HK1218646B

HK1218646B - New morpholinyl anthracycline derivatives

Info

Publication number: HK1218646B
Application number: HK16106598.5A
Authority: HK
Inventors: I．贝里亚; M.卡鲁索; V．卢皮
Original assignee: 内尔维阿诺医学科学有限公司
Priority date: 2013-04-29
Filing date: 2014-04-23
Publication date: 2018-06-08

Description

Novel morpholinyl anthracycline derivatives

Technical Field

The present invention relates to novel morpholinyl anthracycline derivatives, processes for their preparation, pharmaceutical compositions containing them and their use in the treatment of abnormal cell proliferative disorders. As an example, the compounds of the invention may be used for the treatment of tumors.

Background

Anthracyclines are antibiotic compounds that exhibit cytotoxic activity. Several studies have shown that anthracyclines can operate to kill cells by a number of different mechanisms, including: 1) DNA that intercalates into cells, thereby inhibiting DNA-dependent nucleic acid synthesis; 2) the production of free radicals, which subsequently react with cellular macromolecules, leading to cell damage; or 3) interact with cell membranes [ see, e.g., C.Peterson et al, "Transport and storage of anticancer systems and human leukamia" in anticancer Antibiotics anticancer Therapy (1982), pp.132-146; and n.r.bachur, "Free radial Damage" id.pp.97-102 ]. Due to their cytotoxic activity, anthracyclines have been used in the treatment of numerous cancers, such as leukemias, breast cancers, lung cancers, ovarian adenocarcinomas and sarcomas [ see, e.g., P.H-Wiernik, in Anthracyclines: Current Status and New Developments (1980), p 11 ]. Commonly used anthracyclines include doxorubicin, epirubicin, idarubicin and daunorubicin.

In recent years, many new highly cytotoxic anthracycline derivatives have been synthesized.

Anthracycline derivatives with a substituted morpholino ring attached at the C-3' position of the sugar-raising moiety have been shown to be effective against experimental murine tumors [ see: lown, Bioactive Molecules, Vol.6, (1988), pp.55-101 and clinical trials for treatment of hepatocellular carcinoma [ see: crypto-visible anti-tumor activity in c.sessa, o.valota, c.gerni, cardiovacular toxicology, volume 7 (2), (2007), pp.75-79 ].

The patent application WO9802446 filed by Pharmacia & Upjohn SPA has disclosed as anticancer agents novel morpholinyl anthracycline derivatives in which the morpholino ring oxygen atom is bridged at the C-4' position of the sugar residue.

4-amino and 4-fluoroanthracene antibiotic derivatives are also disclosed as antitumor agents in patent applications EP288268 and EP381989 to Farmitalia Carlo Erba Srl.

Despite efforts in anticancer research, cancer remains a hidden and elusive target, and thus, there remains a need for new anticancer drugs.

Disclosure of Invention

The invention relates to morpholinyl anthracycline derivatives of formula (I) or pharmaceutically acceptable salts thereof,

wherein:

r1 is halogen or NR4R 5;

r2 is an optionally substituted group selected from straight or branched chain C₂-C₆Alkyl, NR7R8-C₂-C₆Alkyl, R6O-C₂-C₆Alkyl and COR 9;

r3 is hydrogen, or straight or branched C₁-C₄An alkoxy group;

r4 and R5 are independently hydrogen, mono-substituted benzyl, di-substituted benzyl or an optionally substituted group selected from linear or branched C₁-C₆Alkyl, NR7R8-C₁-C₆Alkyl, R6O-C₁-C₆Alkyl, R7R8N-C₁-C₆Alkylcarbonyl, R6O-C₁-C₆Alkylcarbonyl, R7R8N-C₁-C₆Alkylaminocarbonyl radical, R6O-C₁-C₆Alkylaminocarbonyl radical, R7R8N-C₁-C₆Alkylsulfonyl radical, R6O-C₁-C₆Alkylsulfonyl radical, R7R8N-C₁-C₆Alkoxycarbonyl and R6O-C₁-C₆An alkoxycarbonyl group; or

R4 and R5 together with the bound N atom form a heterocyclic group substituted with R4;

r6, R7 and R8 are independently hydrogen or optionally substituted straight or branched chain C₁-C₆An alkyl group;

r9 is OR6, NR7R8 OR an optionally substituted group selected from linear OR branched C₁-C₄Alkyl, NR7R8-C₁-C₄Alkyl and R6O-C₁-C₄An alkyl group.

The present invention also provides methods for synthesizing morpholinyl anthracycline derivatives of formula (I) and isomers, tautomers, hydrates, solvates, complexes, metabolites, prodrugs, carriers, N-oxides thereof, prepared by a method consisting of standard synthetic transformations.

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) as defined above or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient, carrier or diluent.

The invention also provides pharmaceutical compositions comprising a therapeutically effective amount of a compound of formula (I) and one or more chemotherapeutic agents.

The present invention also provides a pharmaceutical composition comprising an effective amount of a compound of formula (I) in combination with a known anti-cancer treatment, such as radiation therapy or chemotherapy regimen, in combination with: cytostatic or cytotoxic agents, antibiotic-type drugs, alkylating agents, antimetabolites, hormonal drugs, immunological drugs, interferon-type drugs, cyclooxygenase inhibitors (e.g., COX-2 inhibitors), matrix metalloproteinase inhibitors, telomerase inhibitors, tyrosine kinase inhibitors, anti-growth factor receptor drugs, anti-HER 2 drugs, anti-EGFR drugs, anti-angiogenic agents (e.g., angiogenesis inhibitors), farnesyl transferase inhibitors, Ras-Raf signal transduction pathway inhibitors, cell cycle inhibitors, other CDK inhibitors, tubulin binding agents, topoisomerase I inhibitors, topoisomerase II inhibitors, and the like.

In addition, the present invention provides a product comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined above and one or more chemotherapeutic agents as a combined preparation for simultaneous, separate or sequential use in anticancer therapy.

In another aspect, the present invention provides a compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof, for use as a medicament.

The present invention also provides a compound of formula (I) as defined above for use in a method of treating cancer, cell proliferative disorders and viral infections.

Preferably, the compounds of formula (I) as defined above are used in a method of treating a particular type of cancer, such as, but not limited to, cancer, including: bladder cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer, including small cell lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, stomach cancer, cervical cancer, thyroid cancer, prostate cancer, and skin cancer, including squamous cell carcinoma; hematopoietic tumors of lymphoid lineage, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma; hematopoietic tumors of myeloid lineage, including acute and chronic myelogenous leukemias, myelodysplastic syndrome, and promyelocytic leukemia; tumors from mesenchymal cells, including fibrosarcoma and rhabdomyosarcoma; tumors of the central and peripheral nervous system, including astrocytomas, neuroblastomas, gliomas, and schwannomas; and other tumors including melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, thyroid follicular cancer, kaposi's sarcoma, and mesothelioma.

Furthermore, the compounds of formula (I) as defined above are useful in a method for the treatment of: specific cell proliferation disorders, such as benign prostate hyperplasia, Familial Adenomatous Polyposis (FAP), multiple neurofibromatosis, psoriasis, vascular smooth muscle cell proliferation associated with atherosclerosis, pulmonary fibrosis, arthritis, glomerulonephritis, and post-operative stenosis and restenosis.

Furthermore, the compounds of formula (I) as defined above are useful in methods for inhibiting tumor angiogenesis and metastasis, and for treating organ transplant rejection and host versus graft disease.

The present invention also provides a method of treating cancer comprising administering to a mammal in need thereof an effective amount of a compound of formula (I) as defined above. The mammal in need thereof may be, for example, a human.

The invention also provides the use of a compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of cancer.

Detailed Description

The following terms and phrases used herein are intended to have the following meanings unless otherwise indicated.

The term "straight-chain or branched C" as used₁-C₆Alkyl "means any of the following groups: for example, methyl, ethyl, n-propyl, n-butyl, n-pentyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl.

The term "straightChain or branch C₁-C₄Alkoxy "means any of the following groups: for example, methoxy, ethoxy, propoxy, and the like.

The term "halogen" is used to mean fluorine, chlorine, bromine or iodine.

The term "monosubstituted-benzyl" is used to mean any radical, such as 4-methoxybenzyl, 4-methylbenzyl, 4-fluorobenzyl, 3-methoxybenzyl, 3-methylbenzyl, 3-fluorobenzyl, 2-methoxybenzyl, 2-methylbenzyl, 2-fluorobenzyl and the like.

The term "disubstituted-benzyl" is used to mean any group such as 2, 4-dimethoxybenzyl, 2, 4-dimethylbenzyl, 2, 4-difluorobenzyl, 2, 3-dimethoxybenzyl, 2, 3-dimethylbenzyl, 2, 3-difluorobenzyl, 2, 5-dimethoxybenzyl, 2, 5-dimethylbenzyl, 2, 5-difluorobenzyl, 2-fluoro-4-methoxybenzyl, 2-fluoro-4-methylbenzyl and the like.

The term "straight-chain or branched C" as used₁-C₆Alkylcarbonyl "means any group such as methylcarbonyl, ethylcarbonyl, n-butylcarbonyl, isopropylcarbonyl, and the like.

The term "straight-chain or branched C" as used₁-C₆Alkylaminocarbonyl "means any group such as methylaminocarbonyl, ethylaminocarbonyl, n-butylaminocarbonyl, isopropylaminocarbonyl, and the like.

The term "straight-chain or branched C" as used₁-C₆Alkylsulfonyl "means any group such as methylsulfonyl, ethylsulfonyl, n-butylsulfonyl, isopropylsulfonyl, and the like.

The term "straight-chain or branched C" as used₁-C₆Alkoxycarbonyl "means any group such as ethoxycarbonyl, n-butoxycarbonyl, isopropoxycarbonyl, n-propoxycarbonyl, and the like.

The term "heterocyclyl" as used herein means a saturated or unsaturated 5-7 membered carbocyclic ring in which 1-3 carbon atoms may be replaced by heteroatoms, such as nitrogen, oxygen, sulfur, wherein the heteroatoms may be directly connected to each other, nitrogen and sulfur may optionally be oxidized, nitrogen may optionally be quaternized or carry an R4 substituent. Non-limiting examples of heterocyclyl groups are, for example, piperidinyl, piperazinyl, oxazolidinyl, 4-methylpiperazinyl, 4-ethylpiperazinyl, and the like.

The term "aryl" as used herein means a carbocyclic hydrocarbon with 1 to 2 ring portions fused or connected to each other by single bonds, wherein at least 1 ring is aromatic. Examples of aryl groups of the present invention are, for example, phenyl, biphenyl, alpha-or beta-naphthyl, dihydronaphthyl, and the like.

The term "leaving group" means a group that can be substituted with another group in a substitution reaction. Such leaving groups are well known in the art, examples of which include, but are not limited to, halides (fluoride, chloride, bromide, and iodide), azido, sulfonate (e.g., optionally substituted C)₁-C₆Alkanesulfonic acid groups, e.g. methanesulfonic acid and trifluoromethanesulfonic acid groups, or optionally substituted C₇-C₁₂Alkylbenzenesulfonic acid groups such as p-toluenesulfonyl group), succinimide-N-oxide, p-nitrophenoxy, pentafluorophenoxy, tetrafluorophenoxy, carboxylic acid groups, aminocarboxylic acid groups (carbamate groups), and alkoxycarboxylic acid groups (carbonate groups). For substitution on saturated carbons, halide and sulfonate groups are preferred leaving groups. For substitution on the carbonyl carbon, for example, a halide, succinimide-N-oxide, p-nitrophenoxy, pentafluorophenoxy, tetrafluorophenoxy, carboxylic acid group, or alkoxycarboxylic acid group (carbonic acid group) may be used as a leaving group. The term "leaving group" also means a group that can be removed by an elimination reaction (e.g., an electronic cascade or spiro ring reaction). In this case, for example, a halide, a sulfonic acid group, an azide group, an aminocarboxylic acid group (carbamate group), or an alkoxycarboxylic acid group (carbonate group) may be used as the leaving group.

The term "nitrogen protecting group" refers to a group having a nitrogen atom that forms carbamates, amides, cyclic imines, N-alkyl and N-aryl amines. Such Protecting groups are well known in the art (see, e.g., Green T.W., Wuts P.G.M.; protective groups in organic synthesis "). Non-limiting examples of carbamate protecting groups are, for example, methyl and ethyl carbamates, 9-fluorenylmethyl carbamate (Fmoc), 2,2, 2-trichloroethyl carbamate (Troc), t-butyl carbamate (BOC), vinyl carbamate (Voc), allyl carbamate (Alloc), benzyl carbamate (Cbz), p-nitrobenzyl, and the like. Non-limiting examples of amides are, for example, N-trichloroacetamide, N-Trifluoroacetamide (TFA), and the like. Non-limiting examples of cyclic imine protecting groups are, for example, N-phthalimide, N-dithiasuccinimidyl (Dts), and the like. Non-limiting examples of N-alkyl and N-aryl protecting groups are, for example, N-allylamine, N-benzylamine, and the like.

The term "hydroxyl protecting group" refers to a group bearing an oxygen atom to form an ether, ester, cyclic acetal, or ketal. Such Protecting groups are well known in the art (see, e.g., Green t.w., Wuts p.g.m.; protective groups inorganic synthesis). Non-limiting examples of ether protecting groups are, for example, alkyl ethers and benzyl ethers such as methoxymethyl ether (MOM-OR), tetrahydropyranyl ether (THP-OR), Allyl ether (Allyl-OR), benzyl ether (Bn-OR), triphenylmethyl ether (Tr-OR), and the like; OR silyl ethers such as trimethylsilyl ether (TMS-OR), t-butyldimethylsilyl ether (TBS-OR OR TBDMS-OR), t-butyldiphenylsilyl ether (TBDPS-OR), diphenylmethylsilyl ether (DPMS-OR), etc. Non-limiting examples of ester protecting groups are, for example, trifluoroacetate, benzoate (Bz-OR), and carbonates, such as ethyl carbonate and the like. Non-limiting examples of cyclic acetal or ketal protecting groups are, for example, methylene acetal, ethylene acetal, methoxymethylene acetal, and the like.

The term "active ester" means a functional group in which the alkoxy group of the ester moiety is a good leaving group. Examples of such alkoxy groups include, but are not limited to, succinimide-N-oxide, p-nitrophenyloxide, pentafluorobenzene oxide, tetrafluorobenzene oxide, 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, and groups having comparable leaving ability. Unsubstituted alkyl-based alkoxy groups such as methoxy, ethoxy, isopropoxy, and tert-butoxy are not acceptably good leaving groups, and therefore methyl, ethyl, isopropyl, and tert-butyl esters are not considered active esters.

The term "compound of formula (Ia)" or "compound of formula (Ib)" used means a compound as shown below:

wherein R1, R2 and R3 are as defined above.

Pharmaceutically acceptable salts of the compounds of formula (I) also include salts with inorganic or organic bases, for example hydroxides, carbonates or bicarbonates, acyclic or cyclic amines of alkali metals or alkaline earth metals, especially sodium, potassium, calcium, ammonium or magnesium.

If a stereogenic center or another isomeric center form is present in a compound of the invention, all such isomeric or isomeric forms, including enantiomers and diastereomers, are encompassed herein. Compounds containing stereogenic centers may be used in the form of a racemic mixture, an enantiomerically enriched mixture, or the racemic mixture may be separated using known techniques, and the individual enantiomers may be used individually. In the case of compounds having unsaturated carbon-carbon double bonds, all of the cis (Z) and trans (E) isomers are included within the scope of the present invention.

When a compound may exist in tautomeric forms, each form is considered to be included within the scope of the present invention, whether existing in equilibrium or existing predominantly in one form.

Preferred compounds of formula (I) are compounds of formula (Ia) or (Ib):

wherein R1 is fluorine or NR4R5 wherein one of R4 or R5 is hydrogen and the other is hydrogen or an optionally substituted group selected from straight or branched chain C₁-C₆Alkyl, NR7R8-C₁-C₆Alkyl, R6O-C₁-C₆Alkyl, R7R8N-C₁-C₆Alkylcarbonyl, R6O-C₁-C₆Alkylcarbonyl, R7R8N-C₁-C₆Alkylaminocarbonyl radical, R6O-C₁-C₆Alkylaminocarbonyl radical, R7R8N-C₁-C₆Alkoxycarbonyl and R6O-C₁-C₆An alkoxycarbonyl group.

More preferred compounds of formula (I) are compounds of formula (Ia):

wherein R1 is fluorine or NR4R5, wherein R4 and R5 are as defined above, an

R2 is COR9, wherein R9 is as defined above.

Specific non-limiting preferred compounds (compounds), optionally in the form of pharmaceutically acceptable salts, of the present invention are as follows:

1) (8S,10S) -8-acetyl-1-fluoro-6, 8, 11-trihydroxy-10- { [ (1S,3R,4aS,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione;

2) (8S,10S) -1-fluoro-6, 8, 11-trihydroxy-8- (hydroxyacetyl) -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione;

3) (8S,10S) -1-amino-6, 8, 11-trihydroxy-8- (hydroxyacetyl) -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione;

4) (8S,10S) -8-acetyl-1-amino-6, 8, 11-trihydroxy-10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione;

5) (8S,10S) -8-acetyl-6, 8, 11-trihydroxy-1- [ (2-hydroxyethyl) amino ] -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione;

6) (8S,10S) -6,8, 11-trihydroxy-8- (hydroxyacetyl) -1- [ (2-hydroxyethyl) amino ] -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione;

7) (8S,10S) -8-acetyl-1- [ (2-aminoethyl) amino ] -6,8, 11-trihydroxy-10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione; and

8) (8S,10S) -1- [ (2-aminoethyl) amino ] -6,8, 11-trihydroxy-8- (hydroxyacetyl) -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione.

For any particular compound of formula (I) of the invention, optionally in the form of a pharmaceutically acceptable salt, see experimental section and claims.

The present invention also provides a process for the preparation of the compounds of formula (I) as defined above, using the reaction routes and synthetic schemes described below, employing techniques available in the art and readily available starting materials. The preparation of certain embodiments of the present invention is described in the following examples, but one of ordinary skill in the art will recognize that the described preparation can be readily adapted to prepare other embodiments of the present invention. For example, the synthesis of the non-exemplified compounds of the invention can be carried out by modifications that will be apparent to those skilled in the art, for example, by appropriately protecting interfering groups, by modifying other suitable reagents known in the art, or by carrying out routine modifications of reaction conditions. Alternatively, other reactions mentioned herein or known in the art will be considered suitable for preparing other compounds of the invention.

The compounds of formula (I) are prepared according to 5 alternative routes a-E outlined in scheme 1 below; further, outlined in scheme 1 are also the intermediate compounds of formula (V) according to pathway F and the starting materials of formula (II) according to pathway G.

Scheme 1

Wherein "Pathway" represents a Pathway

Route A

Compounds of formula (I) wherein R1 and R3 are as defined above and R2 is COR9 wherein R9 is OR6 OR NR7R8 wherein R6, R7 and R8 are as defined above may be prepared as outlined in scheme 2 below.

Scheme 2

Thus, the method of the invention comprises the following steps:

A1) reacting a compound of the formula (II),

wherein R10 is R1 or a group of formula NR19R20 wherein R19 and R20 are independently a suitable nitrogen protecting group, or one of R19 or R20 is hydrogen and the other is a suitable nitrogen protecting group and R1 is as defined above,

with a compound of the formula (IIa),

wherein R3 is as defined above and X and Y, which are the same or different, are leaving groups, preferably halogen;

A2) the compound of formula (III) obtained is reacted,

wherein R3 and R10 are as defined above,

reacting with ethyl orthoformate and bromine, and then adding HBr;

A3) the compound of formula (IV) obtained is reacted,

wherein R3 and R10 are as defined above,

reacting with a formylating agent;

A4) oxidation to give the compound of the formula (V),

wherein R10 and R3 are as defined above;

A5) the compound of formula (VI) obtained is reacted,

wherein R3 and R10 are as defined above,

with a compound of formula (VIa) or (VIb),

R6-OH(VIa)；R7R8NH(VIb)

wherein R6, R7 and R8 are as defined above;

a6a) reacting the compound of formula (VII) obtained,

wherein R3, R10 and R9 are as defined above,

firstly, reacting with DMDO;

a6b) and then treating the resulting compound of the formula (XX) with cyanuric chloride or an iron (II) salt,

wherein R3 and R10 are as defined above and R2 is COR9, wherein R9 is as defined above and finally, if desired, the nitrogen and/or hydroxyl protecting groups are removed to give the compound of formula (I),

wherein R1 and R3 are as defined above, and R2 is COR9, wherein R9 is as defined above;

optionally converting the first compound of formula (I) to a second compound of formula (I) by known chemical reactions; and/or converting such a compound of formula (I) into a pharmaceutically acceptable salt thereof or converting a salt into the free compound of formula (I), as desired.

Route B

The compounds of formula (I) are prepared as outlined in scheme 3 below, wherein R1 and R3 are as defined above, and R2 is ethyl or COCH₃。

Scheme 3

Thus, the method of the invention comprises the following steps:

B1) reacting a compound of formula (III) as defined above with a hydrazine derivative of formula (IIIa),

R11-NH-NH₂(IIIa)

wherein R11 is aryl, preferably phenyl, 4-methylphenyl or 4-halophenyl, and then reducing the hydrazide;

B2) the compound of formula (VIII) obtained is reacted,

wherein R3 and R10 are as defined above, under the same conditions reported in the above-mentioned steps A6a) and A6b),

to obtain the compound of the formula (I),

wherein R1 and R3 are as defined above, and R2 is ethyl;

or

B3) The compound of formula (III) as defined above is reacted under the same conditions as reported in the above-mentioned steps A6a) and A6b),

to obtain the compound of the formula (I),

wherein R1 and R3 are as defined above, and R2 is COCH_3；

Route C

The compounds of formula (I) may be prepared as outlined in scheme 4 below, wherein R1 and R3 are as defined above, and R2 is selected from linear or branched C₃-C₆Alkyl, NR7R8-C₃-C₆Alkyl, R6O-C₃-C₆Alkyl and COR9, wherein R9 is straight or branched chain C₂-C₄Alkyl, NR7R8-C₂-C₄Alkyl or R6O-C₂-C₄Alkyl, wherein R6, R7, and R8 are as defined above.

Scheme 4

Thus, the method of the invention comprises the following steps:

C1) reacting a compound of formula (III) as defined above with a compound of formula (IIIb),

R12-X (IIIb)

wherein R12 is selected from straight or branched chain C₁-C₄Alkyl, NR7R8-C₁-C₄Alkyl and R6O-C₁-C₄Alkyl, and X is a leaving group, preferably halogen;

C2) (ii) reacting the compound of formula (IX) obtained,

wherein R3, R10 and R12 are as defined above,

under the same conditions reported in the above steps A6a) and A6b),

to obtain the compound of the formula (I),

wherein R1 and R3 are as defined above, and R2 is COR9, wherein R9 is a straight or branched chain C₂-C₄Alkyl, NR7R8-C₂-C₄Alkyl or R6O-C₂-C₄Alkyl, wherein R6, R7, and R8 are as defined above;

or, optionally

C3) Reacting a compound of formula (IX) as defined above under the same conditions as reported in step B1 above;

C4) the compound of formula (X) obtained is reacted,

wherein R3, R10 and R12 are as defined above,

under the same conditions reported in the above steps A6a) and A6b),

to obtain the compound of the formula (I),

wherein R1 and R3 are as defined above, and R2 is selected from straight or branched C₃-C₆Alkyl, NR7R8-C₃-C₆Alkyl and R6O-C₃-C₆A group of alkyl groups, wherein R6, R7 and R8 are as defined above;

Route D

Compounds of formula (I) may be prepared as outlined in scheme 5 below, wherein R1 and R3 are as defined above, and R2 is CH₂-CH₂NR7R8、CH₂-CH₂OR6 OR COR9, wherein R9 is-CH₂NR7R8 or-CH₂OR6, wherein R6, R7 and R8 are as defined above.

Scheme 5

Thus, the method of the invention comprises the following steps:

D1) reacting a compound of formula (IV) as defined above, wherein the carbonyl function may optionally be activated to a phenylhydrazone derivative, with a compound of formula (IVa) or (IVb),

HN-R7R8(IVa)，HOR6(IVb)

wherein R6, R7 and R8 are as defined above and wherein the OH group may optionally be activated to, for example, a tosyl or mesyl derivative and then the hydrazone functionality is removed by hydrolysis if present;

D2) reacting the resulting compound of formula (XI) or (XIa),

wherein R3, R6, R7, R8 and R10 are as defined above

Under the same conditions reported in the above steps A6a) and A6b),

to obtain the compound of the formula (I),

wherein R1 and R3 are as defined above, and R2 is COR9, wherein R9 is-CH₂-NR7R8 or-CH₂-OR6, wherein R6, R7 and R8 are as defined above;

or, optionally

D3) Reacting the resulting compound of formula (XI) or (XIa) as defined above under the same conditions as reported in step B1 above;

D4) the compound of formula (XII) or (XIIa) obtained is reacted with a compound of formula (XII) or (XIIa),

wherein R3, R6, R7, R8 and R10 are as defined above

Under the same conditions reported in the above steps A6a) and A6b),

to obtain the compound of the formula (I),

wherein R1 and R3 are as defined above, and R2 is-CH₂-CH₂-NR7R8 or-CH₂-CH₂-OR6, wherein R6, R7 and R8 are as defined above;

Route E

Compounds of formula (I) may be prepared as outlined in scheme 6 below, wherein R1 and R3 are as defined above, and R2 is CH₂OH or COR9 wherein R9 is CH₂OH。

Scheme 6

Thus, the method of the invention comprises the following steps:

E1) reacting the compound of formula (V) as defined above under the same conditions reported in the above-mentioned steps A6a) and A6b),

to obtain the compound of the formula (I),

wherein R1 and R3 are as defined above, and R2 is COR9, wherein R9 is CH₂OH；

Or, optionally

E2) Reacting a compound of formula (V) as defined above under the same conditions as reported in step B1 above;

E3) the compound of formula (XIII) obtained,

wherein R10 and R3 are as defined above

Under the same conditions reported in the above steps A6a) and A6b),

to obtain the compound of the formula (I),

wherein R1, R3 are as defined above, and R2 is CH₂OH；

According to step A1, the reaction of the compound of formula (II) with the compound of formula (IIa) is carried out in an organic solvent, preferably DMF, at room temperature according to well known methods reported in the art (see, for example, WO91/09046).

According to step a2), the reaction of the compound of formula (III) with ethyl orthoformate and bromine, then with HBr, is carried out in 2 steps according to well known methods reported in the art (see, for example, volume 17 of Doxorubicin Anticancer Antibiotics, 1981, p.168; arcamone et al j.med.chem.1974,17, p.335).

According to step A3), the reaction to obtain the compound of formula (V) is carried out according to well known methods reported in the art (see, for example, Doxorubicin Anticancer Antibiotics Vol.17, 1981, p.168; US 3803124). An example, not limiting to the present process, is the reaction of a compound of formula (IV) with sodium formate. The reaction is carried out in CH₃CN or acetone or mixtures thereof at about 20 c to reflux temperature for a period of about 30 minutes to about 24 hours.

According to step A4), the oxidation of the compound of formula (V) uses an oxidizing agent, preferablyNaIO₄The process is carried out. The reaction is carried out in MeOH or water or mixtures thereof at about 20 ℃ to reflux temperature for about 30 minutes to about 24 hours.

According to step A5), the coupling reaction of a compound of formula (VI) with a compound of formula (VIa) or (VIb) is carried out according to methods well known in the art (for general coupling reagents see, for example, Amino Acids, peptides and Proteins in Organic Chemistry: Building Blocks, Catalysis and coupling Chemistry, Vol.3; andrew B.Hughes, Ayman El-Faham, Fernando Albericio, 2010). An example, not limiting to the present process, is the reaction of a compound of formula (VI) with a compound of formula (VIa) in the presence of a condensing agent such as DCC or EDC. The reaction is carried out in an organic solvent, preferably DMF, at about 20 deg.C to reflux temperature for about 30 minutes to about 24 hours.

According to steps A6a) and 6Ab), the compound of formula (VII) is first reacted with DMDO according to methods well known in the art, and the resulting compound of formula (XX) is then reacted with cyanuric chloride or with an iron (II) salt (see, for example, GB 2296495A; WO 2012073217; WO 9802446).

If necessary, the nitrogen and/or hydroxyl protecting Groups are removed according to well known methods reported in the art (see, e.g., Protective Groups in Organic Synthesis; Theodora W.Green, Peter G.M.Wuts, 4 th edition).

According to step B1, the reaction of the compound of formula (III) with the compound of formula (IIIa) is carried out in an organic solvent, preferably MeOH, at a temperature of about-10 ℃ to about 50 ℃ for about 30 minutes to about 96 hours. Subsequent reduction of the hydrazide derivative to give the compound of formula (VIII) Using NaBH₄Or NaBH₃CN is carried out in the presence of camphorsulfonic acid. The reaction is carried out in an organic solvent, preferably MeOH, at a temperature of about 20 deg.C to reflux for a period of about 30 minutes to about 5 hours (see also Doxorubicin Antibiotics volume 17, 1981, p.165).

According to steps B2 and B3), the reactions were carried out as described in steps A6a) and A6B), respectively, above.

According to step C1), the reaction of the compound of formula (III) with the compound of formula (IIIb) is carried out in an organic solvent, preferably DMF, according to well known methods reported in the art (see, e.g., Smith t.h., Fujiwara a.n., henryd.w.; med chem.1979,22, p.40).

According to step C2), the reaction of the compound of formula (IX) is carried out as described in steps A6a) and A6b) above.

According to step C3), the reaction of the compound of formula (IX) is carried out as described in step B1 above.

According to step C4), the reaction of the compound of formula (X) is carried out as described in steps A6a) and A6b) above.

According to step D1, the reaction of the compound of formula (IV) with the compound of formula (IVa) or (IVb) is optionally carried out in the presence of a base, preferably diethylamine. The reaction is carried out in an organic solvent, preferably acetone, at a temperature of about 20 deg.C to reflux temperature for a period of about 30 minutes to about 24 hours (see, US 4133877).

Such as Baker, t.s.; exley, d.; steroids 1977,29, p.429; sugimoto, k.; sunakawa, n.; ohki, s.; the hydrolysis conditions reported in Chem Pharm Bull 1966,14, p.147 remove the hydrazone function.

The reaction of the compound of formula (XI) or (XIa) according to step D2) is carried out as described in steps A6a) and A6b) above.

According to step D3), the reaction of the compound of formula (XI) or (XIa) is carried out as described in step B1 above.

According to step D4), the reaction of the compound of formula (XII) or (XIIa) is carried out as described in steps A6a) and A6b) above.

According to step E1), the reaction of the compound of formula (V) is carried out as described in steps A6a) and A6b) above.

According to step E2), the reaction of the compound of formula (V) is carried out as described above in step B1.

According to step E3), the reaction of the compound of formula (XIII) is carried out as described in steps A6a) and A6b) above.

Route F

Alternatively, the intermediate compounds of formula (V) as defined above may be prepared according to scheme 7 below.

Scheme 7

Thus, the method of the invention comprises the following steps:

F1) reacting a compound of formula (XIV),

wherein R10 is as defined above, with bromine and potassium acetate;

F2) the compound of formula (XV) obtained,

wherein R10 is as defined above,

with a saccharide of formula (XVa),

wherein R13 and R14 are independently hydrogen or a suitable nitrogen and/or hydroxyl protecting group, for example trifluoroacetyl or benzyl;

F3) the compound of formula (XVI) obtained,

wherein R10, R13 and R14 are as defined above,

reaction with ethyl orthoformate and pyridinium p-toluenesulfonate (PPTS);

F4) the compound of formula (XVII) obtained,

wherein R13 is hydrogen and R10 and R14 are as defined above, with a compound of formula (IIa) as defined above;

F5) deprotecting the resulting compound of formula (XVIII),

wherein R3, R10 and R14 are as defined above, to give a compound of formula (V) as defined above.

According to step F1), the reaction is carried out in an organic solvent, preferably acetone or dioxane, at a temperature of 20 ℃ to reflux for a time of about 30 minutes to about 24 hours.

According to step F2, the glycosylation of the compound of formula (XV) is carried out in the presence of silver triflate according to well known methods reported in the art (see, for example, GB 2225781; GB 2215332A).

The reaction to obtain the compound of formula (XVII) is carried out according to step F3), and the compound of formula (XVI) is reacted with ethyl orthoformate and PPTS. The reaction is carried out in an organic solvent, preferably DCM, at a temperature of 0 ℃ to reflux for a period of about 30 minutes to about 24 hours.

According to step F4), the reaction was carried out as described above in step A1.

According to step F5), the hydroxyl protecting Groups are removed according to well known methods reported in the art (see, for example, Protective Groups in Organic Synthesis; theodora w.greenen, Peter g.m.wuts, 4 th edition).

Route G

Compounds of formula (II) as defined above, wherein R10 is as defined above, except NH, are prepared according to scheme 8 below₂And a halogen.

Scheme 8

Thus, the method of the invention comprises the following steps:

G1) reacting a compound of formula (XIX),

wherein R15 and R16 are independently hydrogen or a suitable hydroxyl protecting group, such as trifluoroacetyl, 9-fluorenylmethyl, di-t-butylmethylsilyl, t-butyldiphenylsilyl, or diphenylmethylsilyl, R20 is R10, wherein R10 is NH-R19, wherein R19 is independently hydrogen or a suitable nitrogen protecting group, and Z is oxygen or a suitable carbonyl protecting group, such as an acetal or ketal, preferably 1-3-dioxane or 1-3-dioxolane,

i) with a compound of formula (XIXa), (XIXb), (XIXc) or (XIXd),

wherein X is a leaving group, preferably halogen; r17 and R18 are the same or different and are independently hydrogen, halogen, straight or branched chain C₁-C₃Alkyl or C₁-C₃An alkoxy group; a is straight or branched chain C₁-C₆An alkyl group; and R6, R7 and R8 are as defined above, if present, after removal of the protecting group, to give the corresponding compound of formula (XIV) as defined above, wherein R10 is a group R1 of formula NR4R5, wherein R4 and R5 are independently hydrogen, mono-substituted-benzyl, di-substituted-benzyl or optionally substituted groups selected from linear or branched C₁-C₆Alkyl, NR7R8-C₁-C₆Alkyl and R6O-C₁-C₆Alkyl, but none of them is hydrogen;

or

ii) with a compound of formula (XIXe) or (XIXf),

wherein Y' is OH or a leaving group, preferably chlorine, and A, R6, R7 and R8 are as defined above, and if present, after removal of the protecting groups, the corresponding compounds of formula (XIV) as defined above are obtained, wherein R10 is a group R1 of formula NR4R5, wherein one of R4 or R5 is hydrogen and the other is a group R7R8N-C₁-C₆Alkylcarbonyl or R6O-C₁-C₆An alkylcarbonyl group;

or

iii) with a compound of formula (XIXg) or (XIXh),

wherein A, R6, R7 and R8 are as defined above, if present, after removal of the protecting groups, the corresponding compounds of formula (XIV) as defined above are obtained, wherein R10 is a group R1 of formula NR4R5, wherein one of R4 or R5 is hydrogen and the other is a group R7R8N-C₁-C₆Alkylaminocarbonyl or R6O-C₁-C₆An alkylamino carbonyl group;

or

iv) with a compound of formula (XIXi) or (XIXm),

wherein Y is OH or a leaving group, preferably chlorine, and A, R6, R7 and R8 are as defined above, if present, after removal of the protecting groups, to give the corresponding compounds of formula (XIV) as defined above, wherein R10 is a group R1 of formula NR4R5, wherein one of R4 or R5 is hydrogen and the other is a group R7R8N-C₁-C₆Alkylsulfonyl or R6O-C₁-C₆An alkylsulfonyl group;

or

v) with a compound of formula (XIXn) or (XIXo),

wherein Y', A, R6, R7 and R8 are as defined above, if present, after removal of the protecting groups, to give the corresponding compounds of formula (XIV) as defined above, wherein R10 is a group R1 of formula NR4R5, wherein one of R4 or R5 is hydrogen and the other is a group R7R8N-C₁-C₆Alkoxycarbonyl or R6O-C₁-C₆An alkoxycarbonyl group;

or

vi) with a compound of formula (XIXp),

wherein W is CH or N and R4 and X are as defined above, if present, after removal of the protecting groups, give the corresponding compounds of formula (XIV) as defined above, wherein R10 is a group R1 of formula NR4R5, wherein R4 or R5 together with the N atom to which they are bound form a 6-membered heterocyclic group substituted by R4;

or

G2) Reacting a compound of formula (XIX),

wherein R15, R16 and Z are as defined above, and R20 is a suitable leaving group, such as mesylate, tosylate or 4-fluoro-benzenesulfonate,

with a compound of formula (XIXq),

wherein R4 and R5 are independently hydrogen, mono-substituted-benzyl, di-substituted-benzyl or an optionally substituted group selected from linear or branched C₁-C₆Alkyl, NR7R8-C₁-C₆Alkyl and R6O-C₁-C₆An alkyl group; or R4 and R5 together with the bound N atom form a substituted heterocyclyl;

if present, after removal of the protecting group, the corresponding compound of formula (XIV) as defined above is obtained, wherein R10 is a group R1 of formula NR4R5, wherein R4 or R5 are as defined above;

G3) the compound of formula (XIV) obtained,

wherein R10 is as defined in step G1 or G2,

reaction with a compound of formula (XVa) as defined above under the same conditions as reported in step F2 above, to give a compound of formula (II) wherein R10 is as defined above.

According to step G1i), the reaction is carried out according to well known methods reported in the art (see, e.g., Ngu, k.; patel, D.V.tetrahedron Lett 1997,38(6), pp.973-976). As an example and not by way of limitation, the reaction is carried out in DCM at a temperature of 20 ℃ to reflux for a period of from 30 minutes to about 24 hours.

According to step G1ii, the coupling reaction between a compound of formula (XIX) and a compound of formula (XIXe) or (XIXf) is carried out according to methods well known in the art (for general coupling reagents see, for example, Amino Acids, Peptides and Proteins in Organic Chemistry: Building Blocks, Catalysis and chemical Chemistry, volume 3; andrew B.Hughes, Ayman El-Faham, Fernando Albericio, 2010). By way of example, but not limitation, in the present process, is the reaction in the presence of condensing agents such as DCC, EDC, sodium formate. The reaction is carried out in an organic solvent. Preferably in DMF at a temperature of 20 ℃ to reflux for a period of 30 minutes to about 24 hours.

According to step G1iii), the reaction is carried out according to well known methods reported in the art (see, e.g., Gopalsamy A. et al Bioorg Med Chem Lett 2005,15(6), pp.1591-1594; lee Y.S. et al Bioorg Med Chem Lett 2004,314, (13), pp.3379-3384). As an example and not by way of limitation, the reaction is carried out in pyridine, DCM at a temperature of 20 ℃ to reflux for a period of 30 minutes to about 24 hours.

According to step G1iv), the reaction is carried out according to well known methods reported in the art (see, for example, Filimonov S.J heterocyclic Chem 2006,43, pp.663-671; rockway, t.w. et al; bioorg MedChem Lett 2006,16, p.3833).

According to step G1v), the reaction is carried out according to well known methods reported in the art (see, e.g., Fukuoka s. et al j. chem. soc. chem. commu. 1984,6, p.399).

According to step G1vi), the reaction is carried out according to well known methods reported in the art (see, e.g., ismalov, v.et al; russ J Org Chem,2004,40(2), pp.284-285; mewshaw r.e. et al; bioorgMed Chem Lett 1998,8(19), pp.2675-2680;mishani e. et al; tetrahedron Lett 1996,37(3), pp.319-322. As an example, without limitation to the present process, the reaction is in DMSO, DCM, MeOH, or mixtures thereof, optionally in a base or Lewis acid (e.g., Al)₂Cl₃) At a temperature of from 20 ℃ to reflux for a time of from 30 minutes to about 24 hours.

According to step G2), the reaction is carried out as described in patent application GB 2215322. As an example, not limiting to the process, the reaction is in CH₃CN, THF or DMF, optionally in the presence of a base, at a temperature of 20 ℃ to reflux for a period of 1 to 72 hours. According to step G3), the reaction is carried out as described above in step F2.

The compounds of formula (II) in which R10 is NH can be prepared as described in patent application EP288268₂。

Compounds of formula (II) wherein R10 is halogen can be prepared as described in patent application WO9802446 and Gary W et al j.o.c 1987,52, p.713.

The compounds of formula (XIX) can be prepared as described in patent application EP 288268.

The compounds of formula (IIa), (IIIa), (IIIb), (IVa), (IVb), (XVa), (XiXa) - (XIXp) are commercially available or can be prepared using known methods.

In summary, it will be apparent to those skilled in the art that when compounds of formula (I) are prepared according to any one of the variants of any of the aforementioned methods, optional functional groups which may cause undesirable side reactions among the starting materials or intermediates thereof need to be suitably protected according to conventional techniques. Likewise, these latter can be converted into the free deprotected compound according to known methods.

It will be readily understood that if the compounds of formula (I) prepared according to the process described above are obtained as a mixture of isomers, their separation into the individual isomers of formula (I) using conventional techniques is also included within the scope of the present invention.

Pharmacology of

The novel morpholinyl anthracycline derivatives of the invention are useful as antitumor agents.

Accordingly, a mammal, such as a human or an animal, may be treated by a method comprising administering thereto a pharmaceutically effective amount of a morpholinyl anthracycline derivative of formula (I).

In this way, the condition of the human or animal can be improved or ameliorated.

Evaluation of cytotoxicity of compounds of formula (I) was evaluated as follows.

In vitro cell proliferation assay

Human cancer cell lines were seeded in white 384-well plates (1250 cells/well) in complete medium (RPMI1640 or EMEM plus 10% fetal bovine serum) and 24h after seeding, treated with compounds dissolved in 0.1% DMSO. At 37 ℃ and 5% CO₂The cells were incubated and after 72h, the plates were treated using the Cell Titer-Glo test (Promega) according to the manufacturer's instructions.

CellTiter-Glo is an indicator of metabolically active cells based on a homologous method for quantifying the presence of ATP. ATP was quantified using a system based on luciferase and D-luciferin, which resulted in light production. Briefly, 25. mu.L/well of reagent solution was added to each well and after shaking for 5 minutes, the microplate was read with a luminometer. The luminescent signal is proportional to the number of viable cells present in the culture.

The antiproliferative activity of a compound was reported as half maximal Inhibitory Concentration (IC) by generating a dose-response curve by insertion of a sigmoidal function of 8 concentration points₅₀)。

Representative compounds of formula (I) of the present invention are determined in the specific in vitro cell proliferation assay described above.

Cell Line IC₅₀nM

As can be appreciated by the person skilled in the art, all these representative compounds are thus particularly advantageous in antitumor therapy.

The compounds of the invention may be administered as a single agent, or, in combination with known anti-cancer therapies such as radiation therapy or chemotherapy regimens, in combination with cytostatic or cytotoxic drugs, antibiotic-type drugs, alkylating agents, antimetabolites, hormonal drugs, immunological drugs, interferon-type drugs, cyclooxygenase inhibitors (e.g., COX-2 inhibitors), matrix metalloproteinase inhibitors, telomerase inhibitors, tyrosine kinase inhibitors, anti-growth factor receptor drugs, anti-HER drugs, anti-EGFR drugs, anti-angiogenic agents (e.g., angiogenesis inhibitors), farnesyl transferase inhibitors, Ras-Raf signal transduction pathway inhibitors, cell cycle inhibitors, other CDKs inhibitors, tubulin binding agents, topoisomerase I inhibitors, topoisomerase II inhibitors, and the like.

If formulated as a fixed dose, such combination products are employed with the compounds of the present invention within the dosage ranges described below and with the other pharmaceutically active agents described within the approved dosage ranges.

When a combined preparation is inappropriate, the compound of formula (I) may be used in sequence with known anticancer agents.

The compounds of formula (I) of the present invention, which are suitable for administration to mammals, e.g. to humans, may be administered by the usual route, the dosage level depending on the age, weight, condition and route of administration of the patient.

For example, a suitable dose of a compound of formula (I) adjusted for oral administration may be about 1 to about 300mg per dose, 1 to 5 times daily. The compounds of the invention may be administered in a variety of dosage forms, for example, orally, in the form of tablets, capsules, sugar or film coated tablets, liquid solutions or suspensions; rectal administration, in the form of suppositories; parenteral administration, for example, subcutaneously, intramuscularly, or by intravenous and/or intrathecal and/or intraspinal injection or infusion.

The invention also includes pharmaceutical compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable excipient, which may be a carrier or diluent.

Pharmaceutical compositions containing the compounds of the invention are generally prepared according to conventional methods and are administered in suitable pharmaceutical forms. For example, solid oral forms can contain, together with the active compound, diluents, e.g., lactose, glucose, sucrose, cellulose, corn starch, or potato starch; lubricants, for example, silica, talc, stearic acid, magnesium or calcium stearate, and/or polyethylene glycol; binding agents, for example, starch, gum arabic, gelatin, methylcellulose, carboxymethylcellulose or polyvinylpyrrolidone; disintegrating agents, for example, starch, alginic acid, alginates or sodium starch glycolate; an effervescent mixture; a dye; a sweetener; wetting agents, such as lecithin, polysorbate, lauryl sulfate; and non-toxic and pharmacologically inert substances commonly used in pharmaceutical formulations. These pharmaceutical formulations can be prepared as follows: in a known manner, for example, by mixing, granulating, tabletting, sugar-coating, or film-coating processes.

Liquid dispersions for oral administration may be, for example, syrups, emulsions and suspensions. As an example, a syrup may comprise sucrose or sucrose containing glycerol and/or mannitol and sorbitol as a carrier.

Suspensions and emulsions may contain natural gums, agar, sodium alginate, pectin, methyl cellulose, carboxymethylcellulose or polyvinyl alcohol as examples of carriers. Suspensions or solutions for intramuscular injections may contain the active compound in association with a pharmaceutically acceptable carrier, for example sterile water, olive oil, ethyl oleate, glycols such as propylene glycol, and appropriate amounts of lidocaine hydrochloride as required. Solutions for intravenous injection or infusion may contain sterile water as the carrier, or preferably they may be in the form of sterile, aqueous, isotonic saline solutions, or they may contain propylene glycol as the carrier. Suppositories may contain the active compound in combination with a pharmaceutically acceptable carrier, for example cocoa butter, polyethylene glycol, a polyoxyethylene sorbitan fatty acid ester surfactant or lecithin.

For the purpose of better illustrating the invention, the following examples are now given, without however limiting the invention in any way.

Examples

The synthetic preparation of certain compounds of formula (I) of the present invention is described in the following examples. The compounds of the invention, as prepared according to the following examples, are also prepared from¹H-NMR and/or by accurate mass data ESI (+).

¹H-NMR spectra were recorded at a constant temperature of 28 ℃ on a Varian INOVA400 chromatograph operating at 400.50MHz and equipped with a 5mm z-axis PFG indirect measurement probe: (¹H{¹⁵N-³¹P})。

Chemical shifts are referenced to residual solvent signal (DMSO-d unless otherwise stated₆: 2.50ppm for¹H) In that respect Data are reported as follows: chemical shift (), multiplicities (s ═ singlet, d ═ doublet, t ═ triplet, q ═ quartet, br.s ═ broad singlet, td ═ doublet of triplet, dd ═ doublet of doublet, ddd ═ doublet of doublet, m ═ multiplet, spt ═ heptaplex), coupling constants (J, Hz) and the number of protons.

Accurate mass data ESI (+) were obtained using a Waters Q-T Ultima mass spectrometer directly connected to the Agilent 1100micro-HPLC system (M.Colombo, F.Ricccardi-Sirtori, V.Rizzo, Rapid Commun. Mass Spectrum.2004, 18, 511-.

The following abbreviations have the following meanings in the examples below and throughout the present application. Terms have their commonly accepted meanings, if not defined.

Example 1

Step A1, step B3 (according to A6a and A6B)

(8S,10S) -8-acetyl-1-fluoro-6, 8, 11-trihydroxy-10- { [ (1S,3R,4aS,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione [ (I) ] (Compound 1)

[R1＝F,R2＝CH₃CO-,R3＝CH₃O-]

Step A1

(1S,3S) -3-acetyl-10-fluoro-3, 5, 12-trihydroxy-6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl 2,3, 6-trideoxy-3- [ (2S) -2-methoxymorpholin-4-yl ] -alpha-L-lysu-hexopyranoside [ (III) ]

(1S,3S) -3-acetyl-10-fluoro-3, 5, 12-trihydroxy-6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl 3-amino-2, 3, 6-trideoxy- α -L-liso-hexopyranoside (70.0mg,0.136mmol) [ preparation reported in WO90/09392 ]]Dissolved in dry DMF (3 mL); a solution of diisopropylethylamine (106mg,0.82mmol) in dry DMF (2mL) and a solution of (1S) -2-iodo-1- (2-iodoethoxy) -1-methoxyethane (IIa) (965mg,2.71mmol) in dry DMF (10mL) were added. The reaction mixture was stirred at room temperature in the dark for 48 hours until no starting material was detected (HPLC analysis). The reaction mixture was then diluted with DCM and washed with water. With anhydrous Na₂SO₄The organic phase was dried, the solvent evaporated in vacuo and the residue purified by flash chromatography on silica gel (230-400 mesh) (eluent: EtOH/DCM; 0.2/9.8) to give the desired product (35mg, red wax).

ESI MS:m/z 616(MH⁺)

¹H NMR(500MHz,CHCl₃-d)ppm 1.39(d,J＝6.71Hz,3H)1.78-1.85(m,2H)2.09-2.14(m,1H)2.46-2.56(m,3H)2.61(dd,J＝11.41,3.97Hz,1H)3.03(d,J＝19.04Hz,1H)3.27(dd,J＝19.10,1.77Hz,1H)3.40(s,3H)3.57(ddd,J＝11.57,5.34,3.11Hz,1H)3.70(s,1H)3.92-3.99(m,1H)4.04(q,J＝6.47Hz,1H)4.48-4.52(m,1H)4.67(s,1H)5.28-5.30(m,1H)5.56(br.s.,1H)7.54(dd,J＝10.44,8.48Hz,1H)7.83(td,J＝7.97,4.58Hz,1H)8.25(d,J＝7.69Hz,1H)13.31(s,1H)13.72(s,1H)

The following compounds were prepared by a similar method:

(1S,3S) -10-fluoro-3, 5, 12-trihydroxy-3- (hydroxyacetyl) -6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl 2,3, 6-trideoxy-3- [ (2S) -2-methoxymorpholin-4-yl ] -a-L-lysu-hexopyranoside [ (III) ]

ESI MS:m/z 632(MH⁺)

Step B3(A6a)

(1S,3S) -3-acetyl-10-fluoro-3, 5, 12-trihydroxy-6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl (3 xi) -2,3, 6-trideoxy-3- [ (2S) -2-methoxy-4-oxomorpholin-4-yl ] -a-L-threo-hexopyranoside [ (XX) ]

(1S,3S) -3-acetyl-10-fluoro-3, 5, 12-trihydroxy-6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl 2,3, 6-trideoxy-3- [ (2S) -2-methoxymorpholin-4-yl ] -a-L-lysu-hexopyranoside (28mg,0.045mmol) [ preparation as reported in step A1 ] was dissolved in DCM (3.0 mL). The solution was treated with 0.1M DMDO in acetone (0.8mL) at room temperature for 30 minutes until no starting material was detected (HPLC analysis). The reaction mixture was then concentrated to dryness in vacuo to afford the desired intermediate (red wax, 24.1 mg).

ESI MS:m/z 632(MH⁺)

¹H NMR(500MHz,CH₃CN-d₃)ppm 1.23(d,J＝6.7Hz,3H)1.96-2.00(m,1H)2.10(m,1H)2.35(s,3H)2.33-2.38(m,1H)2.56-2.64(m,2H)2.94-3.00(m,1H)3.07-3.12(m,1H)3.13-3.16(m,1H)3.23-3.29(m,1H)3.37(s,3H)3.38-3.46(m,2H)3.86-3.95(m,1H)3.99(q,J＝6.7Hz,1H)4.14(s,1H)4.22-4.29(m,1H)4.32(br.s.,1H)4.91(dd,J＝8.1,2.3Hz,1H)5.20(dd,J＝4.6,1.9Hz,1H)5.60(d,J＝3.9Hz,1H)7.62(dd,J_HH＝8.3,J_HF＝10.8Hz,1H)7.91(m,1H)8.20(d,J_HH＝7.7Hz,1H)13.26(br.s.,1H)13.69(br.s.,1H)

The following compounds were prepared by a similar method:

(1S,3S) -10-fluoro-3, 5, 12-trihydroxy-3- (hydroxyacetyl) -6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl (3 xi) -2,3, 6-trideoxy-3- [ (2S) -2-methoxy-4-oxomorpholin-4-yl ] -a-L-threo-hexopyranoside [ (XX) ]

ESI MS:m/z 648(MH⁺)

Step B3(A6B)

The title compound (Compound 1)

To the compound (1S,3S) -3-acetyl-10-fluoro-3, 5, 12-trihydroxy-6, 11-dioxo-1, 2,3,4,6, 11-hexahydronaphthacen-1-yl (3 ξ) -2,3, 6-trideoxy-3- [ (2S) -2-methoxy-4-oxomorpholin-4-yl]- α -L-threo-hexopyranoside [ (XX)](20mg,0.032mmol) in 5.0ml dry CH₃Adding K into CN solution₂CO₃(13.2mg,0.096mmol) and cyanuric chloride (11.8mg,0.064 mmol). The reaction mixture was stirred vigorously in the dark at room temperature for 20 minutes until no starting material was detected. A solution of 3-amino-1, 2-propanediol (17.5mg,0.192mmol) in water (0.84mL) was then added to the reaction mixture and the aqueous phase was extracted with DCM (4X 10 mL). With anhydrous Na₂SO₄The combined organic phases were dried, filtered and evaporated in vacuo. The crude product (AcOEt/toluene; 4/6) was purified by flash column chromatography on silica gel (230-400 mesh) to yield 7.0mg of the title compound as a red solid.

ESI MS:m/z 614(MH⁺)

¹H NMR(500MHz,CH₃CN-d₃)ppm 1.29(d,J＝6.6Hz,3H)1.68-1.73(m,1H)1.86-1.91(m,1H)2.05(dd,J＝14.8,4.3Hz,1H)2.34(s,3H)2.42-2.47(m,1H)2.67-2.81(m,2H)2.93-2.98(m,1H)3.05-3.11(m,1H)3.37(s,3H)3.42-3.47(m,1H)3.52-3.58(m,1H)3.71-3.76(m,1H)4.03(dd,J＝7.1,1.8Hz,1H)4.06-4.12(m,1H)4.26(d,J＝2.8Hz,1H)4.53(d,J＝2.8Hz,1H)4.54(s,1H)5.20(dd,J＝4.3,2.1Hz,1H)5.35(t,J＝5.5Hz,1H)7.60(dd,J_HH＝8.3,J_HF＝11.6Hz,1H)7.89(m,1H)8.19(dd,J_HH＝7.7,J_HF＝0.8Hz,1H)13.25(br.s.,1H)13.61(br.s.,1H)

Similarly, by using the appropriate starting materials, the following compounds were prepared:

(8S,10S) -1-fluoro-6, 8, 11-trihydroxy-8- (hydroxyacetyl) -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5][1,3]Oxazolo [2,3-c][1,4]Oxazin-3-yl]Oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione [ (I)](compound 2) [ R1 ═ F, R2 ═ HOCH₂CO-,R3＝CH₃O-]

ESI MS:m/z 630(MH⁺)

(8S,10S) -8-acetyl-6, 8, 11-trihydroxy-1- [ (2-hydroxyethyl) amino]-10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4, 5)][1,3]Oxazolo [2,3-c][1,4]Oxazin-3-yl]Oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione [ (I)](compound 5) [ R1 ═ HO (CH)₂)₂NH-,R2＝CH₃CO-,R3＝CH₃O-]

ESI MS:m/z 655(MH⁺)

(8S,10S) -6,8, 11-trihydroxy-8- (hydroxyacetyl) -1- [ (2-hydroxyethyl) amino]-10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4, 5)][1,3]Oxazolo [2,3-c][1,4]Oxazin-3-yl]Oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione [ (I)](compound 6) [ R1 ═ HO (CH)₂)₂NH-,R2＝HOCH₂CO-,R3＝CH₃O-]

ESI MS:m/z 671(MH⁺)

(8S,10S) -8-acetyl-1- [ (2-aminoethyl) amino]-6,8, 11-trihydroxy-10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5][1,3]Oxazolo [2,3-c][1,4]Oxazin-3-yl]Oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione [ (I)](compound 7) [ R1 ═ H₂N(CH₂)₂NH-,R2＝CH₃CO-,R3＝CH₃O-]

ESI MS:m/z 654(MH⁺)

(8S,10S) -1- [ (2-aminoethyl) amino group]-6,8, 11-Trihydroxyl-8- (Hydroxyacetyl) -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-Methylooctahydro-1H-pyrano [4',3':4,5][1,3]Oxazolo [2,3-c][1,4]Oxazin-3-yl]Oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione [ (I)](compound 8) [ R1 ═ H₂N(CH₂)₂NH-,R2＝HOCH₂CO-,R3＝CH₃O-]

ESI MS:m/z 670(MH⁺)

Example 2

Step A1, step B3 (according to A6a and A6B)

(8S,10S) -8-acetyl-1-amino-6, 8, 11-trihydroxy-10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione [ (I) ] (Compound 4)

[R1＝NH₂-,R2＝CH₃CO-,R3＝CH₃O-]

Step A1

(1S,3S) -3-acetyl-10-amino-3, 5, 12-trihydroxy-6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl 2,3, 6-trideoxy-3- [ (2S) -2-methoxymorpholin-4-yl ] -a-L-lysu-hexopyranoside [ (III) ]

(1S,3S) -3-acetyl-10-amino-3, 5, 12-trihydroxy-6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl-3-amino-2, 3, 6-trideoxy- α -L-liso-hexopyranoside (165.0mg,0.322mmol) [ preparation as reported in example 3 below ]]Dissolved in dry DMF (3.0 mL); a solution of diisopropylethylamine (221mg,1.71mmol) in dry DMF (3mL) and a solution of (1S) -2-iodo-1- (2-iodoethoxy) -1-methoxyethane (IIa) (2.0g,5.64mmol) in dry DMF (10mL) were added. The reaction mixture was left to stand in toluene at room temperature in the dark for 48 hours until no starting material was detected (HPLC analysis). The reaction mixture was then diluted with DCM and washed with water. With anhydrous Na₂SO₄The organic phase was dried, the solvent evaporated in vacuo and the residue purified by flash chromatography on silica gel (230-400 mesh) (eluent: EtOH/DCM; 0.2/9.8) to give the desired product (105.0mg, red solid).

ESI MS:m/z 613(MH⁺)

¹H NMR(500MHz,CH₃CN-d₃)ppm 1.22-1.28(m,3H)1.65-1.83(m,2H)2.30-2.36(m,4H)2.40(dd,J＝11.22,4.94Hz,3H)2.46-2.54(m,2H)2.87-2.96(m,1H)3.04-3.11(m,1H)3.32(s,3H)3.50(ddd,J＝11.34,6.51,2.83Hz,1H)3.65(br.s.,1H)3.77-3.89(m,1H)4.04(d,J＝6.51Hz,1H)4.44(dd,J＝4.63,2.41Hz,1H)5.16(d,J＝1.99Hz,1H)5.43-5.47(m,1H)7.12-7.16(m,1H)7.24(br.s.,1H)7.50-7.55(m,1H)7.58-7.61(m,1H)

The following compounds were prepared by a similar method:

(1S,3S) -10-amino-3, 5, 12-trihydroxy-3- (hydroxyacetyl) -6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl 2,3, 6-trideoxy-3- [ (2S) -2-methoxymorpholin-4-yl ] -a-L-lysu-hexopyranoside

ESI MS:m/z 629(MH⁺)

(1S,3S) -3-acetyl-3, 5, 12-trihydroxy-10- [ (2-hydroxyethyl) amino ] -6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl 3-amino-2, 3, 6-trideoxy-L-lysu-hexopyranoside

ESI MS:m/z 557(MH⁺)

(1S,3S) -3-acetyl-10- [ (2-aminoethyl) amino ] -3,5, 12-trihydroxy-6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl 3-amino-2, 3, 6-trideoxy-L-lysu-hexopyranoside

ESI MS:m/z 556(MH⁺)

Protection of

N- [ (8S,10S) -8-acetyl-6, 8, 11-trihydroxy-5, 12-dioxo-10- ({2,3, 6-trideoxy-3- [ (2S) -2-methoxymorpholin-4-yl ] -alpha-L-lysu-hexopyranosyl } oxy) -5,7,8,9,10, 12-hexahydrotetracen-1-yl ] -2,2, 2-trifluoroacetamide

Mixing (1S,3S) -3-acetyl-10-amino-3, 5, 12-trihydroxy-6, 11-dioxo-1, 2,3,4,6, 11-hexahydro-tetracen-1-yl 2,3, 6-trideoxy-3- [ (2S) -2-methoxy-morpholin-4-yl]- α -L-lisu-pyriPyranohexaside [ (III)](80.0mg,0.130mmol) was dissolved in dry DCM (11mL), trifluoroacetic anhydride (273.0mg,1.3mmol) was added and the reaction mixture was stirred in the dark at room temperature for 30 min until no starting material was detected (HPLC analysis). The reaction mixture was then diluted with DCM and saturated NaHCO₃Aqueous solution (3X 10mL) then washed with water (1X 10 mL). With anhydrous Na₂SO₄The organic phase was dried, the solvent evaporated in vacuo, the residue thus obtained was treated with MeOH (10mL) at room temperature for 15 min, then evaporated in vacuo to give the desired product (77.0mg, red wax).

ESI MS:m/z 709(MH⁺)

¹H NMR(500MHz,CH₃CN-d₃)ppm 1.25(d,J＝6.59Hz,3H)1.76(dd,J＝8.68,2.61Hz,2H)2.27-2.45(m,8H)2.52(t,J＝10.99Hz,2H)2.96-3.03(m,1H)3.08-3.15(m,1H)3.30-3.33(m,3H)3.50(ddd,J＝11.27,6.57,2.61Hz,1H)3.66(br.s.,1H)3.79-3.87(m,1H)4.05(q,J＝6.62Hz,1H)4.44(dd,J＝4.70,2.35Hz,1H)5.17(d,J＝2.27Hz,1H)5.45(s,1H)7.97(t,J＝8.15Hz,1H)8.24(d,J＝7.50Hz,1H)8.99(d,J＝8.18Hz,1H)

Step B3(A6a)

N- [ (8S,10S) -8-acetyl-6, 8, 11-trihydroxy-5, 12-dioxo-10- ({ (3 xi) -2,3, 6-trideoxy-3- [ (2S) -2-methoxy-4-oxomorpholin-4-yl ] - α -L-threo-hexopyranosyl } oxy) -5,7,8,9,10, 12-hexahydrotetracen-1-yl ] -2,2, 2-trifluoroacetamide [ (XX) ]

N- [ (8S,10S) -8-acetyl-6, 8, 11-trihydroxy-5, 12-dioxo-10- ({2,3, 6-trideoxy-3- [ (2S) -2-methoxymorpholin-4-yl ] - α -L-lysu-hexopyranosyl } oxy) -5,7,8,9,10, 12-hexahydrotetracen-1-yl ] -2,2, 2-trifluoroacetamide (72.0mg,0.102mmol) is dissolved in DCM (6.4 mL). The solution was treated with 0.1M DMDO in acetone (1.7mL) for 30 minutes at room temperature until no starting material was detected (HPLC analysis). The reaction mixture was then concentrated to dryness in vacuo to afford the desired intermediate (red wax, 73.0 mg).

ESI MS:m/z 725(MH⁺)

¹H NMR(500MHz,CH₃CN-d₃)ppm 1.23(d,J＝6.51Hz,3H)2.32-2.39(m,4H)2.57(d,J＝4.54Hz,1H)2.74(d,J＝11.65Hz,1H)2.96-3.02(m,1H)3.08-3.15(m,1H)3.25-3.45(m,7H)3.57(br.s.,1H)3.92(d,J＝12.79Hz,1H)4.04(d,J＝6.88Hz,1H)4.18(s,1H)4.21-4.28(m,1H)4.92(dd,J＝8.21,2.08Hz,1H)5.19(d,J＝2.19Hz,1H)5.61(d,J＝3.71Hz,1H)7.97(t,J＝8.10Hz,1H)8.23(d,J＝7.64Hz,1H)8.98(d,J＝8.32Hz,1H)

By a similar method, the following compounds can be prepared:

(1S,3S) -10-amino-3, 5, 12-trihydroxy-3- (hydroxyacetyl) -6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl (3 xi) -2,3, 6-trideoxy-3- [ (2S) -2-methoxy-4-oxomorpholin-4-yl ] -a-L-threo-hexopyranoside [ (XX) ]

ESI MS:m/z 645(MH⁺)

Step B3(A6B)

N- [ (8S,10S) -8-acetyl-6, 8, 11-trihydroxy-10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -5, 12-dioxo-5, 7,8,9,10, 12-hexahydro-tetracen-1-yl ] -2,2, 2-trifluoroacetamide [ (I) ]

To the compound N- [ (8S,10S) -8-acetyl-6, 8, 11-trihydroxy-5, 12-dioxo-10- ({ (3 ξ) -2,3, 6-trideoxy-3- [ (2S) -2-methoxy-4-oxomorpholin-4-yl]- α -L-threo-hexopyranosylOxy) -5,7,8,9,10, 12-hexahydrotetracen-1-yl]-2,2, 2-trifluoroacetamide [ (XX)](60.0mg,0.083mmol) in 13mL dry CH₃Adding K into CN solution₂CO₃(34.4mg,0.249mmol) and cyanuric chloride (30.6mg,0.166 mmol). The reaction mixture was stirred vigorously in the dark at room temperature for 15 minutes until no starting material was detected. A solution of 3-amino-1, 2-propanediol (45.3mg,0.5mmol) in water (0.22mL) was then added to the reaction mixture, and the aqueous phase was extracted with DCM (4X 10 mL). With anhydrous Na₂SO₄The combined organic phases were dried, filtered and evaporated in vacuo. The crude product (AcOEt/toluene; 4/6) was purified by flash column chromatography on silica gel (230-400 mesh) to yield 12.0mg of the title compound as a red solid.

ESI MS:m/z 707(MH⁺)

¹H NMR(500MHz,CH₃CN-d₃)ppm 1.29(d,J＝6.58Hz,4H)1.70(d,J＝15.21Hz,1H)1.90(d,J＝15.59Hz,2H)2.04-2.08(m,2H)2.45(d,J＝14.98Hz,1H)2.69-2.76(m,1H)2.77-2.83(m,1H)2.97(s,1H)3.08-3.14(m,2H)3.38(s,4H)3.45(d,J＝6.88Hz,2H)3.56(d,J＝5.22Hz,2H)3.74(s,1H)4.04(d,J＝1.89Hz,2H)4.09(d,J＝6.88Hz,1H)4.26(d,J＝2.72Hz,1H)4.52-4.54(m,2H)5.22(br.s.,1H)5.36(t,J＝5.60Hz,1H)7.98(t,J＝8.06Hz,1H)8.26(d,J＝7.87Hz,1H)9.00(d,J＝8.10Hz,1H)

Deprotection of the amino acid

The title compound (Compound 4)

N- [ (8S,10S) -8-acetyl-6, 8, 11-trihydroxy-10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5 ':4][1,3]Oxazolo [2,3-c][1,4]Oxazin-3-yl]Oxy } -5, 12-dioxo-5, 7,8,9,10, 12-hexahydrotetracen-1-yl]The (2, 2, 2-trifluoroacetamide intermediate (4.8mg,0,00679mmol) was cooled at 0 deg.C and 0.1N aqueous NaOH (0.5mL) was added. The reaction mixture was stirred at 0 ℃ for 15 minutes in the dark untilUntil no material was detected. Then using H₂The reaction mixture was diluted with O and extracted with DCM (4 × 5 mL). The combined organic phases were washed with saturated aqueous NaCl (1X 10mL) and anhydrous Na₂SO₄Drying, filtration and evaporation in vacuo gave 4.0mg of the title compound as a red solid.

ESI MS:m/z 611(MH⁺)

¹H NMR(500MHz,CH₃CN-d₃)ppm 1.70(dt,J＝15.06,5.82Hz,2H)1.87(dt,J＝15.17,5.54Hz,1H)2.34(s,3H)2.43(d,J＝14.41Hz,1H)2.68-2.81(m,2H)2.91-2.96(m,1H)3.07(d,J＝18.66Hz,1H)3.37(s,3H)3.44(q,J＝5.87Hz,1H)3.51-3.61(m,2H)3.74(ddd,J＝11.63,8.25,2.96Hz,1H)4.01-4.04(m,1H)4.06-4.13(m,1H)4.26(d,J＝2.66Hz,1H)4.54(d,J＝2.58Hz,1H)5.21(br.s.,1H)5.37(t,J＝5.61Hz,1H)7.16(d,J＝8.57Hz,1H)7.54(t,J＝7.89Hz,1H)7.62(d,J＝7.06Hz,1H)

(8S,10S) -1-amino-6, 8, 11-trihydroxy-8- (hydroxyacetyl) -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione (Compound 3)

[R1＝NH2-,R2＝HOCH₂CO-,R3＝CH₃O-]

ESI MS:m/z 627(MH⁺)

Example 3

Preparation of intermediates of formula (II)

G2 deprotection, protection, G3 deprotection

Synthesis of (1S,3S) -3-acetyl-10-amino-3, 5, 12-trihydroxy-6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl 3-amino-2, 3, 6-trideoxy-L-lyxo-hexopyranoside

Step G2

Synthesis of intermediate (8S,10S) -1- [ (3, 4-dimethoxybenzyl) amino ] -6,8,10, 11-tetrahydroxy-8- (2-methyl-1, 3-dioxolan-2-yl) -7,8,9, 10-tetrahydrotetracene-5, 12-dione (XIV)

To (8S,10S) -4-fluorobenzenesulfonic acid 6,8,10, 11-tetrahydroxy-8- (2-methyl-1, 3-dioxolan-2-yl) -5, 12-dioxo-5, 7,8,9,10, 12-hexahydrotetracen-1-yl ester (400mg,0.682mmol) [ preparation as reported in GB2215332]To a solution in THF (10mL) was added 3, 4-dimethoxybenzylamine (0.532mg,3.1 mmol). The solution was heated at 60 ℃ and stirred in the dark for 24 hours. The solvent was then partially removed under vacuum and the dark purple precipitate collected by filtration with THF (3mL) then Et₂O (10mL) wash. The solid was finally dried in oven under vacuum at 30 ℃ to give the title intermediate (188mg, y 48%).

Similarly, by using the appropriate amines, the following compounds were prepared:

(8S,10S) -6,8,10, 11-tetrahydroxy-1- [ (2-hydroxyethyl) amino ] -8- (2-methyl-1, 3-dioxolan-2-yl) -7,8,9, 10-tetrahydrotetracene-5, 12-dione

ESI MS:m/z 472(MH⁺)

¹H NMR(499.75MHz,DMSO-d₆)ppm 1.33(s,3H),1.82(dd,J＝14.3,4.3Hz,1H),2.20(d,J＝14.3Hz,1H),2.67(d,J＝18.7Hz,1H),3.10(d,J＝18.7Hz,1H),3.45-3.49(m,2H),3.68-3.72(m,2H),3.92-4.01(m,4H),5.00(t,J＝5.1Hz,1H),5.05-5.11(m,1H),5.35(d,J＝7.6Hz,1H),5.44(s,1H),7.35(d,J＝8.7Hz,1H),7.56(d,J＝6.8Hz,1H),7.70(dd,J＝8.7,6.8Hz,1H),9.61(t,J＝5.1Hz,1H),13.52(br.s.,1H),13.74(br.s.,1H)

(8S,10S) -1- [ (2-aminoethyl) amino ] -6,8,10, 11-tetrahydroxy-8- (2-methyl-1, 3-dioxolan-2-yl) -7,8,9, 10-tetrahydrotetracene-5, 12-dione

ESI MS:m/z 471(MH⁺)

Deprotection of the amino acid

Synthesis of intermediate (8S,10S) -8-acetyl-1-amino-6, 8,10, 11-tetrahydroxy-7, 8,9, 10-tetrahydrotetracene-5, 12-dione (XIV)

To cold trifluoroacetic acid (2mL) was added (8S,10S) -1- [ (3, 4-dimethoxybenzyl) amino group]6,8,10, 11-tetrahydroxy-8- (2-methyl-1, 3-dioxolan-2-yl) -7,8,9, 10-tetrahydrotetracene-5, 12-dione (133mg,0.230mmol) and 2 drops of anisole. The solution was stirred at 5 ℃ for 20min and then at room temperature for 2h until no material was detected. The reaction was diluted with water (5mL) and NaHCO₃The solution was neutralized and the aqueous phase was extracted with DCM (3 × 50 mL). With anhydrous Na₂SO₄The combined organic phases were dried, filtered and the solvent evaporated in vacuoUsing Et₂The crude product was treated with O (10 mL). The dark purple precipitate was collected by filtration and dried in oven under vacuum at 30 ℃ to give the desired intermediate (82mg, y 93%).

ESI MS:m/z 384(MH⁺)

¹H NMR(400.5MHz,DMSO-d₆)ppm 1.99(dd,J＝14.4,4.6Hz,1H),2.13-2.19(m,1H),2.30(s,3H),2.88-2.95(m,1H),2.98-3.05(m,1H),5.07(m,1H),5.29(br.s.,1H),6.07(s,1H),7.24(dd,J＝8.3,1.1Hz,1H),7.51(dd,J＝7.3,1.1Hz,,1H),7.55-7.60(m,1H),8.05(br.s.,2H),13.49(br.s.,1H),13.85(br.s.,1H)

Similarly, by using suitable starting materials, the following compounds can be prepared:

(8S,10S) -8-acetyl-6, 8,10, 11-tetrahydroxy-1- [ (2-hydroxyethyl) amino ] -7,8,9, 10-tetrahydrotetracene-5, 12-dione

ESI MS:m/z 428(MH⁺)

¹H NMR(500MHz,DMSO-d₆)ppm 1.98(dd,J＝14.2,4.6Hz,1H),2.16(d,J＝14.2Hz,1H),2.31(s,3H),2.88-2.94(m,1H),2.98-3.04(m,1H),3.46-3.49(m,2H),3.68-3.72(m,2H),5.01(t,J＝5.1Hz,1H),5.05-5.10(m,1H),5.30(d,J＝6.7Hz,1H),6.10(s,1H),7.35(d,J＝8.7Hz,1H),7.56(d,J＝7.1Hz,1H),7.70(dd,J＝8.7,7.1Hz,1H),9.62(t,J＝5.1Hz,1H),13.47(br.s.,1H),13.76(br.s.,1H)

(8S,10S) -8-acetyl-1- [ (2-aminoethyl) amino ] -6,8,10, 11-tetrahydroxy-7, 8,9, 10-tetrahydrotetracene-5, 12-dione

ESI MS:m/z 427(MH⁺)

¹H NMR(500MHz,DMSO-d₆)ppm 1.97(dd,J＝14.2,4.6Hz,1H),2.14(d,J＝14.2Hz,1H),2.31(s,3H),2.88-2.94(m,1H),2.98-3.04(m,1H),3.05-3.22(m,4H),5.05-5.10(m,1H),5.30(d,J＝6.7Hz,1H),6.10(s,1H),7.35(d,J＝8.7Hz,1H),7.54(d,J＝7.1Hz,1H),7.70(dd,J＝8.7,7.1Hz,1H),9.60(t,J＝5.1Hz,1H),13.46(br.s.,1H),13.76(br.s.,1H)

Protection of

Synthesis of intermediate N- [ (8S,10S) -8-acetyl-6, 8,10, 11-tetrahydroxy-5, 12-dioxo-5, 7,8,9,10, 12-hexahydrotetracen-1-yl ] -2,2, 2-trifluoroacetamide

Intermediate (8S,10S) -8-acetyl-1-amino-6, 8,10, 11-tetrahydroxy-7, 8,9, 10-tetrahydrotetracene-5, 12-dione (600.0mg,1.56mmol) was dissolved in dry DCM (120mL) and trifluoroacetic anhydride (1.2mL) was added. The reaction was stirred at rt in the dark for 5 min until no starting material was detected (HPLC analysis). The reaction mixture was then diluted with DCM (100mL) and saturated NaHCO₃Aqueous solution (3X 100mL) then washed with water (1X 100 mL). With anhydrous Na₂SO₄The organic phase was dried and the solvent was removed in vacuo. The residue thus obtained (eluent: CH) was purified by flash chromatography on silica gel (230-400 mesh)₃COCH₃a/DCM; 0.3/9.7) to give the desired product (494.1mg, red solid).

ESI MS:m/z 480(MH⁺)

¹H NMR(500MHz,CHCl₃-d)ppm 2.22(dd,J＝14.5,4.9Hz,1H),2.36-2.41(m,1H),2.45(s,3H),3.01(d,J＝18.7Hz,1H),3.23(dd,J＝18.7,2.2Hz,1H),3.81(d,J＝5.2Hz,1H),4.54(s,1H),5.35(m,1H),7.93(dd,J＝8.4,7.7Hz,1H),8.29(dd,J＝7.7,1.1Hz,1H),9.12(dd,J＝8.4,1.1Hz,1H),13.29(br.s.,1H),13.29(s,1H),13.46(s,1H).

n- (2- { [ (8S,10S) -8-acetyl-6, 8,10, 11-tetrahydroxy-5, 12-dioxo-5, 7,8,9,10, 12-hexahydrotetracen-1-yl ] amino } ethyl) -2,2, 2-trifluoroacetamide

ESI MS:m/z 523(MH⁺)

Step G3

Synthesis of N- [ (8S,10S) -8-acetyl-6, 8, 11-trihydroxy-5, 12-dioxo-10- ({2,3, 6-trideoxy-3- [ (trifluoroacetyl) amino ] -L-lysu-hexopyranosyl } oxy) -5,7,8,9,10, 12-hexahydrotetracen-1-yl ] -2,2, 2-trifluoroacetamide

The intermediate N- [ (8S,10S) -8-acetyl-6, 8,10, 11-tetrahydroxy-5, 12-dioxo-5, 7,8,9,10, 12-hexahydrotetracen-1-yl in a dry 3-necked round-bottomed flask under an argon atmosphere]-2,2, 2-trifluoroacetamide (480.0mg,1.0mmol) is dissolved in dry DCM (110mL) and powdered molecular sieves (b), (d) are added20.0 mg). Cooling the reaction mixture at 10 ℃; simultaneously, silver triflate (334.0mg,1.3mmol) was added in dry Et₂Solution in O (15mL) and 2,3, 6-Trideoxy-4-O- (trifluoroacetyl) -3- [ (trifluoroacetyl) amino]A solution of L-lysu-hexopyranosyl chloride (511.4mg,1.43mmol) in dry DCM (15 mL). The reaction mixture was stirred at 10 ℃ in the dark for 45 minutes until no starting material was detected (HPLC analysis). Adding saturated NaHCO₃Aqueous solution (50mL), the reaction mixture was stirred at room temperature for 30 minutes, then filtered through a pad of celite. The organic phase was separated, washed with water and dried over anhydrous Na₂SO₄And (5) drying. Removing the solvent in vacuo and cooling at 0 deg.CThe resulting residue was washed with MeOH (20mL) and solid NaHCO₃The treatment is carried out for 15 minutes. The solvent was evaporated in vacuo and the residue was purified by flash chromatography on silica gel (230-400 mesh) (eluent: CH)₃COCH₃a/DCM; 0.5/9.5) to give the desired product (320.0mg, red solid).

ESI MS:m/z 705(MH⁺)

¹H NMR(500MHz,CH₃CN-d₃)ppm 1.22(d,J＝6.47Hz,3H)1.77(dd,J＝13.18,4.64Hz,1H)2.00(td,J＝13.15,3.97Hz,1H)2.10(d,J＝10.13Hz,1H)2.33-2.35(m,1H)2.87-3.00(m,1H)3.01-3.13(m,1H)3.22(br.s.,1H)3.61(br.s.,1H)4.09-4.16(m,1H)4.22(q,J＝6.47Hz,1H)4.29(s,1H)5.10(br.s.,1H)5.40(d,J＝3.54Hz,1H)7.32(d,J＝7.81Hz,1H)7.89(t,J＝7.63Hz,1H)8.11(d,J＝7.08Hz,1H)8.89(d,J＝8.30Hz,1H)13.08(br.s.,2H)

n- (2- { [ (8S,10S) -8-acetyl-6, 8, 11-trihydroxy-5, 12-dioxo-10- ({2,3, 6-trideoxy-3- [ (trifluoroacetyl) amino ] -L-lyxol-hexopyranosyl } oxy) -5,7,8,9,10, 12-hexahydrotetracen-1-yl ] amino } ethyl) -2,2, 2-trifluoroacetamide

ESI MS:m/z 748(MH⁺)

(1S,3S) -3-acetyl-3, 5, 12-trihydroxy-10- [ (2-hydroxyethyl) amino ] -6, 11-dioxo-1, 2,3,4,6, 11-hexahydrotetracen-1-yl 2,3, 6-trideoxy-3- [ (trifluoroacetyl) amino ] -L-lysu-hexopyranoside

ESI MS:m/z 653(MH⁺)

Deprotection of the amino acid

The title compound (II)

The intermediate N- [ (8S,10S) -8-acetyl-6, 8, 11-trihydroxy-5, 12-dioxo-10- ({2,3, 6-trideoxy-3- [ (trifluoroacetyl) amino group was cooled at 0 ℃ under an argon atmosphere]- α -L-lysu-hexopyranosyl } oxy) -5,7,8,9,10, 12-hexahydrotetracen-1-yl]2,2, 2-trifluoroacetamide (340.2mg,0.432mmol) is treated with 0.1N aqueous NaOH (12 mL). The reaction mixture was stirred at 0 ℃ for 1 hour in the dark until no starting material was detected (HPLC analysis). The reaction mixture was then diluted with DCM (50mL) and saturated NaHCO₃Aqueous solution (3X 30mL), then water (1X30mL), finally washed with saturated NaCl solution (1X30 mL). With anhydrous Na₂SO₄The organic phase was dried and the solvent removed in vacuo to give the desired product (180.0mg, red solid).

ESI MS:m/z 513(MH⁺)

¹H NMR(500MHz,DMSO-d₆)pm 1.14(d,J＝6.52Hz,2H)1.47(dd,J＝12.61,4.33Hz,1H)1.60(d,J＝3.30Hz,1H)2.06-2.21(m,2H)2.24-2.27(m,3H)2.86(d,J＝12.58Hz,1H)2.88-3.01(m,2H)3.28(br.s.,1H)4.09(d,J＝6.29Hz,1H)4.45(br.s.,1H)4.94(t,J＝4.22Hz,1H)5.19(d,J＝3.53Hz,1H)5.44(s,1H)7.24(d,J＝8.28Hz,1H)7.50(d,J＝7.13Hz,1H)7.51-7.52(m,0H)7.55-7.59(m,1H)8.06(br.s.,2H)

ESI MS:m/z 556(MH⁺)

ESI MS:m/z 557(MH⁺)

Claims

1. A compound of formula (Ia) or a pharmaceutically acceptable salt thereof,

wherein R1 is NR4R5 wherein one of R4 or R5 is hydrogen and the other is hydrogen or selected from straight or branched chain C₁-C₆Alkyl, NR7R8-C₁-C₆Alkyl, R6O-C₁-C₆Alkyl, R7R8N-C₁-C₆Alkylcarbonyl, and R6O-C₁-C₆The group in the alkyl-carbonyl group,

r2 is COR9, wherein R9 is straight or branched chain C₁-C₄Alkyl, or R6O-C₁-C₄An alkyl group, a carboxyl group,

r3 is straight or branched chain C₁-C₄An alkoxy group,

r6, R7 and R8 are independently hydrogen.

2. A compound of formula (Ia) according to claim 1, selected from:

(8S,10S) -1-amino-6, 8, 11-trihydroxy-8- (hydroxyacetyl) -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione;

(8S,10S) -8-acetyl-1-amino-6, 8, 11-trihydroxy-10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione;

(8S,10S) -8-acetyl-6, 8, 11-trihydroxy-1- [ (2-hydroxyethyl) amino ] -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione;

(8S,10S) -6,8, 11-trihydroxy-8- (hydroxyacetyl) -1- [ (2-hydroxyethyl) amino ] -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione;

(8S,10S) -8-acetyl-1- [ (2-aminoethyl) amino ] -6,8, 11-trihydroxy-10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione; and

(8S,10S) -1- [ (2-aminoethyl) amino ] -6,8, 11-trihydroxy-8- (hydroxyacetyl) -10- { [ (1S,3R,4aS,9S,9aR,10aS) -9-methoxy-1-methyloctahydro-1H-pyrano [4',3':4,5] [1,3] oxazolo [2,3-c ] [1,4] oxazin-3-yl ] oxy } -7,8,9, 10-tetrahydrotetracene-5, 12-dione.

3. A pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (Ia) according to claim 1 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient, carrier or diluent.

4. Use of a compound of formula (Ia) according to claim 1 in the manufacture of a medicament for the treatment of cancer.

5. The use according to claim 4, characterized in that the cancer is selected from the group consisting of hematopoietic tumors of lymphoid lineage; hematopoietic tumors of myeloid lineage; tumors from mesenchymal cells; tumors of the central and peripheral nervous systems; and melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, thyroid follicular cancer, Kaposi's sarcoma, and mesothelioma.

6. The use of claim 5, wherein the cancer is selected from the group consisting of bladder, breast, colon, kidney, liver, lung, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin; leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma; myelodysplastic syndrome; fibrosarcoma and rhabdomyosarcoma; astrocytomas, neuroblastomas, gliomas, and schwannoma.

7. Use according to claim 6, characterized in that the leukemia is selected from the group consisting of acute lymphocytic leukemia, acute lymphoblastic leukemia, acute and chronic myelogenous leukemia, and promyelocytic leukemia.

8. A process for the preparation of a compound of formula (Ia) according to claim 1, which process comprises:

firstly, make

A compound of formula (III)

Wherein R3 is as defined in claim 1 and R10 is R1 or a group of the formula NR19R20 wherein R19 and R20 are independently a suitable nitrogen protecting group or one of R19 or R20 is hydrogen and the other is a suitable nitrogen protecting group wherein R1 is as defined in claim 1,

or

A compound of the formula (XIa),

wherein R3 and R10 are as defined above and R6 is as defined in claim 1,

or

A compound of the formula (V),

wherein R3 and R10 are as defined above,

reacting with DMDO;

then, the user can use the device to perform the operation,

treating the resulting compound of formula (XX) with cyanuric chloride or an iron (II) salt,

wherein R3 and R10 are as defined above and R2 is as defined in claim 1, and

finally, if desired, removal of the protecting group affords a compound of formula (Ia) as defined above,

optionally, converting a first compound of formula (Ia) to a second compound of formula (Ia) by known chemical reactions; and/or converting the compound of formula (Ia) to a pharmaceutically acceptable salt thereof or converting a salt to the free compound of formula (Ia), as desired.