3-angelate, are prone to isomerization (double bond) to form tiglinic acid ester, in particular at a basic pH [Beeby, P., Tetrahedron Lett. (1977), 38, 3379-3382, Hoskins, W. M., J. Chem. Soc. Perkin Trans. 1, (1977), 538-544, Bohlmann, F. et al., Chem. Ber. (1970) 103, 561-563].

In addition, it is known that ingenol-3-acylates are unstable, as they undergo rearrangement, thus forming ingenol-5-acylates and ingenol-20-acylates [Sorg, B. et al., Z. Naturforsch., (1982) 37B, 748-756].

WO99 / 08994 describes the isolation of compounds from the Euphorbia plant and their use in the treatment of cancer and other tumor diseases, in senile keratosis or radiation keratosis. WO01 / 93883 describes derivatives of ingenol that are different from the derivatives of ingenol of the present invention, used to prevent a PKC-related condition or disorder in a subject. The diseases referred to in WO01 / 93883 are asthma, atherosclerosis, atopic dermatitis, autoimmune disease, bipolar disorder, blood disease, cardiac hypertrophy, depression, diabetes, hypertension, hyperplastic dermatosis, multiple sclerosis, myocardial ischemia, psoriasis, osteoarthritis, osteoarthritis, osteoarthritis, osteoarthritis transplantation and latent viral disease. WO01 / 93884 describes derivatives of ingenol that are different from the derivatives of ingenol of the present invention and their use in the treatment of inflammatory diseases, such as diseases resulting from infection by pathogenic microorganisms, viruses, yeast, fungi, worms, insects, arachnids, nematodes, amoeba and t .d. WO01 / 93885 describes derivatives of ingenol that are different from the derivatives of ingenol of the present invention and are used for immunostimulation. WO08 / 131491 describes ingenol derivatives that are different from the ingenol derivatives of the present invention and are used to treat infectious diseases caused by the HPV virus. WO06 / 063382 describes derivatives of ingenol that are different from the derivatives of ingenol of the present invention and are used to treat solid tumors. AU 2006201661 describes a method for the treatment of acute myeloid leukemia using ingenol-3-angelate. WO02 / 11743 describes a specific use in the treatment of prostate and bladder cancer. Derivatives of ingenol are described in WO07 / 059584 for use to stimulate wound healing. WO2010 / 091472 describes the use of ingenols and derivatives in other cosmetic applications.

It is believed that ingenol-3-angelate has a double mode of action: 1) inducing cell death by a direct cytotoxic effect or inducing apoptosis, and 2) an immunostimulating effect caused by the recruitment and activation of neutrophils (Rosen, RH, et al., J. Am. Acad . Derm. (2011), e-published Nov 2011; Ersvaer, E., et al., Toxins, (2010), 2, 174-194). Nanomolar concentrations of such an agent cause activation and modulation of protein kinase C (PKC) of classical and new isoforms, especially significantly PKCdelta forms. By activating PKCdelta, the agent induces apoptosis in sensitive cells. (Hampson, P., et al., Blood, (2005), 106, 1362-1368; Cozzi, S.J., et al., Cancer Res., (2006), 66, 10083-10091). Rapid cytotoxic effects on cancer cells are observed at high micromolar concentrations (Ogbourne, S.M., et al., Cancer Res. (2004), 64, 2833-2839). By activating various PKC isoforms, the agent also induces pro-inflammatory actions, including the release of pro-inflammatory mediators (Challacombe, JM, et al., J. Immunol. (2006), 177, 8123-8132, vascular endothelial activation (Hampson, P. et al ., Cancer Immunol Immunother., (2008), 57, 1241-1251); chemo-attraction of neutrophils by inducing interleukin 8 in keratinocytes and the development of certain anti-cancer immune responses to CD8 + cells by adjuvant properties in animal models (Le, TT, et al ., Vacccine, (2009), 27, 3053-3062).

Compounds exhibiting a dual mode of action by inducing cell death by a direct cytotoxic effect or by inducing apoptosis and an immunostimulating effect, including the recruitment and activation of neutrophils, can be used to treat conditions associated with hyperplasia or neoplasia. Compounds causing cell death by primary and / or secondary necrosis and compounds exhibiting a proapoptotic effect can reduce unwanted cell growth and remove unwanted cells, and in addition, stimulating the innate immune response and adjuvant effects can increase the biological response against aberrant or transformed cells .

Compounds inducing cell death by primary and / or secondary necrosis may be useful for treating cosmetic conditions, as these compounds can destroy or remove unwanted tissue or cells.

There is a need for new derivatives of ingenol with similar or improved biological activity compared to ingenol-3-angelate. In addition, there is a need for new derivatives of ingenol that cause cell death by cytotoxicity or apoptosis and / or induce an immunostimulating effect.

The present invention provides branched or substituted 3-O-acylingenol derivatives useful for treating conditions associated with the use of ingenol-3-angelate, or useful for treating conditions that are affected by inducing cell death by cytotoxicity or inducing apoptosis and / or immunostimulating action.

The compounds of the present invention stimulate an oxidative explosion of neutrophils, which is part of an innate immune response.

The compounds of the present invention stimulate the release of IL-8 keratinocytes, thus causing an immunostimulating effect.

Some compounds of the present invention cause rapid necrosis.

The compounds of the present invention exhibit suitable stability.

Some compounds of the present invention exhibit improved activity in the analysis of the oxidative explosion of neutrophils compared to ingenol-3-angelate.

Some compounds of the present invention exhibit improved activity in an IL-8 release assay compared to ingenol-3-angelate.

Some compounds of the present invention exhibit improved activity in the analysis of necrosis compared to ingenol-3-angelate.

SUMMARY OF THE INVENTION

In an embodiment, the invention relates to a compound of general formula I

where R represents (C ₁ -C ₇ ) alkyl, (C ₂ -C ₇ ) alkenyl or (C ₂ -C ₇ ) alkynyl; where R is substituted by one or more substituents independently selected from R1; where R1 represents

(a) fluorine, cyano or hydroxy; or

(b) (C ₁ -C ₄ ) alkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycloalkyl, heterocycloalkenyl, or (C ₃ -C ₆ ) cycloalkylidene, each of which may optionally be substituted with one or more substituents independently selected from R2, where R2 is halogen, cyano, hydroxy, (C ₁ -C ₄ ) alkyl, (C ₂ -C ₄ ) alkenyl, halogen (C ₁ -C ₄ ) alkyl, -NRaCORb, -COORc, -OCORa, -CONRaRb, -OCONRaRb, -NRaCOORb, -NRaCONRaRb, -NRaSO ₂ Rb, -NRaSO ₂ NRaRb, -SO ₂ NRaRb, -SO ₂ Ra, -S (O) Ra, -ORa, -SRa, = N-ORa, = O; or

(c) -NRaCORb, -COORc, -OCORa, -CONRaRb, -OCONRaRb, -NRaCOORb, -NRaCONRaRb, -NRaSO ₂ Rb, -NRaSO ₂ NRaRb, -SO ₂ NRaRb, -SO ₂ Ra, -S (О) , -ORa, -SRa, = N-ORa, -NRaRb or = O;

and where

Ra and Rb independently represent hydrogen, (C ₁ -C ₄ ) alkyl, halogen (C ₁ -C ₄ ) alkyl, (C ₁ -C ₄ ) alkoxy (C ₁ -C ₄ ) alkyl, hydroxy (C ₁ -C ₄ ) alkyl, cyano (C ₁ -C ₄ ) alkyl or aryl;

and where

Rc is (C ₁ -C ₄ ) alkyl, halogen (C ₁ -C ₄ ) alkyl, (C ₁ -C ₄ ) alkoxy (C ₁ -C ₄ ) alkyl, hydroxy (C ₁ -C ₄ ) alkyl or cyano (C ₁ -C ₄ ) alkyl,

and its pharmaceutically acceptable salts, prodrugs, hydrates and solvates;

provided that when R1 is alkyl or alkenyl, the total chain length of R and R1 does not exceed a chain length of 7 carbon atoms,

and provided that R and R1 in combination cannot be a straight unbranched or unsubstituted alkyl chain or a straight unbranched or unsubstituted alkenyl chain,

and provided that R cannot be or R and R1 in combination cannot be trans-2-buten-2-yl.

In an embodiment, the invention relates to a compound of formula I, which is used as a medicine in therapy.

In an embodiment, the invention relates to a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable stereoisomer, salt or in vivo hydrolyzable ester thereof in combination with a pharmaceutically acceptable carrier or excipient.

In an embodiment, the invention relates to a pharmaceutical composition suitable for topical administration and comprising a compound of formula I or a pharmaceutically acceptable stereoisomer, salt or in vivo hydrolyzable ester thereof together with a pharmaceutically acceptable carrier or excipient.

In an embodiment, the invention relates to a compound of formula I, which is used in the treatment, prevention, reduction of symptom intensity or the prevention of physiological disorders or diseases associated with hyperplasia or neoplasia.

In an embodiment, the invention relates to the use of a compound of formula I for the preparation of a medicament for treating, reducing the intensity of symptoms or preventing physiological disorders or diseases associated with hyperplasia or neoplasia.

In an embodiment, the invention relates to a method for preventing, treating, reducing the intensity of symptoms or preventing physiological disorders or diseases associated with hyperplasia or neoplasia by administering to a subject in need thereof a compound of formula I.

In an embodiment, the invention relates to a compound of formula I, useful in treating or reducing the intensity of symptoms of a cosmetic indication.

In an embodiment, the invention relates to the use of a compound of formula I for the preparation of a medicament for treating or reducing the intensity of symptoms of a cosmetic indication.

In an embodiment, the invention relates to a method for treating or reducing the intensity of symptoms of cosmetic indications by administering to a subject in need thereof a compound of formula I.

In an embodiment, the invention relates to a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable stereoisomer, salt or in vivo hydrolyzable ester thereof in combination with one or more other therapeutically active agents.

DETAILED DESCRIPTION OF THE INVENTION

In an embodiment, the invention relates to the above compound of formula I, wherein R is (C ₁ -C ₇ ) alkyl, (C ₂ -C ₇ ) alkenyl or (C ₂ -C ₇ ) alkynyl; where R is substituted by one or more substituents independently selected from R1, where R1 is

(a) fluorine, cyano or hydroxy; or

(b) (C ₁ -C ₄ ) alkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycloalkyl, heterocycloalkenyl, each of which may be optionally substituted with one or more substituents independently selected from R2, where R2 is halogen, cyano, hydroxy , (C ₁ -C ₄ ) alkyl, (C ₂ -C ₄ ) alkenyl, halogen (C ₁ -C ₄ ) alkyl, -NRaCORb, -COORc, -OCORa, -CONRaRb, -OCONRaRb, -NRaCOORb, -NRaCONRaRb, -NRaSO ₂ Rb, -NRaSO ₂ NRaRb, -SO ₂ NRaRb, -SO ₂ Ra, -S (O) Ra, -ORa, -SRa, = N-ORa, = O; or

(c) -NRaCORb, -COORc, -OCORa, -CONRaRb, -OCONRaRb, -NRaCOORb, -NRaCONRaRb, -NRaSO ₂ Rb, -NRaSO ₂ NRaRb, -SO ₂ NRaRb, -SO ₂ Ra, -S (О) , -ORa, -SRa, = N-ORa,

and where

Ra and Rb are hydrogen, (C ₁ -C ₄ ) alkyl, halogen (C ₁ -C ₄ ) alkyl, (C ₁ -C ₄ ) alkoxy (C ₁ -C ₄ ) alkyl, hydroxy (C ₁ -C ₄ ) alkyl, cyano (C ₁ -C ₄ ) alkyl,

and where

Rc is (C ₁ -C ₄ ) alkyl, halogen (C ₁ -C ₄ ) alkyl, (C ₁ -C ₄ ) alkoxy (C ₁ -C ₄ ) alkyl, hydroxy (C ₁ -C ₄ ) alkyl, cyano (C ₁ -C ₄₎ alkyl,

provided that when R1 is alkyl or alkenyl, the total chain length of R and R1 does not exceed a chain length of 7 carbon atoms,

provided that R and R1 in combination cannot be an unbranched or unsubstituted alkyl or alkenyl chain,

provided that R cannot be or R and R1 in combination cannot be trans-2-buten-2-yl.

In an embodiment, the invention relates to a compound of formula I, wherein R1 is located in the alpha or beta position relative to the carbonyl group.

In an embodiment, the invention relates to a compound of formula I, wherein R is substituted with R1 at the alpha or beta position relative to the carbonyl group.

In an embodiment, the invention relates to a compound of formula I, wherein R is substituted with R1 at the alpha position relative to the carbonyl group.

In an embodiment, the invention relates to a compound of formula I, wherein R is (C ₁ -C ₇ ) alkyl.

In an embodiment, the invention relates to a compound of formula I, wherein R is ethyl, propyl, tert-butyl, isopropyl, 2-butyl or 3-pentyl.

In an embodiment, the invention relates to a compound of formula I, wherein R is ethyl, propyl, tert-butyl, isopropyl, 2-butyl, 3-pentyl or isobutyl.

In an embodiment, the invention relates to a compound of formula I, wherein R is (C ₂ -C ₇ ) alkenyl.

In an embodiment, the invention relates to a compound of formula I, wherein R is propenyl, ethenyl.

In an embodiment, the invention relates to a compound of formula I, wherein R is propenyl or ethenyl.

In an embodiment, the invention relates to a compound of formula I, wherein R1 is (C ₁ -C ₄ ) alkyl.

In an embodiment, the invention relates to a compound of formula I, wherein R1 is C ₁ -C ₄ alkyl, aryl, cycloalkyl, cycloalkylidene, = N-ORa, = O, -NRaRb, -COORc or cyano.

In an embodiment, the invention relates to a compound of formula I, wherein R1 is ethyl or methyl.

In an embodiment, the invention relates to a compound of formula I, wherein R1 is ethyl, methyl, phenyl, cyclohexyl, = N-OCH ₃ , —N (CH ₃ ) (C ₆ H ₅ ), —COOCH _3, or cyclohexylidene.

In an embodiment, the invention relates to a compound of formula I, wherein R1 is methyl.

In an embodiment, the invention relates to a compound of formula I, wherein R2 is methyl.

In an embodiment, the invention relates to a compound of formula I, wherein the compound is

ingenol-3- (2-methylacrylate),

ingenol-3- (3-methylbutenoate),

ingenol-3- (2,3-dimethylbutenoate),

ingenol-3- (2-methylene butyrate),

ingenol-3- (2-methylpropanoate),

ingenol-3- (3-methylbutanoate),

ingenol-3- (2RS-methylbutanoate),

ingenol-3- (3,3-dimethylbutanoate),

ingenol-3- (2-ethylbutanoate),

ingenol-3- (2R-methylbutanoate),

ingenol-3-crucible,

ingenol-3- (phenylacetate),

ingenol-3- (2Z- (methoxycarbonyl) acrylate),

ingenol-3- (2-cyclohexylpropanoate),

ingenol-3 - ((2Z) -2-methoxyimino-2-phenylacetate),

ingenol-3 - ((2E) -2-methoxyimino-2-phenylacetate),

ingenol-3 - ((Z) -2-methyl-3- (p-tolyl) prop-2-enoate),

ingenol-3 - ((E) -2-phenylbut-2-enoate),

ingenol-3- (2,2-diphenylacetate),

ingenol-3- (2-cyano-2-cyclohexylidene acetate) or

ingenol-3- (2- (methyl (phenyl) amino) -2-oxoacetate).

In an embodiment, the invention relates to a compound of formula I, wherein said compound is ingenol-3- (2-methylene butyrate).

In an embodiment, the invention relates to a compound of formula I, wherein said compound is ingenol-3 - ((Z) -2-methyl-3- (p-tolyl) prop-2-enoate).

In an embodiment, the invention relates to a compound of formula I, wherein said compound is ingenol-3- (2,3-dimethylbutenoate).

In one embodiment, the invention relates to a compound of formula I, wherein said compound is ingenol-3- (2-ethylbutanoate).

In one embodiment, the invention relates to a compound of formula I, wherein said compound is ingenol-3- (2-cyclohexylpropanoate).

In one embodiment, the invention relates to a compound of formula I, wherein said compound is ingenol-3 - ((E) -2-phenylbut-2-enoate).

Definitions

In this context, the term "(C _a -C _b) alkyl" wherein a and b are integers refers to an alkyl radical, straight or branched chain having from a to b carbon atoms. So, when a is 1 and b is 7, the term, for example, includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, hexyl, isohexyl and heptyl.

The term “carbocyclic” refers to a mono-, bi- or tricyclic radical containing up to 13 ring atoms, all of which are carbon atoms, and including aryl, cycloalkyl and cycloalkenyl.

The term “cycloalkyl” refers to a mono-, bi- or tricyclic saturated cycloalkane radical containing 3 to 13 carbon atoms and including, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclo [2.2.1] heptanyl and adamantyl.

The term "(C _a -C _b) alkenyl" wherein a and b are integers refers to a mono-, di- or tri-unsaturated alkenyl radical with a straight or branched chain having from a to b carbon atoms. So, when a is 1 and b is 7, for example, such a term includes, for example, ethenyl, allyl, propenyl; 1-, 2- or 3-butenyl; 1-, 2-, 3- or 4-pentenyl; 1-, 2-, 3-, 4- or 5-hexenyl.

The term “cycloalkenyl” refers to mono-, di- or tri-unsaturated non-aromatic cyclic hydrocarbon radicals comprising polycyclic radicals containing 3 to 13 carbon atoms and including, for example, cyclopropenyl, cyclobutenyl, cyclopentenyl or cyclohexenyl.

The term "(C _a -C _b) alkynyl" wherein a and b are integers refers to a hydrocarbon radical straight or branched chain having from a to b carbon atoms and containing 1-2 C-C triple bonds. So, when a is 1 and b is 7, the term, for example, includes ethynyl, propynyl, butynyl, pentynyl or hexynyl.

The term “heterocyclic” refers to a carbocyclic radical which has the above meanings, contains 1-4 heteroatoms selected from O, N or S, and includes heteroaryl, heterocycloalkyl and heterocycloalkenyl.

The term “heterocycloalkyl” refers to a cycloalkyl radical having the above meanings, including polycyclic radicals optionally fused to carbocyclic rings, and containing 1-4 heteroatoms selected from O, N or S, for example tetrahydrofuranyl, pyrrolidinyl, dioxolanyl, morpholinyl vinyl, piperidinyl or 5-oxabicyclo [2.2.2] octane.

The term “heterocycloalkenyl” refers to a cycloalkenyl radical as defined above, including polycyclic radicals optionally fused to carbocyclic rings, and containing 1-4 heteroatoms selected from O, N or S, for example dihydropyranyl.

The term “aryl” refers to a radical of aromatic carbocyclic rings containing 6-10 carbon atoms, in particular phenyl, and optionally fused carbocyclic rings with at least one aromatic ring. Thus, the term includes, for example, phenyl, naphthyl, indenyl or indanyl.

The term “heteroaryl” refers to radicals of heterocyclic aromatic rings optionally fused to carbocyclic rings or heterocyclic rings containing 1-4 heteroatoms selected from O, S and N, and 1-12 carbon atoms, in particular 5- or 6-membered rings with 1-4 heteroatoms, or optionally fused bicyclic rings, where at least one ring is aromatic. Thus, the term includes, for example, pyridyl, quinolyl, isoquinolyl, indolyl, tetrazolyl, furyl, thiazolyl, imidazolyl, imidazo [1,2-a] pyrimidinyl, pyrrolyl, pyrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, 1,2,4- triazolyl, thienyl, pyrazinyl, pyrimidinyl, 1,2,3-triazolyl, isothiazolyl, imidazo [2,1-b] thiazolyl, benzimidazolyl, benzofuranyl, benzofuranyl, benzothiophenyl, benzothiazolyl, benzooxazolyl, indazolyl.

The term “halogen” is intended to mean a substituent from the 7th basic group of the periodic table, preferably fluoro, chloro and bromo.

The term “alkoxy” is intended to mean a radical of the formula —OR, wherein R is alkyl having the above meanings, for example, methoxy, ethoxy, n-propoxy, isopropoxy, butoxy, etc.

The term “hydroxyalkyl” is intended to mean a primary, secondary or tertiary radical of the formula —R — OH, where R is alkyl having the above meanings, for example, hydroxymethyl or hydroxyethyl.

The term “cyanoalkyl” is intended to mean a primary, secondary or tertiary radical of the formula —R — CN, where R is alkyl having the above meanings, for example, cyanomethyl or cyanoethyl.

The term “haloalkyl” is intended to mean a primary, secondary or tertiary radical of the formula —RX _(1-3) , where R is alkyl having the above meanings and X is halogen having the above meanings, for example trifluoromethyl, 2 , 2,2-trifluoroethyl or difluoromethyl.

The term “alkoxyalkyl” is intended to mean an alkyl radical having the above meanings, which is substituted with an alkoxy radical having the above meanings, that is, —ROR where each R is alkyl, wherein the alkyls are the same or different and have the meanings given above, for example , methoxymethyl, ethoxymethyl.

In the present context, the term “cycloalkylidene” is intended to mean a divalent radical of cycloalkyl, such as cyclohexylidene, cyclopentylidene or cyclobutylidene, as shown below:

In the present context, the term "trans-2-buten-2-yl" is intended to mean a radical of the structure below.

In this context, substituted alkyl, substituted alkenyl or substituted alkynyl is intended to mean an alkyl, alkenyl or alkynyl moiety which is substituted by one or more substituent groups attached to such an alkyl, alkenyl or alkynyl moiety.

It is intended that the term “substituted,” as used for any part of the context, mean substitution with compatible substituents.

In this context, a straight unbranched alkyl chain is intended to mean an unbranched alkyl moiety such as ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl or n-heptyl.

In this context, a straight unbranched alkenyl chain is intended to mean an unbranched alkenyl moiety such as ethenyl, allyl, propen-1-yl, buten-1-yl, penten-1-yl, hexen-1-yl or hepten-1-yl .

In this context, an unsubstituted alkyl chain is intended to mean an unbranched alkyl moiety without substituents attached to the alkyl moiety, such as ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl or n-heptyl.

In this context, an unsubstituted alkenyl chain is intended to mean an unbranched alkenyl moiety without substituents attached to the alkenyl moiety, such as ethenyl, allyl, propen-1-yl, buten-1-yl, penten-1-yl, hexen-1-yl or hepten-1-yl.

It is assumed that in this context, the Deputy (R1) in the alpha position relative to the carbonyl group means the Deputy (R1) in the alpha position to the carbonyl part, as shown below

It is assumed that in this context, the Deputy (R1) in beta position relative to the carbonyl group means the Deputy (R1) in beta position to the carbonyl part, as shown below

The term “pharmaceutically acceptable salt” is intended to mean salts prepared by reacting a basic compound of formula I with a suitable inorganic or organic acid, such as hydrochloric, hydrobromic, hydroiodic, sulfuric, nitric, phosphoric, formic, acetic, 2.2 - dichloroacetic, choline, adipic, ascorbic, L-aspartic, L-glutamine, galactar, milk, maleic, L-apple, phthalic, lemon, propionic, benzoic, glutaric, gluconic, D-glucuronov I, methanesulfonic, salicylic, succinic, malonic, tartaric, benzenesulfonic, ethane-1,2-disulfonic, 2-hydroxyethanesulfonic, toluenesulfonic, sulfamic or fumaric acid. Pharmaceutically acceptable salts of the compounds of formula I containing an acid moiety can also be prepared by reaction with a suitable base, such as sodium hydroxide, potassium hydroxide, magnesium hydroxide, calcium hydroxide, ammonia, or suitable non-toxic amines, such as lower alkyl amines, e.g. triethylamine, hydroxy ( lower alkyl) amines, for example, 2-hydroxyethylamine, bis- (2-hydroxyethyl) amine, cycloalkylamines, for example dicyclohexylamine, or benzylamines, for example, Ν, Ν'-dibenzylethylenediamine and dibenzylamine, or L-arginine or L-lysine.

The present invention also includes prodrugs of compounds of general formula I, such as esters, acetals, ketals, or other derivatives that undergo in vivo bioconversion before their pharmacological effects.

The term “solvate” is intended to mean the type of compound resulting from the interaction between the compound, for example a compound of formula I, and a solvent, for example alcohol, glycerin or water, wherein said type of compound is in solid form. When the solvent is water, the type of compound is called hydrate.

The compounds of formula I can be obtained in crystalline form either directly by concentration from an organic solvent, or by crystallization or recrystallization from an organic solvent or a mixture of such a solvent and a co-solvent, which may be organic or inorganic, such as water. The crystals can be isolated essentially in a solvent free form or as a solvate such as a hydrate. The invention covers all crystalline modifications and forms, as well as mixtures thereof.

The term “cancer” is intended to include skin cancer, such as non-melanoma skin cancer, malignant melanoma, Merkel cell cancer, squamous cell carcinoma, basal cell carcinoma in the context of the present invention. Basal cell carcinoma also includes surface-localized basal cell carcinoma, as well as nodular basal cell carcinoma. Squamous cell carcinoma covers the intraepithelial form of squamous cell carcinoma (Bowen's disease), invasive squamous cell carcinoma, skin squamous cell carcinoma, squamous cell carcinoma of the mucous membrane, squamous cell carcinoma of the head and neck. Other types of cancer include hematologic cancer, such as myeloid cancers, in particular, such as acute myeloid leukemia and chronic myeloid leukemia; prostate and bladder cancer, including benign prostatic hyperplasia, intraepithelial carcinoma with prostatitis, bladder carcinoma, prostate adenocarcinoma, and kidney cancer. Other cancers include cancer associated with AIDS, auditory nerve neuroma, adenoid cystic cancer, adrenocortical cancer, idiopathic myeloid metaplasia, alopecia, soft alveolar sarcoma, anal canal cancer, angiosarcoma, aplastic anemia, astrocytoma, basal ataxia, and telangiomas cell cancer (bcc), bladder cancer, bone cancer, intestinal cancer, brain stem glioma, cancer of the brain and central nervous system, breast cancer, central nervous system cancer, carcinoid cancer, cervical cancer ki, brain cancers in children, cancer in children, soft tissue sarcomas of children, chondrosarcoma, choriocarcinoma, colorectal cancers, cutaneous T-cell lymphoma, swelling dermatofibrosarcomas, desmoplastic small cell carcinoma, duct cancer, endocrine cancer, endometrial cancer, ependoma , esophageal cancer, Ewing's sarcoma, extrahepatic bile duct cancer, eye cancer, eye diseases: melanoma, retinoblastoma, tubal cancer, Fanconi anemia, fibrosarcoma, gall bladder cancer, stomach cancer, cancer of the stomach of the intestinal tract, gastrointestinal carcinoid cancer, cancers of the genitourinary system, cancers of the germinal cells, gestational trophoblastic disease, glioma, gynecological cancers, hematologic malignancies, including acute myeloid leukemia, head and neck cancer, liver cell cancer, hereditary breast cancer, histiocytosis, Hodgkin's disease, human papillomavirus disease, chorionadenoma, hypercalcemia, carcinoma of the throat, intraocular melanoma, T-cell cancer x islets, Kaposi’s sarcoma, kidney cancer, Langerhans cell histocytosis, larynx cancer, leiomyosarcoma, Fromeni cancer syndrome, lip cancer, liposarcoma, liver cancer, lung cancer, lymphedema, lymphoma, Hodgkin’s lymphoma, non-Hodgkin’s lymphoma, male breast cancer, rhabdoid cancer kidneys, medulloblastoma, mesothelioma, metastatic cancer, oral cancer, multiple endocrine neoplasia, fungoid mycosis, myelodysplastic syndromes, myeloma, myeloproliferative diseases, nose cancer, nasopharyngeal cancer, nephroblastoma, neuroblastoma, neurofibromatosis, Nijmegen’s syndrome, non-small cell lung cancer (nsclc), eye cancer, esophageal cancer, oral cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer of the stoma, pancreatic cancer, paranasal cancer, parathyroid cancer, parotid cancer, penile cancer, peripheral neuroectodermal cancers, pituitary cancer, true polycythemia, prostate cancer, rare cancers and related disorders, retinoblastoma, rhabdomyosarcoma, Rotmund-Thomson syndrome, cancer, sarcoma, schwannomas of the salivary glands, Cesari syndrome, chalk peritoneal lung cancer (sclc), small intestine cancer, soft tissue sarcoma, spinal cord cancer, stomach cancer, synovial sarcoma, testicular cancer, thymus cancer, thyroid cancer, transitional cell (bladder), transient (renal) cancer pelvis / ureter), trophoblastic cancer, urethral cancer, urinary system cancer, uroplatel cancer, uterine sarcoma, uterine cancer, vaginal cancer, vulva cancer, Waldenstrom macroglobulinemia and Wilms cancer. Solid cancer that is treated using the methods of the present invention may be a primary lesion or may result from metastasis of a primary cancer. Furthermore, if the solid cancer is a metastasis of the primary cancer, the primary cancer may be either primary solid cancer, as described above, or may be dispersed primary cancer.

In an embodiment of the present invention, “cancer” is skin cancer. In embodiments of the invention, skin cancer is non-melanoma skin cancer, malignant melanoma, Merkel cell carcinoma, squamous cell carcinoma, squamous cell carcinoma, basal cell carcinoma such as superficial basal cell carcinoma or nodular basal cell carcinoma.

The term “photo-injured skin” is intended to include fine lines, wrinkles, and UV aging in the context of the present invention. UV aging is often manifested by an increase in the thickness of the epidermis or epidermal atrophy and most noticeably by solar elastosis, the accumulation of elastin-containing material directly under the skin-epidermal compound. Collagen and elastic fibers become fragmented and disorganized. In cosmetic cases, this can be observed as redness and / or thickening of the skin, leading in appearance to hard skin, fragility and uneven pigmentation, loss of tone and elasticity, as well as the appearance of wrinkles, dryness, freckles and the formation of deep wrinkles.

The term "viral infections" is intended to include HPV infections in the context of the present invention, leading to the formation of growths (warts) on the body, for example on the skin, genitals and oral cavity. HPV refers to the human papillomavirus. Other viruses are selected from adenoviruses, papovaviruses, herpes viruses (e.g., herpes simplex), chickenpox, Epstein-Barr virus, CMV virus, poxvirus (such as smallpox virus), vaccinia virus, hepatitis A viruses, hepatitis B viruses, hepatitis C, rhinoviruses, polio viruses, rubella, arbovirus, rabies viruses, influenza A and B, measles, mumps and HIV, HTLV I and II. In an embodiment of the invention, HPV infection refers to common warts or genital warts.

The term "bacterial infections" is intended to include prokaryotic and eukaryotic bacterial infections and gram-positive, gram-negative and gram-labile bacteria, and intracellular bacteria in the context of the present invention. Examples of bacteria include Treponema, Borrelia, Neisseria, Legionella, Bordetella, Escherichia, Salmonella, Shigella, Klebsiella, Yersinia, Vibrio, Hemophilus, Rickettsia, Chlamydia, Mycoplasma, Staphylococcus aureus, Streptococcus bacterium, Mycobacterium bacilli, Corynecobacterium bacteria, Corynecrobacterium bacteria, Corynecobacterium bacilli, Icobacterium bacilli, Icobacterium bacilli, bacteria Listeria. In particular, bacteria of the following species: Treponema pallidum (pale spirochete), Borrelia Burgdorferi, Neisseria gonorrhoea, Legionella pneumophila, Bordetella pertussis, Escherichia coli, Salmonella typhi, Salmonella typhimurium, Shigella dysenteriae, Klebsiella fibria pheida phiemis fibria rickettsii, Chlamydia trachomatis, Mycoplasma pneumonia, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Bacillus anthracis, Clostridium botulinum, Clostridium tetani, Clostridium perferium leprebiumerpiobiumerpioberium lepteriberium lepteriberium lepteriberium cholerae, The lower eukaryotic organism includes yeast and fungi, such as Pneumocystis nerinii, Candida albicans, Aspergillus, Histoplasma capsulatum, Blastomyces dermatitidis, Cryptococcus neoformans, Trichophyton and Microsporum. The complex eukaryotic organism includes worms, insects, spiders, nematodes, amoeba, Entamoeba histolytica, Giardia lamblia, Trichonomonas vaginalis, Trypanosoma brucei gembiense, Trypanosoma cruzi, Blantidium coli, Toxoplasma gondii, Cryptosporidium or

The phrase “physiological disorders or diseases associated with hyperplasia or neoplasia” is intended to include, in the context of the present invention, disorders or diseases such as skin warts, including common warts (Verruca vulgaris), plantar warts (Verruca plantaris) and flat warts (verruca plana ); genital warts (condyloma acuminatum), pyogenic granuloma, hemangioma, scleroderma; cancers and precancerous lesions, such as senile keratosis, squamous cell carcinoma including the intraepithelial form of squamous cell carcinoma (Bowen's disease), invasive squamous cell carcinoma, squamous cell carcinoma of the skin, squamous cell carcinoma of the mucous membrane, squamous cell carcinoma of the head and neck; basal cell carcinoma, including superficial basal cell carcinoma and nodal basal cell carcinoma; bladder cancer, lentigo malignant, cervical dysplasia, vulvar dysplasia and anal dysplasia, primary epithelial melanoma, head and neck cancer, skin metastases of any cancer, Kaposi’s sarcoma, keratoacanthoma, Merkel’s tumor cells, prostate cancer, fungoid mycosis, intraoperative including anal cancer, cancer of the cervix, ducts, oral cavity, anus, prostate, penis, vaginal neoplasia and intraepithelial neoplasia of the female external genital organs.

The term “cosmetic indications” is intended to include, in the context of the present invention, indications such as photo-injured skin, seborrheic keratosis, scars, keloids, melasma, Poikiloderma Civatte, tattoo removal, nevus, soft warts.

In the context of the present invention, the term “wound healing” means a reduction or minimization of scar tissue or an improvement in a cosmetic or functional result in reducing wounds and scars when the wound is skin, chronic or, for example, associated with diabetes, and includes cuts and lacerations, surgical incisions , punctures, scratches, compression wounds, abrasions, friction wounds, chronic wounds, ulcers, thermal wounds, chemical wounds, wounds resulting from pathogenic infections, donor skin transplants / grafts and recipient sites, immune response conditions, wounds to the oral cavity, stomach, or intestines, damage to cartilage or bone, amputation of limbs, and damage to the cornea.

The compounds of the present invention are intended to be used in the treatment of cancer, senile keratosis, seborrheic keratosis, viral infections, bacterial infections, wound healing and in the treatment of photo-injured skin.

In an embodiment of the invention, the compounds of the invention are contemplated to be used in the treatment of superficial basal cell carcinoma (BCC), nodular BCC, squamous cell carcinoma, or intracranial squamous cell carcinoma (SCCIS).

In an embodiment of the invention, the compounds of the invention are intended to be used in the treatment of senile keratosis.

In an embodiment of the invention, the compounds of the invention are intended to be used in the treatment of seborrheic keratosis.

In an embodiment of the invention, the compounds of the invention are contemplated for use in the treatment of photo-injured skin.

In an embodiment of the invention, the compounds of the invention are intended to be used in the treatment of injuries caused by HPV infection.

In an embodiment, the lesions are common warts or genital warts.

In an embodiment of the invention, the compounds of the invention are intended to be used in the treatment of an intraepithelial form of squamous cell carcinoma or invasive squamous cell carcinoma.

In an embodiment of the invention, the compounds of the invention are intended to be used in the treatment of squamous cell carcinoma of the skin, squamous cell carcinoma of the mucous membrane, or squamous cell carcinoma of the head and neck.

In an embodiment of the invention, the compounds of the invention are intended to be used in the treatment of superficial basal cell carcinoma or nodular basal cell carcinoma.

In an embodiment of the invention, the compounds of the invention are intended to be used in the treatment of skin warts or genital warts.

In an embodiment of the invention, the compounds of the invention are intended to be used in the treatment of conventional warts, plantar warts and flat warts.

In an embodiment of the invention, the compounds of the invention are intended to be used in the treatment of malignant lentigo.

In an embodiment of the invention, the compounds of the invention are intended to be used in the treatment of cervical intraepithelial neoplasia, anal intraepithelial neoplasia or vulvar intraepithelial neoplasia.

In an embodiment of the invention, the compounds of the invention are intended to be used in the treatment of acute myeloid leukemia.

In an embodiment, the invention relates to a method for treating cancer, senile keratosis, seborrheic keratosis, viral infections, bacterial infections, wound healing and treating photo-injured skin by administering a compound of formula I to a subject in need thereof.

In an embodiment, the invention relates to a method for treating senile keratosis by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to a method for treating seborrheic keratosis by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to a method for treating photo-injured skin by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to a method for treating lesions caused by HPV infection by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to a method for treating common warts or genital warts by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to a method for treating squamous cell carcinoma of the skin, squamous cell carcinoma of the mucous membrane, or squamous cell carcinoma of the head or neck by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to a method for treating common warts, plantar warts and flat warts by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to a method for treating malignant lentigo by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to a method for treating cervical intraepithelial neoplasia, anal intraepithelial neoplasia, or vulva intraepithelial neoplasia by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to a method for treating acute myeloid leukemia by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to the use of the aforementioned compound of formula I for the preparation of a pharmaceutical composition for treating or reducing the intensity of symptoms of a disease, disorder or condition susceptible to stimulation of an oxidative burst of neutrophils.

In an embodiment, the invention relates to the use of the aforementioned compound of formula I for the preparation of a pharmaceutical composition for treating or decreasing the intensity of symptoms of a disease, disorder or condition susceptible to stimulating the release of IL-8 keratinocytes.

In an embodiment, the invention relates to the use of the aforementioned compound of formula I for the preparation of a pharmaceutical composition for treating or reducing the intensity of symptoms of a disease, disorder or condition susceptible to inducing necrosis.

In an embodiment, the invention relates to a method for preventing, treating, reducing the intensity of symptoms or preventing physiological disorders or diseases susceptible to stimulate an oxidative burst of neutrophils by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to a method for preventing, treating, reducing the intensity of symptoms or preventing physiological disorders or diseases susceptible to stimulating the release of IL-8 keratinocytes by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to a method for preventing, treating, reducing the intensity of symptoms or preventing physiological disorders or diseases susceptible to inducing necrosis by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to the aforementioned compound of formula I for use in treating or reducing the intensity of symptoms of a disease, disorder or condition susceptible to stimulation of an oxidative burst of neutrophils.

In an embodiment, the invention relates to the aforementioned compound of formula I for use in treating or reducing the intensity of symptoms of a disease, disorder or condition susceptible to stimulating the release of IL-8 keratinocytes.

In an embodiment, the invention relates to the aforementioned compound of formula I for use in treating or reducing the intensity of symptoms of a disease, disorder or condition susceptible to induce necrosis.

In an embodiment, the invention relates to a method for treating acute myeloid leukemia by administering to a subject in need of the above compound of formula I.

In an embodiment, the invention relates to the aforementioned compound of formula I for use in the treatment, prevention, amelioration of symptoms, or the prevention of physiological disorders or diseases associated with senile keratosis, seborrheic keratosis, cancer, photo-injured skin, or damage caused by HPV infection.

In an embodiment, the invention relates to the use of a compound of formula I for the preparation of a medicament for treating, reducing the intensity of symptoms or preventing physiological disorders or diseases associated with senile keratosis, seborrheic keratosis, cancer, photo-injured skin or damage caused by HPV infection.

In an embodiment, the invention relates to a method for preventing, treating, reducing the intensity of symptoms or preventing physiological disorders or diseases associated with senile keratosis, seborrheic keratosis, cancer, photo-injured skin, or damage caused by HPV infection, by administering to a subject in need thereof a compound formulas I.

Pharmaceutical Compositions

For use in therapy, the compounds of the present invention are usually in the form of a pharmaceutical composition. Thus, the invention relates to a pharmaceutical composition comprising a compound of formula I together with a pharmaceutically acceptable excipient or carrier. The excipient should be “acceptable” in the sense of being compatible with the other ingredients of the composition and should not adversely affect its recipient.

The pharmaceutical compositions of the invention may be in unit dosage form, such as tablets, pills, capsules, powders, granules, elixirs, syrups, emulsions, ampoules, suppositories or parenteral solutions or suspensions, for oral, parenteral, ophthalmic, transdermal, intraarticular, local, pulmonary, nasal, buccal or rectal administration, or any other method suitable for administering the compounds of the invention and in accordance with accepted practices, such as those described in Remingt on: The Science and Practice of Pharmacy, 21 ^st ed., 2000, Lippincott Williams & Wilkins.

For oral administration in the form of a tablet or capsule, the compound of formula I can suitably be mixed with an oral non-toxic pharmaceutically acceptable carrier such as ethanol, glycerin, water or the like. In addition, suitable binders, lubricants, disintegrating agents, flavoring agents and coloring agents can be added to the mixture, if necessary. Suitable binders include, for example, lactose, glucose, starch, gelatin, gum arabic, tragacanth gum, sodium alginate, carboxymethyl cellulose, polyethylene glycol, waxes or the like. Lubricants include, for example, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride or the like. Disintegrating agents include, for example, starch, methyl cellulose, agar, bentonite, xanthan gum or the like. Additional capsule excipients include macrogol or lipids.

To obtain solid compositions such as tablets, the active compound of formula I is mixed with one or more excipients, such as those described above, and other pharmaceutical diluents, such as water, to obtain a solid preformed composition containing a homogeneous mixture of a compound of formula I. It is understood that the term “homogeneous” means that the compound of formula I is uniformly dispersed throughout the composition, so that the composition can be easily divided into equally effective dosage forms Tween forms, such as tablets or capsules.

In the form of a unit dose, the compound can be administered once or several times a day with suitable time intervals between them, always depending, however, on the condition of the patient, and in accordance with a prescription made by a medical practitioner. The unit dose of the preparation advantageously contains from 0.01 mg to 200 mg, preferably from 0.01 to 20 mg, for example, 0.01-5 mg of a compound of formula I.

A suitable dose of a compound of the present invention will depend, inter alia, on the age and condition of the patient, the severity of the disease being treated, and other factors well known to the practitioner. The compound can be administered either orally, parenterally, or topically according to various dosage regimens, for example, daily or at weekly intervals. Typically, a single dose will be in the range of 0.01 to 200 mg / kg body weight. The compound can be administered as a bolus (i.e., the entire daily dose can be administered immediately) or in divided doses two or more times a day.

If the treatment involves administration of another therapeutically active compound it is recommended to receive counseling from a publication Goodman &Gilman's The Pharmacological Basis of Therapeutics, 9 ^{th Ed., JG Hardman and LE} Limbird (Eds.), McGraw-Hill 1995 in the applicable doses of said compounds. Administration of a compound of the present invention with one or more other active compounds can be carried out either simultaneously or sequentially.

Liquid preparations for either oral or parenteral administration of a compound of the invention include, for example, aqueous solutions, syrups, aqueous or oily suspensions, and emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil. Suitable dispersing or suspending agents for aqueous suspensions include synthetic or natural gums such as tragacanth, alginate, gum arabic, dextran, sodium carboxymethyl cellulose, gelatin, methyl cellulose or polyvinylpyrrolidone.

For parenteral administration, for example, for intramuscular, intraperitoneal, subcutaneous or intravenous injection or infusion, the pharmaceutical composition preferably contains a compound of formula I, dissolved or solubilized in a suitable pharmaceutically acceptable solvent. For parenteral administration, the composition of the invention may include a sterile aqueous or non-aqueous solvent, in particular water, isotonic saline, isotonic glucose, buffered saline or other solvent commonly used for parenteral administration of therapeutically active substances. The composition can be sterilized, for example, by filtration through a bacteria-retaining filter, by adding a sterilizing agent to the composition, by irradiating the compositions, or by heating the composition. Alternatively, the compound of the invention may be presented as a sterile solid preparation, for example a lyophilized powder, which is dissolved in a sterile solvent immediately before use. A composition intended for parenteral administration may further comprise conventional additives, such as stabilizers, buffers or preservatives, for example antioxidants, such as methyl hydroxybenzoate or the like.

Formulations for rectal administration may be in the form of a suppository comprising the active ingredient and a carrier, such as cocoa butter, or in the form of an enema. Compositions suitable for intraarticular administration may be in the form of a sterile aqueous preparation of the active ingredient, which may be in microcrystalline form, for example, in the form of an aqueous microcrystalline suspension. Liposome preparations or biodegradable polymer systems can also be used for both intraarticular and ophthalmic administration of the active ingredient.

Compositions suitable for topical administration, including for ophthalmic treatment, include liquid or semi-liquid preparations, such as liniment, lotions, gels, applicators, oil-in-water or water-in-oil emulsions, such as creams, ointments or pastes; or solutions or suspensions, such as drops. Ophthalmic treatment compositions may preferably further comprise cyclodextrin. Compositions suitable for administration to the nasal or oral cavity or for inhalation include powder, self-propelled preparations and nebulized preparations, such as aerosols and atomizers.

Human skin, in particular the outer layer, the stratum corneum of the epidermis, provides an effective barrier against the penetration of microbial pathogens and toxic chemicals. Although this skin property is usually beneficial, it complicates the skin administration of pharmaceutical preparations, so that a large amount, if not a large part, of the active ingredient applied to the skin of a patient suffering from a skin disease cannot penetrate into the viable layers of the skin where it is active .

Penetration into the skin is facilitated by the addition of permeation enhancers, which include isopropyl alcohol, sulfoxides, azones, pyrrolidines, alkanols and glycols. In embodiments of the invention, penetration enhancers include DMSO, laurocapram, 2-pyrrolidone, decanol and propylene glycol. In one embodiment, the penetration enhancer is isopropyl alcohol.

In embodiments of the invention, the therapeutically active compound is dissolved in a suitable solvent. Suitable solvents are glycols, ketones, acetates and ethers. Derivatives of ingenol have been shown to have good stability in alcohols, such as benzyl alcohol and isopropyl alcohol. It was previously shown that, in general, ingenol derivatives have good stability at low pH. In embodiments of the present invention, the pH of the pharmaceutical is below 7. In embodiments of the present invention, the pH of the pharmaceutical is below 6. In embodiments of the present invention, the pH of the pharmaceutical is below 4.5. In embodiments of the present invention, the pH of the pharmaceutical is below 4.0. In embodiments of the present invention, the pH of the pharmaceutical is below 4.5 and not below 2.5. In embodiments of the present invention, the pH of the pharmaceutical is below 4.0 and not below 2.5. A preferred pH range can be obtained by including a suitable buffer in the formulation. In one embodiment, the buffer is an acetate buffer. In embodiments of the invention, citrate buffer is used. In embodiments of the invention, a mixed citrate-phosphate buffer is used.

In one embodiment, the composition is an ointment. According to the current FDA classification, ointment is a semi-solid dosage form, which may contain water and volatile substances in an amount up to 20% of the mass. and which contains more than 50% of the mass. hydrocarbons, waxes or polyols in the filler. Thus, according to the invention, the ointment can be a water-in-oil composition, in which case nanosuspension as such can be added to the lipophilic components of the composition so that the composition contains up to 10% by weight. or preferably up to 5% of the mass. water phase. Alternatively, the composition may be a non-aqueous ointment that contains about less than 2%, preferably less than 1% free water by weight of the composition.

The ointment carrier may suitably contain paraffin selected from paraffins consisting of hydrocarbons with a chain length of C _5-60 and mixtures thereof. An often used ointment carrier is petrolatum or white soft paraffin, which consists of hydrocarbons of various chain lengths and with a maximum chain length of approximately C _40-44 , or a mixture of petroleum jelly and liquid paraffin (consisting of hydrocarbons with various chain lengths with a maximum at C _28-40 ) Although petrolatum provides occlusion of the treated skin surface, reducing transdermal water loss and enhancing the therapeutic effect of the active ingredient in the composition, it tends to give a feeling of a greasy and / or sticky surface that persists for some time after application and is not easy to spread on the surface. Therefore, it may be preferable to use paraffins consisting of hydrocarbons with a slightly shorter chain length, such as paraffins consisting of hydrocarbons with a chain length of maximum C _14-16 , C _18-22 , C _20-26 , or mixtures thereof. It has been found that such paraffins are more cosmetically acceptable in the sense that they are less sticky and / or greasy upon application and more easily distributed on the surface. Therefore, it is anticipated that this will lead to better patient compliance with therapeutic recommendations. Suitable paraffins of this type are manufactured by Sonneborn and sold under the trade name Sonnecone, for example Sonnecone CM, Sonnecone DM1, Sonnecone DM2 and Sonnecone HV. These paraffins are further described and characterized in WO 08/141078, which is incorporated herein by reference. (The hydrocarbon composition of paraffins was determined by gas chromatography).

In order to impart a desired viscosity to the composition, a lipophilic viscosity increasing ingredient such as wax can be included in it accordingly. The wax may be a mineral wax consisting of a mixture of high molecular weight hydrocarbons, for example, saturated C _35-70 alkanes, such as microcrystalline wax. In addition, the wax may be vegetable or animal wax, for example, may consist of esters of C _14-32 fatty acids and C _14-32 fatty alcohols, such as beeswax. The amount of viscosity increasing ingredient may vary depending on the ability of the ingredient to impart viscosity, but typically its content may be in the range of about 1-20% by weight. composition. When the viscosity increasing ingredient is microcrystalline wax, it is usually present in an amount in the range of about 5-15% by weight, for example, about 10% by weight. composition.

To maintain good physical stability of the composition, in particular to avoid separation of the aqueous and lipid phases in it, it may be advisable to include an emulsifier for a water-in-oil emulsion with an HLB value (water-lipophilic balance) of 3-8. Examples of such emulsifiers are polyoxyethylene-C _8-22 alkyl esters, for example polyoxyethylene stearyl ether, polyoxyethylene acetyl ether, polyoxyethylene oleyl ether or polyoxyethylene lauryl ether. The amount of emulsifier is usually in the range of 2-10% w / w. composition. In another embodiment, the composition is a cream that may contain components similar to those of the ointment, but which is usually an oil-in-water emulsion containing a significant amount of water.

The composition may also contain other components commonly used in skin preparations, for example, antioxidants (e.g. alpha tocopherol), preservatives such as benzyl alcohol, sodium edetate, pigments, skin softening agents, skin healing agents and skin conditioning agents such as urea , allantoin or bisabolol, cf. CTFA Cosmetic Ingredients Handbook, 2 ^nd Ed., 1992. In one embodiment, the preservative is benzyl alcohol.

In an embodiment, the composition is a gel. Suitable gelling agents include water soluble cellulose derived polymers such as hydroxyalkyl cellulose polymers. In embodiments of the invention, such polymers are hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose and hydroxypropyl methyl cellulose. Other gelling agents are celluloses such as carboxymethyl cellulose, methyl hydroxyethyl cellulose and methyl cellulose, carbomer such as carbopol and carrageenan. In embodiments of the invention, the gelling agent is obtained from cellulose. In embodiments of the invention, the cellulose is hydroxyalkyl cellulose, such as hydroxyethyl cellulose.

In an embodiment of the invention, the composition comprises an active compound, a penetration enhancer, a preservative, a gelling agent and a buffer at a pH of less than 4 and not less than 2.5. For topical administration, the compound of formula I is usually present in an amount of from 0.001 to 20% of the mass. compositions, for example from 0.01% to about 10%. In embodiments of the present invention, the active compound is present in an amount of 0.05-1%. In an embodiment of the present invention, the active compound is present in an amount of 0.01-0.5%. In an embodiment of the present invention, the active compound is present at a concentration of about 0.1%. In an embodiment of the invention, the composition contains 0.005-0.1% of the active compound, 20-40% of isopropyl alcohol, 0.5-10% of benzyl alcohol, 0.5-5% of hydroxyethyl cellulose and citrate buffer up to 100%.

A topical gel preparation of ingenol is described in WO07 / 068963, which is incorporated by reference.

Production methods

Compounds of formula I can, for example, be prepared using the reactions and procedures described below, together with methods known in the field of synthetic organic chemistry, or variants thereof, used by those skilled in the art. Preferred methods include, but are not limited to, the methods described below. The reactions are carried out in solvents suitable for the reagents and substances used and suitable for the conversion. In addition, in the synthetic methods described below, it should be understood that all proposed reaction conditions, including the choice of solvent, reaction atmosphere, reaction temperature, duration of the experiment and processing procedures, are chosen so that they are standard for the given reaction and can be easily determined by a person skilled in the art. Not all compounds belonging to this class may be compatible with some of the reaction conditions required in some of the described methods. Such restrictions on substituents that are compatible with the reaction conditions will be readily apparent to those skilled in the art, alternative methods may also be used. The compounds of the present invention or any intermediate may be purified if required using standard methods well known to those skilled in synthetic organic chemistry, for example the methods described in the publication W. Armarego "Purification of Laboratory Chemicals" , Butterworth-Heinemann, 6 ^th ed. 2009. The starting materials are either known compounds, commercially available, or can be obtained by conventional synthetic methods well known to those skilled in the art.

The compounds of the invention can, for example, be prepared according to the following non-limiting general methods and examples.

Scheme 1

Scheme 2

Compounds of general formula I can, for example, be synthesized according to Scheme 1 or 2 by reacting ingenol with a hydroxyl protecting agent or dihydroxyl protecting agent to obtain protected ingenol derivatives a or according to the methods described, but not limited to, by “Protective Groups in Organic Synthesis”, 4th ed. P.G.M. Wuts T.W. Greene, John Wiley, 2007 or P.J. Kocienski, "Protecting Groups", 3rd ed. G. Thieme, 2003 and in Publication References.

For example, compound a, where the protecting group (Pg) is triphenylmethyl, can be synthesized by reacting ingenol with a triphenylmethyl-containing reagent, such as triphenylmethylpyridinium fluoroborate or triphenylmethyl chloride, in a suitable solvent such as pyridine, Ν, Ν-dimethylformamide or in the presence of dichloromethane lack of base (e.g., Opferkuch et al., Z. Naturforschung, (1981), 36B, 878). Compound a, where the protecting group (Pg) is silyl, for example, can be synthesized by reacting ingenol with a silyl chloride, such as tert-butyldimethylsilyl chloride, tert-butyldiphenylsilyl chloride or triisopropylsilyl chloride, in a suitable solvent such as N, N-dimethylmethylforminide tetrahydrofuran or acetonitrile, in the presence of a suitable base such as imidazole, triethylamine, N, N-diisopropylethylamine or 4- (N, N-dimethylamino) pyridine (see, for example, Sorg, B. et al., Z. Naturforsch., (1982), 37B, 1640-47), or by the reaction of compound (II) with forces ltriflatom such as t-butyldimethylsilyl trifluoromethanesulfonate in a suitable solvent such as dichloromethane in the presence of a suitable base such as triethylamine.

Compound a, where Pg is 2-tetrahydropyranyl, for example, can be synthesized by reacting ingenol with dihydropyran in a suitable solvent, such as dichloromethane or acetonitrile, in the presence of a suitable acid, such as p-toluenesulfonic acid. A compound c, wherein the protecting group (Pg) is acetal, such as benzylidene acetal, can, for example, be obtained by reacting ingenol with benzaldehyde or benzaldehyde dimethyl acetal in a suitable solvent, such as dichloromethane or N, N-dimethylformamide, in the presence of a suitable acid such as p- toluenesulfonic acid. A compound c, wherein the protecting group (Pg) is a ketal, such as isopropylidene ketal, for example, can be synthesized by reacting ingenol with a ketone, such as acetone, or dimethoxy ketal, such as 2,2-dimethoxypropane, in a suitable solvent, such as dichloromethane or Ν, Ν-dimethylformamide, in the presence of a suitable acid, such as p-toluenesulfonic acid (see, for example, B. Sorg, Z. Naturforsch. (1982), 37b, 748-756). Acetone and 2,2-dimethoxypropane may also act as solvents.

As shown in Schemes 1 and 2, protected derivatives of ingenol a or c can be esterified to give compounds of general formula b or d according to the methods for the esterification of hydroxyl groups described, but not limited to, by Esterification, J. Otera, Wiley-VCH, 2003 and in the links in it. Compound b or d can, for example, be synthesized by reacting compound a or c with an activated derivative of an acid, such as an acid halide, such as an acid chloride. The esterification by reaction with an acid chloride can be carried out in a suitable solvent, such as dichloromethane or toluene, without an activator, or it can be carried out in the presence of a base such as pyridine, triethylamine or 4- (N, N-dimethylamino) pyridine (see, for example, B. Sorg, Z. Naturforsch, (1982), 37b, 748-756).

Compound b or d can, for example, be synthesized by reacting compound a or c with an activated acid derivative such as acid anhydride. The esterification by reaction with an acid anhydride can be carried out without a catalyst (see, for example, Opferkuch et al., Z. Naturforschung, (1981), 36B, 878) or in the presence of an acid catalyst, which is used as an acid such as perchloric acid or acid Lewis, such as scandium (III) triflate or bismuth (III) triflate, or in the presence of a base such as sodium bicarbonate or triethylamine.

Compound b or d can, for example, be synthesized by reacting compound a or c with an activated derivative of an acid, such as a mixed acid anhydride, such as trichlorobenzoic acid. The esterification by reaction with mixed anhydride can be carried out in a suitable solvent without a catalyst or in the presence of an acid catalyst using an acid such as perchloric acid or Lewis acid, such as scandium (III) triflate or bismuth (III) triflate, or in the presence of a base such like sodium bicarbonate or triethylamine. Compound b or d can, for example, be synthesized by reacting compound a or c with an acid in the presence of a coupling reagent such as carbodiimide, such as dicyclohexylcarbodiimide or N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide, in the presence or absence of a base such like 4- (N, N-dimethylamino) pyridine, and without catalysts or with catalysts such as 4- (N, N-dimethylamino) pyridine in a suitable solvent such as dichloromethane (see, for example, Appendino et al., Eur. J. Org. Chem. (1999), 3413). Solid-state coupling reagents can also be used at the esterification stage [Nam, N.-H., Journal of Combinatorial Chemistry, (2003), 5, 479-545 or “Esterification” by J. Otera, Wiley-VCH, 2003].

Compounds of formula I can be prepared by selectively removing Pg protecting groups on compounds of general structure b or d according to methods for protecting groups on hydroxyl or dihydroxyl protected groups as described in Protective Groups in Organic Synthesis, 4th ed. P.G.M. Wuts T.W. Greene, John Wiley, 2007 or P.J. Kocienski, "Protecting Groups", 3rd ed. G. Thieme, 2003 and references therein, but are not limited to these described methods.

Compounds of general formula I can, for example, be prepared from compounds of general formula d, where Pg is acetal, such as benzylidene acetal, or ketal, such as isopropylidene ketal, by cleaving the protecting group in the presence of a suitable acid, such as aqueous hydrogen chloride, acetic acid, trifluoroacetic acid acid or p-toluenesulfonic acid, in a suitable solvent, such as methanol or aqueous tetrahydrofuran. Compounds of general formula I can, for example, be prepared from compounds of general formula b, where Pg is alkoxyalkyl, such as 2-tetrahydropyranyl, by cleaving the acetal moiety, for example, by acid-catalyzed cleavage in the presence of a suitable acid, such as p-toluenesulfonic acid, in a suitable solvent such as methanol. Compounds of general formula I can, for example, be prepared from compounds of general formula b, where Pg is silyl, such as tert-butyldimethylsilyl, by reacting compound b with a suitable acid, such as hydrogen chloride, in a suitable solvent, such as methanol, or by reaction with a fluoride source, such as tetra-n-butylammonium fluoride or tetrafluorosilane, in a suitable solvent, such as tetrahydrofuran or acetonitrile. Compounds of general formula I can, for example, be prepared from compounds of general formula b, where Pg is triphenylmethyl, by reacting compound b with a suitable acid, such as formic acid or trifluoroacetic acid, in a suitable solvent, such as ether, methanol or dichloromethane.

Compounds of formulas b, d or I of the above scheme 1 or 2 can, for example, be synthesized by enzymatic esterification by reacting compound a, c or ingenol with an acyl donor such as acid anhydride, an ester such as a vinyl ester or thioester, in the presence of an enzyme such as lipase or esterase.

Examples

General Provisions

All starting materials used are commercially available, unless otherwise described. For ¹ H nuclear magnetic resonance (NMR) spectra, chemical shifts (δ) (in ppm) are indicated; and tetramethylsilane (δ = 0.00) is the standard. The values of a specific doublet (d), triplet (t), quartet (sq.) Or multitude (m) are indicated. Chemical shifts of interchangeable protons (often broadened singlets (br. S)) are sometimes difficult to detect in the spectrum. All organic solvents used were anhydrous unless otherwise indicated. Flash chromatography was performed on silica gel. Suitable mixtures of ethyl acetate and heptane were used as eluents, unless otherwise indicated. Compounds were detected on TLC plates with the development of an aqueous solution of potassium permanganate.

Ingenol-5.20-acetonide

Ingenol (1.00 g, 2.30 mmol) was dissolved in a solution of p-toluenesulfonic acid monohydrate in acetone (0.47 mg / ml, 22.5 ml). The resulting solution was stirred at room temperature for 25 minutes. To this solution was added a saturated aqueous solution of NaHCO ₃ (0.2 ml). The resulting mixture was concentrated in vacuo. The residue was dissolved in brine and extracted with ethyl acetate. The combined organic phases were dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash chromatography (heptane / ethyl acetate 19: 1 → heptane / ethyl acetate 1: 0) to give the title compound as a white solid (616 mg, 69%). (See also Opferkuch, HJ et al., Z. Naturforsch., (1981), 86b, 878-887).

¹ H NMR (300 MHz, CDCl ₃ ) δ 5.91 (q, J = 1.5 Hz, 1H), 5.79 (m, 1H), 4.25 (d, J = 4.5 Hz, 1H), 4.20-4.07 (m, 3H), 3.93 (s, 1H), 3.51 (s, 1H), 2.57-2.41 (m, 2H), 2.25 (ddd, J = 15.7, 8.4, 2.9 Hz, 1H), 1.85 (d, J = 1.5 Hz, 3H), 1.77 (dt, J = 15.8, 5 , 9 Hz, 1H), 1.41 (s, 3H), 1.35 (s, 3H), 1.13 (s, 3H), 1.05 (s, 3H), 1.00-0.87 (m, 4H), 0.70 (td, J = 8.4, 6.4 Hz, 1H).

General procedures for the preparation of compounds of general formula II

Procedure a

A mixture of carboxylic acid (0.100 mmol) of dicyclohexylcarbodiimide (0.100 mmol), 4- (N, N-dimethylamino) pyridine (0.0025 mmol) and ingenol-5.20-acetonide (0.050 mmol) was stirred at room temperature in dichloromethane for 20 -24 hours This mixture was mixed with ethyl acetate, filtered and washed with a saturated aqueous solution of sodium chloride. The organic phase was dried with sodium sulfate, concentrated in vacuo and purified by flash chromatography (heptane → heptane / ethyl acetate 7: 3) to give the title compound as a white solid.

Procedure b

A mixture of acyl chloride (0.0625 mmol), diisopropylethylamine (0.075 mmol), 4- (N, N-dimethylamino) pyridine (0.070 mmol) and ingenol-5.20-acetonide (0.050 mmol) was stirred at 55 ° C in tetrahydrofuran for 6-20 hours This mixture was mixed with ethyl acetate, filtered and washed with a saturated aqueous solution of sodium chloride. The organic phase was dried with sodium sulfate, concentrated in vacuo and purified by flash chromatography (heptane → heptane / ethyl acetate 7: 3) to give the title compound as a white solid.

Procedure with

A mixture of carboxylic acid (0.100 mmol), dicyclohexylcarbodiimide (0.100 mmol), 4- (N, N-dimethylamino) pyridine (0.025 mmol) and ingenol-5.20-acetonide (0.050 mmol) was stirred in a microwave oven at 150 ° C in acetonitrile within 5 minutes This mixture was mixed with ethyl acetate, filtered and washed with a saturated aqueous solution of sodium chloride. The organic phase was dried over sodium sulfate, concentrated in vacuo and purified by flash chromatography (heptane → heptane / ethyl acetate, 7: 3), to thereby give the title compound as a white solid.

Procedure d

A mixture of acyl chloride (0.125 mmol), diisopropylethylamine (0.250 mmol), 4- (N, N-dimethylamino) pyridine (0.025 mmol) and ingenol-5.20-acetonide (0.050 mmol) was stirred in a microwave oven at 150 ° C in acetonitrile in within 20 minutes The mixture was mixed with ethyl acetate, filtered and washed with a saturated aqueous solution of sodium chloride. The organic phase was dried with sodium sulfate, concentrated in vacuo and purified by flash chromatography (heptane → heptane / ethyl acetate, 7: 3), to thereby give the title compound as a white solid.

General Procedure for the Preparation of Compounds of General Formula I

Procedure e

Ingenol-5,20-acetonide-3-acylate (0.10 mmol) was dissolved in tetrahydrofuran (0.47 ml) in an argon atmosphere. An aqueous HCl solution (4 M, 4.7 μl) was added. The solution was stirred at room temperature for 20-27 hours. The solution was concentrated in vacuo. The residue was purified by flash chromatography (heptane / ethyl acetate 5: 1 → heptane / ethyl acetate, 3: 7) to give the title compound.

Getting connection 201

Ingenol-5,20-acetonide-3- (2-methyl acrylate) (compound 201)

Compound 201 was prepared according to procedure a.

Starting material: 2-methylacrylic acid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.12-6.11 (m, 1H), 6.05 (m, 1H), 5.79-5-77 (m, 1H), 5.62 ( s, 1H), 5.60 (m, 1H), 4.25-4.11 (m, 3H), 4.02 (s, 1H), 3.14 (s, 1H), 2.61-2 54 (m, 1H), 2.31-2.22 (m, 1H), 1.97 (m, 3H), 1.81-1.72 (m, 4H), 1.47 (s, 3H) ), 1.42 (s, 3H), 1.09 (s, 3H), 1.05 (s, 3H), 0.99 (d, 3H), 0.94-0.88 (m, 1H) 0.73-0.65 (m, 1H).

Getting connection 202

Ingenol-5,20-acetonide-3- (3-methylbutenoate) (compound 202)

Compound 202 was obtained according to procedure a.

Starting material: 3-methylbutenic acid.

Preparation of Compound 203

Ingenol-5,20-acetonide-3- (2,3-dimethylbutenoate) (compound 203)

Compound 203 was obtained according to procedure a.

Starting material: 2,3-dimethylbutenic acid.

Getting connection 204

Ingenol-5,20-acetonide-3- (methylenebutanoate) (compound 204)

Compound 204 was obtained according to procedure a.

Starting material: 2-methylenebutanoic acid.

Getting connection 205

Ingenol-5,20-acetonide-3- (2-methylpropanoate) (compound 205)

Compound 205 was obtained according to procedure a.

Starting material: 2-methylpropanoic acid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.05-6.04 (m, 1H), 5.78-5.76 (m, 1H), 5.54 (s, 1H), 4.24- 4.10 (m, 3H), 4.00 (s, 1H), 3.09 (s, 1H), 2.65-2.54 (m, 2H), 2.31-2.22 (m, 1H), 1.80-1.71 (m, 4H), 1.45 (s, 3H), 1.41 (s, 3H), 1.21 (d, 3H), 1.19 (d, 3H) ), 1.09 (s, 3H), 1.05 (s, 3H), 0.98 (d, 3H), 0.93-0.87 (m, 1H), 0.73-0.65 ( m, 1H).

Preparation of Compound 206

Ingenol-5,20-acetonide-3- (3-methylbutanoate) (compound 206)

Compound 206 was obtained according to procedure a.

Starting material: 3-methylbutanoic acid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.04-6.03 (m, 1H), 5.79-5.77 (m, 1H), 5.55 (s, 1H), 4.24- 4.10 (m, 3H), 4.00 (s, 1H), 3.15 (s, 1H), 2.60-2.52 (m, 1H), 2.32-2.04 (m, 4H), 1.80-1.71 (m, 4H), 1.45 (s, 3H), 1.41 (s, 3H), 1.09 (s, 3H), 1.05 (s, 3H ), 0.99 (d, 6H), 0.98 (d, 3H), 0.94-0.87 (m, 1H), 0.73-0.65 (m, 1H).

Getting connection 207

Ingenol-5,20-acetonide-3- (2 (RS) -methylbutanoate) (compounds 207a and 207b)

Compounds 207a and 207b were prepared according to procedure a.

Starting material: (RS) -3-methylbutanoic acid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.05-6.03 (m, 1H), 5.79-5.76 (m, 1H), 5.55 (s, 1H), 4.24- 4.10 (m, 3H), 4.00 (s, 1H), 3.12 (s, 1H), 2.61-2.53 (m, 1H), 2.48-2.40 (m, 1H), 2.31-2.22 (m, 1H), 1.80-1.66 (m, 5H), 1.56-1.47 (m, 1H), 1.46 (s, 3H) , 1.41 (s, 3H), 1.20-1.16 (2 × d, 3H), 1.09 (s, 3H), 1.05 (s, 3H), 0.99-0.87 (m, 7H); 0.73-0.65 (m, 1H).

Getting connection 208

Ingenol-5,20-acetonide-3- (3,3-dimethylbutanoate) (compound 208)

Compound 208 was prepared according to Procedure a.

Starting material: 3,3-dimethylbutanoic acid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.04-6.02 (m, 1H), 5.78-5.77 (m, 1H), 5.55 (s, 1H), 4.24- 4.10 (m, 3H), 4.00 (s, 1H), 3.15 (s, 1H), 2.62-2.55 (m, 1H), 2.30-2.20 (m, 1H), 2.27 (s, 2H), 1.78-1.69 (m, 4H), 1.45 (s, 3H), 1.41 (s, 3H), 1.10 (s, 3H) ), 1.06 (s, 9H), 1.05 (s, 3H), 0.97 (d, 3H), 0.93-0.87 (m, 1H), 0.73-0.65 ( m, 1H).

Getting connection 209

Ingenol-5,20-acetonide-3- (2-ethylbutanoate) (compound 209)

Compound 209 was obtained according to procedure a.

Starting material: 2-ethylbutanoic acid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.05-6.03 (m, 1H), 5.78-5.76 (m, 1H), 5.56 (s, 1H), 4.25- 4.10 (m, 3H), 4.01 (s, 1H), 3.16 (s, 1H), 2.64-2.54 (m, 1H), 2.31-2.21 (m, 2H), 1.78-1.48 (m, 8H), 1.46 (s, 3H), 1.41 (s, 3H), 1.09 (s, 3H), 1.04 (s, 3H ), 0.98 (d, 3H), 0.94 (t, 3H), 0.93 (t, 3H), 0.93-0.87 (m, 1H), 0.72-0.65 ( m, 1H).

Getting connection 210

Ingenol-5,20-acetonide-3- (2R-methylbutanoate) (compound 210)

Compound 210 was obtained according to procedure a.

Starting material: (R) -2-methylbutanoic acid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.05-6.03 (m, 1H), 5.79-5.76 (m, 1H), 5.55 (s, 1H), 4.24- 4.10 (m, 3H), 4.00 (s, 1H), 3.12 (s, 1H), 2.61-2.53 (m, 1H), 2.50-2.39 (m, 1H), 2.31-2.22 (m, 1H), 1.79-1.66 (m, 5H), 1.59-1.47 (m, 1H), 1.45 (s, 3H) 1.41 (s, 3H), 1.19-1.16 (d, 3H), 1.09 (s, 3H), 1.05 (s, 3H), 0.99-0.87 (m , 7H), 0.73-0.65 (m, 1H).

Getting connection 211

Ingenol-5,20-acetonide-3-crucible (compound 211)

Compound 211 was obtained according to procedure a.

Starting material: tiglinic acid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.90-6.82 (m, 1H), 6.05-6.04 (m, 1H), 5.78-5.76 (m, 1H), 5.62 (s, 1H), 4.24-4.11 (m, 3H), 4.01 (s, 1H), 3.17 (s, 1H), 2.61-2.56 (m, 1H), 2.31-2.22 (m, 1H), 1.86-1.72 (m, 10H), 1.45 (s, 3H), 1.42 (s, 3H), 1.09 (s, 3H), 1.05 (s, 3H), 1.00 (d, 3H), 0.94-0.88 (m, 1H), 0.73-0.63 (m, 1H).

Getting connection 212

Ingenol-5.20-acetonide-3- (phenylacetate) (compound 212)

Compound 212 was obtained according to procedure a.

Starting material: phenylacetic acid.

Getting connection 213

Ingenol-5,20-acetonide-3- (2Z- (methoxycarbonyl) acrylate) (compound 213)

Compound 213 was obtained according to procedure a.

Starting material: fumaric acid monomethyl ether.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.93 (d, 1H), 6.85 (d, 1H), 6.09 (m, 1H), 6.80-6.78 (m, 1H) 5.62 (s, 1H), 4.26-4.09 (m, 3H), 4.02 (s, 1H), 3.82 (s, 3H), 3.59-3.53 (m , 1H), 2.29-2.20 (m, 1H), 2.82-2.73 (m, 4H), 1.47 (s, 3H), 1.43 (s, 3H), 1, 27 (br s, 1H), 1.09 (s, 3H), 1.05 (s, 3H), 1.00 (d, 3H), 0.94-0.85 (m, 1H), 0 73-0.65 (m, 1H).

Getting connection 214

Ingenol-5,20-acetonide-3- (2-cyclohexylpropanoate) (compound 414)

Compound 214 was prepared according to procedure c.

Starting material: 2-cyclohexylpropanoic acid.

Getting connection 215

Ingenol-5.20-acetonide-3 - ((2Z) -2-methoxyimino-2-phenylacetate) (compound 215)

Compound 215 was prepared according to procedure c.

Starting material: (2Z) -2-methoxyimino-2-phenylacetic acid.

Getting connection 216

Ingenol-5.20-acetonide-3 - ((2E) -2-methoxyimino-2-phenylacetate) (compound 216)

Compound 216 was prepared according to procedure c.

Starting material: (2E) -2-methoxyimino-2-phenylacetic acid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.47-7.36 (m, 5H), 6.03-6.02 (m, 1H), 5.78-5.76 (m, 1H), 5.70 (s, 1H), 4.23-4.09 (m, 3H), 4.05 (s, 3H), 4.01 (s, 1H), 3.32 (s, 1H), 2 , 28-2.13 (m, 2H), 1.77 (d, 3H), 1.68-1.57 (m, 1H), 1.45 (s, 3H), 1.41 (s, 3H) ), 1.10 (s, 3H), 1.05 (s, 3H), 0.92-0.86 (m, 1H), 0.81 (d, 3H), 0.70-0.62 ( m, 1H).

Getting connection 217

Ingenol-5,20-acetonide-3 - ((Z) -2-methyl-3- (p-tolyl) prop-2-enoate) (compound 217)

Compound 217 was obtained according to procedure c.

Starting material: (Z) -2-methyl-3- (p-tolyl) prop-2-enoic acid.

Getting connection 218

Ingenol-5,20-acetonide-3 - ((E) -2-phenylbut-2-enoate) (compound 218)

Compound 218 was prepared according to procedure c.

Starting material: (E) -2-phenylbut-2-enoic acid.

Getting connection 219

Ingenol-5.20-acetonide-3- (2,2-diphenylacetate) (compound 219)

Compound 219 was prepared according to procedure c.

Starting material: 2,2-diphenylacetic acid.

Getting connection 220

Ingenol-5,20-acetonide-3- (2-cyano-2-cyclohexylidene acetate) (compound 220)

Compound 220 was prepared according to procedure d, but with an extension of the reaction time to 40 minutes.

Starting material: 2-cyano-2-cyclohexylideneacetyl chloride obtained from 2-cyano-2-cyclohexylideneacetic acid by reaction with 1.25 eq. oxalyl chloride in dichloromethane and a drop of N, N-dimethylformamide at room temperature for 30 minutes, followed by evaporation of the volatile components in vacuo.

Getting connection 221

Ingenol-5,20-acetonide-3- (2- (methyl (phenyl) amino) -2-oxoacetate) (compound 221)

Compound 221 was prepared according to procedure c.

Starting material: 2- (methyl (phenyl) amino) -2-oxoacetic acid.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.41-7.24 (m, 5H), 5.99-5.98 (m, 1H), 5.76-5.73 (m, 1H), 5.39 (s, 1H), 4.19-4.02 (m, 3H), 3.87-3.86 (m, 1H), 3.36 (s, 3H), 2.75 (s, 1H), 2.46-2.41 (m, 1H), 2.24-2.15 (m, 1H), 1.79-1.67 (m, 1H), 1.51 (d, 3H) 1.36 (s, 3H), 1.32 (s, 3H), 1.09 (s, 3H), 1.06 (s, 3H), 0.94 (d, 3H), 0.91- 0.84 (m, 1H); 0.72-0.64 (m, 1H).

Example 101

Ingenol-3- (2-methylacrylate) (compound 101)

Compound 101 was prepared according to procedure e.

Starting material: compound 201.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.15 (s, 1H), 6.06-6.04 (m, 2H), 5.65-5.64 (m, 1H), 5.56 ( s, 1H), 4.20-4.11 (m, 3H), 4.05 (s, 1H), 3.49 (br s, 1H), 2.9-2.7 (br s, 2H), 2.55-2.50 (m, 1H), 2.31-2.22 (m, 1H), 1.99 (s, 3H), 1.79 (d, 3H), 1.80 -1.72 (m, 1H), 1.09 (s, 3H), 1.05 (s, 3H), 0.98 (d, 3H), 0.98-0.91 (m, 1H), 0.74-0.65 (m, 1H).

Example 102

Ingenol-3- (3-methylbutenoate) (compound 102)

Compound 102 was obtained according to procedure e.

Starting material: compound 202.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.05 (d, 1H), 6.02-6.01 (m, 1H), 5.77-5.76 (m, 1H), 5.51 ( s, 1H), 4.15-4.09 (m, 3H), 4.03 (s, 1H), 3.48 (br s, 1H), 2.79 (br s, 2H), 2 54-2.49 (m, 1H), 2.30-2.20 (m, 1H), 2.20 (d, 3H), 1.94 (d, 3H), 1.88-1.72 (m, 1H), 1.78 (d, 3H), 1.09 (s, 3H), 1.05 (s, 3H), 0.98-0.91 (m, 4H), 0.73- 0.65 (m, 1H).

Example 103

Ingenol-3- (2,3-dimethylbutenoate) (compound 103)

Compound 103 was obtained according to procedure e.

Starting material: compound 203.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.07-6.05 (d, 1H), 6.03-6.02 (m, 1H), 5.50 (s, 1H), 4.15- 4.09 (m, 3H), 4.05 (s, 1H), 3.47 (br s, 1H), 2.55-2.48 (m, 1H), 2.30-2.21 ( m, 1H), 2.07 (m, 3H), 1.89 (m, 3H), 1.85 (s, 3H), 1.80 (m, 3H), 1.99-1.55 (m , 3H), 1.09 (s, 3H), 1.05 (s, 3H), 0.96-0.88 (d, 4H), 0.74-0.65 (m, 1H).

Example 104

Ingenol-3- (2-methylenebutyrate) (compound 104)

Compound 104 was obtained according to procedure e.

Starting material: compound 204.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.19 (s, 1H), 6.06-6.04 (m, 2H), 5.61 (m, 1H), 5.57 (s, 1H) 4.20-4.11 (m, 3H), 4.05 (s, 1H), 3.5 (br s, 1H), 2.9 (br s, 2H), 2.55-2 50 (m, 1H), 2.39-2.22 (m, 3H), 1.79 (d, 3H), 1.80-1.72 (m, 1H), 1.11 (t, 3H) ), 1.09 (s, 3H), 1.05 (s, 3H), 0.98 (d, 3H), 0.97-0.91 (m, 1H), 0.74-0.65 ( m, 1H).

Example 105

Ingenol-3- (2-methylpropanoate) (compound 105)

Compound 105 was obtained according to procedure e.

Starting material: compound 205.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.05-6.02 (m, 2H), 5.45 (s, 1H), 4.19-4.11 (m, 3H), 4.03 ( s, 1H), 3.45 (broad s, 1H), 2.65 (m, 1H), 2.54-2.49 (m, 1H), 2.5 (broad s, 1H), 2 31-2.22 (m, 1H), 1.81-1.71 (m, 1H), 1.77 (d, 3H), 1.23 (d, 3H), 1.21 (d, 3H) ), 1.09 (s, 3H), 1.05 (s, 3H), 0.97 (d, 3H), 0.97-0.86 (m, 2H), 0.73-0.65 ( m, 1H).

Example 106

Ingenol-3- (3-methylbutanoate) (compound 106)

Compound 106 was obtained according to procedure e.

Starting material: compound 206.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.05-6.02 (m, 2H), 5.43 (s, 1H), 4.19-4.11 (m, 3H), 4.03 ( s, 1H), 3.49 (broad s, 1H), 2.6-2.4 (broad s, 1H), 2.53-2.45 (m, 1H), 2.31-2, 23 (m, 3H), 2.17-2.04 (m, 1H), 1.81-1.71 (m, 1H), 1.78 (d, 3H), 1.27 (br s, 1H), 1.09 (s, 3H), 1.05 (s, 3H), 1.00 (d, 6H), 0.97 (d, 3H), 0.97-0.86 (m, 1H ), 0.73-0.66 (m, 1H).

Example 107

Ingenol-3- (RS-2-methylbutanoate) (compound 107a and compound 107b)

Compounds 107a and 107b were prepared according to procedure e.

Starting material: Compounds 207a and 207b.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.06-6.02 (m, 2H), 5.46 (s, 1H), 4.19-4.11 (m, 3H), 4.03 ( s, 1H), 3.45 (broad s, 1H), 2.6 (broad s, 1H), 2.54-2.41 (m, 2H), 2.31-2.22 (m, 1H), 1.78 (d, 3H), 1.80-1.64 (m, 2H), 1.57-1.47 (m, 1H), 1.28 (br s, 1H), 1 , 21-1.18 (m, 3H), 1.09 (s, 3H), 1.05 (s, 3H), 0.98-0.85 (m, 7H), 0.73-0.65 (m, 1H).

Example 108

Ingenol-3- (3,3-dimethylbutanoate) (compound 108)

Compound 108 was obtained according to procedure e.

Starting material: compound 208.

¹ H NMR (300 MHz, CDCl ₃₎ δ 6,06-6,02 (m, 2H), 5,47 (s, 1H), 4,19-4,11 (m, 3H), 4,03 ( s, 1H), 3.51 (s, 1H), 2.6-2.4 (br s, 1H), 2.54-2.49 (m, 1H), 2.30 (s, 2H) , 2.30-2.21 (m, 1H), 1.78 (d, 3H), 1.77-1.70 (m, 1H), 1.27 (br s, 1H), 1.09 (s, 3H), 1.06 (s, 9H), 1.05 (s, 3H), 0.96 (d, 3H), 0.98-0.88 (m, 1H), 0.73- 0.65 (m, 1H).

Example 109

Ingenol-3- (2-ethylbutanoate) (compound 109)

Compound 109 was prepared according to procedure e.

Starting material: compound 209.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.06-6.03 (m, 2H), 5.43 (s, 1H), 4.19-4.11 (m, 3H), 4.04 ( s, 1H), 3.50 (s, 1H), 2.6-2.4 (br s, 1H), 2.54-2.49 (m, 1H), 2.34-2.22 ( m, 2H), 1.78 (d, 3H), 1.79-1.51 (m, 5H), 1.27 (br s, 1H), 1.09 (s, 3H), 1.05 (s, 3H), 0.98-0.89 (m, 10H), 0.73-0.65 (m, 1H).

Example 110

Ingenol-3- (2R-methylbutanoate) (compound 110)

Compound 110 was obtained according to procedure e.

Starting material: compound 210.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.06-6.02 (m, 2H), 5.46 (s, 1H), 4.19-4.11 (m, 3H), 4.03 ( s, 1H), 3.46 (broad s, 1H), 2.6-2.4 (broad s, 1H), 2.54-2.44 (m, 2H), 2.31-2, 22 (m, 1H), 1.78 (d, 3H), 1.80-1.66 (m, 2H), 1.57-1.48 (m, 1H), 1.27 (br s, 1H), 1.19 (d, 3H), 1.09 (s, 3H), 1.05 (s, 3H), 0.98-0.86 (m, 7H), 0.73-0.65 (m, 1H).

Example 111

Ingenol-3-crucible (compound 111)

Compound 111 was prepared according to procedure e.

Starting material: compound 211.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.95-6.88 (m, 1H), 6.05-6.03 (m, 2H), 5.51 (s, 1H), 4.14- 4.11 (m, 3H), 4.05 (s, 1H), 3.47 (br s, 1H), 2.77 (br s, 2H), 2.54-2.49 (m, 1H), 2.30-2.21 (m, 1H), 1.87-1.72 (m, 10H), 1.09 (s, 3H), 1.05 (s, 3H), 0.98 (d, 3H), 0.98-0.91 (m, 1H), 0.73-0.65 (m, 1H).

Example 112

Ingenol-3- (phenylacetate) (compound 112)

Compound 112 was prepared according to procedure e.

Starting material: compound 212.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.36-7.25 (m, 5H), 6.00-5.98 (m, 2H), 5.53 (s, 1H), 4.16- 4.00 (m, 3H), 3.94 (s, 1H), 3.72 (s, 2H), 3.14 (br s, 1H), 2.3-1.9 (br s, 1H), 2.23-2.18 (m, 1H), 2.09-1.99 (m, 1H), 1.74 (d, 3H), 1.69-1.59 (m, 1H) 1.27 (br s, 1H); 1.05 (s, 6H); 0.93-0.85 (m, 4H); 0.70-0.62 (m, 1H).

Example 113

Ingenol-3- (2Z- (methoxycarbonyl) acrylate) (compound 113)

Compound 113 was obtained according to procedure e.

Starting material: compound 213.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.95 (d, 1H), 6.88 (d, 1H), 6.09 (m, 1H), 6.05 (d, 1H), 5.65 (s, 1H), 4.54 (d, 1H), 4.20-4.13 (m, 3H), 4.05 (d, 1H), 3.83 (s, 3H), 3.57 ( s, 1H), 2.54-2.43 (m, 2H), 2.31-2.22 (m, 1H), 1.81-1.72 (m, 4H), 1.07 (s, 3H), 1.05 (s, 3H), 0.97 (d, 3H), 0.95-0.89 (m, 1H), 0.74-0.65 (m, 1H).

Example 114

Ingenol-3- (2-cyclohexylpropanoate) (compound 114)

Compound 114 was obtained according to procedure e.

Starting material: compound 214.

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.06-6.02 (m, 2H), 5.46 (d, 1H), 3.32 (t, 1H), 4.19-4.11 ( m, 3H), 4.04-4.03 (m, 1H), 3.52 (s, 1H), 2.55-2.47 (m, 1H), 2.39-2.23 (m, 2H), 1.80-1.60 (m, 10H), 1.30-0.86 (m, 19H), 0.73-0.65 (m, 1H).

Example 115

Ingenol-3 - ((2Z) -2-methoxyimino-2-phenylacetate) (compound 115)

Compound 115 was obtained according to procedure e.

Starting material: compound 215.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.70-7.66 (m, 2H), 7.45-7.38 (m, 3H), 6.12-6.11 (m, 1H), 6.09 (s, 1H), 6.07-6.05 (m, 1H), 4.22-4.12 (m, 4H), 4.07 (s, 3H), 4.00 (d, 1H), 3.58 (d, 1H), 2.41-2.36 (m, 1H), 2.29-2.18 (m, 2H), 1.83 (d, 3H), 1.80 -1.73 (m, 1H), 1.12 (s, 3H), 1.07 (s, 3H), 0.96-0.86 (m, 4H), 0.75-0.67 (m , 1H).

Example 116

Ingenol-3 - ((2E) -2-methoxyimino-2-phenylacetate) (compound 116)

Compound 116 was obtained according to procedure e.

Starting material: compound 216.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.70-7.66 (m, 2H), 7.45-7.38 (m, 3H), 6.12-6.05 (m, 3H), 4.22-4.09 (m, 4H), 4.07 (s, 3H), 4.00 (d, 1H), 3.60 (d, 1H), 2.41-2.36 (m, 1H), 2.29-2.20 (m, 2H), 1.83 (d, 3H), 1.80-1.73 (m, 1H), 1.12 (s, 3H), 1.07 (s, 3H), 0.99-0.92 (m, 4H), 0.75-0.67 (m, 1H).

Example 117

Ingenol-3 - ((Z) -2-methyl-3- (p-tolyl) prop-2-enoate (compound 117)

Compound 117 was obtained according to procedure e.

Starting material: compound 217.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.70 (s, 1H), 7.32 (d, 2H), 7.21 (d, 2H), 6.07-6.05 (m, 2H) 5.60 (s, 1H), 4.44 (d, 1H), 4.17-4.08 (m, 4H), 3.55 (s, 1H), 2.61-2.49 (m , 2H), 2.38 (s, 3H), 2.31-2.23 (m, 1H), 2.15 (d, 3H), 1.83 (d, 3H), 1.80-1, 73 (m, 1H), 1.09 (s, 3H), 1.04 (s, 3H), 1.01 (d, 3H), 0.98-0.92 (m, 1H), 0.74 -0.66 (m, 1H).

Example 118

Ingenol-3 - ((E) -2-phenylbut-2-enoate) (compound 118)

Compound 118 was obtained according to procedure e.

Starting material: compound 218.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.39-7.30 (m, 3H), 7.24 (q, 1H), 7.20-7.15 (m, 2H), 5.99- 5.96 (m, 1H), 5.95-5.93 (m, 1H), 5.57 (s, 1H), 4.16-3.93 (m, 4H), 3.80 (d, 1H), 3.19 (s, 1H), 2.34 (t, 1H), 1.95-1.83 (m, 2H), 1.78 (s, 3H), 1.77 (d, 3H) ), 1.57-1.46 (m, 1H), 1.05 (s, 3H), 1.04 (s, 3H), 0.93-0.85 (m, 1H), 0.76 ( d, 3H), 0.65-0.58 (m, 1H).

Example 119

Ingenol-3- (2,2-diphenylacetate) (compound 119)

Compound 119 was obtained according to procedure e.

Starting material: compound 219.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.35-7.25 (m, 10H), 6.01-5.96 (m, 2H), 5.62 (s, 1H), 5.12 ( s, 1H), 4.13-3.93 (m, 5H), 3.14 (s, 1H), 2.34 (t, 1H), 2.12-2.05 (m, 1H), 2 04-1.93 (m, 1H), 1.72 (d, 3H), 1.61-1.52 (m, 1H), 1.05 (s, 3H), 1.04 (s, 3H) ), 0.91-0.83 (m, 4H), 0.67-0.59 (m, 1H).

Example 120

Ingenol-3- (2-cyano-2-cyclohexylidene acetate) (compound 120)

Compound 120 was obtained according to procedure e.

Starting material: compound 220.

¹ H NMR (300 MHz, CDCl ₃₎ δ 6,12-6,10 (m, 1H), 6,08-6,06 (m, 1H), 5.67 (s, 1H), 4,20 4.02 (m, 5H), 3.56 (s, 1H), 3.02 (t, 2H), 2.71-2.63 (m, 3H), 2.32-2.23 (m, 1H), 2.13-2.08 (m, 1H), 1.85-1.65 (m, 10H), 1.09 (s, 3H), 1.06 (s, 3H), 1.00 (d, 3H), 0.96-0.88 (m, 1H), 0.75-0.67 (m, 1H).

Example 121

Ingenol-3- (2- (methyl (phenyl) amino-2-oxoacetate) (compound 121)

Compound 121 was obtained according to procedure e.

Starting material: compound 221.

¹ H NMR (300 MHz, CDCl ₃₎ δ 7,43-7,33 (m, 3H), 7,28-7,24 (m, 2H), 6,00-5,98 (m, 2H), 5.52 (s, 1H), 4.16-4.02 (m, 3H), 3.84 (s, 2H), 3.37 (s, 3H), 3.29 (s, 1H), 2 , 38-2.32 (m, 2H), 2.24-2.14 (m, 1H), 1.81-1.69 (m, 1H), 1.50 (d, 3H), 1.08 (s, 3H), 1.06 (s, 3H), 0.96 (d, 3H), 0.93-0.87 (m, 1H), 0.73-0.65 (m, 1H).

Example 1

Neutrophil oxidative burst analysis

PMNs (polymorphonuclear leukocytes) were isolated and purified from fresh leukocyte films by sequential precipitation, density gradient centrifugation and lysis of contaminating red blood cells. Leukocyte films were incubated with 2% metocel for 30-45 min for differential erythrocyte sedimentation. The leukocyte-enriched supernatant was transferred to lymphatic tubes to remove mononuclear cells by density gradient centrifugation (400 × g, 30 min). The pellet was resuspended and any remaining red blood cells were lysed using a 0.2% NaCl solution for 30 seconds before isotonicity was restored by adding 1.2% NaCl. This step was repeated until the cell pellet was relatively free of red blood cells. Cells were resuspended in DPBS (Dulbecco phosphate buffered saline) (without Ca ²⁺ , Mg ²⁺ ) and the concentration was adjusted to 1.4 × 10 ⁶ cells / ml in HBSS (Hanks balanced salt solution) (with Ca ²⁺ , Mg ²⁺ ) containing 0.1% BSA (bovine serum albumin) and 5 mM glucose, just before the start of the analysis. The titrated standard and test compounds were pre-mixed with HE (hydroethidine) (to a final assay concentration of 10 μM) before being added to 96-well plates containing 2.5 × 10 ⁵ cells. After a 40 min incubation at room temperature, changes in the respiratory burst were evaluated by measuring fluorescence at 579 nm (excitation: 485 nm) using an Envision plate reader.

The titration curves of the test compounds were adjusted to a 4-parameter sigmoid curve after normalizing the action of the test compound relative to the positive control (5 × 10 ^-7 M PEP0005). Rel EC ₅₀ means the concentration of test compound producing effect, which is the average between the upper and lower fitted effect. Abs EC ₅₀ is the concentration of the test compound, which causes a response corresponding to 50% of the maximum effect associated with the positive control (5 × 10 ^-7 M PEP0005).

Example 2

HeKa Cytokine Release Assay (IL-8)

Human primary epidermal keratinocytes, HeKa, were plated (10,000 cells / well) in 96-well plates the day before analysis. The test compounds were diluted in DMSO (dimethyl sulfoxide) and then diluted in assay medium and pipetted into wells of 96-well plates containing HeKa cells. The plates were incubated for 6 hours at 37 ° C in humidified air with 5% CO ₂ . The plates were briefly centrifuged to pellet the cells at 4 ° C, the supernatant was removed and analyzed using a 4-spot cytokine assay using the Meso Detection Scale (MSD) (Pro-inflammatory Ultrasensitive Kit II, MSD, MD, USA). In the MSD assay, a sandwich immunoassay format was used where capture antibodies were coated in an array of a specific pattern at the bottom of the wells of a 4-Spot-Multi-MSD plate. Standard samples were also incubated in MULTI-SPOT plates and the cytokine (IL-8) bound to its corresponding antibody capture spot. The cytokine level was quantified on a SECTOR ^™ visualizer using cytokine-specific detection of antibodies labeled with MSD SULFO-TAGTM reagent.

The titration curves of the test compound were adjusted to a 4-parameter sigmoid curve after normalizing the action of the test compound relative to the positive control (1.5 × 10 ^-7 M PEP0005). Rel EC ₅₀ means the concentration of test compound producing effect, which is the average between the upper and lower fitted effect. Abs EC ₅₀ is the concentration of the test compound that elicits a response corresponding to 50% of the maximum effect associated with the positive control (1.5 × 10 ^-7 M PEP0005).

Example 3

Necrosis analysis

HeLa cells (ATCC CCL-002) were grown in minimal essential medium (Invitrogen catalog No. 42360) containing 10% fetal calf serum, 100 IU / ml penicillin and 100 μg / ml streptomycin. 4000-6000 cells were plated in 96-well black transparent bottom ViewPlates (Perkin Elmer) in 100 μl of medium and incubated overnight. Compounds were dissolved and pre-diluted in DMSO in 96-well polypropylene plates (Greiner) at a concentration range of 15 μM to 600 μM. During the experiment, cell plates were placed on heating blocks at 37 ° C, the medium was removed and 40 μl of fresh, pre-heated medium was added to each well. Cells were incubated for 15 min before adding compounds. In parallel, 3 μl of compounds was diluted with 197 μl of culture medium on a Tecan freedom-EVO pipetting apparatus using a pipetting rate of 250 μl / sec to ensure efficient mixing of highly concentrated solutions of the compounds with the aqueous phase. These pre-diluted mixtures of the plates were then balanced by heating on heating blocks at 37 ° C for 10 minutes. 80 μl of the pre-diluted compound were transferred manually to the corresponding wells containing HeLa cells to obtain a compound concentration of 10 μΜ to 400 μΜ. The control conditions were 1% DMSO in culture medium (100% viability) and 400 μΜ ingenol mebutate in culture medium (0% viability). The plates were incubated on heating blocks at 37 ° C for 30 minutes. At the end of incubation, 10 μl of PrestoBlue reagent (Invitrogen) was added to each well, the plates were sealed with a black seal, followed by incubation at 37 ° C for 10 min with gentle shaking (150 rpm). Then the tablets were kept at room temperature for 20-30 minutes. The tablets were immediately read on a fluorescence apparatus for reading Envision plates (Perkin Elmer) with excitation at 535 nm and radiation at 630 nm. The titration curves of the test compounds were adjusted to a sigmoid curve with four parameters after normalizing the action of the test compound with respect to the positive control (4 × 10 ^-4 M PEP0005 / ingenol mebutate). AbsEC ₅₀ means the concentration of the test compound, producing 50% action.

The compounds of the present invention were tested in an oxidative burst analysis of neutrophils as described in Example 1, in an analysis of the release of HeKa cytokines as described in Example 2, and in a necrosis analysis as described in Example 3.

Ranges of values of Rel EC ₅₀ oxidative explosion of neutrophils:

* indicates that Rel EC ₅₀ values are ≥100 nM,

** indicates that Rel EC ₅₀ values are ≥20 nM and <100 nM,

*** indicates that Rel EC ₅₀ values <20 nM.

Ranges of Rel EC ₅₀ (IL-8) HeKa Cytokine Release Ranges

* Indicates that the value Rel EC ₅₀ ≥100 nM

** indicates that the value Rel EC ₅₀ ≥20 nM and <100 nM

*** indicates that the value of Rel EC ₅₀ <20 nM.

Ranges of values of the EU ₅₀ HeLa necrosis

* indicates that EC ₅₀ values are ≥350 μM,

** indicates that EC ₅₀ values are ≥150 μM and <350 μM,

*** indicates that the values of the EC ₅₀ <150 μm.

Name and number of connection Rel EC range of ₅₀ oxidative neutrophil explosions Rel EC Range ₅₀ HeKa Cytokine Release (IL-8) EU Range ₅₀ HeLa Necrosis Ingenol-3- (2-methylacrylate) (compound 101) *** * ** Ingenol-3- (3-methylbutenoate) (compound 102) *** ** ** Ingenol 3- (2,3-dimethylbutenoate) (compound 103) *** *** ** Ingenol-3- (2-methylenebutyrate) (compound 104) *** ** ** Ingenol-3- (2-methylpropanoate) (compound 105) *** ** - Ingenol-3- (3-methylbutanoate) (compound 106) *** ** - Ingenol-3- (RS-2-methylbutanoate) (compounds 107a and compound 107b) *** ** - Ingenol-3- (3,3-dimethylbutanoate) (compound 108) *** ** ** Ingenol-3- (2-ethylbutanoate) (compound 109) *** *** ** Ingenol-3- (2R-methylbutanoate) (compound 110) *** ** - Ingenol 3-crucible (compound 111) *** ** * Ingenol-3- (phenylacetate) (compound 112) *** * ** Ingenol-3- (2Z- (methoxycarbonyl) acrylate (compound 113) * - - Ingenol-3- (2-cyclohexylpropanoate) (compound 114) *** *** ** Ingenol-3 - ((2Z) -2-methoxyimino-2-phenylacetate) (compound 115) ** *** *** Ingenol-3 - ((2E) -2-methoxyimino-2-phenylacetate) (compound 116) ** ** - Ingenol-3 - ((Z) -2-methyl-3- (p-tolyl) prop-2-enoate) (compound 117) *** *** *** Ingenol-3 - ((E) -2-phenylbut-2-enoate) (compound 118) *** *** *** Ingenol-3- (2,2-diphenylacetate) (compound 119) *** *** *** Ingenol-3- (2-cyano-2-cyclohexylidene acetate) (compound 120) *** * - Ingenol-3- (2- (methyl (phenyl) amino) -2-oxoacetate) (compound 121) ** * * PEP005, Ingenol-3-Angelate *** *** **

Claims (24)

1. The compound of General formula I

where R represents (C ₁ -C ₇ ) alkyl or (C ₂ -C ₇ ) alkenyl, where R is substituted by one or more substituents independently selected from R1; where R1 represents
(a) cyano; or
(b) (C ₁ -C ₄ ) alkyl, (C ₆ ) cycloalkyl or (C ₆ ) aryl, each of which may optionally be substituted with one or more substituents independently selected from R2, where R2 is (C ₁ -C ₄ ) alkyl or —COORc; or
(c) = N-ORa, -NRaRb or = O;
and where Ra and Rb independently represent (C ₁ -C ₄ ) alkyl or (C ₆ ) aryl;
and where Rc is (C ₁ -C ₄ ) alkyl, and pharmaceutically acceptable salts thereof;
provided that when R1 is alkyl or alkenyl, the total chain length of R and R1 does not exceed a chain length of 7 carbon atoms,
and provided that R and R1 in combination cannot be a straight unbranched or unsubstituted alkyl chain or a straight unbranched or unsubstituted alkenyl chain,
and provided that R cannot be or R and R1 in combination cannot be trans-2-buten-2-yl. 2. The compound of claim 1, wherein R is substituted with R1 at the alpha or beta position relative to the carbonyl group. 3. The compound of claim 1, wherein R is substituted with R1 at the alpha position relative to the carbonyl group. 4. The compound of claim 1, wherein R is C ₁ -C ₇ alkyl. 5. The compound of claim 4, wherein R is ethyl, propyl, tert-butyl, isopropyl, 2-butyl, 3-pentyl or isobutyl. 6. The compound of claim 1, wherein R is C ₂ -C ₇ alkenyl. 7. The compound of claim 6, wherein R is propenyl or ethenyl. 8. The compound according to any one of paragraphs. 1-7, where R1 represents C ₁ -C ₄ alkyl, (C ₆ ) aryl, (C ₆ ) cycloalkyl or cyano. 9. The compound according to any one of paragraphs. 1-7, where R1 represents ethyl, methyl, phenyl, cyclohexyl. 10. The compound according to any one of paragraphs. 1-7, where R1 represents methyl. 11. The compound according to any one of paragraphs. 1-7, where R2 is methyl. 12. The compound according to any one of paragraphs. 1-7, representing
ingenol-3- (2-methylacrylate),
ingenol-3- (3-methylbutenoate),
ingenol-3- (2,3-dimethylbutenoate),
ingenol-3- (2-methylene butyrate),
ingenol-3- (2-methylpropanoate),
ingenol-3- (3-methylbutanoate),
ingenol-3- (2RS-methylbutanoate),
ingenol-3- (3,3-dimethylbutanoate),
ingenol-3- (2-ethylbutanoate),
ingenol-3- (2R-methylbutanoate),
ingenol-3-crucible,
ingenol-3- (phenylacetate),
ingenol-3- (2Z- (methoxycarbonyl) acrylate),
ingenol-3- (2-cyclohexylpropanoate),
ingenol-3 - ((2Z) -2-methoxyimino-2-phenylacetate),
ingenol-3 - ((2E) -2-methoxyimino-2-phenylacetate),
ingenol-3 - ((Z) -2-methyl-3- (p-tolyl) prop-2-enoate),
ingenol-3 - ((E) -2-phenylbut-2-enoate),
ingenol-3- (2,2-diphenylacetate),
ingenol-3- (2-cyano-2-cyclohexylidene acetate) or
ingenol-3- (2- (methyl (phenyl) amino) -2-oxoacetate)
and their pharmaceutically acceptable salts. 13. The compound according to any one of paragraphs. 1-7 for use as a medicine in therapy for preventing, treating, or reducing the intensity of symptoms of a disease or condition susceptible to stimulating an oxidative burst of neutrophils, susceptible to stimulating the release of IL-8 keratinocytes, or susceptible to inducing necrosis. 14. The use of compounds according to any one of paragraphs. 1-12 for the preparation of a pharmaceutical composition for treating or reducing the intensity of symptoms of a disease, disorder or condition susceptible to stimulation of an oxidative burst of neutrophils. 15. The use of compounds according to any one of paragraphs. 1-12 for the preparation of a pharmaceutical composition for treating or reducing the intensity of symptoms of a disease, disorder or condition susceptible to stimulating the release of IL-8 keratinocytes. 16. The use of compounds according to any one of paragraphs. 1-12 for the preparation of a pharmaceutical composition for treating or reducing the intensity of symptoms of a disease, disorder or condition susceptible to induce necrosis. 17. A method of preventing, treating, reducing the intensity of symptoms or preventing physiological disorders or diseases susceptible to stimulation of an oxidative explosion of neutrophils by administering to a subject in need thereof a compound according to any one of claims. 1-12. 18. A method of preventing, treating, reducing the intensity of symptoms or preventing physiological disorders or diseases susceptible to stimulating the release of IL-8 keratinocytes by administering to a subject in need thereof a compound according to any one of claims. 1-12. 19. A method of preventing, treating, reducing the intensity of symptoms or preventing physiological disorders or diseases susceptible to inducing necrosis by administering to a subject in need thereof a compound according to any one of claims. 1-12. 20. The compound according to any one of paragraphs. 1-7 for use in the treatment or reduction of the intensity of symptoms of a disease, disorder or condition susceptible to stimulation of an oxidative explosion of neutrophils. 21. The compound according to any one of paragraphs. 1-7 for use in treating or decreasing the intensity of symptoms of a disease, disorder or condition susceptible to stimulating the release of IL-8 keratinocytes. 22. The compound according to any one of paragraphs. 1-7 for use in treating or reducing the intensity of symptoms of a disease, disorder, or condition susceptible to inducing necrosis. 23. A pharmaceutical composition for preventing, treating, or reducing the intensity of symptoms of a disease or condition susceptible to stimulating an oxidative burst of neutrophils, susceptible to stimulating the release of IL-8 keratinocytes, or susceptible to inducing necrosis, comprising a compound according to any one of claims. 1-12 or a pharmaceutically acceptable stereoisomer or salt thereof together with a pharmaceutically acceptable excipient or excipient. 24. The pharmaceutical composition of claim 23, wherein the composition is suitable for topical administration.