WO2013026453A1 - Treatment of inflammatory disorders with anthracyclines - Google Patents

Treatment of inflammatory disorders with anthracyclines Download PDF

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Publication number
WO2013026453A1
WO2013026453A1 PCT/DK2012/050303 DK2012050303W WO2013026453A1 WO 2013026453 A1 WO2013026453 A1 WO 2013026453A1 DK 2012050303 W DK2012050303 W DK 2012050303W WO 2013026453 A1 WO2013026453 A1 WO 2013026453A1
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alkyl
group
composition according
monoglucosides
aryl
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PCT/DK2012/050303
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French (fr)
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Elisabeth de DARKÓ
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Valderm Aps
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/08—Drugs for disorders of the urinary system of the prostate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/10—Drugs for disorders of the urinary system of the bladder
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/10—Anti-acne agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis

Definitions

  • the present invention relates to pharmaceutical formulations comprising an
  • Anthracyclines are antibiotics having potent antineoplastic activity, and accordingly they have been used in the treatment of a variety of cancers. Anthracyclines are amongst the most utilised antitumor drugs ever developed. Anthracyclines mediate their anticancer effect in part by targeting topisomerase II, which leads to DNA damage. Anthracyclines are essential components of several drug combinations for the therapy of a number of cancer diseases. It is a drug of primary use in the treatment of patients with lymphoma, breast cancer and sarcoma.
  • the group of anthracyclines for example comprises doxorubicin, daunorubicin, carcinomycin, valrubicin, epirubicin, and idarubicin.
  • Doxorubicin, daunorubicin, idarubicin and epirubicin are usually administered systemically by intravenous injection or infusion. Systemic administration of these anthracyclines, however, results in a number of undesirable side effects such as cardiotoxicity and bone marrow suppression
  • Anthracyclines are in general known to be very tissue toxic. For example, paravenous extravasation of doxorubicin results in severe necrosis and immediate measures have to be undertaken to avoid severe local toxicity. It is also known that several
  • Valrubicin is a semisynthetic analogue of doxorubicin and it was developed for the treatment of superficial bladder cancer and approved for such use in the United States. Usually a total of 800 mg is administered by intravesical instillation of two hours +- duration for a total of 6 times once a week. (FDA. ValstarTM (Valrubicin) Sterile
  • Valrubicin for the treatment of hyperproliferative conditions of body surfaces. It is described that Valrubicin has antiproliferative effects and that it is suitable for topical treatment of hyperproliferative skin disordes, e.g. plaque psoriasis, non-melanoma skin tumors and carcinoma in situ.
  • anthracyclines and in particular lipophilic anthracyclines are useful for local/topical treatment of inflammatory diseases and in particular for local treatment of skin diseases with an inflammatory component of the pathogenesis.
  • antracyclines which do not induce necrosis and/or lasting tissue damage are useful for local/topical treatment of diseases, wherein both infection and inflammation are significant factors of pathogenesis.
  • the infection may be any infection, but in one embodiment of the invention the infection is a bacterial infection, e.g. infection by Propionibacterium acnes.
  • CAELYX® is a medicament containing pegylated liposomal doxorubicin hydrochloride it is stated that in ovarian cancer patients treated with CAELYX® one common dermatological adverse effect (>1 % and ⁇ 10%) is acne.
  • FIG. 1 shows that Topical valrubicin treatment reduces ear thickness in an acute 12- O-tetradecanoylphorbol 13-acetate (TPA) induced dermatitis model.
  • TPA O-tetradecanoylphorbol 13-acetate
  • FIG. 3 shows that valrubicin treatment decreases the mRNA expression levels of the proinflammatory cytokines IL- ⁇ (a) and IL-6 (b).
  • the mRNA levels were determined by qRT-PCR and normalized to glyceraldehyde-3-phosphate
  • dehydrogenase GPDH
  • the columns show indexed values of mRNA expression compared with control mice. Results are presented as means + SEM. Significant differences are marked, * p ⁇ 0.001. Note that the y-axis have different scales in each sub-figure.
  • Figure 4 shows that valrubicin treatment prevents the TPA induced increase of I L-1 ⁇ and IL-6 at protein levels. Protein expressions were determined by sandwich enzyme- linked immunoabsorbant assay (ELISA). Columns represent cytokine expression in pg per mg total protein, shown as means + SEM,* p ⁇ 0.0001. Note that the y-axis have different scales in each sub-figure.
  • FIG. 5 shows that topical application of valrubicin reduced ear thickness in a 12-O- tetradecanoylphorbol 13-acetate (TPA)-induced chronic inflammation mouse model.
  • TPA 12-O- tetradecanoylphorbol 13-acetate
  • Mice were challenged with TPA on day 1 , 3, 5, 8 and 10 (TPA), or TPA on day 1 , 3, 5, 8 and 10 and valrubicin on day 5, 6, 7, 8, 9, 10, 1 1 and 12 (valrubicin) or acetone on day 1 , 3, 5, 8 and 10 (control).
  • Ear thickness was measured on day 1 , 3, 5, 6, 7, 8, 9, 10, 1 1 and 12.
  • TPA challenge significantly increased ear thickness compared to control (**, p ⁇ 0.01).
  • Valrubicin treatment significantly reduced ear thickness compared to TPA challenge alone (*, p ⁇ 0.01).
  • FIG. 6 shows that topical application of valrubicin reduced 12-O-tetradecanoylphorbol 13-acetate (TPA)-increased neutrophil cell infiltration in mice ears in a chronic inflammation mouse model.
  • Mice were challenged with TPA on day 1 , 3, 5, 8 and 10 (TPA), or TPA on day 1 , 3, 5, 8 and 10 and valrubicin on day 5, 6, 7, 8, 9, 10, 11 and 12 (valrubicin) or acetone on day 1 , 3, 5, 8 and 10 (control).
  • TPA 12-O-tetradecanoylphorbol 13-acetate
  • FIG. 7 shows one mode of action of Valrubicin.
  • the enzyme SphK1 catalyses formation of GlcCer, S1 P and C1 P, which have several negative effects in relation to cancer.
  • Valrubicin reduces the activity of SphKl
  • Figure 8 shows the level of IL8 secreted by RWPE-1 cells infected with P. acnes after 24 h of treatment with valrubicin.
  • the anthracyclines to be used with the present invention may be any anthracycline, and in particular an anthracycline of formula I as described herein below. It is furthermore preferred that in general the anthracyclines to be used with the present invention do not induce necrosis and/or lasting tissue damage upon application to skin of human beings. Even more preferably anthracyclines according to the present invention are not locally toxic, when applied topically to a body surface of an individual, more preferably, the anthracyclines are not, or are only mildly irritant, when applied to a body surface of an individual in an effective dose.
  • a compound such as an anthracycline is not, or only mildly irritant, is preferably determined as described herein below in the section "Skin irritation".
  • anthracyclines widely used in cancer chemotherapy are highly toxic and extravasion of these anthracyclines into surrounding tissues during chemotherapy may lead to lasting tissue damage and necrosis (see e.g. Ener et al, Annals of Oncology 15: 858-862, 2004) .
  • the anthracyclines to be used with the present invention are neither doxorubicin, daunorubicin, idarubicin nor epirubicin.
  • the anthracycline to be used with the present invention is not toxic to or only mildly toxic to cells surrounding the site of inflammation, which are not desirable to target.
  • the anthracycline is not toxic to or only mildly toxic to sebaceous cells.
  • anthracycline to be used with the present invention are in general anthracyclines of the general formula I:
  • Ri is selected from the group consisting of -H, Ci_ 6 -carboxyl, Ci -6 -alkyl and Ci_ 6 -alkoxy;
  • R 2 is selected from the group consisting of
  • Y is selected from the group consisting of O, S and N;
  • R 8 is only present in the event Y is N and R 8 is selected from the group consisting of -NH-(CH 2 )p-NH-CH3, -NH-(CH 2 )p-CH3, -NH-(CH 2 ) P -aryl, -NH-
  • R 4 is selected from the group consisting of Ci -6 -alkyl and -H;
  • R 5 is selected from the group consisting of -O-Ri 0 and -N-R 10 , wherein
  • R 10 is selected from the group consisting of -H, Ci. 6 -alkyl,
  • R 6 is selected from the group consisting of -N-(Rn)(R 12 ) and -O-Rn, wherein
  • R 7 is selected from the group consisting of -OH, Ci. 6 -alkoxy and -H; and wherein the ring indicates an aliphatic 5 to 6 membered heterocyclic ring, and wherein R 4 , R 5 and R 6 may be attached at any useful position on said ring.
  • Said aliphatic 5 to 6 membered heterocyclic ring is preferably a 6 membered heterocyclic ring, more preferably a 6 membered hetero-cyclic ring of one oxygen and 5 carbon atoms, substituted with R 4 , R 5 and R 6, wherein R 4 , R 5 and R 6 are as define herein above and preferably as described herein below in this section.
  • the C5 carbonyl group of formula I is desirable for optimal function of the claimed anthracyclines and it is thus preferred that said C5 carbonyl group is present in the anthracyclines to be used with the present invention.
  • anthracycline is a compound of the formula I I
  • R ⁇ R 2 , R3, R 4 , R5, R6 and R 7 are as defined herein above in relation to formula I .
  • R 4 of the compound of either formula I or formula II is selected from the group consisting of Ci -6 -alkyl and -H, preferably from the group consisting of Ci. 3 -alkyl and - H, more preferably from the group consisting of -CH 3 and -H, even more preferably - CH 3 .
  • R 5 of the compound of either formula I or formula II is selected from the group consisting of -O-Ri 0 and -N-R 10 , however preferably R 5 is -O-R1 0 , wherein R 10 is as defined herein above in relation to compounds of formula I .
  • R 6 of the compound of either formula I or formula II is selected from the group consisting of -N-(Rn)(R 12 ) and -O-Rn , however preferably R6 is -N-(Rn)(R 12 ), wherein Rn and R 12 are as defined herein above in relation to compounds of formula I.
  • anthracyclines to be used are compounds of formula III:
  • R ⁇ R 2 , R3, R 10 , Rn and R 12 are as defined herein above in relation to compounds of formula I.
  • R ⁇ and R 2 comprises a carbonyl or a hydrazine group and more preferably that either R ⁇ or R 2 , but not both comprises a carbonyl or a hydrazine group.
  • Ci-e-acyl for example when is then R 2 is selected from the group consisting of -OH, -Ci_ 6 -alkyl-OH and Ci -6 -alkyl, more preferably R 2 is selected from the group consisting of Ci -6 -alkyl, even more preferably from the group consisting of d-3-alkyl. rred that when R 2 is selected from the group consisting of
  • R 2 of formula I, II and III may be selected from the group consisting of R 9 ,
  • Preferab 2 of formula I, II and III is selected from the group consisting of
  • Y is selected from the group consisting of O and N, more preferably Y is O. It follows that in the very preferred embodiment, where Y is O, then R 8 is not present.
  • p is an integer in the range of 1 to 4, for example p is an integer in the range of 2 to 4.
  • R 3 , R 7 , R 9 , R 10 , Rn and R 12 are as defined herein above in relation to formula I.
  • R 3 of the compound according to formula I, formula II, formula III or formula IV is selected from the group consisting of -OH, -SH and NH 2 ; preferably R 3 is selected from the group consisting of -NH 2 and -OH, even more preferably R 3 is -OH.
  • R 7 of compounds of formula I, formula II, formula III or formula IV is are is selected from the group consisting of -OH, Ci_ 6 -alkoxy and -H, more preferably R 7 is selected from the group consisting of -OH, -H and Ci_ 3 -alkoxy, even more preferably R 7 is selected from the group consisting -OH, -H and methoxy. In particular R 7 may be methoxy.
  • R 9 of formula I, II or III is preferably Ci -6 -alkyl, more preferably Ci -3 -alkyl, even more preferably methyl.
  • R 9 of formula I, II, III or IV is preferably selected from the group consisting of Ci_ 6 alkyl, Ci_ 6 alkyl-OH, -(CH 2 ) n -0-
  • R 9 of formula I, II, II or IV is alkyl
  • R 9 is Ci_ 6 alkyl, preferably Ci. 3 alkyl, more preferably methyl.
  • R 9 of formula I, II, II or IV is an alcohol
  • R 9 is Ci_ 6 alkyl-OH, preferably Ci_ 3 alkyl-OH, more preferably -CH 2 -OH.
  • X is preferably selected from the group consisting of -COOH, -OH and - CH 3 , for example X may be -CH 3 .
  • n is an integer in the range of 0 to 6, more preferably n is an integer in the range of 0 to 3, even more preferably n is an integer in the range of 0 to 1 , for example n may be 1.
  • m is an integer in the range of 0 to 6, preferably in the range of 1 to 5, more preferably in the range of 2 to 4, such as 3.
  • said alkyl may preferably be Ci-6 alkyl, more preferably Ci_ 3 alkyl.
  • q is an integer in the range of 0 to 6, preferably in the range of 0 to 3, more preferably in the range of 0 to 1 , even more preferably q is 1.
  • R 10 of the compounds of formula I, II, III and IV r is an integer in the range of 1 to 7, preferably in the range of 2 to 6, more preferably in the range of 3 to 5, even more preferably r is 4.
  • the monoglucoside may be any monoglucoside as defined herein below.
  • the monoglucoside is a furanoside (i.e. 5 atom ring), a pyranoside (i.e. 6 atom ring) or septanoside (i.e. 7 atoms ring). More preferably, the monoglucoside is a pyranoside. In particular, it is preferred that the monoglucoside is selected from the group consisting of
  • the amino-monoglucoside may be any monoglucoside as defined herein below, wherein one or more -OH has been replaced by a -NH 2 group or wherein the -O- group has been replaced by a - NH- group.
  • the amino-monoglucoside is an amino-furanoside (i.e. 5 atom ring), an amino-pyranoside (i.e. 6 atom ring) or an amino-septanoside (i.e. 7 atoms ring). More preferably, the amino-monoglucoside is an amino-pyranoside.
  • amino-monoglucoside is selected from the group consisting of amino-glucopyranoside, amino-altropyranoside, amino-mannopyranoside, amino- gulopyranoside, amino-idopyranoside, amino-galactopyranoside and amino- talopyranoside.
  • R 10 of the compounds of formula I, II, III and IV comprises a monoglucoside or - (CHOH) r -(CH 2 OH), for example any of the monoglucoside or -(CHOH) r -(CH 2 OH) described herein above, it is preferred that said monoglucoside or -(CHOH) r -(CH 2 OH) is either unsubstituted or substituted at in the range of 1 to 3 positions, preferably at 1 position. Said monoglucoside or -(CHOH) r -(CH 2 OH) is preferably substituted with one or more substituents selected from the group consisting of monoglucoside or -(CHOH) r - (CH 2 OH).
  • aryl is as defined herein below.
  • the aryl is a 5 or 6 membered aromatic or heteroaromtic ring, more preferably the aryl is a 5 or 6 membered aromatic or heteroaromatic ring comprising at the most 1 heteroatom.
  • said aryl is selected from the group consisting of furanyl, pyrrolyl, thiophenyl, phenyl and pyridinyl, however preferably said aryl is selected from the group consisting of phenyl and pyridinyl.
  • R 10 of the compounds of formula I, II, III and IV comprises aryl, i.e.
  • any of the aryls described herein above it is preferred that said aryl is either unsubstituted or substituted at in the range of 1 to 3 positions.
  • Said aryl is preferably substituted with one or more substituents selected from the group consisting of halogen, -NH 2 and N0 2 .
  • substituents selected from the group consisting of halogen, -NH 2 and N0 2 .
  • halogen is as defined herein below, however preferably said halogen is selected from the group consisting of chloro and fluoro, more preferably said halogen is flouro.
  • said alkyl may preferably be Ci-6 alkyl, more preferably Ci_ 3 alkyl.
  • q is an integer in the range of 0 to 6, preferably in the range of 0 to 3, more preferably in the range of 0 to 1 , even more preferably q is 1.
  • I I, II I and IV r is an integer in the range of 1 to 7, preferably in the range of 2 to 6, more preferably in the range of 3 to 5, even more preferably r is 4.
  • the monoglucoside may be any monoglucoside as defined herein below.
  • the monoglucoside is a furanoside (i.e. 5 atom ring), a pyranoside (i.e. 6 atom ring) or septanoside (i.e. 7 atoms ring). More preferably, the monoglucoside is a pyranoside. In particular, it is preferred that the monoglucoside is selected from the group consisting of
  • the amino-monoglucoside may be any monoglucoside as defined herein below, wherein one or more -OH has been replaced by a -NH 2 group or wherein the -O- group has been replaced by a - NH- group.
  • the amino-monoglucoside is an amino-furanoside (i.e. 5 atom ring), an amino-pyranoside (i.e. 6 atom ring) or an amino-septanoside (i.e. 7 atoms ring). More preferably, the amino-monoglucoside is an amino-pyranoside.
  • amino-monoglucoside is selected from the group consisting of amino-glucopyranoside, amino-altropyranoside, amino-mannopyranoside, amino- gulopyranoside, amino-idopyranoside, amino-galactopyranoside and amino- talopyranoside.
  • Rn of the compounds of formula I, II, III and IV comprises a monoglucoside or - (CHOH) r -(CH 2 OH), for example any of the monoglucoside or -(CHOH) r -(CH 2 OH) described herein above, it is preferred that said monoglucoside or -(CHOH) r -(CH 2 OH) is either unsubstituted or substituted at in the range of 1 to 3 positions, preferably at 1 position. Said monoglucoside or -(CHOH) r -(CH 2 OH) is preferably substituted with one or more substituents selected from the group consisting of monoglucoside or -(CHOH) r - (CH 2 OH).
  • aryl is as defined herein below.
  • the aryl is a 5 or 6 membered aromatic or heteroaromtic ring, more preferably the aryl is a 5 or 6 membered aromatic or heteroaromatic ring comprising at the most 1 heteroatom.
  • said aryl is selected from the group consisting of furanyl, pyrrolyl, thiophenyl, phenyl and pyridinyl, however preferably said aryl is selected from the group consisting of phenyl and pyridinyl.
  • Rn of the compounds of formula I, II, III and IV comprises aryl, i.e. any of the aryls described herein above, it is preferred that said aryl is either unsubstituted or substituted at in the range of 1 to 3 positions. Said aryl is preferably substituted with one or more substituents selected from the group consisting of halogen, -NH 2 and N0 2 .
  • halogen is as defined herein below, however preferably said halogen is selected from the group consisting of chloro and fluoro, more preferably said halogen is flouro.
  • Rn is not a bulky group and vice versa.
  • R 12 is selected from the group consisting of -H and Ci -6 -alkyl.
  • R 12 is selected from the group consisting of -H and d-3-alkyl, more preferably from the group consisting of -H and methyl, yet more preferably R 12 is -H.
  • the anthracycline is not a highly toxic anthracycline. Accordingly, it is preferred that the anthracycline is a compound of formula I, II, III and IV, wherein when R 7 is methoxy, then at least one of R 5 or R 6 is not -OH or -NH 2 .
  • anthracycline to be used with the present invention may be a compound of formula I, II, III or IV substituted as described herein above in this section or a pharmaceutically acceptable salt thereof.
  • the anthracycline is a lipophilic anthracycline, in particular such lipophilic anthracyclines, which are capable of passing the cell membrane and enter the cytoplasm of cells in a fast manner.
  • the lipophilic anthracyclines according to the invention do not enter nucleus of cells to any significant degree.
  • the lipophilic anthracyclines may in some instances be characterised by their partition coefficient (P) and log P.
  • the partition coefficient is the ratio of a given compound partitioned between two solvents, traditionally the solvent system is octanol/water.
  • the log P oc tanoi can be measured in a range from -2 to 6, and compounds having a log P oc tanoi/water of about 0,5 to about 2 may be considered moderately lipophilic, compounds having a log P oc tanoi/water above 2 may be considered increasingly lipophilic.
  • lipophilic anthracyclines according to the present invention may preferably have a log P oc tanoi/water value greater than 0.5, preferably greater than 1.0, more preferably greater than 1.5, even more preferably greater than 2.0, yet even more preferably greater than 3.0.
  • the pH may have an influence on the determination of the partition coefficient, due to possible ionization of the compound in question, it may be preferred to determine the value at a fixed pH.
  • a buffer system such as e.g. PBS (wherein PBS is phosphate buffered saline, preferably 0.01 M phosphate in 8.5% NaCI, pH 7.2.).
  • the lipophilic anthracycline has a log Poctanoi water value greater than 0.5, preferably greater than 1.0, more preferably greater than 1.5, even more preferably greater than 2.0, yet even more preferably greater than 3.0 when determined in octanol and PBS, preferably PBS as specified above.
  • the lipophilicity is characterised by the partition coefficient (P) where the lipophilic anthracycline preferably have an octanol/buffer partition coefficient (such as e.g. octanol PBS partition coefficient) greater than 40, more preferably greater than 60, even more preferably greater than 80, and yet even more preferably greater than 100.
  • an octanol/buffer partition coefficient such as e.g. octanol PBS partition coefficient
  • Preferred lipophilic anthracyclines which can be used in the present invention, may have the general formula V:
  • R 2 is selected from the group consisting of-C(0)CH 2 -0-(CrC 6 -acyl), - C(0)CH 2 -0-(C C 6 -alkyl), -C(0)CH 2 -0-(C C 6 -alkoxy), -C(0)CH 2 -0-(C C 6 -acyl)- (C C 6 -alkoxy) and -C(0)CH 2 -0-C(0)(CH 2 ) n X,
  • n is an interger in the range of 1 to 10 and X is selected from the group consisting of -CH 3 , -OH and COOH;
  • R 6 is selected from the group consisting of-NH-(CrC 6 -acyl), -NH-(CrC 6 -alkyl), -NH- (d-Ce-alkoxy), -N(C C6-acyl)(CrC 6 -acyl), -N(C C6-acyl)(CrC 6 -alkyl), -N(C C 6 -acyl) (d-Ce-alkoxy), -N(CrC 6 -alkyl)(CrC 6 -alkyl), -N(CrC 6 -alkyl)(CrC 6 -alkoxy), -N(C C 6 - alkoxy)(C C 6 -alkoxy), -heterocyclyl, -C(0)CH 2 -0-(C C 6 -alkyl), -(C C 6 -acyl), -NH 2 , - OH, and H; wherein any alkyl, acyl,
  • the lipophilic anthracycline is a compound of the general formula V as outlined herein above or a pharmaceutical acceptable salt, solvate or prodrug thereof; more preferably the lipophilic anthracycline is a compound of the general formula V or a pharmaceutically acceptable salt thereof.
  • R 2 may preferably be -C(0)CH 2 -0-(C C 6 - acyl); and more preferably R 2 may be -COCH 2 OCO(CH 2 ) 3 CH 3 .
  • a preferred group of R 2 may be defined as -C(0)CH 2 -0- C(0)(CH 2 ) n X, wherein n is an integer in the range of 1 to 10, preferably in the range of 2 to 5, more preferably in the range of 3 to 4, yet more preferably 3, and X is selected from the group consisting of -CH 3 , -OH and COOH , preferably X is -CH 3 .
  • R 6 may preferably be selected from the group consisting of-NH-(C C 6 -acyl), -NH-(C C 6 -alkyl), -NH-(C C 6 - alkoxy), -N(C C6-acyl)(CrC 6 -acyl), -N(C C6-acyl)(CrC 6 -alkyl), -N(C C 6 -acyl) (C C 6 - alkoxy), -N(C C6-alkyl)(CrC 6 -alkyl), -N(C C6-alkyl)(CrC 6 -alkoxy), -N(C C 6 - alkoxy)(C C 6 -alkoxy), -heterocyclyl, -C(0)CH 2 -0-(C C 6 -alkyl), and -(C C 6 -acyl); wherein any alkyl, acyl, alkoxy, or hetero
  • R 2 may be selected from the group consisting of-NH-(C C 6 -acyl), -NH-(C C 6 -alkyl), -NH-(C C 6 -alkoxy), -N(C C 6 - acyl)(C C 6 -acyl), -N(Ci-C e -acyl)(Ci-C e -alkyl), -N(C C 6 -acyl) (C C 6 -alkoxy), -N(C C 6 - alkyl)(C C 6 -alkyl), -N(C C6-alkyl)(CrC 6 -alkoxy), -N(C C6-alkoxy)(CrC 6 -alkoxy), - heterocyclyl, and -(d-C 6 -acyl); wherein any alkyl, acyl, alkoxy, or heterocyclyl moiety of R 2 optionally is substituted with one or more of methyl, e
  • R 6 is selected from the group consisting of -COCH 2 OCO(CH 2 ) 3 CH 3 , -COCH 2 OH, -COCH 3 , -NH 2 , -H, -OH, and - NHCOCF 3 ; wherein any alkyl, acyl, or alkoxy moiety of R 2 optionally is substituted with one or more of CrC 3 -alkyl, C C 2 -alkoxy, -OH, halogen, -NH 2 , -NH-(CrC 4 -alkyl), or - N(Ci-C 4 -alkyl)(CrC 4 -alkyl).
  • R 2 is selected from the group consisting of -COCH 2 OCO(CH 2 ) 3 CH 3 , -COCH 2 OH, -COCH 3 ,
  • any alkyl, acyl, or alkoxy moiety of R 2 optionally is substituted with one or more of C C 3 -alkyl, C C 2 -alkoxy, -OH, halogen, -NH 2 , -NH-(C C 4 -alkyl), or -N(C C 4 -alkyl)(C C 4 -alkyl).
  • the lipophilic anthracyclines may be selected from the group consisting of OctADR (adriamycin octanoyl-hydrazone), MRA-CN (3'-deamino-3'-(3-cyano-4- morpholinyl)adriamycin), AD32 (valrubicin), AD41 , AD143, AD194, AD198, AD199, AD201 , AD202, and AD288, or mixtures thereof, or pharmaceutical acceptable salts, solvates or prodrugs thereof.
  • the lipophilic anthracyclines may also be selected from the group consisting of derivatives of OctADR, MRA-CN, AD32, AD41 , AD143, AD194, AD198, AD199, AD201 , AD202, AD288, and mixtures thereof.
  • the lipophilic anthracycline is valrubicin, which may also be denoted AD32.
  • derivatives as used herein is meant a compound, in which one atom or a group of atoms is replaced with another atom or a group of atoms.
  • N-alkyladriamycin (AD41 , AD194, AD198, AD199, AD201 , AD202, AD288) analogues and N-trifluoro-acetyladriamycin-14-O-hemiadipate (AD143) may be synthesised and characterised as described by Israel and colleagues (Israel, M. et al.; J. Med. Chem.; 28; p. 1223-1228; and Israel, M. et al.; US patent 4,610,977).
  • valrubicin is very lipophilic due to the less ionisation compared to other anthracyclines. Accordingly, valrubicin may pass the cell membrane and enter the cytoplasm of cells in a fast manner.
  • alkyl includes saturated monovalent hydrocarbon radicals having straight or branched moieties.
  • alkyl moieties include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and neopentyl.
  • Alkyl is preferably C C 6 alkyl, i.e. groups containing from 1 to 6 carbon atoms, and for some embodiments of the present invention, more preferably C C 4 alkyl, such as e.g. C C 3 alkyl.
  • acyl refers to formyl as well as other alkyl substituted carbonyl groups, wherein “alkyl” is as defined above.
  • acyl includes groups such as (CrC 6 )alkanoyl (e.g., formyl, acetyl, propionyl, butyryl, valeryl, caproyl, t-butylacetyl, etc.).
  • alkoxy means an -O-alkyl group wherein “alkyl” is as defined above. Examples include, but are not limited to methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, pentoxy, 2-pentyloxy, isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, and 3-methylpentoxy.
  • Alkoxy is preferably Ci-C 6 alkoxy, i.e. groups containing from 1 to 6 carbon atoms, and for some
  • C C 4 alkoxy such as e.g. C C 2 alkoxy.
  • aryl refers to a substituent, which is a simple aromatic or heteroaromatic ring.
  • heterocyclyl refers to non-aromatic cyclic groups containing one or more heteroatoms selected from O, S and N. Preferably from one to four heteroatoms, more preferably from one to two heteroatoms. Heterocyclyl groups also include groups that are substituted with one or more oxo moieties. Examples of heterocyclyl include, but are not limited to morpholinyl, piperidinyl, piperazinyl, 1 ,2,3,6- tetrahydropyridinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl,
  • tetrahydrothiopyranyl and thiomorpholinyl.
  • morpholiny as e.g. in MRA-CN.
  • Halogen includes fluoro, chloro, bromo and iodo.
  • any alkyl, acyl, alkoxy, or heterocyclyl moiety are “optionally substituted"
  • the moiety in question may be unsubstituted or optionally substituted with one of more substituents (typically, one to three substituents) independently selected from the group of substituents listed.
  • substituents typically, one to three substituents
  • substituted used as herein in relation to organic compounds refers to that one hydrogen atom of the "substituted" compound has been replaced with another chemical moiety.
  • the term “monoglucoside” refers to the cyclic form of a monosaccharide in which the hydroxyl -OH at the anomeric centre is replaced by an oxygen-bridged group linking the glucoside to the rest of the anthracycline.
  • Linear monosaccharides form the cyclic form through a nucleophilic addition reaction between the carbonyl group and one of the hydroxyls of the same molecule.
  • amino-monoglucoside refers to a monoglucoside, wherein one or more -OH groups has been replaced by a -NH 2 group.
  • pharmaceutical acceptable salt, solvate or prodrug refers to those acid and base additions salts, solvates, and prodrugs of the compounds of the present invention which are, within the scope of sound medical judgment, suitable for use without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention.
  • Pharmaceutically acceptable acid and base addition salts refers to the relatively non- toxic, inorganic and organic addition salts of compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds, or by subsequently reacting the purified compound in its free acid or base form with a suitable organic or inorganic compound and isolating the salt thus formed.
  • the compounds of formula (I) of this invention are basic compounds, they are all capable of forming a wide variety of different salts with various inorganic and organic acids.
  • the pharmaceutically acceptable acid addition salts of the basic compounds are prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt in the conventional manner.
  • the free base form may be
  • the free base forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free base for purposes of the present invention.
  • Pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali and alkaline earth metal hydroxides, or of organic amines.
  • metals used as cations are sodium, potassium, magnesium, calcium, and the like.
  • suitable amines are ⁇ , ⁇ '-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine, and procaine.
  • the base addition salts of acidic compounds are prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt in the conventional manner.
  • the free acid form may be regenerated by contacting the salt form with an acid and isolating the free acid in a conventional manner.
  • the free acid forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free acid for purposes of the present invention.
  • Salts may be prepared from inorganic acids sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic, phosphorus, and the like.
  • Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate mesylate, glucoheptonate, lactobionate, laurylsulphonate and isethionate salts, and the like.
  • Salts may also be prepared from organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. and the like.
  • organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. and the like.
  • Representative salts include acetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, methanesulfonate, and the like.
  • Pharmaceutically acceptable salts may include cations based on the alkali and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations including, but not limited to, ammonium, tetramethylammonium,
  • the compounds of the present invention may exist in unsolvated forms as well as in solvated forms, including hydrated forms.
  • the solvated forms, including hydrated forms are equivalent to unsolvated forms and are intended to be
  • prodrug refers to compounds that are rapidly transformed in vivo to yield the parent compound of the above formulae, for example, by hydrolysis. A thorough discussion is provided in T. Higuchi and V Stella, "Pro-drugs as Novel Delivery
  • prodrugs include pharmaceutically acceptable, non-toxic esters of the compounds of the present invention, including C C 6 alkyl esters wherein the alkyl group is a straight or branched chain. Acceptable esters also include C 5 -C 7 cycloalkyl esters as well as arylalkyl esters such as, but not limited to benzyl. C C 4 alkyl esters are preferred. Esters of the compounds of the present invention may be prepared according to conventional methods "March's Advanced Organic Chemistry, 5 th Edition". M. B. Smith & J. March, John Wiley & Sons, 2001.
  • Compounds of formula (I) may contain chiral centers and therefore may exist in different enantiomeric and diastereomeric forms.
  • This invention relates to all optical isomers and all stereoisomers of compounds of the formula (I), both as racemic mixtures and as individual enantiomers and diastereoismers ((+)- and (-)-optically active forms) of such compounds, and mixtures thereof, and to all pharmaceutical compositions and methods of treatment defined below that contain or employ them, respectively.
  • Individual isomers can be obtained by known methods, such as optical resolution, optically selective reaction, or chromatographic separation in the preparation of the final product or its intermediate.
  • the pharmaceutical composition according to the present invention may comprise the lipophilic anthracycline in an amount of at least 0.1 %, preferably at least 0.5%, more preferably at least 1 % of said lipophilic anthracycline (w/w %).
  • the pharmaceutical composition according to the present invention may comprise the lipophilic anthracycline in an amount of 0.1 to 10 w/w %, such as e.g., from 0.1 to 8 w/w %, from 0.1 to 5 w/w %, from 1 to 5 w/w % ,from 0.1 to 2.5 w/w %, from 0.1 to 1.5 w/w %, from 0.25 to 1.25 w/w %, from 0.5 to 2.5 w/w %, from 0.5 to 2.0 w/w %, from 0.5 to 1.5 w/w %, or of about 1.0 w/w %. More preferably in an amount of from 0.25 to 1.25 w/w %, and more preferably in an amount of 1.0 w/w %.
  • compositions comprising an anthracycline, preferably any of the anthracyclines described herein above in the section "Anthracycline” for treatment of a disease where inflammation is a factor of the pathogenesis, preferably any of the inflammatory diseases described herein below in the section "Inflammatory disease”.
  • composition may comprise one or more pharmaceutically acceptable excipients.
  • Appropriate dosage forms for such administration may be prepared by conventional techniques.
  • the anthracycline may be encapsulated in a suitable carrier, for example the anthracycline may be encapsulated in a nanoparticle or a liposome.
  • suitable nanoparticles are known to the skilled person and may for example be as described by V Labhasetwar in Advanced Drug Delivery Reviews (1997) Volume: 24, Issue: 1 , Pages: 63-85.
  • the nanoparticles may also be prepared by the cholate dialysis method and are preferably characterised by a mean diameter of in the range of 30 to 60 nm.
  • a non-limiting example of such nanoparticle incorporating valrubicin were presented by Nirupama Sabnis at the nineteenth annual Research Appreciation Day (RAD) on Friday, April 1 , 201 1 at UNT Health Science Center.
  • compositions of the invention are formulated for local treatment.
  • the agent may be applied to the skin or mucosa directly, or the agent may be injected into the site of action, for example into the diseased tissue, e.g. a diseased joint or a diseased prostate.
  • the composition comprising anthracycline(s) to be used with the present invention are preferably formulated as a lotion, cream, ointment, foam or gel. Appropriate lotions, creams, ointments, foams or gels for local
  • administration to the skin may be prepared by conventional techniques known to the person skilled in the art.
  • compositions comprising the anthracycline for local administration to the skin may in a preferred embodiment be in a form selected from the group consisting of creams, gels, lotions and foams .
  • the composition for local administration to the skin comprises: i) an anthracycline as described herein above in the section "Anthracyclines", preferably a lipophilic anthracycline;
  • composition for local administration to the skin may for example be encapsulated in a nanoparticle or a liposome as described herein above.
  • composition for local administration to the skin may comprise: i) an anthracycline as described herein above in the section "Anthracyclines", preferably a lipophilic anthracycline;
  • composition further comprises an aqueous phase comprising at least one buffering system having a pH from pH 2.2 to pH 6.
  • aqueous phase comprising at least one buffering system having a pH from pH 2.2 to pH 6.
  • the water content of the pharmaceutical composition may be less than 30%.
  • the composition may preferably be in a form selected from the group consisting of a water in oil emulsion, an emulsified gel, a gel, and an oil in water emulsion; more preferably a water in oil emulsion or an emulsified gel.
  • the composition may for example be a composition comprising only a lipid phase, such as an ointment.
  • the composition may for example be a water in oil emulsion.
  • the composition is an emulsified gel.
  • the composition is a, gel or an oil in water emulsion.
  • Such compositions may be as described in international patent application WO
  • useful compositions may be as described in WO 2011/006504, p. 18, I. 21 to p. 41 , I. 23.
  • the one or more oil phase carriers, one or more pharmaceutical acceptable solubilizers, one or more co-surfactants and said aqueous phase comprising at least one buffering system may be as described in more detail in international patent application WO 2011/006504 on p. 18, I. 21 to p. 41 , I. 23 .
  • Non-limiting examples of specific useful compositions for use with the present invention, in particular for use in treatment of inflammatory skin disorder are described in WO 2011/006504 in Examples 4, 5, 6, 7, 9, 10, 15 and 16.
  • compositions comprising the anthracycline may also be formulated for local administration to other sites than skin. Depending on the particular site of inflammation the compositions may be formulated in a manner useful for local administration to said site.
  • the composition may be in a form selected from the group consisting of creams, ointments, gels, lotions, foams, pastes, sticks, patches, membranes, suppositories, sprays, and solutions or suspensions for injection.
  • the composition may be in a form selected from the group consisting of creams, gels, lotions, foams and pastes.
  • the composition may be formulated for injection, in particular for intra-articular injection.
  • composition for intra-articular injection must be sterile.
  • composition for injection is typically a liquid formulation comprising one or more solvents.
  • the solvent may for example be selected from the group consisting of Ethanol, Polyethylene glycol (PEG400), Dimethylene glycol monoethyl ether,
  • Phenoxyethanol Propylene glycol, water and mixtures thereof.
  • composition for injection frequently comprises water, however for storage purposes the composition may be devoid of water or comprise only low levels of water. Sterile water may then be added immediately prior to administration.
  • composition may comprise other excipients, such as emulsifiers, preservatives or pH adjuster.
  • the composition may also comprise salts, for example sodium chloride.
  • the emulsifier may for example be polysorbates.
  • a composition for intra-articular injection may be formulated as a dosage unit, typically a dosage unit comprising in the range of 0.1 to 10 ml, preferably in the range of 0.5 to 5 ml., more preferably in the range of 0.5 to 2 ml of a sterile liquid solution comprising in range of 0.1 to 10% w/w of said lipophilic anthracycline.
  • the composition comprises said anthracycline and in particular a lipophilic anthracycline as the only active ingredient.
  • the anthracycline, and in particular the lipophilic anthracycline is in a free form, and thus it is preferred that the anthracycline, and in particular the lipophilic anthracycline is not coupled to another compound or encapsulated in another compound. In this embodiment the anthracycline, and in particular the lipophilic anthracycline is thus not covalently linked to another compound. Furthermore, it is preferred that the anthracycline, and in particular the lipophilic anthracycline is not encapsulated, e.g. it is preferred that the anthracycline, and in particular the lipophilic anthracycline is not encapsulated by lipid encapsulation or into nanoparticles.
  • compositions containing an anthracycline according to the present invention may be prepared by conventional techniques, e.g. as described in Remington: The Science and Practice of Pharmacy 1995, edited by E. W. Martin, Mack Publishing Company, 19 th edition, Easton, Pa, unless otherwise described herein.
  • compositions for treatment of an inflammatory disease preferably only comprise one particular kind of anthracycline, however they may comprise more than one different anthracycline, such as 2, for example 3, such as 4, for example 5, such as more than 5 different anthracyclines, wherein said anthracyclines are one of the anthracyclines described herein above in the section "Anthracyclines".
  • all of said anthracyclines are lipophilic anthracyclines.
  • compositions may comprise any suitable amount of said anthracycline. It is however preferred that all of the anthracycline within the compositions is in solution, accordingly the composition should preferably not contain more anthracycline than what is soluble in the composition.
  • the composition may comprise at least 0.1 %, preferably at least 0.5%, more preferably at least 1 % of said lipophilic anthracycline.
  • the composition may comprise in the range of 0.1 % to 30%, such as in the range of 0.1 % to 20%, for example in the range of 0.1 % to 10%, such as in the range of 0.1 % to 5%, for example in the range of 0.5% to 30%, such as in the range of 0.5% to 20%, for example in the range of 0.5% to 10%, such as in the range of 0.5% to 5%, for example in the range of 0.5% to 3%, such as in the range of 0.5% to 1.5%, for example in the range of 1 % to 30%, such as in the range of 1 % to 20%, for example in the range of 1 % to 10%, such as in the range of 1 % to 5%, for example in the range of 1 % to 3%, such as in the range of 1 % to 2%, and preferably in the range of 0.25 to 1.25%
  • the % is given as w/w%.
  • said solution may comprise at least 0.1 %, preferably at least 0.5%, more preferably at least 1 % of said lipophilic anthracycline.
  • said solution may comprise in the range of 0.1 % to 30%, such as in the range of 0.1 % to 20%, for example in the range of 0.1 % to 10%, such as in the range of 0.1 % to 5%, for example in the range of 0.5% to 30%, such as in the range of 0.5% to 20%, for example in the range of 0.5% to 10%, such as in the range of 0.5% to 5%, for example in the range of 0.5% to 3%, such as in the range of 0.5% to 1.5%, for example in the range of 1 % to 30%, such as in the range of 1 % to 20%, for example in the range of 1 % to 10%, such as in the range of 1 % to 5%, for example in the range of 1 % to 3%, such as in the range of 1 % to 2%.
  • % is presented as w/w %.
  • said solution may comprise at least 0.1 %, preferably at least 0.5%, more preferably at least 1 % of said lipophilic anthracycline.
  • said gel, cream or lotion may comprise in the range of 0.1 % to 10%, such as in the range of 0.1 % to 5%, for example in the range of 0.5% to 10%, such as in the range of 0.5% to 5%, for example in the range of 0.5% to 3%, such as in the range of 0.5% to 1.5%, for example in the range of 1 % to 10%, such as in the range of 1 % to 5%, for example in the range of 1 % to 3%, such as in the range of 1 % to 2%.
  • % is presented as w/w %.
  • compositions according to the invention may also comprise one or more additional active compounds in addition to said anthracycline.
  • additional active compounds may be selected from the group consisting of steroid hormones, Benzoyl peroxide, antibiotics, estrogen, progesterone, retinoids, nicotinamide, Ibuprofen and tetracycline, preferably the additional active compounds may be selected from the group consisting of benzoyl peroxide, retinoids, azelaic acid and antibiotics.
  • the additional active compound may be selected from the aforementioned group.
  • compositions according to the invention may also comprise one or more additional active
  • Said additional active compound may for example be one or more selected from the group consisting of antibiotics, lauric acid, azelaic acid and benxoyl peroxide.
  • compositions comprising an anthracycline, preferably a lipophilic anthracycline, such as any of the anthracyclines described in the section "Anthracyclines" herein above for treatment of a clinical condition, where inflammation is a factor of the pathogenesis.
  • anthracycline preferably a lipophilic anthracycline, such as any of the anthracyclines described in the section "Anthracyclines" herein above for treatment of a clinical condition, where inflammation is a factor of the pathogenesis.
  • the clinical condition may be an anthracycline, preferably a lipophilic anthracycline, such as any of the anthracyclines described in the section "Anthracyclines" herein above for treatment of a clinical condition, where inflammation is a factor of the pathogenesis.
  • the clinical condition may be an anthracycline, preferably a lipophilic anthracycline, such as any of the anth
  • inflammation is a factor of the pathogenesis
  • diseases according to the invention are such diseases wherein inflammation is a significant factor of the pathogenesis.
  • inflammation may be the only or at least the primary factor of the pathogenesis.
  • inflammation is one significant factor of the pathogenesis, which may however also involve other factors.
  • the clinical condition is a clinical condition associated with local inflammation. It is furthermore preferred that the clinical condition is a clinical condition, where both inflammation and infection are factors of the pathogenesis. Even more preferably the clinical condition is a condition, where both inflammation and infection are primary factors of the pathogenesis.
  • the clinical condition to be treated according to the present invention is preferably a clinical condition where inflammation is a factor of the pathogenesis and in particular it may be a clinical condition associated with local inflammation.
  • inflammation is a factor of pathogenesis
  • Diseases wherein inflammation is a primary factor of pathogenesis include for example autoimmune diseases.
  • the clinical condition according to the present invention may be an acute inflammation.
  • Acute inflammation is a protective immediate innate immune response of vascularized tissue activated by tissue injury or intrusion by foreign substances.
  • the clinical condition may also be an autoimmune disease.
  • Autoimmune diseases are characterized by that the afflicted individual's own immune response being directed against its own tissues, causing prolonged inflammation and subsequent tissue destruction.
  • the clinical condition according to the invention may be a clinical condition, where inflammation is a factor of pathogenesis and wherein said inflammation may be caused or induced by an agent selected from the group consisting of a pathogenic organism, a foreign body and a physical trauma.
  • Acute inflammation is normally initiated by the release of inflammatory cytokines, chemokines and lipid mediators from mast cells and macrophages in the damaged or assaulted tissue (Lawrence and Gilroy 2007) followed by vascular events such as increased vascular flow and permeability with extravasation of leukocytes from the microcirculation and subsequent migration to- and accumulation at the focus of injury.
  • the recruited polymorphonuclear leukocytes are activated by the chemical mediators of inflammation produced and released locally by neutrophils, macrophages and mast cells as a response to the injury.
  • Cytokine cascades (TNF-a, IL-1 , IL-6, IL-12 and IFN) typically begin immediately after the injury.
  • Cytokines are key modulators of inflammation. They participate in both acute and chronic inflammation in a complex network of interactions If the agent causing or inducing the inflammation cannot be quickly eliminated, for example by repeated injuries, persistent infections or an autoimmune (auto- inflammatory) disease, the result may be chronic inflammation.
  • Chronic inflammation is predominantly associated with the presence of lymphocytes, plasma cells and macrophages, although infiltration with neutrophilic cells is also seen (Underwood 2009) Likewise proliferation of blood vessels and connective tissue is characteristic for chronic inflammation.
  • the dominating inflammatory cytokines in chronic inflammation are mainly IL-2, IL-5, IFN- ⁇ , and IL-4 (Janeway 1997). Some cytokines, such as IL-1 , significantly contribute to both acute and chronic inflammation.
  • the inflammatory skin disease may be selected from the group consisting of atopic dermatitis, eczema, acne and rosacea.
  • the inflammatory skin disease may be selected from the group consisting of eczema, acne and rosacea.
  • the inflammatory skin disease is acne.
  • the compositions comprising an anthracycline preferably are prepared for topical application to the skin, for example as described herein above in the section
  • the autoimmune disease may be an inflammatory joint condition.
  • the inflammatory joint disease may preferably be a reactive joint diseases.
  • the inflammatory joint disease may be selected from the group consisting of rheumatoid arthritis, ankylosing spondylitis, gout and bursitis.
  • the compositions comprising an anthracycline preferably are prepared for local administration to a joint, preferably they are prepared for administration to a joint by intra-articular injection, for example as described herein above in the section
  • compositions comprising an anthracycline, preferably a lipophilic anthracycline, which may be any of the
  • anthracyclines described herein above in the section "Anthracyclines" and notably the lipophilic anthracycline may be valrubicin, wherein said composition is for treatment of atopic dermatitis.
  • Atopic dermatitis is a chronic inflammatory skin disease characterized by dry thickened, scaly, inflamed, and itchy rashes on the skin and flexure surfaces and is often seen in individuals with familial history of allergic conditions.
  • a variety of topical Over The Counter (OTC) and prescription (Rx) therapies and systemic treatments are available for the management of atopic dermatitis mainly targeting inflammation and dry skin.
  • compositions comprising a anthracycline preferably are prepared for topical application to the skin, for example as described herein above in the section "Pharmaceutical compositions”.
  • compositions comprising an anthracycline, preferably a lipophilic anthracycline, which may be any of the
  • anthracyclines described herein above in the section "Anthracyclines”, and notably the anthracycline may be valrubicin, wherein said composition is for treatment of eczema.
  • Said eczema may preferably be selected from the group consisting of Contact
  • the compositions comprising a anthracycline preferably are prepared for topical application to the skin, for example as described herein above in the section "Pharmaceutical compositions".
  • the disease to be treated with the compositions of the invention is a disease wherein both infection and inflammation are significant factors of pathogenesis.
  • the infection may be any infection, but in one embodiment of the invention the infection is infection by a bacterium, preferably a Gram-positive bacterium.
  • Said bacterium may in one embodiment be a bacterium, which is found in human tissue, for example on skin of human beings, and preferably said bacterium may be found in both healthy human beings as well as in human beings suffering from said clinical disease.
  • the infection is an infection by Propionibacterium acnes.
  • Said Propionibacterium acnes may be
  • Propionibacterium acnes of any serotype e.g. Propionibacterium acnesTypel or Propionibacterium acnes Type II. Said Propionibacterium acnes may also be
  • Propionibacterium acnes of any biotype such as any of the biotypes of
  • compositions comprising an anthracycline, such as a lipoohilic anthracycline for treatment of a clinical condition wherein infection by Propionibacterium acnes and inflammation both are factors of the pathogenesis.
  • anthracycline such as a lipoohilic anthracycline for treatment of a clinical condition wherein infection by Propionibacterium acnes and inflammation both are factors of the pathogenesis.
  • compositions comprising an anthracycline, preferably a lipophilic anthracycline, which may be any of the
  • Acne is a common skin disorder characterized by lesions, which frequently are inflammatory lesions.
  • Acne has a multifactorial pathogenesis and thus acne is an example of a clinical condition, where inflammation is a significant factor of the pathogenesis, but not the only factor.
  • Contributors to acne development include for example increased sebum secretion, colonization with Propionibacterium acnes, inflammation of hair follicles and sebaceous glands preceding follicular hyperkeratinisation and abnormal desquamation.
  • Propionibacterium acnes activate toll like receptors (TLR-2) on the cell membranes of inflammatory cells thus stimulating the secretion of cytokines such as interleukin (IL)-6 and IL-8 by follicular keratinocytes and IL-8 and -12 in macrophages giving rise to inflammation.
  • TLR-2 toll like receptors
  • Acne to be treated with the compositions according to the invention may be any kind of acne, but preferably is Acne Vulgaris.
  • compositions comprising an anthracycline, preferably a lipophilic anthracycline, preferably are prepared for topical application to the skin, for example as described herein above in the section "Pharmaceutical compositions”.
  • the invention relates to
  • compositions comprising an anthracycline, preferably a lipophilic anthracycline as described herein above in the section "Anthracyclines" for treatment of rosacea.
  • Rosacea is a common chronic inflammatory skin disease characterized by symptoms of facial flushing and a spectrum of clinical signs, including erythema, telangiectasia, coarseness of skin, and an inflammatory papulopustular eruption resembling acne.
  • the aetiology of rosacea is still unknown. Recent molecular studies suggest that an altered innate immune response is involved in the pathogenesis of the vascular and inflammatory disease seen in patients with rosacea.
  • rosacea Current management of rosacea depends on the clinical findings/ subtype and comprise topical and systemic treatment with products such as mitronidazole, azeleic acid, tetracyclines and erythromycin.
  • the antibiotics are mainly used due to their anti- inflammatory effect and not for their antibacterial effect.
  • compositions comprising a lipophilic anthracycline preferably are prepared for topical application to the skin, for example as described herein above in the section "Pharmaceutical compositions”.
  • the invention in oneembodiment relates to compositions comprising lipophilic anthracyclines for treatment of inflammatory joint diseases, preferably arthritis.
  • Said arthritis is preferably selected from the group consisting of Psoriatic arthritis,
  • the present invention relates to compositions comprising anthracyciines, preferably lipophilic anthracyciines for treatment of rheumatoid arthritis.
  • Rheumatoid arthritis is the most common autoimmune inflammatory arthritis, affecting approximately 1 % of the population. It is a chronic, frequently progressive disease caused by a dysfunction of the immune system, triggered by external factors such as e.g. stress. Inflammatory cells accumulating in synovial membranes activate synovial cells.
  • cytokines- TNF-alpha tumor necrosis factor-a
  • IL-6 IL-6 are upregulated in rheumatoid arthritis causing erosion and destruction of articular cartilage and adjacent bone causing damage to the joint capsule and the articular (joint) cartilage as these structures are replaced by scarlike tissue called pannus.
  • Rheumatoid arthritis may be associated with infection by Propionibacterium acnes.
  • the clinical condition is rheumatoid arthritis associated with infection by Propionibacterium acnes.
  • rheumatoid arthritis Treatment of rheumatoid arthritis is in general aimed at reducing inflammation in the joints, relieving pain, preventing or reducing joint damage and reducing disability in order to improve the quality of life of patients.
  • the following treatments are current standard therapies; non-steroidal anti-inflammatory drugs (NSAIDs), corticosteroids, disease modifying anti-rheumatic drugs (DMARDs) such as methotrexate,
  • NSAIDs non-steroidal anti-inflammatory drugs
  • corticosteroids corticosteroids
  • DMARDs disease modifying anti-rheumatic drugs
  • compositions comprising an anthracycline preferably a lipophilic anthracycline preferably are prepared for local administration to a joint, preferably they are prepared for administration to a joint by intra-articular injection, for example as described herein above in the section "Pharmaceutical compositions”.
  • the clinical condition where inflammation is a factor of pathogenesis may also be chronic juvenile arthritis and in particular chronic juvenile arthritis associated with infection by with Propionibacterium acnes.
  • the invention in one embodiment also relates to compositions comprising
  • anthracyclines preferably lipophilic anthracyclines for treatment of bursitis.
  • Bursae are filled with synovial fluid and are found around most joints of the body. Irritation from pressure, friction or trauma of a bursa may lead to bursitis (non-infected inflammation of a bursa).
  • the bursae most commonly involved in bursitis are the prepatellar bursa, the infrapatellar bursa and the anserine bursa and thus the invention in particular relate to compositions comprising a lipophilic anthracycline for treatment of bursitis of the prepatellar bursa, the infrapatellar bursa and/or the anserine bursa.
  • Bursitis is common amongst athletes and in certain occupations e.g. plumbers, carpet layers etc.
  • the current medical treatment is painkillers and anti-inflammatory drugs such as aspirin, NSAIDs and injections with corticosteroids.
  • compositions comprising a lipophilic anthracycline preferably are prepared for local administration to a joint, preferably they are prepared for administration to a joint by intra-articular injection, for example as described herein above in the section "Pharmaceutical compositions".
  • the invention relates to compositions comprising an anthracycline, preferably a lipophilic anthracycline for treatment of gout, which herein also may be referred to as gouty arthritis or podagra.
  • Gouty arthritis is a characteristically acute inflammatory reaction that erupts in response to articular deposits of monosodium urate (MSU) crystals.
  • MSU monosodium urate
  • MSU crystals promote inflammation directly by stimulating cells via Toll-like receptor signalling and by providing a surface for cleavage of C5 and formation of complement membrane attack complex (C5b-9), culminating in secretion of cytokines, chemokines, and other inflammatory mediators with a dramatic influx of neutrophils into the joint.
  • C5b-9 complement membrane attack complex
  • compositions comprising a lipophilic anthracycline preferably are prepared for local administration to a joint, preferably they are prepared for administration to a joint by intra-articular injection, for example as described herein above in the section "Pharmaceutical compositions”.
  • a clinical condition associated with hyperproliferation i.e. cell hyperproliferation, preferably epithelial hyperproliferation and/or abnormal cell differentiation.
  • compositions comprising anthracyclines, preferably lipophilic anthracyclines may be prepared for treatment of inflammation associated with lichen planus.
  • Lichen planus is an inflammatory autoimmune papulosquamous disorder of unknown aetiology that involves the skin and mucous membranes. It is characterised by generalized or localized small, purplish, itchy, papules having a predilection for the trunk and flexor surfaces. The lesions may be discrete or coalesce to form plaques that are flattened on the skin or, in the case of the oral cavity, a milky-white colour.
  • Treatment of lichen planus is as described for eczema in general.
  • compositions comprising an anthracycline, preferably a lipophilic anthracycline of the invention may also for example be prepared for treatment of inflammation associated with psoriasis.
  • Psoriasis is a chronic inflammatory autoimmune disease characterized by inflammatory cell infiltration and vascular dilatation of epidermis and dermis, epidermal hyperproliferation, abnormal cell differentiation with reduced programmed cell death (apoptosis) and parakeratosis resulting in the characteristic clinical symptoms erythema, thickening of the skin and scaling.
  • compositions comprising an anthracycline, preferably a lipophilic anthracycline are for treatment of an inflammatory disease, with the proviso that when said inflammatory disease is not associated with infection then said clinical condition is also not associated with hyperproliferation, i.e. cell hyperproliferation, preferably epithelial hyperproliferation and/or abnormal cell differentiation.
  • hyperproliferation i.e. cell hyperproliferation, preferably epithelial hyperproliferation and/or abnormal cell differentiation.
  • compositions comprising at least one anthracycline and preferably a lipophilic anthracycline for the treatment of a) a clinical condition wherein both inflammation and infection are factors of the pathogenesis, preferably both inflammation and infection are significant factors of the pathogenesis, for example inflammation and infection are primary factors of the pathogenesis; or
  • compositions comprising an anthracycline, preferably a lipophilic anthracycline according to the invention are for treatment of an inflammatory disease, with the proviso that said inflammatory disease is not psoriasis or lichen planus.
  • the individual to be treated according to the present invention is preferably an individual suffering from an inflammatory disease, preferably one of the inflammatory diseases described herein above in the section "Inflammatory disease".
  • the individual may be any animal, however, preferably the individual is a mammal, more preferably a human being.
  • the treatment may be ameliorating treatment and/or the treatment may be curative treatment.
  • the treatment may abolish or relieve some or all of the symptoms of the condition during treatment and/or for a specific period of time after cessation of treatment, but then one or more symptoms may reappear.
  • the symptoms may reappear about 1 day, such as about 2 days, for example about 3 days, such as about 3 to 5 days, for example about 5 to 7 days, such as about 7 to 10 days, for example about 10 to 15 days, such as about 15 to 20 days, for example about 20 to 30 days, such as about 30 to 60 days, for example about 60 to 120 days, such as more than 120 days after cessation of treatment.
  • the symptoms may preferably resumed.
  • the pharmaceutical formulations according to the present invention are preferably formulated for local administration, such as e.g. to the skin
  • Local administration should be understood as local administration directly to the site of disease.
  • Local administration may be a topical administration.
  • local administration results in that the majority of the active compound, i.e. anthracycline, is not systemically absorbed and hence substantially only capable of exerting its effect locally at the site of application.
  • systemic absorption is less than 10%, such as less than 8%, for example less than 6%, such as less than 5% for example less than 4%, such as less than 3%, for example less than 2%, such as less than 1 %.
  • Even more preferably systemic uptake is less than 10 ng/ml, more preferably less than 5 ng/ml, even more preferably less than 1 ng/ml as measured in blood of a patient receiving treatment.
  • administration of the pharmaceutical compositions according to the present invention does not result in any severe malaise or any severe irritation, more preferably administration does not result in any significant nuisance to the individual to be treated, most preferably, administration does result in only mild and/or no malaise, irritation and/or nuisance.
  • Administration frequency will depend on the particular clinical condition to be treated and the particular formulation of the pharmaceutical composition.
  • the pharmaceutical composition as described herein is formulation for administration once or twice daily, preferably once daily.
  • the treatment is normally continued until the symptoms of the disease, wherein inflammation is a factor of the pathogenesis have disappeared or at least are significantly reduced.
  • the treatment may be continued for in the range of 1 day to several months, such as in the range of 1 day to 1 month, for example for 1 day to 2 weeks. For example, it may be in the range of 1 day to 2 months.
  • the treatment may be repeated.
  • compositions according to the invention are not or only mildly irritant when applied to skin of a subject.
  • anthracyclines such as the lipophilic anthracyclines comprised in the compositions according to the invention are not or only mildly irritant when applied to skin or mucosa of a subject.
  • irritation is determined by visual assessment by an observer
  • a score of skin irritation is provided on a scale from 0 to 9
  • the visual assessment is preferably carried out by one or more trained observer(s).
  • a score of 0 on such a scale indicates that the pharmaceutical formulation or the anthracycline is not irritant
  • a score of in the range of 1 to 3 on such a scale indicates that the pharmaceutical formulation or the anthracycline is mildly irritant.
  • the assessment is made in the range of 23 to 76 hours after application of the pharmaceutical formulation or the anthracycline onto the skin of said subject, more preferably in the range of 23 to 48 hours after application of the pharmaceutical formulation or the anthracycline or other compound onto the skin of said subject.
  • the subject may be any mammal, for example mini pigs, preferably the subject is a human being.
  • the scale used is as follows:
  • the composition when applied to skin has a score for skin irritation of at the most 3 on a scale from 0 to 9, wherein 0 is no reaction and 9 is the strongest reaction.
  • skin irritation is determined as described in Basketter et al., Contact Dermatitis, 1997, 37:218-220.
  • skin irritation may be determined by a standard human 4-h patch test as described in D. A. Basketter et al, Contact Dermatitis, 2004, 51 :1-4: "Determination of skin irritation potential in the human 4-h patch test". The assessment of the skin reaction is graded as follows:
  • the pharmaceutical compositions according to the present invention is preferably not, or only mildly irritant when applied to skin, as corresponding to a grading of 0 or + on the human 4-h patch scale.
  • the pharmaceutical compositions, when applied to skin has a grading in the human 4-h patch test of at the most +, on a scale from 0 to +++, wherein 0 is no reaction and +++ is the strongest reaction.
  • mice C57BL/6J mice (6-10 weeks-old) (can e.g. be purchased from Taconic (Ry, Denmark). Mice were fed a standard rodent laboratory diet and given water ad libitum. Animals were housed under standard conditions with a 12 hour light/dark cycle and controlled temperature. Mice were anesthetized with isoflurane prior TPA application, measurement of ear thickness, and collection of punch biopsies from the ear. Post treatment mice were sacrificed by cervical dislocation.
  • mice were divided into 3 treatment groups.
  • Group 1 and 2 were challenged by a single application of TPA (0.125 ug/uL dissolved in acetone, Sigma-Aldrich, St. Louis, MO) in a volume of 10 uL per dorsal and 10 uL per ventral surface of each ear.
  • Group 2 was furthermore treated with a single application of valrubicin (10 ug/uL dissolved in acetone, provided by Sicor, Italy in a volume of 10 uL per dorsal and 10 uL per ventral surface of each ear 30 min. post challenge.
  • Group 3 served as control, and received 10 ⁇ _ of acetone on the dorsal and 10 uL on the ventral surface of each ear.
  • ear thickness was measured by a Mitutoyo digimatic indicator and punch biopsies taken. Punch biopsies were snap frozen in liquid nitrogen and stored at -80°C until further processed for MPO, mRNA, or protein purification. Neutrophil cell infiltration and cytokine levels were determined as described herein below.
  • Valrubicin significantly reduced ear thickness compared to TPA challenge alone (p ⁇ 0,001) measured by mitutyo digimatic indicator (see figure 1). Valrubicin treatment significantly reduced neutrophil cell infiltration compared to TPA challenge alone (*, p ⁇ 0, 00001 ) assessed by MPO assay (see figure 2 and 3). Further valrubicin explicated its effect by significantly decreasing the expression levels of the pro-inflammatory cytokines I L-1 ⁇ and IL-6 measured by protein level, ELISA, (p ⁇ 0,0001) and messenger RNA level, qRT-PCR (p ⁇ 0,001) (see figure 4 and 5)
  • mice were divided into 3 treatment groups.
  • Group 1 and 2 were challenged by topical TPA application (0.1 ug/uL dissolved in acetone, Sigma-Aldrich, St. Louis, MO) in a volume of 10 uL per dorsal and 10 uL per ventral surface of each ear on day 1 , 3, 5, 8, and 10.
  • Group 2 was furthermore treated with applications of valrubicin (10 ug/uL dissolved in acetone, provided by Valderm ApS, Lyngby, Denmark) in a volume of 10 uL per dorsal and 10 uL per ventral surface of each ear on day 5, 6, 7, 8, 9, 10, and 1 1 ; valrubicin was applied 30 min. post challenge.
  • Group 3 served as control, and received 10 of acetone on the dorsal and 10 uL on the ventral surface of each ear on day 1 , 3, 5, 8, and 10. Ear thickness was measured on day 1 , 3, 5, 6, 7, 8, 9, 10, and 1 1 and biopsies obtained at indicated time points after the last valrubicin application.
  • Neutrophil cell infiltration and cytokine levels were determined as described herein below.
  • Valrubicin significantly decreased the ear thickness compared to TPA challenge alone (p ⁇ 0.001) measured by mitutyo digimatic indicator (see figure 6).
  • Valrubicin treatment significantly reduced neutrophil cell infiltration compared to TPA challenge alone (*, p ⁇ 0.002) assessed by MPO assay (see figure 7 and 8).
  • MPO Myeloperoxidase assay
  • Biopsies were placed in 1 mL 0.5% hexadecyltrimethylammonium bromide (HTAB, Sigma, St. Louis, MO), homogenized (TissueLyser, Qiagen, Haan, Germany), and afterwards incubated at 37°C for 1 hour. The supernatant was collected and centrifuged (4000 rpm, 5 min, 20°C). 20 of each sample and 100 TMB ONE, Ready-to-use Substrate (Kem-En-Tec Diagnostics A/S, Copenhagen, Denmark) were added to 96-well plates in triplicates. Plates were processed in the dark for 10 min.
  • HTAB hexadecyltrimethylammonium bromide
  • the reaction was stopped by adding 100 ⁇ _ of 0.2 M H 2 S0 4 and the absorbance was measured at 450 nm (iEMS reader MF, Laboratory Systems, Copenhagen, Denmark).
  • a standard curve was created based on human neutrophilic cells collected from blood samples from a healthy volunteer. Cells were isolated using PolymorphrepTM (Axis- Shield PoC AS, Oslo, Norway) according to the product instructions.
  • RNA laterOice (Ambion Inc., Austin, TX) was added to each mouse ear biopsy. Biopsies remained at -80°C for at least 20 minutes before they were transferred to -20°C and kept at this temperature overnight. Biopsies were cut into smaller pieces and transferred to tubes containing 175 RNA Lysis Buffer including ⁇ -mercaptoethanol (SV Total RNA Isolation System, Promega, Madison, Wl) and homogenized 2 x 2 min, 25 Hz. The further handling was in accordance with the SV Total RNA Isolation System vacuum protocol. RNA contents in purified samples were measured (NanoDrop 2000, Thermo Fisher Scientific Inc, Wilmington, DE).
  • cDNA was synthesized using Taqman Reverse Transcription Reagents (Applied Biosystems, Foster City, CA) and a Peltier Thermal Cycler-200 (MJ Research, Inc., Waltham, MA). qRT-PCR was performed in 20 in a StepOnePlusTM Real-Time PCR system machine (Applied Biosystems, San Francisco, CA) using Universal PCR Master Mix, No Amp-Erase (Applied Biosystems, Foster City, CA). Expression of I L-1 ⁇ and IL-6 were determined by TaqMan® Gene Expression Assays (Assay ID: IL-1 ⁇ : Mm01336189_m1 , IL-6: Mm00446190_m1 , Applied Biosystems, San Francisco, CA).
  • GAPDH was used as internal control (Assay ID: GAPDH: M m 99999915_g 1 , Applied Biosystems, San Francisco, CA). The expression of each gene was analyzed in triplicates. PCR conditions were: 2 min at 50°C, 10 min at 95°C followed by 40 cycles of 15 s at 95°C, and 60 s at 60°C.
  • I L-1 ⁇ and IL-6 protein expression levels were measured by use of DuoSet® ELISA development kits (IL-1 ⁇ : DY401 , IL-6: DY406, R&D Systems, Oxon, UK) and substrates (Substrate reagent Pack: DY999, R&D Systems, Oxon, UK) on 96-well Maxisorb plates (Invitrogen, Carlsbad, CA, USA) according to manufacturer's instructions. Protein expression was determined in duplicate by an ELISA reader (Laboratory Systems iEMS Reader MF, Copenhagen, Denmark) at 450 nm. Statistics
  • P. acnes is intradermally injected into ears of mice and valrubicin is given as either topical or systemic treatment.
  • valrubicin is given as either topical or systemic treatment.
  • Sebocytes will be cultured in the presence of p. acnes. Following the expression levels of inflammatory cytokines (e.g. TNFa, IFNy, IL-1 a, IL-6, IL-8, IL-17A, and IL-20) will be evaluated on both mRNA and protein expression level.
  • inflammatory cytokines e.g. TNFa, IFNy, IL-1 a, IL-6, IL-8, IL-17A, and IL-20
  • Sphingosine kinase-1 (SphK1) is a key enzyme in the sphingolipid biology, as it regulates the levels of ceramide and sphingosine 1 -phosphate (S1 P).
  • SphKl phosphorylates phingosine to produce sphingosine 1 -phosphate (S1 P), a pro- survival, anti-apoptotic and pro-angiogenic second messenger.
  • SphK1 is an oncogenic enzyme whose expression is found significantly higher in various cancers including prostate cancer than the normal counterparts. SphK1 overexpression makes cancer cells more resistant to apoptosis in response to various chemotherapeutic agents and is considered as a sensor of chemotherapy effect.
  • sphingolipids are mediators of inflammation; changes in their cellular concentration modulate specific cellular functions.
  • SPHK 1 is a potential therapeutic target for the control of cancer and inflammation.
  • a SphK1 -selective compound, 5c has been shown to be efficacious in bacterial sepsis Science. 2010;328: 1290-1294.
  • Test item Valrubicin (N-trifluroacetyladriamycin-14-valerate)(Diluted Valstar, Endo Pharmaceuticals, USA).
  • Prostate cancer cell line was plated at a density ranging from 1 X 106 to 2 X 106 in 90 mm culture dish. The cells were incubated overnight and subsequently treated with Valrubicin. Untreated cells were used as control. Valrubicin was dissolved in DMSO to get main stock of 20 mM. This stock was used for preparation of further dilutions ranging from 0.1 ⁇ - 10 ⁇ in serum free media for subsequent dilutions. Valrubicin was administered once at the beginning of the experiment. After the administration of valrubicin, the cells were incubated at 37°C for 48 h.
  • SphK1 inhibitory potential of Valrubicin in human prostate cancer cell lines, LNCaP or PC-3 was detected with respect to untreated control cells by determination of SphK1 activity in cell lysate using fluorescence based spingosine kinase 1 inhibitor screening assay kit.
  • valrubicin inhibits SphK1 activity by 19-23%.
  • doxorubicin enhances SphK1 activity in Jurkat cells several fold (see figure 4D in Gude et al., 2008, FASEB, vol. 22, p. 2629-2638).
  • the human prostate epithelial cell line, RWPE-1 was grown to a density of 0.1x10 6 cells/well.
  • P. acnes (P6:ST33) were inititally cultured in blood agar at 37°C under anaerobic conditions and maintained in reinformced Clostridium borth at 37°C under anaerobic conditions.
  • the bacteria were grown to a density of 10 9 per ml, washed in PBS and added to the RWPE-1 cells at different dilutions for 24 h.
  • the levels of IL-6 and IL-8 were determined in the culture supernatants by ELISA and the multiplicity of infection (MOI) of P. acnes for RWPE-1 cells was determined to be 12.5:1.
  • 0.1x106 RWPE-1 cells/well were then treated with P. acnes (P6:ST33) at MOI and together with valrubicin for 24 h.
  • Valrubicin was added in the concentration 0.01 ⁇ , 0.05 ⁇ and 0.1 ⁇ .
  • IL-8 The level of IL-8 was determined in the culture supernatants by ELISA and the results are shown in figure 8.
  • Treatment of RWPE-1 cells with valrubicin at all the tested concentrations resulted in inhibition of IL-8 as compared to control.
  • IL-8 is a marker for inflammation.
  • the example shows that valrubicin is capable of reducing inflammation induced by prostate cells infected with P. acnes.

Abstract

The invention relates to pharmaceutical compositions comprising an anthracycline for the treatment of an inflammatory skin disease, wherein both inflammation and infection are factors of the pathogenesis. The inflammatory skin disease may for example be acne. The compositions are in general prepared for direct administration to the skin. The anthracycline is an anthracycline, which does not induce necrosis and/or lasting tissue damage upon application to skin of human beings, for example valrubicin.

Description

Treatment of inflammatory disorders with anthracyclines Field of invention
The present invention relates to pharmaceutical formulations comprising an
anthracycline and in particular a lipophilic anthracycline and the uses thereof in the treatment of clinical disorders where inflammation is a factor of the pathogenesis.
Background of invention Anthracyclines
Anthracyclines are antibiotics having potent antineoplastic activity, and accordingly they have been used in the treatment of a variety of cancers. Anthracyclines are amongst the most utilised antitumor drugs ever developed. Anthracyclines mediate their anticancer effect in part by targeting topisomerase II, which leads to DNA damage. Anthracyclines are essential components of several drug combinations for the therapy of a number of cancer diseases. It is a drug of primary use in the treatment of patients with lymphoma, breast cancer and sarcoma.
The group of anthracyclines for example comprises doxorubicin, daunorubicin, carcinomycin, valrubicin, epirubicin, and idarubicin.
Doxorubicin, daunorubicin, idarubicin and epirubicin are usually administered systemically by intravenous injection or infusion. Systemic administration of these anthracyclines, however, results in a number of undesirable side effects such as cardiotoxicity and bone marrow suppression
Anthracyclines are in general known to be very tissue toxic. For example, paravenous extravasation of doxorubicin results in severe necrosis and immediate measures have to be undertaken to avoid severe local toxicity. It is also known that several
anthracyclines are toxic, when applied directly to the skin. Valrubicin is a semisynthetic analogue of doxorubicin and it was developed for the treatment of superficial bladder cancer and approved for such use in the United States. Usually a total of 800 mg is administered by intravesical instillation of two hours +- duration for a total of 6 times once a week. (FDA. Valstar™ (Valrubicin) Sterile
Solution. 2009). International patent application WO 2003/015759 describes valrubicin for the treatment of hyperproliferative conditions of body surfaces. It is described that Valrubicin has antiproliferative effects and that it is suitable for topical treatment of hyperproliferative skin disordes, e.g. plaque psoriasis, non-melanoma skin tumors and carcinoma in situ.
It has been described in an article by Cecilia R.Kjeldsen et al. (C. Rosada et al., Valrubicin in a topical formulation treats psoriasis in a xenograft transplantation model, J Invest Dermatol. 2010 Feb; 130(2):455-63), that valrubicin in a topical formulation treats psoriasis in a xenograft transplantation model.
It has been described in an article by Andersen, S. M et al. (Andersen, S. M et al., Topical application of Valrubicin has a beneficial effect on developing skin tumors. Carcinogenesis(2010) 31 (8): 1483-1490 that valrubicin in a topical formulation significantly inhibited tumor formation in a mouse skin carcinogenesis model and has a beneficial effect on developing skin tumors.
Summary of invention
Surprisingly the present inventors have found that anthracyclines, and in particular lipophilic anthracyclines are useful for local/topical treatment of inflammatory diseases and in particular for local treatment of skin diseases with an inflammatory component of the pathogenesis.
In particular, the present inventors have very surprisingly found that antracyclines, which do not induce necrosis and/or lasting tissue damage are useful for local/topical treatment of diseases, wherein both infection and inflammation are significant factors of pathogenesis. The infection may be any infection, but in one embodiment of the invention the infection is a bacterial infection, e.g. infection by Propionibacterium acnes.
In contrast other anthracyclines, such as doxorubicin may in fact cause acne. Thus, in the datasheet of CAELYX®, which is a medicament containing pegylated liposomal doxorubicin hydrochloride it is stated that in ovarian cancer patients treated with CAELYX® one common dermatological adverse effect (>1 % and <10%) is acne. Figures
Figure 1 shows that Topical valrubicin treatment reduces ear thickness in an acute 12- O-tetradecanoylphorbol 13-acetate (TPA) induced dermatitis model. C57BL/6J mice were divided into 3 groups. Mice in group 1 were challenged by a single application of TPA. Group 2 received valrubicin (acetone solution) treatment 30 min. post TPA challenge. Group 3 received a single application of acetone and served as control. Methods and treatments are the same for figures 1 to 5. Before and at various time points after ear challenge, ear thickness was measured by a
mitutoyodigimaticindicator. Results are presented as means ±SEM. * Indicates significant difference (p < 0.001 between TPA challenged group and valrubicin treated group). Figure 2 shows results of myeloperoxidase assays. The assays were performed to quantify the degree of neutrophilic cell infiltration in the inflamed mice ears. 4 mm punch biopsies were obtained from TPA challenged (n = 8), valrubicin treated (n = 8) and control mice (n = 3) 24 h post TPA challenge. Valrubicin significantly decreased neutrophilic infiltration. Columns indicate means + SEM, * p < 0.00001.
Figure 3 shows that valrubicin treatment decreases the mRNA expression levels of the proinflammatory cytokines IL-^(a) and IL-6 (b). TPA challenged mice (n = 8), valrubicintreated mice (n = 8) and control mice (n = 3). The mRNA levels were determined by qRT-PCR and normalized to glyceraldehyde-3-phosphate
dehydrogenase(GAPDH). The columns show indexed values of mRNA expression compared with control mice. Results are presented as means + SEM. Significant differences are marked, * p < 0.001. Note that the y-axis have different scales in each sub-figure. Figure 4 shows that valrubicin treatment prevents the TPA induced increase of I L-1 β and IL-6 at protein levels. Protein expressions were determined by sandwich enzyme- linked immunoabsorbant assay (ELISA). Columns represent cytokine expression in pg per mg total protein, shown as means + SEM,* p < 0.0001. Note that the y-axis have different scales in each sub-figure. Figure 5 shows that topical application of valrubicin reduced ear thickness in a 12-O- tetradecanoylphorbol 13-acetate (TPA)-induced chronic inflammation mouse model. Mice were challenged with TPA on day 1 , 3, 5, 8 and 10 (TPA), or TPA on day 1 , 3, 5, 8 and 10 and valrubicin on day 5, 6, 7, 8, 9, 10, 1 1 and 12 (valrubicin) or acetone on day 1 , 3, 5, 8 and 10 (control). Ear thickness was measured on day 1 , 3, 5, 6, 7, 8, 9, 10, 1 1 and 12. TPA challenge significantly increased ear thickness compared to control (**, p < 0.01). Valrubicin treatment significantly reduced ear thickness compared to TPA challenge alone (*, p < 0.01). Statistical results showing significant difference between TPA-challenged and valrubicin-treated ears at a 95 % level are marked with *.
Figure 6 shows that topical application of valrubicin reduced 12-O-tetradecanoylphorbol 13-acetate (TPA)-increased neutrophil cell infiltration in mice ears in a chronic inflammation mouse model. Mice were challenged with TPA on day 1 , 3, 5, 8 and 10 (TPA), or TPA on day 1 , 3, 5, 8 and 10 and valrubicin on day 5, 6, 7, 8, 9, 10, 11 and 12 (valrubicin) or acetone on day 1 , 3, 5, 8 and 10 (control). On day 12, 24 hours post- treatment of the valrubicin-group, 4-mm punch biopsies were taken from the left ear of all mice. The number of neutrophil cells per 4-mm punch biopsy is shown. Valrubicin treatment significantly reduced neutrophil cell infiltration compared to TPA challenge alone (*, p < 0.002).
Figure 7 shows one mode of action of Valrubicin. The enzyme SphK1 catalyses formation of GlcCer, S1 P and C1 P, which have several negative effects in relation to cancer. Valrubicin reduces the activity of SphKl
Figure 8 shows the level of IL8 secreted by RWPE-1 cells infected with P. acnes after 24 h of treatment with valrubicin.
Detailed description of the invention Anthracyclines
The anthracyclines to be used with the present invention may be any anthracycline, and in particular an anthracycline of formula I as described herein below. It is furthermore preferred that in general the anthracyclines to be used with the present invention do not induce necrosis and/or lasting tissue damage upon application to skin of human beings. Even more preferably anthracyclines according to the present invention are not locally toxic, when applied topically to a body surface of an individual, more preferably, the anthracyclines are not, or are only mildly irritant, when applied to a body surface of an individual in an effective dose.
Whether a compound, such as an anthracycline is not, or only mildly irritant, is preferably determined as described herein below in the section "Skin irritation". Several anthracyclines widely used in cancer chemotherapy are highly toxic and extravasion of these anthracyclines into surrounding tissues during chemotherapy may lead to lasting tissue damage and necrosis (see e.g. Ener et al, Annals of Oncology 15: 858-862, 2004) . Thus, preferably the anthracyclines to be used with the present invention are neither doxorubicin, daunorubicin, idarubicin nor epirubicin.
It is also preferred that the anthracycline to be used with the present invention is not toxic to or only mildly toxic to cells surrounding the site of inflammation, which are not desirable to target. Thus, in one embodiment of the invention the anthracycline is not toxic to or only mildly toxic to sebaceous cells.
The anthracycline to be used with the present invention are in general anthracyclines of the general formula I:
Figure imgf000006_0001
wherein Ri is selected from the group consisting of -H, Ci_6-carboxyl, Ci-6-alkyl and Ci_6-alkoxy; and
R2 is selected from the group consisting of
Figure imgf000007_0001
R9 , -OH, -Ci-6-alkyl-OH and Ci-6-alkyl, wherein
Y is selected from the group consisting of O, S and N; and
R8 is only present in the event Y is N and R8 is selected from the group consisting of -NH-(CH2)p-NH-CH3, -NH-(CH2)p-CH3, -NH-(CH2)P-aryl, -NH-
(C=0)-aryl, wherein p is an integer in the range of 1 to 6 and said aryl may be substituted with one or more substituents selected from the group consisting of -NH2, N02 and halogen; and R9 is selected from the group consisting of Ci_6 alkyl, Ci_6 alkyl-OH, -
(CH2)n-0-(C=0)-(CH2)m-X, -(CH2)n-0-(CH2)m-X and -(CH2)n-(C=0)-(CH2)m- X, wherein X is selected from the group consisting of -COOH, -OH and - CH3 and n and m individually are integers in the range of 0 to 6; and R3 is selected from the group consisting of -OH, -SH and NH2; and
R4 is selected from the group consisting of Ci-6-alkyl and -H; and
R5 is selected from the group consisting of -O-Ri0 and -N-R10, wherein
R10 is selected from the group consisting of -H, Ci.6-alkyl,
monoglucosides, -O-monoglucoside, amino-monoglucosides, -O-amino- monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)-CH3, -(CH2)q- (CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, wherein said -H, Ci-6-alkyl, monoglucosides, -O-monoglucoside, amino- monoglucosides, -O-amino-monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)-CH3, -(CH2)q-(CHOH)r(CH2OH), -0-(CH2)q-aryl and - (CH2)q-aryl, may optionally be substituted with one or more substituents selected from the group consisting of -(CHOH)r-(CH2OH), monoglucosides, amino-monoglucosides, halogen, -NH2 and -N02; and wherein q is an integer in the range of 0 to 6 and r is an integer in the range of 1 to 7; and
R6 is selected from the group consisting of -N-(Rn)(R12) and -O-Rn, wherein
Rii is selected from the group consisting of -H, Ci.6-alkyl, monoglucosides, -O-monoglucoside, amino-monoglucosides, -O-amino- monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)-CH3, -(CH2)q- (CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, wherein said -H, Ci-6-alkyl, monoglucosides, -O-monoglucoside, amino- monoglucosides, -O-amino-monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)-CH3, -(CH2)q-(CHOH)r(CH2OH), -0-(CH2)q-aryl and - (CH2)q-aryl may optionally be substituted with one or more selected from the group consisting of -(CHOH)r-(CH2OH), monoglucosides, amino- monoglucosides, halogen, -NH2 and -N02; and wherein R12 is selected from the group consisting of -H and Ci-6-alkyl; and wherein q is an integer in the range of 0 to 6 and r is an integer in the range of 1 to 7; and
R7 is selected from the group consisting of -OH, Ci.6-alkoxy and -H; and wherein the ring indicates an aliphatic 5 to 6 membered heterocyclic ring, and wherein R4, R5 and R6 may be attached at any useful position on said ring. Said aliphatic 5 to 6 membered heterocyclic ring is preferably a 6 membered heterocyclic ring, more preferably a 6 membered hetero-cyclic ring of one oxygen and 5 carbon atoms, substituted with R4, R5 and R6, wherein R4, R5 and R6 are as define herein above and preferably as described herein below in this section.
The C5 carbonyl group of formula I is desirable for optimal function of the claimed anthracyclines and it is thus preferred that said C5 carbonyl group is present in the anthracyclines to be used with the present invention.
Thus, in one preferred embodiment of the invention the anthracycline is a compound of the formula I I
Figure imgf000009_0001
wherein each of R^ R2, R3, R4, R5, R6 and R7 are as defined herein above in relation to formula I .
R4 of the compound of either formula I or formula II is selected from the group consisting of Ci-6-alkyl and -H, preferably from the group consisting of Ci.3-alkyl and - H, more preferably from the group consisting of -CH3 and -H, even more preferably - CH3.
R5 of the compound of either formula I or formula II is selected from the group consisting of -O-Ri0 and -N-R10, however preferably R5 is -O-R10, wherein R10 is as defined herein above in relation to compounds of formula I . R6 of the compound of either formula I or formula II is selected from the group consisting of -N-(Rn)(R12) and -O-Rn , however preferably R6 is -N-(Rn)(R12), wherein Rn and R12 are as defined herein above in relation to compounds of formula I.
Thus, in a preferred embodiment of the present invention the anthracyclines to be used are compounds of formula III:
Figure imgf000010_0001
wherein R^ R2, R3, R10, Rn and R12 are as defined herein above in relation to compounds of formula I. R of formula I, formula II or formula III may be selected from the group consisting of - H, Ci-e-acyl, Ci-6-alkyl and Ci.6-alkoxy; preferably R^ is selected from the group consisting of -H and
Figure imgf000010_0002
is selected from the group consisting of -(C=0)-0-CH3 and -H.
It is preferred that at least one of R^ and R2 comprises a carbonyl or a hydrazine group and more preferably that either R^ or R2, but not both comprises a carbonyl or a hydrazine group. It is preferred that when is Ci-e-acyl, for example when is
Figure imgf000011_0001
then R2 is selected from the group consisting of -OH, -Ci_6-alkyl-OH and Ci-6-alkyl, more preferably R2 is selected from the group consisting of Ci-6-alkyl, even more preferably from the group consisting of d-3-alkyl. rred that when R2 is selected from the group consisting of
Figure imgf000011_0002
Figure imgf000011_0003
R2 of formula I, II and III may be selected from the group consisting of R9 ,
OH, -Ci-6-alkyl-OH and Ci-6-alkyl, wherein Y, R8 and R9 are as defined herein above. Preferab 2 of formula I, II and III is selected from the group consisting of
Figure imgf000011_0004
9 , wherein Y, R8 and R9 are as defined herein above. Even more preferably, Y is selected from the group consisting of O and N, more preferably Y is O. It follows that in the very preferred embodiment, where Y is O, then R8 is not present.
However, in other embodiments of the invention, wherein Y is N, then R8 of formula I, II and III is selected from the group consisting of -NH-(CH2)p-NH-CH3, -NH-(CH2)p-CH3, - NH-(CH2)p-aryl, -NH-(C=0)-aryl, wherein p is an integer in the range of 1 to 6.
Preferably, p is an integer in the range of 1 to 4, for example p is an integer in the range of 2 to 4. Preferably, when Y is N, then R8 of formula I, II and III is selected from the group consisting of -NH-(CH2)p-NH-CH3 and -NH-(C=0)-aryl, wherein p is an integer in the range of 2 to 4, more preferably R8 is selected from the group consisting of -N(CH2CH2)2NCH3 and -NH-(C=0)-phenyl.
Figure imgf000012_0001
wherein R3, R7, R9, R10, Rn and R12 are as defined herein above in relation to formula I.
R3 of the compound according to formula I, formula II, formula III or formula IV is selected from the group consisting of -OH, -SH and NH2; preferably R3 is selected from the group consisting of -NH2 and -OH, even more preferably R3 is -OH. R7 of compounds of formula I, formula II, formula III or formula IV is are is selected from the group consisting of -OH, Ci_6-alkoxy and -H, more preferably R7 is selected from the group consisting of -OH, -H and Ci_3-alkoxy, even more preferably R7 is selected from the group consisting -OH, -H and methoxy. In particular R7 may be methoxy. R9 of the compounds of formula I, II, III or IV is selected from the group consisting of d-e alkyl, Ci.e alkyl-OH, -(CH2)n-0-(C=0)-(CH2)m-X, -(CH2)n-0-(CH2)m-X and -(CH2)n- (C=0)-(CH2)m-X, wherein X is selected from the group consisting of -COOH, -OH and -CH3 and n and m individually are integers in the range of 0 to 6. In embodiments of the invention, wherein Y is N, then R9 of formula I, II or III is preferably Ci-6 -alkyl, more preferably Ci-3-alkyl, even more preferably methyl.
In embodiments of the invention where Y is O, then R9 of formula I, II, III or IV is preferably selected from the group consisting of Ci_6 alkyl, Ci_6 alkyl-OH, -(CH2)n-0-
(C=0)-(CH2)m-X, -(CH2)n-0-(CH2)m-X and -(CH2)n-(C=0)-(CH2)m-X, wherein X, n and m is as defined herein above.
When R9 of formula I, II, II or IV is alkyl, it is preferred that R9 is Ci_6 alkyl, preferably Ci. 3 alkyl, more preferably methyl.
When R9 of formula I, II, II or IV is an alcohol, it is preferred that R9 is Ci_6 alkyl-OH, preferably Ci_3 alkyl-OH, more preferably -CH2-OH. In certain preferred embodiments of the invention wherein Y, then R9 of formula I, II, III or IV is preferably selected from the group consisting of -(CH2)n-0-(C=0)-(CH2)m-X, - (CH2)n-0-(CH2)m-X and -(CH2)n-(C=0)-(CH2)m-X, wherein X, n and m is as defined herein above. X is preferably selected from the group consisting of -COOH, -OH and - CH3, for example X may be -CH3. n is an integer in the range of 0 to 6, more preferably n is an integer in the range of 0 to 3, even more preferably n is an integer in the range of 0 to 1 , for example n may be 1. m is an integer in the range of 0 to 6, preferably in the range of 1 to 5, more preferably in the range of 2 to 4, such as 3.
R10 of the compounds of formula I, II, III and IV may be selected from the group consisting of -H, Ci-6-alkyl, monoglucosides, -O-monoglucoside, amino- monoglucosides, -O-amino-monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)- CH3, -(CH2)q-(CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, preferably from the group consisting of -H, monoglucosides, -O-monoglucoside, amino-monoglucosides, - O-amino-monoglucosides, -(CH2)q-(CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, and even more preferably from the group consisting of -H, monoglucosides, -O- monoglucoside, -(CH2)q-monoglucoside, (C=0)-(CH2)q)-CH3, -(CH2)q-(CHOH)r- (CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, wherein said monoglucoside, -O-monoglucoside, amino-monoglucosides, -O-amino- monoglucosides, -(CH2)q-(CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, may optionally be substituted with one or more substituents selected from the group consisting of -(CHOH)r-(CH2OH), monoglucosides, amino-monoglucosides, halogen, - NH2 and -N02, preferably from the group consisting of halogen, monoglucoside and - NH2.
In relation to R10 of the compounds of formula I, II, III and IV, said alkyl may preferably be Ci-6 alkyl, more preferably Ci_3 alkyl. Said (C=0)-(CH2)q)-CH3 may preferably be (C=0)-CH3, (C=0)-(CH2)-CH3 or (C=0)-(CH2)2-CH3. Each of said alkyl or. (C=0)- (CH2)q)-CH3 is preferably unsubstitued or substituted within the range of 1 to 3 substituents, wherein said substituents are selected from the group consisting of halogen and monoglucosides.
In relation to R10 of the compounds of formula I, II, III and IV q is an integer in the range of 0 to 6, preferably in the range of 0 to 3, more preferably in the range of 0 to 1 , even more preferably q is 1.
In relation to R10 of the compounds of formula I, II, III and IV r is an integer in the range of 1 to 7, preferably in the range of 2 to 6, more preferably in the range of 3 to 5, even more preferably r is 4.
In relation to R10 of the compounds of formula I, II, III and IV the monoglucoside may be any monoglucoside as defined herein below. Preferably, the monoglucoside is a furanoside (i.e. 5 atom ring), a pyranoside (i.e. 6 atom ring) or septanoside (i.e. 7 atoms ring). More preferably, the monoglucoside is a pyranoside. In particular, it is preferred that the monoglucoside is selected from the group consisting of
glucopyranoside, altropyranoside, mannopyranoside, gulopyranoside, idopyranoside, galactopyranoside and talopyranoside.
In relation to R10 of the compounds of formula I, II, III and IV the amino-monoglucoside may be any monoglucoside as defined herein below, wherein one or more -OH has been replaced by a -NH2 group or wherein the -O- group has been replaced by a - NH- group. Preferably, the amino-monoglucoside is an amino-furanoside (i.e. 5 atom ring), an amino-pyranoside (i.e. 6 atom ring) or an amino-septanoside (i.e. 7 atoms ring). More preferably, the amino-monoglucoside is an amino-pyranoside. In particular, it is preferred that the amino-monoglucoside is selected from the group consisting of amino-glucopyranoside, amino-altropyranoside, amino-mannopyranoside, amino- gulopyranoside, amino-idopyranoside, amino-galactopyranoside and amino- talopyranoside. When R10 of the compounds of formula I, II, III and IV comprises a monoglucoside or - (CHOH)r-(CH2OH), for example any of the monoglucoside or -(CHOH)r-(CH2OH) described herein above, it is preferred that said monoglucoside or -(CHOH)r-(CH2OH) is either unsubstituted or substituted at in the range of 1 to 3 positions, preferably at 1 position. Said monoglucoside or -(CHOH)r-(CH2OH) is preferably substituted with one or more substituents selected from the group consisting of monoglucoside or -(CHOH)r- (CH2OH).
In relation to R10 of the compounds of formula I, II, III and IV aryl is as defined herein below. Preferably, the aryl is a 5 or 6 membered aromatic or heteroaromtic ring, more preferably the aryl is a 5 or 6 membered aromatic or heteroaromatic ring comprising at the most 1 heteroatom. Thus, preferably said aryl is selected from the group consisting of furanyl, pyrrolyl, thiophenyl, phenyl and pyridinyl, however preferably said aryl is selected from the group consisting of phenyl and pyridinyl. When R10 of the compounds of formula I, II, III and IV comprises aryl, i.e. any of the aryls described herein above, it is preferred that said aryl is either unsubstituted or substituted at in the range of 1 to 3 positions. Said aryl is preferably substituted with one or more substituents selected from the group consisting of halogen, -NH2 and N02. In relation to R10 of the compounds of formula I, II, III and IV halogen is as defined herein below, however preferably said halogen is selected from the group consisting of chloro and fluoro, more preferably said halogen is flouro.
Rii of the compounds of formula I, II, III and IV may be selected from the group consisting of -H, monoglucosides, -O-monoglucoside, amino-monoglucosides, -O- amino-monoglucosides, -(CH2)q-(CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, preferably R11 is selected from the group consisting of -H, Ci-6-alkyl, -(CH2)q- monosaccharide, -(C=0)-(CH2)qCH3 and -(CH2)q-aryl, wherein said monoglucoside, -O-monoglucoside, amino-monoglucosides, -O-amino- monoglucosides, -(CH2)q-(CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, may optionally be substituted with one or more substituents selected from the group consisting of -(CHOH)r-(CH2OH), monoglucosides, amino-monoglucosides, halogen- NH2 and N02, preferably from the group consisting of monoglucosides, halogen, -NH2 and N02.
In relation to Rn of the compounds of formula I, I I, II I and IV, said alkyl may preferably be Ci-6 alkyl, more preferably Ci_3 alkyl. Said (C=0)-(CH2)q)-CH3 may preferably be (C=0)-CH3, (C=0)-(CH2)-CH3 or (C=0)-(CH2)2-CH3. Each of said alkyl or. (C=0)- (CH2)q)-CH3 is preferably unsubstitued or substituted with in the range of 1 to 3 substituents, wherein said substituents are selected from the group consisting of halogen and monoglucosides. In relation to Rn of the compounds of formula I, II , II I and IV q is an integer in the range of 0 to 6, preferably in the range of 0 to 3, more preferably in the range of 0 to 1 , even more preferably q is 1.
In relation to Rn of the compounds of formula I, I I, II I and IV r is an integer in the range of 1 to 7, preferably in the range of 2 to 6, more preferably in the range of 3 to 5, even more preferably r is 4.
In relation to Rn of the compounds of formula I, II , II I and IV the monoglucoside may be any monoglucoside as defined herein below. Preferably, the monoglucoside is a furanoside (i.e. 5 atom ring), a pyranoside (i.e. 6 atom ring) or septanoside (i.e. 7 atoms ring). More preferably, the monoglucoside is a pyranoside. In particular, it is preferred that the monoglucoside is selected from the group consisting of
glucopyranoside, altropyranoside, mannopyranoside, gulopyranoside, idopyranoside, galactopyranoside and talopyranoside.
In relation to Rn of the compounds of formula I , I I, I I I and IV the amino-monoglucoside may be any monoglucoside as defined herein below, wherein one or more -OH has been replaced by a -NH2 group or wherein the -O- group has been replaced by a - NH- group. Preferably, the amino-monoglucoside is an amino-furanoside (i.e. 5 atom ring), an amino-pyranoside (i.e. 6 atom ring) or an amino-septanoside (i.e. 7 atoms ring). More preferably, the amino-monoglucoside is an amino-pyranoside. In particular, it is preferred that the amino-monoglucoside is selected from the group consisting of amino-glucopyranoside, amino-altropyranoside, amino-mannopyranoside, amino- gulopyranoside, amino-idopyranoside, amino-galactopyranoside and amino- talopyranoside.
When Rn of the compounds of formula I, II, III and IV comprises a monoglucoside or - (CHOH)r-(CH2OH), for example any of the monoglucoside or -(CHOH)r-(CH2OH) described herein above, it is preferred that said monoglucoside or -(CHOH)r-(CH2OH) is either unsubstituted or substituted at in the range of 1 to 3 positions, preferably at 1 position. Said monoglucoside or -(CHOH)r-(CH2OH) is preferably substituted with one or more substituents selected from the group consisting of monoglucoside or -(CHOH)r- (CH2OH). In relation to Rn of the compounds of formula I, II, III and IV aryl is as defined herein below. Preferably, the aryl is a 5 or 6 membered aromatic or heteroaromtic ring, more preferably the aryl is a 5 or 6 membered aromatic or heteroaromatic ring comprising at the most 1 heteroatom. Thus, preferably said aryl is selected from the group consisting of furanyl, pyrrolyl, thiophenyl, phenyl and pyridinyl, however preferably said aryl is selected from the group consisting of phenyl and pyridinyl.
When Rn of the compounds of formula I, II, III and IV comprises aryl, i.e. any of the aryls described herein above, it is preferred that said aryl is either unsubstituted or substituted at in the range of 1 to 3 positions. Said aryl is preferably substituted with one or more substituents selected from the group consisting of halogen, -NH2 and N02.
In relation to Rn of the compounds of formula I, II, III and IV halogen is as defined herein below, however preferably said halogen is selected from the group consisting of chloro and fluoro, more preferably said halogen is flouro.
In relation to compounds of formula I, II, III or IV, it is preferred that, when R10 is a bulky group, then Rn is not a bulky group and vice versa. Thus, preferably, when R10 is selected from the group consisting of monoglucosides, -O-monoglucoside, amino- monoglucosides, -O-amino-monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)- CH3, -(CH2)q-(CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl as defined herein above, then preferably Rn is -H. Similarly, it is preferred that when Rn is selected from the group consisting of monoglucosides, -O-monoglucoside, amino-monoglucosides, - O-amino-monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)-CH3, -(CH2)q- (CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl as defined herein above, then R10 is -H.
In relation to compounds of formula I, II, III or IV then R12 is selected from the group consisting of -H and Ci-6-alkyl. Preferably, R12 is selected from the group consisting of -H and d-3-alkyl, more preferably from the group consisting of -H and methyl, yet more preferably R12 is -H.
As mentioned herein above it is preferred that the anthracycline is not a highly toxic anthracycline. Accordingly, it is preferred that the anthracycline is a compound of formula I, II, III and IV, wherein when R7 is methoxy, then at least one of R5 or R6 is not -OH or -NH2.
Thus, the anthracycline to be used with the present invention may be a compound of formula I, II, III or IV substituted as described herein above in this section or a pharmaceutically acceptable salt thereof.
In certain embodiments of the invention, it is preferred that the anthracycline is a lipophilic anthracycline, in particular such lipophilic anthracyclines, which are capable of passing the cell membrane and enter the cytoplasm of cells in a fast manner.
Preferably, the lipophilic anthracyclines according to the invention do not enter nucleus of cells to any significant degree.
In connection with the present invention the lipophilic anthracyclines may in some instances be characterised by their partition coefficient (P) and log P. The partition coefficient is the ratio of a given compound partitioned between two solvents, traditionally the solvent system is octanol/water. The log Poctanoi can be measured in a range from -2 to 6, and compounds having a log Poctanoi/water of about 0,5 to about 2 may be considered moderately lipophilic, compounds having a log Poctanoi/water above 2 may be considered increasingly lipophilic. Hence lipophilic anthracyclines according to the present invention may preferably have a log Poctanoi/water value greater than 0.5, preferably greater than 1.0, more preferably greater than 1.5, even more preferably greater than 2.0, yet even more preferably greater than 3.0.
As the pH may have an influence on the determination of the partition coefficient, due to possible ionization of the compound in question, it may be preferred to determine the value at a fixed pH. This may for example be done by the use of octanol and a buffer system, such as e.g. PBS (wherein PBS is phosphate buffered saline, preferably 0.01 M phosphate in 8.5% NaCI, pH 7.2.). Thus preferably, the lipophilic anthracycline has a log Poctanoi water value greater than 0.5, preferably greater than 1.0, more preferably greater than 1.5, even more preferably greater than 2.0, yet even more preferably greater than 3.0 when determined in octanol and PBS, preferably PBS as specified above.
In an alternative embodiment of the present invention, the lipophilicity is characterised by the partition coefficient (P) where the lipophilic anthracycline preferably have an octanol/buffer partition coefficient (such as e.g. octanol PBS partition coefficient) greater than 40, more preferably greater than 60, even more preferably greater than 80, and yet even more preferably greater than 100. One useful method for determining the octanol/buffer partition coefficient is described in Panayiotis et al., 1989, Chemistry and Physics of Lipids, 51 : 105-1 18.
Preferred lipophilic anthracyclines, which can be used in the present invention, may have the general formula V:
Figure imgf000019_0001
wherein R2 is selected from the group consisting of-C(0)CH2-0-(CrC6-acyl), - C(0)CH2-0-(C C6-alkyl), -C(0)CH2-0-(C C6-alkoxy), -C(0)CH2-0-(C C6-acyl)- (C C6-alkoxy) and -C(0)CH2-0-C(0)(CH2)nX,
wherein n is an interger in the range of 1 to 10 and X is selected from the group consisting of -CH3, -OH and COOH; and
R6 is selected from the group consisting of-NH-(CrC6-acyl), -NH-(CrC6-alkyl), -NH- (d-Ce-alkoxy), -N(C C6-acyl)(CrC6-acyl), -N(C C6-acyl)(CrC6-alkyl), -N(C C6-acyl) (d-Ce-alkoxy), -N(CrC6-alkyl)(CrC6-alkyl), -N(CrC6-alkyl)(CrC6-alkoxy), -N(C C6- alkoxy)(C C6-alkoxy), -heterocyclyl, -C(0)CH2-0-(C C6-alkyl), -(C C6-acyl), -NH2, - OH, and H; wherein any alkyl, acyl, alkoxy, or heterocyclyl moiety of R2 optionally is substituted with one or more of CrC3-alkyl, C C2-alkoxy, -OH, halogen, -NH2, -NH- (C C4-alkyl), or -N(C C4-alkyl)(C C4-alkyl).
In a preferred embodiment the lipophilic anthracycline is a compound of the general formula V as outlined herein above or a pharmaceutical acceptable salt, solvate or prodrug thereof; more preferably the lipophilic anthracycline is a compound of the general formula V or a pharmaceutically acceptable salt thereof.
In relation to the compounds of formula V, R2 may preferably be -C(0)CH2-0-(C C6- acyl); and more preferably R2 may be -COCH2OCO(CH2)3CH3.
In one embodiment, a preferred group of R2 may be defined as -C(0)CH2-0- C(0)(CH2)nX, wherein n is an integer in the range of 1 to 10, preferably in the range of 2 to 5, more preferably in the range of 3 to 4, yet more preferably 3, and X is selected from the group consisting of -CH3, -OH and COOH , preferably X is -CH3.
Furthermore, in relation to the compounds of formula V, R6 may preferably be selected from the group consisting of-NH-(C C6-acyl), -NH-(C C6-alkyl), -NH-(C C6- alkoxy), -N(C C6-acyl)(CrC6-acyl), -N(C C6-acyl)(CrC6-alkyl), -N(C C6-acyl) (C C6- alkoxy), -N(C C6-alkyl)(CrC6-alkyl), -N(C C6-alkyl)(CrC6-alkoxy), -N(C C6- alkoxy)(C C6-alkoxy), -heterocyclyl, -C(0)CH2-0-(C C6-alkyl), and -(C C6-acyl); wherein any alkyl, acyl, alkoxy, or heterocyclyl moiety of R2 optionally is substituted with one or more of C C3-alkyl, C C2-alkoxy, -OH, halogen, -NH2, -NH-(C C4-alkyl), or -N(Ci-C4-alkyl)(CrC4-alkyl). More preferably R2may be selected from the group consisting of-NH-(C C6-acyl), -NH-(C C6-alkyl), -NH-(C C6-alkoxy), -N(C C6- acyl)(C C6-acyl), -N(Ci-Ce-acyl)(Ci-Ce-alkyl), -N(C C6-acyl) (C C6-alkoxy), -N(C C6- alkyl)(C C6-alkyl), -N(C C6-alkyl)(CrC6-alkoxy), -N(C C6-alkoxy)(CrC6-alkoxy), - heterocyclyl, and -(d-C6-acyl); wherein any alkyl, acyl, alkoxy, or heterocyclyl moiety of R2 optionally is substituted with one or more of methyl, ethyl, isopropyl, methoxy, ethoxy, isopropoxy, -OH, -Br, -F, -CI, -NH2, -NH-(C C3-alkyl), or -N(CrC3-alkyl)(C C3- alkyl).
In a specific embodiment of the compounds of formula V R6 is selected from the group consisting of -COCH2OCO(CH2)3CH3, -COCH2OH, -COCH3, -NH2, -H, -OH, and - NHCOCF3; wherein any alkyl, acyl, or alkoxy moiety of R2 optionally is substituted with one or more of CrC3-alkyl, C C2-alkoxy, -OH, halogen, -NH2, -NH-(CrC4-alkyl), or - N(Ci-C4-alkyl)(CrC4-alkyl). In an even more specific embodiment R2 is selected from the group consisting of -COCH2OCO(CH2)3CH3, -COCH2OH, -COCH3,
and -NHCOCF3; wherein any alkyl, acyl, or alkoxy moiety of R2 optionally is substituted with one or more of C C3-alkyl, C C2-alkoxy, -OH, halogen, -NH2, -NH-(C C4-alkyl), or -N(C C4-alkyl)(C C4-alkyl).
In particular, the lipophilic anthracyclines may be selected from the group consisting of OctADR (adriamycin octanoyl-hydrazone), MRA-CN (3'-deamino-3'-(3-cyano-4- morpholinyl)adriamycin), AD32 (valrubicin), AD41 , AD143, AD194, AD198, AD199, AD201 , AD202, and AD288, or mixtures thereof, or pharmaceutical acceptable salts, solvates or prodrugs thereof. The lipophilic anthracyclines may also be selected from the group consisting of derivatives of OctADR, MRA-CN, AD32, AD41 , AD143, AD194, AD198, AD199, AD201 , AD202, AD288, and mixtures thereof. Preferably, the lipophilic anthracycline is valrubicin, which may also be denoted AD32. By the term "derivatives" as used herein is meant a compound, in which one atom or a group of atoms is replaced with another atom or a group of atoms.
The structure of OctADR, MRA-CN, AD32, AD41 , AD143, AD194, AD198, AD199, AD201 , AD202, and AD288 and the lipofilicity is described by Constantinides et al.
(Constantinides, P.P. et al.; Chemistry and Physics of Lipids, 51 (1989) p. 105-118). The above mentioned N-alkyladriamycin (AD41 , AD194, AD198, AD199, AD201 , AD202, AD288) analogues and N-trifluoro-acetyladriamycin-14-O-hemiadipate (AD143) may be synthesised and characterised as described by Israel and colleagues (Israel, M. et al.; J. Med. Chem.; 28; p. 1223-1228; and Israel, M. et al.; US patent 4,610,977). One example of a preferred lipophilic anthracycline is valrubicin, which is very lipophilic due to the less ionisation compared to other anthracyclines. Accordingly, valrubicin may pass the cell membrane and enter the cytoplasm of cells in a fast manner.
The term "alkyl", as used herein, unless otherwise indicated, includes saturated monovalent hydrocarbon radicals having straight or branched moieties. Examples of alkyl moieties include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and neopentyl. Alkyl is preferably C C6 alkyl, i.e. groups containing from 1 to 6 carbon atoms, and for some embodiments of the present invention, more preferably C C4 alkyl, such as e.g. C C3 alkyl.
The term "acyl", as used herein, means an alkyl group as defined above containing at least one oxo moiety ( -C=0 ).
Preferably, the term "acyl", as used herein, refers to formyl as well as other alkyl substituted carbonyl groups, wherein "alkyl" is as defined above. For example, acyl includes groups such as (CrC6)alkanoyl (e.g., formyl, acetyl, propionyl, butyryl, valeryl, caproyl, t-butylacetyl, etc.).
The term "alkoxy", as used herein, means an -O-alkyl group wherein "alkyl" is as defined above. Examples include, but are not limited to methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, pentoxy, 2-pentyloxy, isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, and 3-methylpentoxy. Alkoxy is preferably Ci-C6 alkoxy, i.e. groups containing from 1 to 6 carbon atoms, and for some
embodiments of the present invention, more preferably C C4 alkoxy, such as e.g. C C2 alkoxy.
The term "aryl" as used herein refers to a substituent, which is a simple aromatic or heteroaromatic ring.
The terms "heterocyclyl", as used herein, refer to non-aromatic cyclic groups containing one or more heteroatoms selected from O, S and N. Preferably from one to four heteroatoms, more preferably from one to two heteroatoms. Heterocyclyl groups also include groups that are substituted with one or more oxo moieties. Examples of heterocyclyl include, but are not limited to morpholinyl, piperidinyl, piperazinyl, 1 ,2,3,6- tetrahydropyridinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl,
tetrahydrothiopyranyl, and thiomorpholinyl. Preferably morpholiny, as e.g. in MRA-CN. The term "Halogen", as used herein, includes fluoro, chloro, bromo and iodo.
When it is indicated that any alkyl, acyl, alkoxy, or heterocyclyl moiety are "optionally substituted", the moiety in question may be unsubstituted or optionally substituted with one of more substituents (typically, one to three substituents) independently selected from the group of substituents listed. The term "substituted" used as herein in relation to organic compounds refers to that one hydrogen atom of the "substituted" compound has been replaced with another chemical moiety.
The term "monoglucoside" refers to the cyclic form of a monosaccharide in which the hydroxyl -OH at the anomeric centre is replaced by an oxygen-bridged group linking the glucoside to the rest of the anthracycline. A linear monosaccharide is a compound of the general formula H-(CH(OH))x-(C=0)-(CH(OH))y-H, wherein x+y typically is an integer in the range of 4 to 7, preferably 5 to 6. Linear monosaccharides form the cyclic form through a nucleophilic addition reaction between the carbonyl group and one of the hydroxyls of the same molecule. The term "amino-monoglucoside" refers to a monoglucoside, wherein one or more -OH groups has been replaced by a -NH2 group.
The term "pharmaceutical acceptable salt, solvate or prodrug" as used herein refers to those acid and base additions salts, solvates, and prodrugs of the compounds of the present invention which are, within the scope of sound medical judgment, suitable for use without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention.
Pharmaceutically acceptable acid and base addition salts refers to the relatively non- toxic, inorganic and organic addition salts of compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds, or by subsequently reacting the purified compound in its free acid or base form with a suitable organic or inorganic compound and isolating the salt thus formed. In so far as the compounds of formula (I) of this invention are basic compounds, they are all capable of forming a wide variety of different salts with various inorganic and organic acids. Although such salts must be pharmaceutically acceptable for administration to animals, it is often desirable in practice to initially isolate the base compound from the reaction mixture as a pharmaceutically unacceptable salt and then simply convert to the free base compound by treatment with an alkaline reagent and thereafter convert the free base to a pharmaceutically acceptable acid addition salt.
The pharmaceutically acceptable acid addition salts of the basic compounds are prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt in the conventional manner. The free base form may be
regenerated by contacting the salt form with a base and isolating the free base in the conventional manner. The free base forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free base for purposes of the present invention.
Pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali and alkaline earth metal hydroxides, or of organic amines. Examples of metals used as cations are sodium, potassium, magnesium, calcium, and the like. Examples of suitable amines are Ν,Ν'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine, and procaine. The base addition salts of acidic compounds are prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt in the conventional manner. The free acid form may be regenerated by contacting the salt form with an acid and isolating the free acid in a conventional manner. The free acid forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free acid for purposes of the present invention.
Salts may be prepared from inorganic acids sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic, phosphorus, and the like. Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate mesylate, glucoheptonate, lactobionate, laurylsulphonate and isethionate salts, and the like. Salts may also be prepared from organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. and the like. Representative salts include acetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, methanesulfonate, and the like. Pharmaceutically acceptable salts may include cations based on the alkali and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations including, but not limited to, ammonium, tetramethylammonium,
tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. Also contemplated are the salts of amino acids such as arginate, gluconate, galacturonate, and the like. (See, for example, Berge S.M. et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977;66: 1-19 which is incorporated herein by reference.)
The compounds of the present invention may exist in unsolvated forms as well as in solvated forms, including hydrated forms. In general, the solvated forms, including hydrated forms, are equivalent to unsolvated forms and are intended to be
encompassed within the scope of the present invention.
The term "prodrug" refers to compounds that are rapidly transformed in vivo to yield the parent compound of the above formulae, for example, by hydrolysis. A thorough discussion is provided in T. Higuchi and V Stella, "Pro-drugs as Novel Delivery
Systems," Vol. 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and
Pergamon Press, 1987, both of which are hereby incorporated by reference. Examples of prodrugs include pharmaceutically acceptable, non-toxic esters of the compounds of the present invention, including C C6 alkyl esters wherein the alkyl group is a straight or branched chain. Acceptable esters also include C5-C7 cycloalkyl esters as well as arylalkyl esters such as, but not limited to benzyl. C C4 alkyl esters are preferred. Esters of the compounds of the present invention may be prepared according to conventional methods "March's Advanced Organic Chemistry, 5th Edition". M. B. Smith & J. March, John Wiley & Sons, 2001. Compounds of formula (I) may contain chiral centers and therefore may exist in different enantiomeric and diastereomeric forms. This invention relates to all optical isomers and all stereoisomers of compounds of the formula (I), both as racemic mixtures and as individual enantiomers and diastereoismers ((+)- and (-)-optically active forms) of such compounds, and mixtures thereof, and to all pharmaceutical compositions and methods of treatment defined below that contain or employ them, respectively. Individual isomers can be obtained by known methods, such as optical resolution, optically selective reaction, or chromatographic separation in the preparation of the final product or its intermediate. The pharmaceutical composition according to the present invention may comprise the lipophilic anthracycline in an amount of at least 0.1 %, preferably at least 0.5%, more preferably at least 1 % of said lipophilic anthracycline (w/w %).
Preferably the pharmaceutical composition according to the present invention may comprise the lipophilic anthracycline in an amount of 0.1 to 10 w/w %, such as e.g., from 0.1 to 8 w/w %, from 0.1 to 5 w/w %, from 1 to 5 w/w % ,from 0.1 to 2.5 w/w %, from 0.1 to 1.5 w/w %, from 0.25 to 1.25 w/w %, from 0.5 to 2.5 w/w %, from 0.5 to 2.0 w/w %, from 0.5 to 1.5 w/w %, or of about 1.0 w/w %. More preferably in an amount of from 0.25 to 1.25 w/w %, and more preferably in an amount of 1.0 w/w %.
Pharmaceutical compositions
The present invention relates to compositions comprising an anthracycline, preferably any of the anthracyclines described herein above in the section "Anthracycline" for treatment of a disease where inflammation is a factor of the pathogenesis, preferably any of the inflammatory diseases described herein below in the section "Inflammatory disease".
In addition to said anthracycline(s) the composition may comprise one or more pharmaceutically acceptable excipients. Appropriate dosage forms for such administration may be prepared by conventional techniques.
In one embodiment of the invention the anthracycline may be encapsulated in a suitable carrier, for example the anthracycline may be encapsulated in a nanoparticle or a liposome. Suitable nanoparticles are known to the skilled person and may for example be as described by V Labhasetwar in Advanced Drug Delivery Reviews (1997) Volume: 24, Issue: 1 , Pages: 63-85. The nanoparticles may also be prepared by the cholate dialysis method and are preferably characterised by a mean diameter of in the range of 30 to 60 nm. A non-limiting example of such nanoparticle incorporating valrubicin were presented by Nirupama Sabnis at the nineteenth annual Research Appreciation Day (RAD) on Friday, April 1 , 201 1 at UNT Health Science Center.
It is preferred that the compositions of the invention are formulated for local treatment. Accordingly, the agent may be applied to the skin or mucosa directly, or the agent may be injected into the site of action, for example into the diseased tissue, e.g. a diseased joint or a diseased prostate. For local administration to the skin the composition comprising anthracycline(s) to be used with the present invention are preferably formulated as a lotion, cream, ointment, foam or gel. Appropriate lotions, creams, ointments, foams or gels for local
administration to the skin may be prepared by conventional techniques known to the person skilled in the art.
The compositions comprising the anthracycline for local administration to the skin may in a preferred embodiment be in a form selected from the group consisting of creams, gels, lotions and foams . In one preferred embodiment of the invention the composition for local administration to the skin comprises: i) an anthracycline as described herein above in the section "Anthracyclines", preferably a lipophilic anthracycline;
ii) one or more oil phase carriers;
iii) one or more pharmaceutical acceptable solubilizers; iv) one or more co-surfactants.
In said composition for local administration to the skin the anthracycline may for example be encapsulated in a nanoparticle or a liposome as described herein above. Thus for example the composition for local administration to the skin may comprise: i) an anthracycline as described herein above in the section "Anthracyclines", preferably a lipophilic anthracycline;
ii) one or more oil phase carriers;
iii) one or more pharmaceutical acceptable solubilizers;
iv) one or more co-surfactants; and
wherein the composition further comprises an aqueous phase comprising at least one buffering system having a pH from pH 2.2 to pH 6. For example the water content of the pharmaceutical composition may be less than 30%.
In this embodiment the composition may preferably be in a form selected from the group consisting of a water in oil emulsion, an emulsified gel, a gel, and an oil in water emulsion; more preferably a water in oil emulsion or an emulsified gel. In one embodiment, the composition may for example be a composition comprising only a lipid phase, such as an ointment. In one embodiment, the composition may for example be a water in oil emulsion. In another preferred embodiment of the present invention the composition is an emulsified gel. In alternative embodiments of the present invention the composition is a, gel or an oil in water emulsion. Such compositions may be as described in international patent application WO
201 1/006504, which is hereby incorporated by reference. In particular useful compositions may be as described in WO 2011/006504, p. 18, I. 21 to p. 41 , I. 23. In particular the one or more oil phase carriers, one or more pharmaceutical acceptable solubilizers, one or more co-surfactants and said aqueous phase comprising at least one buffering system may be as described in more detail in international patent application WO 2011/006504 on p. 18, I. 21 to p. 41 , I. 23 . Non-limiting examples of specific useful compositions for use with the present invention, in particular for use in treatment of inflammatory skin disorder are described in WO 2011/006504 in Examples 4, 5, 6, 7, 9, 10, 15 and 16. The compositions comprising the anthracycline may also be formulated for local administration to other sites than skin. Depending on the particular site of inflammation the compositions may be formulated in a manner useful for local administration to said site. For example, the composition may be in a form selected from the group consisting of creams, ointments, gels, lotions, foams, pastes, sticks, patches, membranes, suppositories, sprays, and solutions or suspensions for injection. Preferably, the composition may be in a form selected from the group consisting of creams, gels, lotions, foams and pastes.
For example, if the inflammation is an inflammatory joint disease, then the composition may be formulated for injection, in particular for intra-articular injection.
Typically, a composition for intra-articular injection must be sterile. Furthermore, the composition for injection is typically a liquid formulation comprising one or more solvents. The solvent may for example be selected from the group consisting of Ethanol, Polyethylene glycol (PEG400), Dimethylene glycol monoethyl ether,
Phenoxyethanol, Propylene glycol, water and mixtures thereof.
The composition for injection frequently comprises water, however for storage purposes the composition may be devoid of water or comprise only low levels of water. Sterile water may then be added immediately prior to administration.
In addition to water the composition may comprise other excipients, such as emulsifiers, preservatives or pH adjuster. The composition may also comprise salts, for example sodium chloride. The emulsifier may for example be polysorbates.
A composition for intra-articular injection may be formulated as a dosage unit, typically a dosage unit comprising in the range of 0.1 to 10 ml, preferably in the range of 0.5 to 5 ml., more preferably in the range of 0.5 to 2 ml of a sterile liquid solution comprising in range of 0.1 to 10% w/w of said lipophilic anthracycline.
In one embodiment of the invention it is preferred that the composition comprises said anthracycline and in particular a lipophilic anthracycline as the only active ingredient.
In another embodiment it is preferred that the anthracycline, and in particular the lipophilic anthracycline is in a free form, and thus it is preferred that the anthracycline, and in particular the lipophilic anthracycline is not coupled to another compound or encapsulated in another compound. In this embodiment the anthracycline, and in particular the lipophilic anthracycline is thus not covalently linked to another compound. Furthermore, it is preferred that the anthracycline, and in particular the lipophilic anthracycline is not encapsulated, e.g. it is preferred that the anthracycline, and in particular the lipophilic anthracycline is not encapsulated by lipid encapsulation or into nanoparticles.
It is even more preferred that the composition comprises said anthracycline and in particular a lipophilic anthracycline as the only active ingredient and that said anthracycline, and in particular the lipophilic anthracycline is in a free form Pharmaceutical compositions containing an anthracycline according to the present invention may be prepared by conventional techniques, e.g. as described in Remington: The Science and Practice of Pharmacy 1995, edited by E. W. Martin, Mack Publishing Company, 19th edition, Easton, Pa, unless otherwise described herein. The compositions for treatment of an inflammatory disease preferably only comprise one particular kind of anthracycline, however they may comprise more than one different anthracycline, such as 2, for example 3, such as 4, for example 5, such as more than 5 different anthracyclines, wherein said anthracyclines are one of the anthracyclines described herein above in the section "Anthracyclines". Preferably, all of said anthracyclines are lipophilic anthracyclines.
The compositions may comprise any suitable amount of said anthracycline. It is however preferred that all of the anthracycline within the compositions is in solution, accordingly the composition should preferably not contain more anthracycline than what is soluble in the composition.
In general, the composition may comprise at least 0.1 %, preferably at least 0.5%, more preferably at least 1 % of said lipophilic anthracycline. Thus, the composition may comprise in the range of 0.1 % to 30%, such as in the range of 0.1 % to 20%, for example in the range of 0.1 % to 10%, such as in the range of 0.1 % to 5%, for example in the range of 0.5% to 30%, such as in the range of 0.5% to 20%, for example in the range of 0.5% to 10%, such as in the range of 0.5% to 5%, for example in the range of 0.5% to 3%, such as in the range of 0.5% to 1.5%, for example in the range of 1 % to 30%, such as in the range of 1 % to 20%, for example in the range of 1 % to 10%, such as in the range of 1 % to 5%, for example in the range of 1 % to 3%, such as in the range of 1 % to 2%, and preferably in the range of 0.25 to 1.25%. The % is given as w/w%. When the pharmaceutical composition is a solution or when the pharmaceutical composition comprises a solution, then said solution may comprise at least 0.1 %, preferably at least 0.5%, more preferably at least 1 % of said lipophilic anthracycline. Thus, said solution may comprise in the range of 0.1 % to 30%, such as in the range of 0.1 % to 20%, for example in the range of 0.1 % to 10%, such as in the range of 0.1 % to 5%, for example in the range of 0.5% to 30%, such as in the range of 0.5% to 20%, for example in the range of 0.5% to 10%, such as in the range of 0.5% to 5%, for example in the range of 0.5% to 3%, such as in the range of 0.5% to 1.5%, for example in the range of 1 % to 30%, such as in the range of 1 % to 20%, for example in the range of 1 % to 10%, such as in the range of 1 % to 5%, for example in the range of 1 % to 3%, such as in the range of 1 % to 2%. Again % is presented as w/w %.
When the pharmaceutical composition is a gel, cream, lotion, ointment, foam or spray or when the pharmaceutical composition comprises a gel, cream, lotion, ointment, foam or spray then said solution may comprise at least 0.1 %, preferably at least 0.5%, more preferably at least 1 % of said lipophilic anthracycline. Preferably, said gel, cream or lotion may comprise in the range of 0.1 % to 10%, such as in the range of 0.1 % to 5%, for example in the range of 0.5% to 10%, such as in the range of 0.5% to 5%, for example in the range of 0.5% to 3%, such as in the range of 0.5% to 1.5%, for example in the range of 1 % to 10%, such as in the range of 1 % to 5%, for example in the range of 1 % to 3%, such as in the range of 1 % to 2%. % is presented as w/w %.
The compositions according to the invention may also comprise one or more additional active compounds in addition to said anthracycline. For example said additional active compounds may be selected from the group consisting of steroid hormones, Benzoyl peroxide, antibiotics, estrogen, progesterone, retinoids, nicotinamide, Ibuprofen and tetracycline, preferably the additional active compounds may be selected from the group consisting of benzoyl peroxide, retinoids, azelaic acid and antibiotics. In particular, when the clinical condition is acne, then the additional active compound may be selected from the aforementioned group. In embodiments of the invention, wherein the clinical condition is a disease, wherein both inflammation and infection are significant factors of pathogenesis, in particular in clinical conditions where both a bacterial infection (e.g. infection by Propionibacterium acnes) and inflammation are significant factors of pathogenesis, then the compositions according to the invention may also comprise one or more additional active
compounds, which have an anti-bacterial effect. Said additional active compound may for example be one or more selected from the group consisting of antibiotics, lauric acid, azelaic acid and benxoyl peroxide. Clinical conditions
The invention relates to compositions comprising an anthracycline, preferably a lipophilic anthracycline, such as any of the anthracyclines described in the section "Anthracyclines" herein above for treatment of a clinical condition, where inflammation is a factor of the pathogenesis. In particular the clinical condition may be an
inflammatory skin disease.
Diseases where inflammation is a factor of the pathogenesis may be any disease which is associated with inflammation. However, preferably the diseases according to the invention are such diseases wherein inflammation is a significant factor of the pathogenesis. In some embodiments of the invention inflammation may be the only or at least the primary factor of the pathogenesis. However, in other embodiments of the invention inflammation is one significant factor of the pathogenesis, which may however also involve other factors.
In particular it is preferred that the clinical condition, is a clinical condition associated with local inflammation. It is furthermore preferred that the clinical condition is a clinical condition, where both inflammation and infection are factors of the pathogenesis. Even more preferably the clinical condition is a condition, where both inflammation and infection are primary factors of the pathogenesis.
Inflammatory disease The clinical condition to be treated according to the present invention is preferably a clinical condition where inflammation is a factor of the pathogenesis and in particular it may be a clinical condition associated with local inflammation. Several clinical conditions exist where inflammation is a factor of pathogenesis including for example acute inflammations and chronic inflammations. Diseases wherein inflammation is a primary factor of pathogenesis include for example autoimmune diseases. The clinical condition according to the present invention may be an acute inflammation. Acute inflammation is a protective immediate innate immune response of vascularized tissue activated by tissue injury or intrusion by foreign substances.
The clinical condition may also be an autoimmune disease. Autoimmune diseases are characterized by that the afflicted individual's own immune response being directed against its own tissues, causing prolonged inflammation and subsequent tissue destruction.
The inflammatory response is similar, regardless of the nature of the agent causing or inducing the inflammation. Thus the clinical condition according to the invention may be a clinical condition, where inflammation is a factor of pathogenesis and wherein said inflammation may be caused or induced by an agent selected from the group consisting of a pathogenic organism, a foreign body and a physical trauma. Acute inflammation is normally initiated by the release of inflammatory cytokines, chemokines and lipid mediators from mast cells and macrophages in the damaged or assaulted tissue (Lawrence and Gilroy 2007) followed by vascular events such as increased vascular flow and permeability with extravasation of leukocytes from the microcirculation and subsequent migration to- and accumulation at the focus of injury. The recruited polymorphonuclear leukocytes are activated by the chemical mediators of inflammation produced and released locally by neutrophils, macrophages and mast cells as a response to the injury. Cytokine cascades (TNF-a, IL-1 , IL-6, IL-12 and IFN) typically begin immediately after the injury. Cytokines are key modulators of inflammation. They participate in both acute and chronic inflammation in a complex network of interactions If the agent causing or inducing the inflammation cannot be quickly eliminated, for example by repeated injuries, persistent infections or an autoimmune (auto- inflammatory) disease, the result may be chronic inflammation.
Chronic inflammation is predominantly associated with the presence of lymphocytes, plasma cells and macrophages, although infiltration with neutrophilic cells is also seen (Underwood 2009) Likewise proliferation of blood vessels and connective tissue is characteristic for chronic inflammation.
The dominating inflammatory cytokines in chronic inflammation are mainly IL-2, IL-5, IFN-γ, and IL-4 (Janeway 1997). Some cytokines, such as IL-1 , significantly contribute to both acute and chronic inflammation. The inflammatory skin disease may be selected from the group consisting of atopic dermatitis, eczema, acne and rosacea. For example the inflammatory skin disease may be selected from the group consisting of eczema, acne and rosacea. Preferably, the inflammatory skin disease is acne. For treatment of inflammatory skin diseases the compositions comprising an anthracycline preferably are prepared for topical application to the skin, for example as described herein above in the section
"Pharmaceutical compositions".
The autoimmune disease may be an inflammatory joint condition. The inflammatory joint disease may preferably be a reactive joint diseases. In particular the inflammatory joint disease may be selected from the group consisting of rheumatoid arthritis, ankylosing spondylitis, gout and bursitis. For treatment of inflammatory joint diseases, the compositions comprising an anthracycline preferably are prepared for local administration to a joint, preferably they are prepared for administration to a joint by intra-articular injection, for example as described herein above in the section
"Pharmaceutical compositions".
In one embodiment the present invention relates to compositions comprising an anthracycline, preferably a lipophilic anthracycline, which may be any of the
anthracyclines described herein above in the section "Anthracyclines" , and notably the lipophilic anthracycline may be valrubicin, wherein said composition is for treatment of atopic dermatitis. Atopic dermatitis is a chronic inflammatory skin disease characterized by dry thickened, scaly, inflamed, and itchy rashes on the skin and flexure surfaces and is often seen in individuals with familial history of allergic conditions. A variety of topical Over The Counter (OTC) and prescription (Rx) therapies and systemic treatments are available for the management of atopic dermatitis mainly targeting inflammation and dry skin.
For treatment of atopic dermatitis the compositions comprising a anthracycline preferably are prepared for topical application to the skin, for example as described herein above in the section "Pharmaceutical compositions".
In another embodiment the present invention relates to compositions comprising an anthracycline, preferably a lipophilic anthracycline, which may be any of the
anthracyclines described herein above in the section "Anthracyclines", and notably the anthracycline may be valrubicin, wherein said composition is for treatment of eczema. Said eczema may preferably be selected from the group consisting of Contact
Dermatitis, Hand Eczema, Nummular eczema and Seborrhoic eczema.
Contact dermatitis (irritant/allergic), hand eczema, nummular eczema and seborrhoic eczema all display the clinical signs of an inflammatory response and as a
consequence current treatment is focused on relieving the inflammation and dryness of the skin by treatment with standard topical anti-inflammatory therapies such as e.g. corticosteroids, immunosuppressants, retinoids and antibiotics often in combination with moisturizers.
For treatment of eczema the compositions comprising a anthracycline preferably are prepared for topical application to the skin, for example as described herein above in the section "Pharmaceutical compositions". In certain embodiments of the invention the disease to be treated with the compositions of the invention is a disease wherein both infection and inflammation are significant factors of pathogenesis. The infection may be any infection, but in one embodiment of the invention the infection is infection by a bacterium, preferably a Gram-positive bacterium. Said bacterium may in one embodiment be a bacterium, which is found in human tissue, for example on skin of human beings, and preferably said bacterium may be found in both healthy human beings as well as in human beings suffering from said clinical disease. In one preferred embodiment of the invention the infection is an infection by Propionibacterium acnes. Said Propionibacterium acnes may be
Propionibacterium acnes of any serotype, e.g. Propionibacterium acnesTypel or Propionibacterium acnes Type II. Said Propionibacterium acnes may also be
Propionibacterium acnes of any biotype, such as any of the biotypes of
Propionibacterium acnes described in "Handbook of hydrocarbon and lipid
microbiology", ed. K.N. Timnis, Springer-Verlag Berlin Heidelberg, 2010, Chapter 66, H. Bruggemann "Skin: Acne and Propionibacterium acnes Genomics. Thus in one embodiment the invention relates to compositions comprising an anthracycline, such as a lipoohilic anthracycline for treatment of a clinical condition wherein infection by Propionibacterium acnes and inflammation both are factors of the pathogenesis.
Examples of such clinical conditions include Acne, such as acne vulgaris. In a very preferred embodiment the invention relates to compositions comprising an anthracycline, preferably a lipophilic anthracycline, which may be any of the
anthracyclines described herein above in the section "Anthracyclines", and notably the anthracycline may be valrubicin, wherein said composition is for treatment of acne. Acne is a common skin disorder characterized by lesions, which frequently are inflammatory lesions.
Acne has a multifactorial pathogenesis and thus acne is an example of a clinical condition, where inflammation is a significant factor of the pathogenesis, but not the only factor. Contributors to acne development include for example increased sebum secretion, colonization with Propionibacterium acnes, inflammation of hair follicles and sebaceous glands preceding follicular hyperkeratinisation and abnormal desquamation. Propionibacterium acnes activate toll like receptors (TLR-2) on the cell membranes of inflammatory cells thus stimulating the secretion of cytokines such as interleukin (IL)-6 and IL-8 by follicular keratinocytes and IL-8 and -12 in macrophages giving rise to inflammation.
There is a variety of agents available for the treatment of acne, including topical and systemic retinoids, antibiotics and antibacterials, such as benzoyl peroxide (BPO). Decades of clinical experience have shown unequivocally that acne responds best to a combination of agents that act upon different areas of pathophysiology, and drug development has increasingly focused on formulating combination products that can more effectively address multiple pathogenetic targets.
Acne to be treated with the compositions according to the invention may be any kind of acne, but preferably is Acne Vulgaris.
For treatment of acne the compositions comprising an anthracycline, preferably a lipophilic anthracycline, preferably are prepared for topical application to the skin, for example as described herein above in the section "Pharmaceutical compositions".
In a further preferred embodiment of the invention, the invention relates to
compositions comprising an anthracycline, preferably a lipophilic anthracycline as described herein above in the section "Anthracyclines" for treatment of rosacea. Rosacea is a common chronic inflammatory skin disease characterized by symptoms of facial flushing and a spectrum of clinical signs, including erythema, telangiectasia, coarseness of skin, and an inflammatory papulopustular eruption resembling acne. The aetiology of rosacea is still unknown. Recent molecular studies suggest that an altered innate immune response is involved in the pathogenesis of the vascular and inflammatory disease seen in patients with rosacea.
Current management of rosacea depends on the clinical findings/ subtype and comprise topical and systemic treatment with products such as mitronidazole, azeleic acid, tetracyclines and erythromycin. The antibiotics are mainly used due to their anti- inflammatory effect and not for their antibacterial effect.
For treatment of rosacea the compositions comprising a lipophilic anthracycline preferably are prepared for topical application to the skin, for example as described herein above in the section "Pharmaceutical compositions".
The invention in oneembodiment relates to compositions comprising lipophilic anthracyclines for treatment of inflammatory joint diseases, preferably arthritis. Said arthritis is preferably selected from the group consisting of Psoriatic arthritis,
Rheumatoid arthritis (RA), Juvenile rheumatoid arthritis (JRA) and Ankylosing spondylitis (AS), Gout and Bursitis. Thus, in oneembodiment the present invention relates to compositions comprising anthracyciines, preferably lipophilic anthracyciines for treatment of rheumatoid arthritis. Rheumatoid arthritis (RA) is the most common autoimmune inflammatory arthritis, affecting approximately 1 % of the population. It is a chronic, frequently progressive disease caused by a dysfunction of the immune system, triggered by external factors such as e.g. stress. Inflammatory cells accumulating in synovial membranes activate synovial cells. The inflammatory biomarkers, cytokines- TNF-alpha (tumor necrosis factor-a) and IL-6 are upregulated in rheumatoid arthritis causing erosion and destruction of articular cartilage and adjacent bone causing damage to the joint capsule and the articular (joint) cartilage as these structures are replaced by scarlike tissue called pannus.
Rheumatoid arthritis may be associated with infection by Propionibacterium acnes. Thus in one embodiment of the invention the clinical condition is rheumatoid arthritis associated with infection by Propionibacterium acnes.
Treatment of rheumatoid arthritis is in general aimed at reducing inflammation in the joints, relieving pain, preventing or reducing joint damage and reducing disability in order to improve the quality of life of patients. The following treatments are current standard therapies; non-steroidal anti-inflammatory drugs (NSAIDs), corticosteroids, disease modifying anti-rheumatic drugs (DMARDs) such as methotrexate,
hydroxychloroquine and sulfasalazin and biological treatments with TNF-alpha inhibitors, rituximab and tocilizumab.
For treatment of rheumatoid arthritis, the compositions comprising an anthracycline, preferably a lipophilic anthracycline preferably are prepared for local administration to a joint, preferably they are prepared for administration to a joint by intra-articular injection, for example as described herein above in the section "Pharmaceutical compositions".
The clinical condition where inflammation is a factor of pathogenesis may also be chronic juvenile arthritis and in particular chronic juvenile arthritis associated with infection by with Propionibacterium acnes. The invention in one embodiment also relates to compositions comprising
anthracyclines, preferably lipophilic anthracyclines for treatment of bursitis. Bursae are filled with synovial fluid and are found around most joints of the body. Irritation from pressure, friction or trauma of a bursa may lead to bursitis (non-infected inflammation of a bursa).
The bursae most commonly involved in bursitis are the prepatellar bursa, the infrapatellar bursa and the anserine bursa and thus the invention in particular relate to compositions comprising a lipophilic anthracycline for treatment of bursitis of the prepatellar bursa, the infrapatellar bursa and/or the anserine bursa. Bursitis is common amongst athletes and in certain occupations e.g. plumbers, carpet layers etc. The current medical treatment is painkillers and anti-inflammatory drugs such as aspirin, NSAIDs and injections with corticosteroids. For treatment of bursitis, the compositions comprising a lipophilic anthracycline preferably are prepared for local administration to a joint, preferably they are prepared for administration to a joint by intra-articular injection, for example as described herein above in the section "Pharmaceutical compositions". In one embodiment the invention relates to compositions comprising an anthracycline, preferably a lipophilic anthracycline for treatment of gout, which herein also may be referred to as gouty arthritis or podagra. Gouty arthritis is a characteristically acute inflammatory reaction that erupts in response to articular deposits of monosodium urate (MSU) crystals. The innate immune inflammatory response is critically involved in the pathology of gout. Specifically, MSU crystals promote inflammation directly by stimulating cells via Toll-like receptor signalling and by providing a surface for cleavage of C5 and formation of complement membrane attack complex (C5b-9), culminating in secretion of cytokines, chemokines, and other inflammatory mediators with a dramatic influx of neutrophils into the joint.
Current treatment options for gouty arthritis are NSAIDs, COX-2inhibitors, systemic corticosteroids and colchicine.
For treatment of gout, the compositions comprising a lipophilic anthracycline preferably are prepared for local administration to a joint, preferably they are prepared for administration to a joint by intra-articular injection, for example as described herein above in the section "Pharmaceutical compositions".
In certain specific embodiments of the invention it relates to methods of treating inflammation associated with a clinical condition associated with hyperproliferation, i.e. cell hyperproliferation, preferably epithelial hyperproliferation and/or abnormal cell differentiation.
For example the compositions comprising anthracyclines, preferably lipophilic anthracyclines may be prepared for treatment of inflammation associated with lichen planus.
Lichen planus is an inflammatory autoimmune papulosquamous disorder of unknown aetiology that involves the skin and mucous membranes. It is characterised by generalized or localized small, purplish, itchy, papules having a predilection for the trunk and flexor surfaces. The lesions may be discrete or coalesce to form plaques that are flattened on the skin or, in the case of the oral cavity, a milky-white colour.
Histologically, there is a "saw-tooth" pattern of epidermal hyperplasia and vacuolar alteration of the basal layer of the epidermis along with an intense upper dermal inflammatory infiltrate composed predominantly of T-cells.
Treatment of lichen planus is as described for eczema in general.
The compositions comprising an anthracycline, preferably a lipophilic anthracycline of the invention may also for example be prepared for treatment of inflammation associated with psoriasis. Psoriasis is a chronic inflammatory autoimmune disease characterized by inflammatory cell infiltration and vascular dilatation of epidermis and dermis, epidermal hyperproliferation, abnormal cell differentiation with reduced programmed cell death (apoptosis) and parakeratosis resulting in the characteristic clinical symptoms erythema, thickening of the skin and scaling.
However, in very preferred embodiments of the invention the compositions comprising an anthracycline, preferably a lipophilic anthracycline are for treatment of an inflammatory disease, with the proviso that when said inflammatory disease is not associated with infection then said clinical condition is also not associated with hyperproliferation, i.e. cell hyperproliferation, preferably epithelial hyperproliferation and/or abnormal cell differentiation.
Thus in oneembodiment the invention relates to compositions comprising at least one anthracycline and preferably a lipophilic anthracycline for the treatment of a) a clinical condition wherein both inflammation and infection are factors of the pathogenesis, preferably both inflammation and infection are significant factors of the pathogenesis, for example inflammation and infection are primary factors of the pathogenesis; or
b) a clinical condition wherein inflammation is a factor of pathogenesis, preferably inflammation is a significant factor of pathogenesis, more preferably inflammation is the primary factor of pathogenesis, with the proviso that the clinical condition is not associated with hyperproliferation. Accordingly, preferably the compositions comprising an anthracycline, preferably a lipophilic anthracycline according to the invention are for treatment of an inflammatory disease, with the proviso that said inflammatory disease is not psoriasis or lichen planus. Administration forms
The individual to be treated according to the present invention is preferably an individual suffering from an inflammatory disease, preferably one of the inflammatory diseases described herein above in the section "Inflammatory disease". The individual may be any animal, however, preferably the individual is a mammal, more preferably a human being.
The treatment may be ameliorating treatment and/or the treatment may be curative treatment. In some embodiments of the present invention, the treatment may abolish or relieve some or all of the symptoms of the condition during treatment and/or for a specific period of time after cessation of treatment, but then one or more symptoms may reappear. For example, the symptoms may reappear about 1 day, such as about 2 days, for example about 3 days, such as about 3 to 5 days, for example about 5 to 7 days, such as about 7 to 10 days, for example about 10 to 15 days, such as about 15 to 20 days, for example about 20 to 30 days, such as about 30 to 60 days, for example about 60 to 120 days, such as more than 120 days after cessation of treatment. In cases, where the symptoms reappear the treatment is preferably resumed.
The pharmaceutical formulations according to the present invention are preferably formulated for local administration, such as e.g. to the skin
Local administration according to the present invention should be understood as local administration directly to the site of disease. Local administration may be a topical administration. Preferably, local administration results in that the majority of the active compound, i.e. anthracycline, is not systemically absorbed and hence substantially only capable of exerting its effect locally at the site of application. Preferably, systemic absorption is less than 10%, such as less than 8%, for example less than 6%, such as less than 5% for example less than 4%, such as less than 3%, for example less than 2%, such as less than 1 %. Even more preferably systemic uptake is less than 10 ng/ml, more preferably less than 5 ng/ml, even more preferably less than 1 ng/ml as measured in blood of a patient receiving treatment.
Preferably, administration of the pharmaceutical compositions according to the present invention does not result in any severe malaise or any severe irritation, more preferably administration does not result in any significant nuisance to the individual to be treated, most preferably, administration does result in only mild and/or no malaise, irritation and/or nuisance.
Administration frequency will depend on the particular clinical condition to be treated and the particular formulation of the pharmaceutical composition. In general however, the pharmaceutical composition as described herein is formulation for administration once or twice daily, preferably once daily.
The treatment is normally continued until the symptoms of the disease, wherein inflammation is a factor of the pathogenesis have disappeared or at least are significantly reduced. Thus, the treatment may be continued for in the range of 1 day to several months, such as in the range of 1 day to 1 month, for example for 1 day to 2 weeks. For example, it may be in the range of 1 day to 2 months. In the event that the symptoms of the disease reappears, the treatment may be repeated.
Skin irritation As described herein above, it is preferred that the pharmaceutical compositions according to the invention are not or only mildly irritant when applied to skin of a subject. Furthermore, it is preferred that the anthracyclines, such as the lipophilic anthracyclines comprised in the compositions according to the invention are not or only mildly irritant when applied to skin or mucosa of a subject.
Several methods for determining irritation are available, for example a visual assessment or a more recent human 4-h patch test. When the irritation is determined by visual assessment by an observer, a score of skin irritation is provided on a scale from 0 to 9 The visual assessment is preferably carried out by one or more trained observer(s). A score of 0 on such a scale indicates that the pharmaceutical formulation or the anthracycline is not irritant, whereas a score of in the range of 1 to 3 on such a scale indicates that the pharmaceutical formulation or the anthracycline is mildly irritant. Preferably, the assessment is made in the range of 23 to 76 hours after application of the pharmaceutical formulation or the anthracycline onto the skin of said subject, more preferably in the range of 23 to 48 hours after application of the pharmaceutical formulation or the anthracycline or other compound onto the skin of said subject.
The subject may be any mammal, for example mini pigs, preferably the subject is a human being.
Preferably, the scale used is as follows:
0 no reaction
1 marginal reaction
2 slight perceptible erythema
3 a greater than slight reaction which is not sufficient to be classed as
distinct
4 distinct erythema
5 a greater than distinct reaction which is insufficient to be classed as well developed
6 well developed, possibly spreading erythema
7 a greater reaction which is not sufficient to be classed as strong
8 strong, deep erythema, which may extend beyond the treatment site
9 a more intense reaction than above In a preferred embodiment of the invention, the composition when applied to skin has a score for skin irritation of at the most 3 on a scale from 0 to 9, wherein 0 is no reaction and 9 is the strongest reaction. In one embodiment of the present invention skin irritation is determined as described in Basketter et al., Contact Dermatitis, 1997, 37:218-220.
Alternatively, skin irritation may be determined by a standard human 4-h patch test as described in D. A. Basketter et al, Contact Dermatitis, 2004, 51 :1-4: "Determination of skin irritation potential in the human 4-h patch test". The assessment of the skin reaction is graded as follows:
0 No reaction
+ Weakly positive reaction (usually characterized by mild erythema or dryness across most of the treatment site)
++ Moderately positive reaction (usually distinct erythema possibly spreading
beyond the treatment site)
+++ Strongly positive reaction (strong, often spreading erythema with oedema)
Accordingly, the pharmaceutical compositions according to the present invention is preferably not, or only mildly irritant when applied to skin, as corresponding to a grading of 0 or + on the human 4-h patch scale. In one embodiment of the present invention the pharmaceutical compositions, when applied to skin has a grading in the human 4-h patch test of at the most +, on a scale from 0 to +++, wherein 0 is no reaction and +++ is the strongest reaction.
Examples
The following examples illustrate preferred embodiments of the invention and should not be regarded as limiting for the invention.
Example 1
Inhibition of inflammation by valrubicin
Mice C57BL/6J mice (6-10 weeks-old) (can e.g. be purchased from Taconic (Ry, Denmark). Mice were fed a standard rodent laboratory diet and given water ad libitum. Animals were housed under standard conditions with a 12 hour light/dark cycle and controlled temperature. Mice were anesthetized with isoflurane prior TPA application, measurement of ear thickness, and collection of punch biopsies from the ear. Post treatment mice were sacrificed by cervical dislocation.
Acute TPA-induced irritant contact dermatitis model
Mice were divided into 3 treatment groups. Group 1 and 2 were challenged by a single application of TPA (0.125 ug/uL dissolved in acetone, Sigma-Aldrich, St. Louis, MO) in a volume of 10 uL per dorsal and 10 uL per ventral surface of each ear. Group 2 was furthermore treated with a single application of valrubicin (10 ug/uL dissolved in acetone, provided by Sicor, Italy in a volume of 10 uL per dorsal and 10 uL per ventral surface of each ear 30 min. post challenge. Group 3 served as control, and received 10 μΙ_ of acetone on the dorsal and 10 uL on the ventral surface of each ear. At indicated time points, after ear challenge (0, 4, 8, 24, 48, 72 hrs), ear thickness was measured by a Mitutoyo digimatic indicator and punch biopsies taken. Punch biopsies were snap frozen in liquid nitrogen and stored at -80°C until further processed for MPO, mRNA, or protein purification. Neutrophil cell infiltration and cytokine levels were determined as described herein below.
In the validated acute murine TPA-induced skin inflammation model (irritant contact dermatitis) it was demonstrated that valrubicin alleviates the induced acute immune response.
Valrubicin significantly reduced ear thickness compared to TPA challenge alone (p< 0,001) measured by mitutyo digimatic indicator (see figure 1). Valrubicin treatment significantly reduced neutrophil cell infiltration compared to TPA challenge alone (*, p <0, 00001 ) assessed by MPO assay (see figure 2 and 3). Further valrubicin explicated its effect by significantly decreasing the expression levels of the pro-inflammatory cytokines I L-1 β and IL-6 measured by protein level, ELISA, (p<0,0001) and messenger RNA level, qRT-PCR (p<0,001) (see figure 4 and 5)
Chronic TPA-induced irritant contact dermatitis model
Mice were divided into 3 treatment groups. Group 1 and 2 were challenged by topical TPA application (0.1 ug/uL dissolved in acetone, Sigma-Aldrich, St. Louis, MO) in a volume of 10 uL per dorsal and 10 uL per ventral surface of each ear on day 1 , 3, 5, 8, and 10. Group 2 was furthermore treated with applications of valrubicin (10 ug/uL dissolved in acetone, provided by Valderm ApS, Lyngby, Denmark) in a volume of 10 uL per dorsal and 10 uL per ventral surface of each ear on day 5, 6, 7, 8, 9, 10, and 1 1 ; valrubicin was applied 30 min. post challenge. Group 3 served as control, and received 10 of acetone on the dorsal and 10 uL on the ventral surface of each ear on day 1 , 3, 5, 8, and 10. Ear thickness was measured on day 1 , 3, 5, 6, 7, 8, 9, 10, and 1 1 and biopsies obtained at indicated time points after the last valrubicin application.
Neutrophil cell infiltration and cytokine levels were determined as described herein below.
In the validated chronic murine TPA-induced skin inflammation model (irritant contact dermatitis) it was demonstrated that valrubicin reduces the TPA-induced chronic inflammation (chronic immune response).
Valrubicin significantly decreased the ear thickness compared to TPA challenge alone (p < 0.001) measured by mitutyo digimatic indicator (see figure 6).
Valrubicin treatment significantly reduced neutrophil cell infiltration compared to TPA challenge alone (*, p < 0.002) assessed by MPO assay (see figure 7 and 8).
Myeloperoxidase assay (MPO)
Biopsies were placed in 1 mL 0.5% hexadecyltrimethylammonium bromide (HTAB, Sigma, St. Louis, MO), homogenized (TissueLyser, Qiagen, Haan, Germany), and afterwards incubated at 37°C for 1 hour. The supernatant was collected and centrifuged (4000 rpm, 5 min, 20°C). 20 of each sample and 100 TMB ONE, Ready-to-use Substrate (Kem-En-Tec Diagnostics A/S, Copenhagen, Denmark) were added to 96-well plates in triplicates. Plates were processed in the dark for 10 min. The reaction was stopped by adding 100 μΙ_ of 0.2 M H2S04 and the absorbance was measured at 450 nm (iEMS reader MF, Laboratory Systems, Copenhagen, Denmark). A standard curve was created based on human neutrophilic cells collected from blood samples from a healthy volunteer. Cells were isolated using Polymorphrep™ (Axis- Shield PoC AS, Oslo, Norway) according to the product instructions. qRT-PCR
One day prior to RNA extraction 500 μΙ RNA laterOice (Ambion Inc., Austin, TX) was added to each mouse ear biopsy. Biopsies remained at -80°C for at least 20 minutes before they were transferred to -20°C and kept at this temperature overnight. Biopsies were cut into smaller pieces and transferred to tubes containing 175 RNA Lysis Buffer including β-mercaptoethanol (SV Total RNA Isolation System, Promega, Madison, Wl) and homogenized 2 x 2 min, 25 Hz. The further handling was in accordance with the SV Total RNA Isolation System vacuum protocol. RNA contents in purified samples were measured (NanoDrop 2000, Thermo Fisher Scientific Inc, Wilmington, DE). cDNA was synthesized using Taqman Reverse Transcription Reagents (Applied Biosystems, Foster City, CA) and a Peltier Thermal Cycler-200 (MJ Research, Inc., Waltham, MA). qRT-PCR was performed in 20 in a StepOnePlus™ Real-Time PCR system machine (Applied Biosystems, San Francisco, CA) using Universal PCR Master Mix, No Amp-Erase (Applied Biosystems, Foster City, CA). Expression of I L-1 β and IL-6 were determined by TaqMan® Gene Expression Assays (Assay ID: IL-1 β: Mm01336189_m1 , IL-6: Mm00446190_m1 , Applied Biosystems, San Francisco, CA). GAPDH was used as internal control (Assay ID: GAPDH: M m 99999915_g 1 , Applied Biosystems, San Francisco, CA). The expression of each gene was analyzed in triplicates. PCR conditions were: 2 min at 50°C, 10 min at 95°C followed by 40 cycles of 15 s at 95°C, and 60 s at 60°C.
ELISA
I L-1 β and IL-6 protein expression levels were measured by use of DuoSet® ELISA development kits (IL-1 β: DY401 , IL-6: DY406, R&D Systems, Oxon, UK) and substrates (Substrate reagent Pack: DY999, R&D Systems, Oxon, UK) on 96-well Maxisorb plates (Invitrogen, Carlsbad, CA, USA) according to manufacturer's instructions. Protein expression was determined in duplicate by an ELISA reader (Laboratory Systems iEMS Reader MF, Copenhagen, Denmark) at 450 nm. Statistics
For statistical analysis the 2-tailed Student's t-test was employed. P values≤ 0.001 were considered significant. Example 2
Treatment of acne with valrubicin
Investigation of valrubicin's effects on inflammatory acne vulgaris in vivo:
P. acnes is intradermally injected into ears of mice and valrubicin is given as either topical or systemic treatment. One or more of the following is investigated:
• Ear swelling as measured by a Mitutoyo digimatic indicator
• The number of p. acnes is counted by plating homogenized punch biopsies on agar plates
• Neutrophil infiltrate is assayed in H&E stained cross sectioned punch biopsies
• Macrophage infiltrate is assayed in toluidin blue stained cross sectioned punch biopsies
• Neutrophil infiltrate is assayed by myeloperoxidase assay of punch biopsies
• mRNA expression levels of innate inflammatory cytokines is assayed by qRT- PCR
• Protein expression levels of innate inflammatory cytokines is assayed by ELISA
• Effect on the draining lymph nodes is assayed by their excision and weighing Investigation of valrubicin's effects on inflammatory acne vulgaris in vitro: Sebocytes will be cultured in the presence of p. acnes. Following the expression levels of inflammatory cytokines (e.g. TNFa, IFNy, IL-1 a, IL-6, IL-8, IL-17A, and IL-20) will be evaluated on both mRNA and protein expression level.
Investigation of valrubicin's anti-bacterial effect in vitro: P. acnes, S. aureus, and S. epidermidis are grown in the presence of valrubicin and the effect on growth is measured by absorbance at 600 nm.
Example 3
Sphingosine kinase 1 (SphK 1) inhibitory potential of Valrubicin in human prostate cancer cells Sphingosine kinase-1 (SphK1) is a key enzyme in the sphingolipid biology, as it regulates the levels of ceramide and sphingosine 1 -phosphate (S1 P).
SphKl phosphorylates phingosine to produce sphingosine 1 -phosphate (S1 P), a pro- survival, anti-apoptotic and pro-angiogenic second messenger. SphK1 is an oncogenic enzyme whose expression is found significantly higher in various cancers including prostate cancer than the normal counterparts. SphK1 overexpression makes cancer cells more resistant to apoptosis in response to various chemotherapeutic agents and is considered as a sensor of chemotherapy effect. Furthermore, sphingolipids are mediators of inflammation; changes in their cellular concentration modulate specific cellular functions. SPHK 1 is a potential therapeutic target for the control of cancer and inflammation.
It has been shown that inhibition of SphK1 activity resulted in decreased expression of inflammatory mediators and decreased activation of inflammatory pathways in TNFa stimulated murine Fibroblast-like synoviocytes (Baker et al. Inflammation & Allergy - Drug Targets, Volume 10, Number 6, December 201 1 , pp. 464-471 (8)
A SphK1 -selective compound, 5c, has been shown to be efficacious in bacterial sepsis Science. 2010;328: 1290-1294.
Test item: Valrubicin (N-trifluroacetyladriamycin-14-valerate)(Diluted Valstar, Endo Pharmaceuticals, USA).
Name of the cell line 2: Human prostate cancer cell line (PC-3)
Media : F12K + 10 % FBS
Source : American Type Culture Collection, USA
Growth Conditions : 37°C, 5% C02 and 95 % Humidity
Name of the Vehicle : DMSO (Merck, USA)
The SphK1 inhibitory potential of Valrubicin in human prostate cell line was
investigated by analyzing relative decrease in SphK1 activity as compared to control cells (untreated) using the fluorescence based Sphingosine Kinase 1 Inhibitor Screening Assay Kit, Item no. 700430 (Cayman chemical company, USA). Prostate cancer cell line was plated at a density ranging from 1 X 106 to 2 X 106 in 90 mm culture dish. The cells were incubated overnight and subsequently treated with Valrubicin. Untreated cells were used as control. Valrubicin was dissolved in DMSO to get main stock of 20 mM. This stock was used for preparation of further dilutions ranging from 0.1 μΜ - 10 μΜ in serum free media for subsequent dilutions. Valrubicin was administered once at the beginning of the experiment. After the administration of valrubicin, the cells were incubated at 37°C for 48 h.
SphK1 inhibitory potential of Valrubicin in human prostate cancer cell lines, LNCaP or PC-3, was detected with respect to untreated control cells by determination of SphK1 activity in cell lysate using fluorescence based spingosine kinase 1 inhibitor screening assay kit.
The result is provided in Table 1.
Table 1
Figure imgf000050_0001
*Percentage Inhibition with respect to vehicle treated cells
As shown valrubicin inhibits SphK1 activity by 19-23%. In contrast doxorubicin enhances SphK1 activity in Jurkat cells several fold (see figure 4D in Gude et al., 2008, FASEB, vol. 22, p. 2629-2638).
Example 4
The human prostate epithelial cell line, RWPE-1 was grown to a density of 0.1x106 cells/well. P. acnes (P6:ST33) were inititally cultured in blood agar at 37°C under anaerobic conditions and maintained in reinformced Clostridium borth at 37°C under anaerobic conditions. The bacteria were grown to a density of 109 per ml, washed in PBS and added to the RWPE-1 cells at different dilutions for 24 h.
The levels of IL-6 and IL-8 were determined in the culture supernatants by ELISA and the multiplicity of infection (MOI) of P. acnes for RWPE-1 cells was determined to be 12.5:1.
0.1x106 RWPE-1 cells/well were then treated with P. acnes (P6:ST33) at MOI and together with valrubicin for 24 h. Valrubicin was added in the concentration 0.01 μΜ, 0.05 μΜ and 0.1 μΜ.
The level of IL-8 was determined in the culture supernatants by ELISA and the results are shown in figure 8. Treatment of RWPE-1 cells with valrubicin at all the tested concentrations resulted in inhibition of IL-8 as compared to control. IL-8 is a marker for inflammation. The example shows that valrubicin is capable of reducing inflammation induced by prostate cells infected with P. acnes.
REFERENCER;
Rosada, C, K. Stenderup, E. de Darko, F. Dagnaes-Hansen, S. Kamp and T. N. Dam (2009). "Valrubicin in a Topical Formulation Treats Psoriasis in a Xenograft Transplantation Model." J Invest Dermatol 130(2): 455-463.
Lawrence, T. and D. W. Gilroy (2007). "Chronic inflammation: a failure of resolution?" International Journal of Experimental Pathology 88(2): 85-94.
Nakatsuji T et al.Antimicrobial property of Laurie acid against Propionibacterium Acnes:
Its Therapeutic potential for inflammatory acne vulgaris. Journal of Investigative Dermatology (2009) 129, 2480-2488;
Andersen, S. M., C. Rosada, F. Dagnaes-Hansen, I. G. Laugesen, E. de Darko, T. N.
Dam and K. Stenderup (2010). "Topical application of valrubicin has a beneficial effect on developing skin tumors." Carcinogenesis 31 (8): 1483-1490.

Claims

Claims
1. A composition comprising at least one anthracycline for the treatment of an inflammatory skin disease, wherein both inflammation and infection are factors of the pathogenesis, wherein said anthracycline is an anthracycline, which does not induce necrosis and/or lasting tissue damage upon application to skin of human beings and wherein the composition is prepared for topical administration to the skin.
2. The composition according to any one of the preceding claims, wherein the inflammatory skin disease is a clinical condition where both inflammation and a bacterial infection are factors of the pathogenesis.
3. The composition according to any one of the preceding claims, wherein the inflammatory skin disease is a clinical condition associated with both inflammation and infection by Propionibacterium acnes.
4. The composition according to any one ofclaims 1 to 3, wherein the inflammatory skin disease is Acne. 5. The composition according to any one or the preceding claims, wherein the inflammatory skin disease is Acne vulgaris.
6. The composition according to any one of the preceding claims, wherein the composition is prepared for local administration to the site of inflammation. 7. The composition according to any one of claims 1 to 6, wherein the composition is in the form of a cream, gel, lotion, foam or spray
8. The composition according to any one of claims 1 to 7 , wherein the composition comprises
i) a lipophilic anthracycline;
ii) one or more oil phase carriers;
iii) one or more pharmaceutical acceptable solubilizers;
iv) one or more co-surfactants; and
wherein the composition further comprises an aqueous phase comprising at least one buffering system having a pH from pH 2.2 to pH 6,.
9. The composition according to any one of the preceding claims, wherein the anthracycline is a compound of formula I,
Figure imgf000053_0001
or a pharmaceutically acceptable salt thereof, wherein is selected from the group consisting of -H, Ci_6-carboxyl, Ci-6-alkyl and Ci.6-alkoxy; and
R2 is selected from the group consisting of
Figure imgf000053_0002
R9 , -OH, -Ci-6-alkyl-OH and Ci-6-alkyl, wherein
Y is selected from the group consisting of O, S and N; and
R8 is only present in the event Y is N and R8 is selected from the group consisting of -NH-(CH2)p-NH-CH3, -NH-(CH2)p-CH3, -NH-(CH2)P-aryl, -NH- (C=0)-aryl, wherein p is an integer in the range of 1 to 6 and said aryl may be substituted with one or more substituents selected from the group consisting of -NH2, N02 and halogen; and R9 is selected from the group consisting of Ci_6 alkyl, Ci_6 alkyl-OH, - (CH2)n-0-(C=0)-(CH2)m-X, -(CH2)n-0-(CH2)m-X and -(CH2)n-(C=0)-(CH2), X, wherein X is selected from the group consisting of -COOH, -OH and - CH3 and n and m individually are integers in the range of 0 to 6; and from the group consisting of -OH, -SH and NH2; and from the group consisting of Ci-6-alkyl and -H; and from the group consisting of -O-Ri0 and -N-R10, wherein
R10 is selected from the group consisting of -H, Ci.6-alkyl,
monoglucosides, -O-monoglucoside, amino-monoglucosides, -O-amino- monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)-CH3, -(CH2)q- (CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, wherein said -H, Ci-6-alkyl, monoglucosides, -O-monoglucoside, amino- monoglucosides, -O-amino-monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)-CH3, -(CH2)q-(CHOH)r(CH2OH), -0-(CH2)q-aryl and - (CH2)q-aryl, may optionally be substituted with one or more substituents selected from the group consisting of -(CHOH)r-(CH2OH),
monoglucosides, amino-monoglucosides, halogen, -NH2 and -N02; and wherein q is an integer in the range of 0 to 6 and r is an integer in the range of 1 to 7; and from the group consisting of -N-(Rn)(R12) and -O-Rn, wherein
Rii is selected from the group consisting of -H, Ci.6-alkyl,
monoglucosides, -O-monoglucoside, amino-monoglucosides, -O-amino- monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)-CH3, -(CH2)q- (CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, wherein said -H, Ci-6-alkyl, monoglucosides, -O-monoglucoside, amino- monoglucosides, -O-amino-monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)-CH3, -(CH2)q-(CHOH)r(CH2OH), -0-(CH2)q-aryl and - (CH2)q-aryl may optionally be substituted with one or more selected from the group consisting of -(CHOH)r-(CH2OH), monoglucosides, amino- monoglucosides, halogen, -NH2 and -N02; and wherein R12 is selected from the group consisting of -H and Ci-6-alkyl; and wherein q is an integer in the range of 0 to 6 and r is an integer in the range of 1 to 7; and
R7 is selected from the group consisting of -OH, Ci.6-alkoxy and -H; and wherein the ring indicates an aliphatic 5 to 6 membered heterocyclic ring, and wherein R4, R5 and R6 may be attached at any useful position on said ring.
10. The composition according to claim 9, wherein said said aliphatic ring is a 6 membered hetero-cyclic ring, more preferably a 6 membered hetero-cyclic ring of one oxygen and 5 carbon atoms.
1 1. The composition according to any one of the preceding claims, wherein the anthracycline is a compound of the formula II
Figure imgf000056_0001
or a pharmaceutically acceptale salt thereof, wherein each of R2, R3, R4, R5, R6 and R7 are as defined in claim 9.
12. The composition according to any one of claims 9 to 1 1 , wherein R4 is selected from the group consisting of Ci.3-alkyl and -H, more preferably from the group consisting of -CH3 and -H, even more preferably -CH3. 13. The composition according to any one of claims 9 to 1 1 , wherein R5 is selected from the group consisting of -O-Ri0, wherein R10 is as defined in claim 20.
14. The composition according to any one of claims 9 to 1 1 , wherein R6 is -N- (R11XR12), wherein Rn and R12 are as defined in claim 9.
15. The composition according to any one of the preceding claims, wherein the anthracycline is a compound of formula III:
Figure imgf000057_0001
or a pharmaceutically acceptable salt thereof, wherein R2, R3, R10, Rn and R12 are as defined in claim 9.
16. The composition according to any one of claims 9 to 15, wherein R^ is selected from the group consisting of -H and
Figure imgf000057_0002
is selected from the group consisting of -(C=0)-0-CH3 and -H.
17. The composition according to any one of claims 9 to 16, wherein R^ is Ci-e-acyl, for example when
Figure imgf000057_0003
and R2 is selected from the group consisting of -OH, -d.e-alkyl-OH and Ci.e-alkyl.
18. The composition according to any one of claim 9 to 16, wherein R2 is selected from sisting of
Figure imgf000057_0004
and R is -H.
19. The composition accordi of claims 9 to 18, wherein R2 is selected
Figure imgf000058_0001
from the group consisting of R9 , wherein Y is N and R8 is selected from the group consisting of -NH-(CH2)p-NH-CH3 and -NH-(C=0)-aryl, wherein p is an integer in the range of 2 to 4, more preferably R8 is selected from the group consisting of - N(CH2CH2)2NCH3 and -NH-(C=0)-phenyl.
20. The composition according to claim 19, wherein R9 is Ci-6 -alkyl, more preferably d-3-alkyl, even more preferably methyl.
Figure imgf000058_0002
or a pharmaceutically acceptable salt thereof, wherein R3, R7, R9, R10, Rn and R12 are as defined in claim 9.
22. The composition according to any one of claims 9 to 21 , wherein R3 is selected from the group consisting of -NH2 and -OH, even more preferably R3 is -OH.
23. The composition according to any one of claims 9 to 22, wherein R7 is selected from the group consisting of -OH, -H and Ci.3-alkoxy, even more preferably R7 is selected from the group consisting -OH, -H and methoxy.
24. The composition according to any one of claims 9 to 19 and 21 to 23, wherein R9 is selected from the group consisting of Ci_6 alkyl, Ci_6 alkyl-OH, -(CH2)n-0-(C=0)-(CH2)m- X, -(CH2)n-0-(CH2)m-X and -(CH2)n-(C=0)-(CH2)m-X, wherein X is as defined in claim 22 and n and m individually are integers in the range of 0 to 6.
25. The composition according to any one of claims 9 to 24, wherein X is selected from the group consisting of -COOH, -OH and -CH3, for example X may be -CH3.
26. The composition according to any one of claims 9 to 25, wherein
R10 is selected from the group consisting of monoglucosides, -O-monoglucoside, amino-monoglucosides, -O-amino-monoglucosides, -(CH2)q-monoglucoside, (C=0)- (CH2)q)-CH3, -(CH2)q-(CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, and Rn is -H.
27. The composition according to any one of claims 9 to 25, wherein Rn is selected from the group consisting of monoglucosides, -O-monoglucoside, amino- monoglucosides, -O-amino-monoglucosides, -(CH2)q-monoglucoside, (C=0)-(CH2)q)- CH3, -(CH2)q-(CHOH)r(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl and R10 is -H.
28. The composition according to any one of claims 9 to 26, wherein R10 is selected from the group consisting of -H, monoglucosides, -O-monoglucoside, amino- monoglucosides, -O-amino-monoglucosides, -(CH2)q-(CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl, wherein said -H, monoglucosides, -O-monoglucoside, amino- monoglucosides, -O-amino-monoglucosides, -(CH2)q-(CHOH)r-(CH2OH), -0-(CH2)q-aryl and -(CH2)q-aryl may optionally be substituted with one or more substituents selected from the group consisting of -(CHOH)r-(CH2OH), monoglucosides, amino- monoglucosides, halogen, -NH2 and -N02.
29. The composition according to any one of claims 9 to 26, wherein R10 is Ci_6 alkyl, more preferably Ci_3 alkyl.
30. The composition according to any one of claims 9 to 26, wherein R10 is selected from the group consisting of (C=0)-CH3, (C=0)-(CH2)-CH3 and (C=0)-(CH2)2-CH3.
31. The composition according to any one of claims 29 and 30, wherein said alkyl or (C=0)-(CH2)q)-CH3 is unsubstitued or substituted with in the range of 1 to 3
substituents, wherein said substituents are selected from the group consisting of halogen and monoglucosides.
32. The composition according to any one of claims 9 to 26, wherein R10 comprises a monoglucoside selected from the group consisting of furanosides (i.e. 5 atom ring), pyranosides (i.e. 6 atom ring) and septanosides (i.e. 7 atoms ring) or -(CHOH)r- (CH2OH) and said monoglucoside or -(CHOH)r-(CH2OH) is either unsubstituted or substituted at in the range of 1 to 3 positions with one or more substituents selected from the group consisting of monoglucoside or -(CHOH)r-(CH2OH).
33. The composition according to any one of claims 9 to 26, wherein R10 comprises an aryl, which is a 5 or 6 membered aromatic or heteroaromatic ring comprising at the most 1 heteroatom, preferably an aryl selected from the group consisting of furanyl, pyrrolyl, thiophenyl, phenyl and pyridinyl, more preferably an aryl selected from the group consisting of phenyl and pyridinyl, wherein said aryl is either unsubstituted or substituted at in the range of 1 to 3 positions with one or more substituents selected from the group consisting of halogen, -NH2 and N02.
34. The composition according to any one of claims 9 to 25 and 27 to 32, wherein Rn is selected from the group consisting of -H, Ci-6-alkyl, -(CH2)q-monosaccharide, - (C=0)-(CH2)qCH3 and -(CH2)q-aryl, wherein said Ci-6-alkyl, -(CH2)q-monosaccharide, - (C=0)-(CH2)qCH3 and -(CH2)q-aryl may optionally be substituted with one or more substituents selected from the group consisting of -(CHOH)r-(CH2OH),
monoglucosides, amino-monoglucosides, halogen-NH2 and N02.
35. The composition according to any one of claims 9 to 25 and 27 to 32, wherein Rn is Ci-6 alkyl, more preferably Ci_3 alkyl.
36. The composition according to any one of claims 9 to 25 and 27 to 32, wherein Rn is selected from the group consisting of (C=0)-CH3, (C=0)-(CH2)-CH3 and (C=0)- (CH2)2-CH3.
37. The composition according to any one of claims 35 and 36, wherein said alkyl or (C=0)-(CH2)q)-CH3 is unsubstitued or substituted with in the range of 1 to 3
substituents, wherein said substituents are selected from the group consisting of halogen and monoglucosides.
38. The composition according to any one of claims 9 to 25 and 27 to 32, wherein Rn comprises a monoglucoside selected from the group consisting of furanosides (i.e. 5 atom ring), pyranosides (i.e. 6 atom ring) and septanosides (i.e. 7 atoms ring) or - (CHOH)r-(CH2OH) and said monoglucoside or -(CHOH)r-(CH2OH) is either
unsubstituted or substituted at in the range of 1 to 3 positions with one or more substituents selected from the group consisting of monoglucoside or -(CHOH)r- (CH2OH).
39. The composition according to any one of claims 9 to 25 and 27 to 32, wherein Rn comprises an aryl, which is a 5 or 6 membered aromatic or heteroaromatic ring comprising at the most 1 heteroatom, preferably an aryl selected from the group consisting of furanyl, pyrrolyl, thiophenyl, phenyl and pyridinyl, more preferably an aryl selected from the group consisting of phenyl and pyridinyl, wherein said aryl is either unsubstituted or substituted at in the range of 1 to 3 positions with one or more substituents selected from the group consisting of halogen, -NH2 and N02.
40. The composition according to any one of claims 9 to 36, wherein R12 is selected from the group consisting of -H and Ci_3-alkyl, more preferably from the group consisting of -H and methyl, yet more preferably -H.
41. The compositions comprising an anthracyclin according to any one of the preceding claims, wherein said anthracycline is not, or is only mildly irritant, when applied to a body surface of an individual in an effective dose.
42. The composition comprising an anthracycline according to anyone of the preceding claims, with the proviso that said anthracycline is neither doxorubicin, daunorubicin nor epirubicin, 43. The composition according any one of the preceding claims, wherein the lipophilic anthracycline has an log P octanol/water value greater than 0.5, preferably greater than 1.0, more preferably greater than 1.5, even more preferably greater than 2.0, yet even more preferably greater than 3.0. 44. The pharmaceutical composition according to any one of the preceding claims, wherein the lipophilic anthracycline has the general formula V:
Figure imgf000062_0001
wherein R2 is selected from the group consisting of -C(0)CH2-0-(CrC6-acyl), - C(0)CH2-0-(C C6-alkyl), -C(0)CH2-0-(C C6-alkoxy), -C(0)CH2-0-(C C6-acyl)- (C C6-alkoxy) and -C(0)CH2-0-C(0)(CH2)nX,
wherein n is an interger in the range of 1 to 10 and X is selected from the group consisting of -CH3, -OH and COOH; and
R6 is selected from the group consisting of -NH-(CrC6-acyl), -NH-(CrC6-alkyl), -NH- (d-Ce-alkoxy), -N(C C6-acyl)(CrC6-acyl), -N(C C6-acyl)(CrC6-alkyl), -N(C C6-acyl) (d-Ce-alkoxy), -N(CrC6-alkyl)(CrC6-alkyl), -N(CrC6-alkyl)(CrC6-alkoxy), -N(C C6- alkoxy)(C C6-alkoxy), -heterocyclyl, -C(0)CH2-0-(C C6-alkyl), -(C C6-acyl), -NH2, - OH, and H; wherein any alkyl, acyl, alkoxy, or heterocyclyl moiety of R2 optionally is substituted with one or more of CrC3-alkyl, C C2-alkoxy, -OH, halogen, -NH2, -NH- (CrC4-alkyl), or -N(C C4-alkyl)(C C4-alkyl).
45. The composition according to claim 44, wherein R2 is -C(0)CH2-0-(CrC6-acyl); more preferably R2 is -COCH2OCO(CH2)3CH3. 46. The composition according to claim 44, wherein R2 is -C(0)CH2-0-C(0)(CH2)nX and X is -CH3.
47. The composition according to any one of claims 44 to 45, wherein R2 is -C(0)CH2- 0-C(0)(CH2)nX and n is an integer in the range of 1 to 5, preferably 2 to 4, more preferably 3.
48. The composition according to any one of claims 44 to 47, wherein R6 is selected from the group consisting of -NH-(C C6-acyl), -NH-(C C6-alkyl), -NH-(C C6- alkoxy), -N(Ci-Ce-acyl)(Ci-Ce-acyl), -N(Ci-Ce-acyl)(Ci-Ce-alkyl), -N(C C6-acyl) (C C6- alkoxy), -N(Ci-Ce-alkyl)(Ci-Ce-alkyl), -N(Ci-Ce-alkyl)(Ci-Ce-alkoxy), -N(C C6- alkoxy)(C C6-alkoxy), -heterocyclyl, -C(0)CH2-0-(C C6-alkyl), and -(C C6-acyl); wherein any alkyl, acyl, alkoxy, or heterocyclyl moiety of R2 optionally is substituted with one or more of CrC3-alkyl, C C2-alkoxy, -OH, halogen, -NH2, -NH-(CrC4-alkyl), or -N(C C4-alkyl)(C C4-alkyl).
49. The composition according to any one of claim 44 to 47, wherein R6 is selected from the group consisting of -NH-(C C6-acyl), -NH-(C C6-alkyl), -NH-(C C6- alkoxy), -N(C C6-acyl)(CrC6-acyl), -N(C C6-acyl)(CrC6-alkyl), -N(C C6-acyl) (C C6- alkoxy), -N(C C6-alkyl)(CrC6-alkyl), -N(C C6-alkyl)(CrC6-alkoxy), -N(C C6- alkoxy)(CrC6-alkoxy), -heterocyclyl, and -(CrC6-acyl); wherein any alkyl, acyl, alkoxy, or heterocyclyl moiety of R2 optionally is substituted with one or more of methyl, ethyl, isopropyl, methoxy, ethoxy, isopropoxy, -OH, -Br, -F, -CI, -NH2, -NH-(CrC3-alkyl), or - N(C C3-alkyl)(CrC3-alkyl). 50. The composition according to any one of claims 44 to 47, wherein R6 is selected from the group consisting of -COCH2OCO(CH2)3CH3, -COCH2OH, -COCH3, -NH2, -H, - OH, and -NHCOCF3; wherein any alkyl, acyl, or alkoxy moiety of R2 optionally is substituted with one or more of C C3-alkyl, C C2-alkoxy, -OH, halogen, -NH2, -NH- (C C4-alkyl), or -N(C C4-alkyl)(C C4-alkyl).
51. The composition according to any one of claims 44 to 47, wherein R6 is selected from the group consisting of -COCH2OCO(CH2)3CH3, -COCH2OH, -COCH3, and -NHCOCF3; wherein any alkyl, acyl, or alkoxy moiety of R2 optionally is substituted with one or more of CrC3-alkyl, C C2-alkoxy, -OH, halogen, -NH2, -NH-(CrC4-alkyl), or -N(Ci-C4-alkyl)(CrC4-alkyl).
52. The composition according to any one of the preceding claims, wherein the lipophilic anthracycline is selected from the group consisting of OctADR, MRA-CN, AD32 (valrubicin), AD41 , AD143, AD194, AD198, AD199, AD201 , AD202, and AD288, or mixtures thereof, or pharmaceutical acceptable salt, solvates or prodrugs thereof.
53. The composition according to any one of the preceding claims, wherein the anthracycline is valrubicin. 54. The composition according to any of the preceding claims, wherein the composition comprises at least 0.1 %, preferably at least 0.5%, more preferably at least 1 % of said anthracycline.
66. The composition according to any of the preceding claims, wherein the composition comprises said anthracyline in an amount of 0.1 to 10 w/w %, such as e.g., from 0.1 to 8 w/w %, from 0.1 to 5 w/w %, from 0.25 to 1.25 w/w %, from 1 to 5 w/w %, from 0.1 to 2.5 w/w %, from 0.1 to 1.5 w/w %, from 0.5 to 2.5 w/w %, from 0.5 to 2.0 w/w %, from 0.5 to 1.5 w/w %, or of about 1.0 w/w %.
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