CA2015398A1 - Rhodomycin derivatives with cytostatic activity - Google Patents
Rhodomycin derivatives with cytostatic activityInfo
- Publication number
- CA2015398A1 CA2015398A1 CA002015398A CA2015398A CA2015398A1 CA 2015398 A1 CA2015398 A1 CA 2015398A1 CA 002015398 A CA002015398 A CA 002015398A CA 2015398 A CA2015398 A CA 2015398A CA 2015398 A1 CA2015398 A1 CA 2015398A1
- Authority
- CA
- Canada
- Prior art keywords
- nra
- hydrogen
- derivatives
- formula
- anthracycline
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/24—Condensed ring systems having three or more rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/24—Condensed ring systems having three or more rings
- C07H15/252—Naphthacene radicals, e.g. daunomycins, adriamycins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Saccharide Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
Abstract of the disclosure Rhodomycin derivatives with cytostatic activity The invention relates to new anthracycline derivatives with cytostatic activity and the formula I
Description
-` 20~3~8 BE~RINGWERKE AKTIENGESELLSCHAFT 89/B 018 - Ma 772/773 Dr. Ha/Sd Description Rhodomycin derivatives with cytostatic activity The invention relates to new anthracycline derivatives with cytostatic activity and the formula I, and the physiologically acceptable salts thereof, a process for the preparation thereof and the use thereof as pharma-ceuticals. -~.
R O OH OH
R O OH OH
2 ~ I
OH O GH G
H"Cj~J o In formula I:
Rl i~ hydrogen or a hydroxyl group and R2 i8 NRA-(CHz)~-NR~*
with n = 0-12, preferably with n = O or 2-5, where Rn and * can each be hydrogen, C~-C4-alkyl or benzyl, or R2 i8 NRA-CHR3-CH2-R4, where R~ has the said meaning, and R3 is hydrogen or ~-a carboxyl group and R4 is the residue of an L-amino acid or of a biogenic amine, preferably the residue of an aromatic L-amino acid or of an aromatic biogenic amine and, particularly preferably, the residue of the biogenic amine tyramine, tryptamine, histamine, dopamine or 5-hydroxydopamine, in which case the L-amino acid i8 defined by R2 with R3 = carboxyl and the biogenic amine i~ defined by R2 with R3 = hydrogen.
2~39~
Currently about 7 to 10% of malignant tumors which are no longer curable by surgical or radiotherapeutic measure~
can be cured by chemotherapy. ~n particular, rapidly proliferating tumors such as, for example, acute lym-phatic leukemia in children, Hodgkin~s disease, testicu-lar tumors and Wilms~ tumors display high sensitivity to chemotherapy with remission rates of 60 to 90%.
Furthermore, it is possible by chemotherapy to achieve a slowing of tumor growth or partial or complete remission of the oncoses in about 30~ of all tumor patients.
However, even when the chemotherapy is continued there i8, after time intervals which vary in length, recurrence of the tumor, which is then uaually resistant to the initially success~ul chemotherapy. This so-called ~secon-dary resistance occurring during chemotherapy thenlimits the possibility of a further influence on the tumor by chemotherapy. This group of tumors include~
tumors of the breast and ovary, small-cell lung car-cinomas and some lymphomas.
:
At present, the influence on many malignant diseases possible by chemotherapy is, however, zero or only marginal, so that it must be assumed in these cases that there is a primary resistance of the tumor to the sub-stances currently known. This group of tumors includes non-small-cell lung tumors and many gastrointestinal tumors. When this group of tumors with primary resistance is considered in terms of their rate of proliferation, it is ~een that the tumors which can be influenced by chemotherapy with difficulty or not at all are precisely the ones which have a particularly ~low proliferation rate.
Thus, it is a matter of the utmost priority to look for new substances which are effective precisely on human tumors which proliferate slowly and have primary resist-ance. Moreover, future chemotherapeutics should, wherepossible, have no cross-resi~tance with substances ~ .
:,1 , . .. . .
''~' ' : .'- . .- -, : ........ .: .
2Q~539~
already known, in order to be utilizable with prospects of success even where there is secondary resistance to already established substances.
In the anthracycline class, particularly doxorubicin (adriamycin), epirubicin or daunomycin have therapeutic efficacy on a large number of oncoses. However, repeated treatment is followed by a distinct reduction in the antitumor activity, as far as to ineffectiveness, caused by the development of ~secondary resistance" of the tumor cells to the anthracycline.
Another considerable problem in the use of these known anthracyclines for tumor therapy is that, besides the desired cytostatic activity, these compounds also have undesired side effects such as, for example, hemato-logical or cardiac toxicity.
~ 'The activity of daunomycin and adriamycin in tumor therapy is mainly attributed to the ability of these anthracyclines to intercalate into the DNA, which sup-presses growth of the tumor cell and brings about death of the cell.
Based on this state of the art, the ob~ect of the present invention is to provide new anthracycline derivatives which are, where possible, not cross-resistant with adriamycin and which are distinguished by a new spectrum of action and a lower toxicity than adriamycin, and thus can be used advantageously in tumor therapy.
It has already been proposed for this purpose to synthe-size rhodosaminylanthracyclinones of the formula I in which R1 has the said meaning, and R2 is hydroxyl, Cl-C4-alkoxy, branched or unbranched or ~ubstituted or unsub-stituted, or NR~2 with R~ = hydrogen or C1-C4-alkyl.
The anthracycline derivatives of the formula I have now been found, in which the ~olubility in water and/or :",, ~ ", ~ .
4 20~398 reactivity and/or toxicity of the anthracycline is altered from the anthracyclines of the state of the art by changing the radical R2 in the manner indicated by the definition of R2.
For example, it has been found that when R2 is NH-(CH2)n-NH2 with n = 2 or 4, the anthracycline molecule has amphoteric properties and it~ solubility in water is higher than that of the anthracyclines of the state of the art.
It has furthermore been found that when R2 is NH-CHR3C~2-R4, in which R3 is hydrogen and R4 is the residue of the biogenic amine histamine, tyramine or tryptamine, the anthracycline i8 able - besides its ability to inter-calate into DNA - also to interact with certain enzymes, which alters its reactivity by comparison with the anthracyclines of the state of the art. -~
Hence the invention relates to compounds of the formula I with the stated meanings.
Preferred meanings:
2 0 R2 i8 NRa--( CH2) n--NRA*;
R2 is NR~-NR~Rb~ NR~- ( CH2)2-NR~Rb or NR~- ( CH2)4-NRH* ~ where derivatives with R~ = Rb = hydrogen are preferred;
R2 is NRn-CHR3-CH2-R4~ where compounds in which R3 is hydro~en are ~referred;
R4 iS the residue of an aromatic L-amino acid or of an aromatic biogenic amine;
R4 is the residue of the biogenic amine tyramine, trypt-amine, histamine, dopamine or 5-hydroxydopamine.
The process for the preparation of the new anthracycline derivatives according to the invention which have cyto-static activity and the formula I comprises reacting a compound of the formula I in which R1 has the said mean-ing, and R2 i8 Cl-C4-alkoxy, branched or unbranched, or benzyloxy, with a compound of the formula HR~N-(CH2)n--; . . . . . . . . . : .
2 ~ 9 8 NRaRb in which n, R~ and * have the said meaning, or with a biogenic amine or an amino acid in a suitable solvent such as, for example, dimethylformamide or ethanol, at a temperature between, for example, ODC and the boiling point of the solvent, and isolating and purifying the compound of the formula I obtained in this way, in which R1 has the said meaning, and R2 is NR~-(CH2)n-NRa* or NH-C~R3-C-~R4 where n, R~, *, R3 and R4 have the said meaning.
The n0w anthracycline derivatives obtained by the process according to the invention are distinguished by cyto-static activity, and they can therefore be processed -together with the customary pharmaceutical formulating agents and/or diluents to give pharmaceuticals which are used in cancer therapy. The mode of dosage and adminis-tration thereof e~sentially corresponds to that for the known substances adriamycin, daunomycin, aclacinomycin, 4'-epi-adriamycin, 4~-methoxyadriamycin or 4~-deoxyadria-mycin.
The pharmaceuticals prepared in this way can additionally also contain other active compounds as long as these do not show any undesired side effects together with the compounds according to the invention.
The cytostatic activity of the compounds according to the invention was tested using L1210 mouse leukemia cells.
The formation of colonies of L1210 leukemia cells in agar plates was used for this purpose. This method iB employed to detect the effect of test 6ubstances on the growth behavior of the cells over 1 hour or over several genera-tions. At a cell cycle time of 10-12 hours, this entails about 14 consecutive generations being observed in the 7 days the test lasts. In this test, the substances with cytostatic activity according to the invention bring about a reduction in the number of colonies to be observed by comparison with an untreated control sample.
Details of the test method employed are evident from the - 6 - 2~3 g8 following descriptions of the procedures for determining the formation of colonies.
To explain the preparation process according to the invention, Examples 1 to 5 in which preferred compound~
according to the invention were prepared by the claimed process are detailed hereinafter.
Characterization of comPounds of the formula I
The progress of the reactions and the resulting compounds were investigated by thin-layer chromatography or with the HPLC technique. Thin-layer chromatography was carried out, unless noted otherwise, on precoated silica gel plates (Merck). Column chromatography was carried out on silica gel 60 (Merck) of particle size 0.040-0.063 mm.
The yields are not optimized.
The following solvent mixtures were used for the thin-layer and column chromatography (data in percent by volume in each case):
Solvent mixtures A B C
Composition ` 20 Chloroform (%) 70 89 77 Methanol (%) 18 7.4 14 Acetic acid (%) 8.5 3 7 Water (%) 3.5 0.6 2 The ~tructures of the prepared compounds were determined by lH-NMR and MS spectroscopy.
.
.
~ .;; , . , ~
2~a3~
EXAMPL~S
Preparation of the startina com~ounds:
Preparation of the startina com~ound of the formula II in which R1 is hydroaen and R2 is ethoxy:
7-0-(3'-N-Ethoxycarbonylmethyl-3'-N-meth~l-u-L-daunos-aminvl)-~-rhodomycinone 100 mg (0.189 mmol) of 7-0-(3~-N-methyl-~-L-daunosam-inyl)-~-rhodomycinone and 75 ~1 (113 mg = 0.677 mmol =
OH O GH G
H"Cj~J o In formula I:
Rl i~ hydrogen or a hydroxyl group and R2 i8 NRA-(CHz)~-NR~*
with n = 0-12, preferably with n = O or 2-5, where Rn and * can each be hydrogen, C~-C4-alkyl or benzyl, or R2 i8 NRA-CHR3-CH2-R4, where R~ has the said meaning, and R3 is hydrogen or ~-a carboxyl group and R4 is the residue of an L-amino acid or of a biogenic amine, preferably the residue of an aromatic L-amino acid or of an aromatic biogenic amine and, particularly preferably, the residue of the biogenic amine tyramine, tryptamine, histamine, dopamine or 5-hydroxydopamine, in which case the L-amino acid i8 defined by R2 with R3 = carboxyl and the biogenic amine i~ defined by R2 with R3 = hydrogen.
2~39~
Currently about 7 to 10% of malignant tumors which are no longer curable by surgical or radiotherapeutic measure~
can be cured by chemotherapy. ~n particular, rapidly proliferating tumors such as, for example, acute lym-phatic leukemia in children, Hodgkin~s disease, testicu-lar tumors and Wilms~ tumors display high sensitivity to chemotherapy with remission rates of 60 to 90%.
Furthermore, it is possible by chemotherapy to achieve a slowing of tumor growth or partial or complete remission of the oncoses in about 30~ of all tumor patients.
However, even when the chemotherapy is continued there i8, after time intervals which vary in length, recurrence of the tumor, which is then uaually resistant to the initially success~ul chemotherapy. This so-called ~secon-dary resistance occurring during chemotherapy thenlimits the possibility of a further influence on the tumor by chemotherapy. This group of tumors include~
tumors of the breast and ovary, small-cell lung car-cinomas and some lymphomas.
:
At present, the influence on many malignant diseases possible by chemotherapy is, however, zero or only marginal, so that it must be assumed in these cases that there is a primary resistance of the tumor to the sub-stances currently known. This group of tumors includes non-small-cell lung tumors and many gastrointestinal tumors. When this group of tumors with primary resistance is considered in terms of their rate of proliferation, it is ~een that the tumors which can be influenced by chemotherapy with difficulty or not at all are precisely the ones which have a particularly ~low proliferation rate.
Thus, it is a matter of the utmost priority to look for new substances which are effective precisely on human tumors which proliferate slowly and have primary resist-ance. Moreover, future chemotherapeutics should, wherepossible, have no cross-resi~tance with substances ~ .
:,1 , . .. . .
''~' ' : .'- . .- -, : ........ .: .
2Q~539~
already known, in order to be utilizable with prospects of success even where there is secondary resistance to already established substances.
In the anthracycline class, particularly doxorubicin (adriamycin), epirubicin or daunomycin have therapeutic efficacy on a large number of oncoses. However, repeated treatment is followed by a distinct reduction in the antitumor activity, as far as to ineffectiveness, caused by the development of ~secondary resistance" of the tumor cells to the anthracycline.
Another considerable problem in the use of these known anthracyclines for tumor therapy is that, besides the desired cytostatic activity, these compounds also have undesired side effects such as, for example, hemato-logical or cardiac toxicity.
~ 'The activity of daunomycin and adriamycin in tumor therapy is mainly attributed to the ability of these anthracyclines to intercalate into the DNA, which sup-presses growth of the tumor cell and brings about death of the cell.
Based on this state of the art, the ob~ect of the present invention is to provide new anthracycline derivatives which are, where possible, not cross-resistant with adriamycin and which are distinguished by a new spectrum of action and a lower toxicity than adriamycin, and thus can be used advantageously in tumor therapy.
It has already been proposed for this purpose to synthe-size rhodosaminylanthracyclinones of the formula I in which R1 has the said meaning, and R2 is hydroxyl, Cl-C4-alkoxy, branched or unbranched or ~ubstituted or unsub-stituted, or NR~2 with R~ = hydrogen or C1-C4-alkyl.
The anthracycline derivatives of the formula I have now been found, in which the ~olubility in water and/or :",, ~ ", ~ .
4 20~398 reactivity and/or toxicity of the anthracycline is altered from the anthracyclines of the state of the art by changing the radical R2 in the manner indicated by the definition of R2.
For example, it has been found that when R2 is NH-(CH2)n-NH2 with n = 2 or 4, the anthracycline molecule has amphoteric properties and it~ solubility in water is higher than that of the anthracyclines of the state of the art.
It has furthermore been found that when R2 is NH-CHR3C~2-R4, in which R3 is hydrogen and R4 is the residue of the biogenic amine histamine, tyramine or tryptamine, the anthracycline i8 able - besides its ability to inter-calate into DNA - also to interact with certain enzymes, which alters its reactivity by comparison with the anthracyclines of the state of the art. -~
Hence the invention relates to compounds of the formula I with the stated meanings.
Preferred meanings:
2 0 R2 i8 NRa--( CH2) n--NRA*;
R2 is NR~-NR~Rb~ NR~- ( CH2)2-NR~Rb or NR~- ( CH2)4-NRH* ~ where derivatives with R~ = Rb = hydrogen are preferred;
R2 is NRn-CHR3-CH2-R4~ where compounds in which R3 is hydro~en are ~referred;
R4 iS the residue of an aromatic L-amino acid or of an aromatic biogenic amine;
R4 is the residue of the biogenic amine tyramine, trypt-amine, histamine, dopamine or 5-hydroxydopamine.
The process for the preparation of the new anthracycline derivatives according to the invention which have cyto-static activity and the formula I comprises reacting a compound of the formula I in which R1 has the said mean-ing, and R2 i8 Cl-C4-alkoxy, branched or unbranched, or benzyloxy, with a compound of the formula HR~N-(CH2)n--; . . . . . . . . . : .
2 ~ 9 8 NRaRb in which n, R~ and * have the said meaning, or with a biogenic amine or an amino acid in a suitable solvent such as, for example, dimethylformamide or ethanol, at a temperature between, for example, ODC and the boiling point of the solvent, and isolating and purifying the compound of the formula I obtained in this way, in which R1 has the said meaning, and R2 is NR~-(CH2)n-NRa* or NH-C~R3-C-~R4 where n, R~, *, R3 and R4 have the said meaning.
The n0w anthracycline derivatives obtained by the process according to the invention are distinguished by cyto-static activity, and they can therefore be processed -together with the customary pharmaceutical formulating agents and/or diluents to give pharmaceuticals which are used in cancer therapy. The mode of dosage and adminis-tration thereof e~sentially corresponds to that for the known substances adriamycin, daunomycin, aclacinomycin, 4'-epi-adriamycin, 4~-methoxyadriamycin or 4~-deoxyadria-mycin.
The pharmaceuticals prepared in this way can additionally also contain other active compounds as long as these do not show any undesired side effects together with the compounds according to the invention.
The cytostatic activity of the compounds according to the invention was tested using L1210 mouse leukemia cells.
The formation of colonies of L1210 leukemia cells in agar plates was used for this purpose. This method iB employed to detect the effect of test 6ubstances on the growth behavior of the cells over 1 hour or over several genera-tions. At a cell cycle time of 10-12 hours, this entails about 14 consecutive generations being observed in the 7 days the test lasts. In this test, the substances with cytostatic activity according to the invention bring about a reduction in the number of colonies to be observed by comparison with an untreated control sample.
Details of the test method employed are evident from the - 6 - 2~3 g8 following descriptions of the procedures for determining the formation of colonies.
To explain the preparation process according to the invention, Examples 1 to 5 in which preferred compound~
according to the invention were prepared by the claimed process are detailed hereinafter.
Characterization of comPounds of the formula I
The progress of the reactions and the resulting compounds were investigated by thin-layer chromatography or with the HPLC technique. Thin-layer chromatography was carried out, unless noted otherwise, on precoated silica gel plates (Merck). Column chromatography was carried out on silica gel 60 (Merck) of particle size 0.040-0.063 mm.
The yields are not optimized.
The following solvent mixtures were used for the thin-layer and column chromatography (data in percent by volume in each case):
Solvent mixtures A B C
Composition ` 20 Chloroform (%) 70 89 77 Methanol (%) 18 7.4 14 Acetic acid (%) 8.5 3 7 Water (%) 3.5 0.6 2 The ~tructures of the prepared compounds were determined by lH-NMR and MS spectroscopy.
.
.
~ .;; , . , ~
2~a3~
EXAMPL~S
Preparation of the startina com~ounds:
Preparation of the startina com~ound of the formula II in which R1 is hydroaen and R2 is ethoxy:
7-0-(3'-N-Ethoxycarbonylmethyl-3'-N-meth~l-u-L-daunos-aminvl)-~-rhodomycinone 100 mg (0.189 mmol) of 7-0-(3~-N-methyl-~-L-daunosam-inyl)-~-rhodomycinone and 75 ~1 (113 mg = 0.677 mmol =
3.58 equiv.) of ethyl bromoacetate were reacted in the presence of 80 ~1 (58 mg = 0.574 mmol = 3.0 equiv.) of triethylamine for 2 h. The ~-product mixture after concentration was dissolved without delay in a little chloroform, loaded onto a silica gel column made up in ether (15 g of silica gel) and eluted with about 100 ml of ether to remove the excess bromoace-tate. The desired compound was then eluted in chloroform/
ethan~l (20/1) (RF 0.32).
Yield 70 mg (0.114 mmol) = 60%
MS-FAB (M+Ht) m/e = 616 `-lH NMR (300 NHz, CDC13) ~ 1.12 (t, 3H, Jl314 = 7.4 Hz, Me-14), 1.16 (t, 3H, Jbc = 7.1 Hz, Me-c), 1.40 (d, 3H, J5 6~
= 6.8 Hz, Me-6'), 1.7-19 (m, 4H, CH2-2' and CH2-13), 2.11 (dd, lH, J7~, = 4 Hz, J8.8b = 15 Hz, H-8a), 2.26 (d, lH, J8. 6b = 15 Hz, H-8b), 2.38 (8~ 3H, N-CH3), 2.72 (d, lH, J1o ~-1o = 4 Hz, OH-10), 2.82 (m, lH, H-3'), 3.14 (bs, lH, OH-4'), 3.31 (d, lH, J,,. = 17 Hz, H-a), 3.33 (d, lH, J~
= 17 Hz, H-a'), 3.65 (bs, lH, H-4'), 4.03 (s, lH, OH-9), 4.08 (m, 3H, CH2-b and H-5'), 4-92 (d, lH~ J1oo~-1o = 4 Hz~
H-10), 5.15 (m, lH, H-7), 5.51 (bd, lH~ J1 2~ = 2-5 Hz, H-1'), 7.33 (dd, lH, J13 = 1 Hz, J23 = 8.4 Hz, H-3), 7.73 (t~ lH~ J12 = J23 = 8-4 Hz, H-2), 7.90 (dd, lH, J12 = 8.4 Hz, J13 = 1 Hz, H-l), 12.16 (bs, lH, OH-4), 12.84 (bs, lH, OH-6), 13.63 (bs, lH, OH-ll).
~,, 2 ~
, Example 1:
Preparation of a compound of the formula I in which R1 i8 H and R2 is NH-NH2 7-0-(3'-N-Hydrazinocarbonylmethyl-3'-N-methyl-~-L-daunos-S aminyl)-~-rhodomycinone (compound 1) A solution of7-0-(3~-N-ethoxycarbonylmethyl-3~-N-methyl-~-L-daunosaminyl)-~-rhodomycinone (154 mg = 0.25 mmol) in ethanol (38 ml) was mixed with 100% hydrazine hydrate (17.5 ~1 = 18 mg = O.36 mmol = 1.44 equiv.) and stirred at room temperature in the dark for 60 h. The reaction mixture was then evaporated to dryness in a rotary evaporator and chromatographed on 16 g of ~ilica gel in solvent mixture A ( RF . 26). Water wa~ added to the combined fractions to ~eparate the phases, the pH was ad~usted to 7 with 10 percent (w/v) sodium hydroxide solution and then ad~usted to pH 8 by addition of satura-ted aqueous sodium bicarbonate solution. The phases were then separated in a separating funnel, the aqueous phase was extracted several times with chloroform, and the combined organic phases were evaporated to dryness in a rotary evaporator.
Yield 51 mg (0.085 mmol ) = 34%
MS-FAB (M~H+) m/e = 602 lH-NMR (300 MHz, CDC13)~D6-DMSO (6/1)) ~ 1-10 (t, 3H~ Jl3 14 = 7.4 Hz, Me-14), 1.35 (d, 3H, J5~ 6~ = 6.5 Hz, Me-6'), 1.6-2.0 (m, 4H, CH2-2'and CH2-13), 2.~ (bs, 2H, CH2-8), 2.26 (s, 3H, N-CH3), 2.52 (bd, lH, J2~ 3. = 10-5 Hz, H-3')~
3.06 (d, lH, J~ A~ = 16.6 Hz, H-a), 3.19 (d, lH, JA ~
16.6 Hz, H-a'), 3.70 (bs, lH, H-4'), 3.90 (bs, lH, OH-9), 4.06 (q, lH, J5~6~ = 6.6 Hz, H-5'), 4.84 (8, lH, H-10), 5.12 (m, lH, H-7), 5.48 (bd, lH, J1' 2. = 3.2 Hz, H-1'), 7.31 (dd, lH, Jl3 = 1 Hz, J2 3 = 8-4 Hz, H-3), 7-72 (t~
lH~ Jl2 = J23 = 8-3 Hz, H-2), 7.87 (dd, lH, Jl 2 = 7.7 Hz, J13 = 1 Hz, H-l), 9.03 (8~ lH, hydrazide-NH).
Exam~le 2:
Preparation of the compound of the formula I in which R1 is H and R2 is NH-(CHæl7-NH~:
7-0-(3'-N-Ethylenediaminocarbonvlmethyl-3'-N-methyl-~-L-daunosaminvl~-~-rhodomvcinone (compound 2) A solution of7-0-(3'-N-ethoxycarbonylmethyl-3'-N-methyl-~-L-daunosaminyl)-~-rhodomycinone (150 mg = 0.24 mmol) in dimethylformamide (20 ml) was mixed with 1 ml of ethyl-enediamine and stirred at room temperature in the dark for 3 h. The reaction mixture was then evaporated to dryness in a rotary evaporator and chromatographed on 15 g of silica gel in solvent mixture A (Rp 0.15), and the fractions were worked up in analogy to Example 1.
Yield: 120 mg ~0.19 mmol) = 79%
MS-FAB (M+H~) m/e = 630 1H-NMR (300 MHz, CDCl3/D6-DMSO) (4/1)) ~ 1.09 (t, 3H, Jl3 14 = 7.4 Hz, Me-14), 1.30 (d, 3H, J5 6. = 6.5 Hz, ~e-6'), 1.6-1.9 (m, 4H, CH2-2' and CH2-13), 2.19 (bs, 2H, CH2-8), 2.28 (s, 3H, N-CH3), 2.55 (m, lH, H-3'), 2.84 (m, 2H, CH2-c), 3.01 (d, lH, J~ = 18 Hz, H-a), 3.15 (d, lH, J,~.
= 18 Hz, H-a'), 3.25 (m, lH, H-b), 3.38 (m, lH, H-b'), 3.62 (bs, lH, H-4'), 4.05 (q, lH, J56 = 6.6 Hz, H-5'), 4.82 (s, lH, H-10), 5.12 (m, lH, H-7), 5.48 (bd, lH, Jl-2 = 3 Hz~ H-1'), 7-32 (dd, lH, J13 = 1 Hz, J23 = 8-4 Hz, H-3), 7.73 (t, lH, J12 s J23 = 8 Hz, H-2), 7.87 (dd, lH, Jl2 = 7.5 Hz, J~3 = l Hz, H-l), 8.06 (t, lH, J~ b = 6 Hz, -CO-NH-).
ExamDle 3:
Preparation of a compound of the formula I in which Rl is H and R2 is NH-(CH2~2-N(CH?)2 7-0-(3'-N-(Dimethylaminoethylaminocarbonyl)-3'-N-methyl-~-L-daunosaminyl~-~-rhodomycinone (com~ound 3) A solution of 7-0-(3'-N-ethoxycarbonylmethyl-3'-N-methyl-~-L-daunosaminyl)-~-rhodomycinone (16 mg = 0.026 mmol) in methanol (8 ml) was mixed with 100 ~1 (81 mg = 0.92 mmol) of N,N-dimethylaminoethylamine and 6tirred at room temperature in the dark for 16 h. The mixture was then evaporated to dryness in a rotary evaporator and dried under high vacuum. The reaction mixture was chromato-graphed on 4 g of silica gel in solvent mixture A (Rp 0.14), and the fractions were worked up in analogy to Example 1.
Yield 13 mg (0.020 mmol) = 77%
H-NNR (300 MHz, CDCl3) ~ 1-12 (t, 3H, Jl314 = 7-4 Hz, Me-14), 1.30 (d, 3H, J5.6. = 6.4 Hz, Me-6'), 1.7-2.05 (m, 4H, CH2-2' and CH2-13), 2.11 (dd, lH, J78~ = 4 Hz, J~b = 15 Hz, H-8a), 2.25 (d, lH, J~ ~b = 15 Hz, H-8b), 2.28 (8, 6H, N(CH3)2), 2.31 (~l 3H, N-CH3), 2.51 (m, 3H, CH2-c and H-3~)l 2.98 (d, lH, J~a = 18 Hz, H-a), 3.19 (m, lH, H-b), 3.20 (d, lH, J~ n~ = 18 Hz, H-a'), 3.68 (m, lHI H-b'), 5.04 (q, lH J5.6 = 6.5 Hz, H-5~), 4.10 (8l lH, OH-9), 4.91 (8l lH, H-10)~ 5.15 (m, lH~ H-7), 5.52 (bd, lH, J1.2. = 3 Hz, H-1~), 7-33 (d~ lH~ J23 = 8 Hz, H-3)~ 7.72 (t, lH, Jl2 =
J23 = 8 Hz, H-2), 7-89 (d, lH, Jl2 = 7 Hz, H-1), 8.14 (m, lH, NH).
Example 4:
Pre~aration of a com~ound of the formula I in which Rl is H and R2 i8 NH-(CH?)~
7-0-(3'-N-(4-Aminobutylaminocarbonyl)-3 ~ -N-methyl-~-L-daunosaminvl)--~-rhodomycinone (com~ound 4):
A solution of7-0-(3'-N-ethoxycarbonylmethyl-3'-N-methyl-~-L-daunosaminyl)-~-rhodomycinone (16 mg = 0.026 mmol) in acetonitrile (3 ml) was mixed with 200 ~1 of 1,4-diamino-butane and stirred at room temperature in the dark for 3 h. The reaction mixture was then evaporated to dryness in a rotary evaporator, and chromatographed on 3 g of silica gel in solvent mixture C (RF 0.16), and the fractions were worked up in analogy to Example 1.
Yield: 11 mg (0.017 mmol) = 65%
MS-FAB (M+H+) m/e = 658 - 11 - 2~398 lH-NMR (300 MHz, CDCl3/D6-DMSO (4/1)) ~ 1.10 (~, 3H, J13,14 = 7.4 Hz, Me-14), 1.32 (d, 3H, J5~ 6~ = 6.5 Hz, Me-6'), 1.4-1.6 (m, 4H, CH2-c and CH2-d), 1.6-2.0 (m, 4H, CH2-2' and CH2-13), 2.17 (bs, 2H, CH2-8), 2.29 (s, 3H, N-CH3), 2.55 (m, lH, H-3'), 2.69 (bt, 2H, Jd u = 6.5 Hz, CH2-e)~
3.06 (d, lH, J~a~ = 17 Hz, H-a), 3-08 (d, lH, Ja a' = 17 Hz, H-a'), 3.20 (bt, 2H, Jb c = 6.5 Hz, CH2-b), 3.71 (bs, lH, H-4'), 4.06 (q, lH, J5~ 6~ = 6.6 Hz, H-5'), 4.82 (s, lH, H-10), 5.12 (m, lH, H-7), 5.48 (bd, lH, Jl' 2~ = 3 Hz~
H-l'), 7-29 (dd, lH, Jl 3 = 1 Hz, J23 = 8.4 Hz, H-3), 7.70 (t~ lH~ Jl2 = J23 = 8 Hz, H-2), 7.85 (dd, lH, Jl2 = 7.5 Hz, H-l, Jl3 = 1 Hz, H-l).
Example 5:
7-0-(3'-N-Hi6taminocarbonylmethyl-3'-N-methvl-~-L-daunos-aminyl)-~-rhodomycinone (compound 5) A solution of 7-0-(3~-N-ethoxycarbonylmethyl-3'-N-methyl-~-L-daunosaminyl)-~-rhodomycinone (25 mg = 0.04 mmol) in dimethylformamide (7 ml) was mixed with 20 mg (0.18 mmol) of histamine and stirred at 50C in the dark for 30 h.
The reaction mixture was then evaporated to dryness in a rotary evaporator and chromatographed in solvent mixture C (RF 0.14), and the fractions were worked up in analogy to Example 1.
Yield: 21 mg (0.03 mmol) = 75 NS-FAB (M+Ht) m/e = 681 H-NMR (300 MHz, CDCl3) ~, 1.13 (t, 3H, Jl31~ = 7.4 Hz, Me-14), 1.40 (d, 3H, J5. 6' = 6.5 Hz, Me-6'), 1.6-2.0 (m, 4H, CHz-2' and CH2-13), 2-11 (dd, lH, J7Ba = 4 Hz, J8~Bb = 15 Hz, H-8a), 2.26 (d, lH, JôuBb = 15 Hz, H-8b), 2.29 (s, 3H, N-CH3), 2.30 (bd, lH, J2.3. = 11 Hz, H-3'), 2.78 (bt, 2H, Jbc = 5.6 Hz, CH2-c), 2.96 ~d, lH, Ja- = 17 Hz, H-a), 3.14 (d, lH, J,~. = 17 Hz, H-a'), 3.49 (m, 2H, CH2-b), 3.73 (8~ lH, H-4'), 4.10 (q, lH, J5 6~ = 6-3 Hz, H-5')~
4.92 (8~ lH, H-~0), 5.16 (m, lH, H-7), 5-51 (bd, lH, Jl.2.
= 3.5 Hz, H-l'), 6.81 (8, lH, H-d), 7.33 (dd, lH, Jl 3 =
1 Hz, J23 = 8.4 Hz, H-3), 7.56 (s, lH, H-e), 7.73 (t, lH, I
` - 12 - 2~ 98 Jl,2 = J2,3 = 8 Hz, H-2), 7.89 (dd, lH, Jl 2 = 7.5 Hz, Jl 3 =
1 Hz, H-1), 8.57 (bs, lH, NH).
Example 6:
7-0-(3'-N-Methvl-3'-N-tyraminocarbonylmethyl-~-~-daunos-aminvl)-B-rhodomycinone (compound 6) The reaction was carried out in analogy to Example 5 with 20 mg (0.032 mmol) of startin~ compound and 20 mg (0.15 mmol) of tyramine in dimethylformamide (3 ml), but with stirring at 50-60C for 50 h. Purification was carried out in solvent mixture C ( RF . 5)-Yield 12 mg (0.017 mmol) = 53%
MS-FAB (M+H~) m/e = 707 lH-NMR (300 NHz, CDCl3/D6-DMSO (4/1)) ~ 1.10 (t, 3H, Jl314 = 7.4 Hz, Me-14), 1.32 (d, 3H, J5. ~. = 6.5 Hz, Me-6'), 1.6-2.0 (m, 4H, CH2-2' and CH2-13), 2.19 (bs, 2H, CH2-8), 2.20 (s, 3H, N-CH3), 2.54 (bd, lH, J2' 3~ = 11 Hz~ H-3')~
2.69 (bt, 2H, Jb c = 7 Hz, CH2-c), 3.03 (d~ lH~ J~ a~ = 17 H~, H-a), 3.05 (d, lH, J~ a~ = 17 Hz, H-a'), 3.40 (m, 2H, CH2-b), 3.65 (bs, lH, H-4'), 3.89 (bs, lH, OH-9), 4.05 (q, lH, J5~ 6~ = 6.7 Hz, H-5'), 4.85 (s, lH, H-10), 5.12 (m, lH, H-7), 5.47 (bd, lH, Jl' 2~ = 3 Hz, H-l'), 6.74 (d, 2H, Jd ~ = 8.5 Hz, CH2-d), 6.97 (d, 2H, Jd ~ = 8.5 Hz, CH2-e), 7.32 (dd, lH, Jl 3 = 1 Hz, J2 3 = 8-4 Hz~ H-3)~ 7-49 (bt, lH, J~,b = 6 Hz, NH), 7-72 t (lH, Jl,2 = J2.3 = 8 Hz~
H-2), 7.87 (dd, lH, Jl 2 = 7-5 Hz, Jl 3 = 1 Hz~ H-l)-Example 7:
7-0-(3'-N-Methyl-3'-N-tryptaminocarbonylmethyl-~-L-daunosaminyl~-g-rhodomycinone (compound 7) The reaction was carried out in analogy to Example 5 with ~ --20 mg (0.032 mmol) of starting compound and 20 mg (0.12 mmol) of tryptamine in dimethylformamide (3 ml);
but with stirring at 50-60~C for 50 h. Purification was ~ '':'' ~,-,~, . .. . . .
13 2~ ~3~8 carried out in solvent mixture C (RF 0.46).
Yield 17 mg (O.023 mmol) = 72%
Ms-FAs (M+H+) m/e = 730 lH-NMR (300 MHz, CDCl3) ~, 1.13 (t, 3H, Jl314 = 7-4 Hz, Me S 14), 1.23 (d, 3H, J5~ 6' = 6.5 Hz, Me-6~), 1.5-2.0 ~m, 4H, CH2-2' and CH2-13), 2.12 (dd, lH, J7,8n = 4 Hz, J8~,~b = 15 Hz, H-8a), 2.17 (8, 3H, N-CH3), 2.20 (d, lH, J8~,8b = 15 Hz, H-8a~), 2.48 (m, lH, H-3~), 2.96 (bt, 2H, Jb c = 6.5 Hz, CH2-c), 3.46 (bs, lH, H-4~), 3.59 (m, 2H, CH2-b), 3.96 (s~ lH, OH-9), 4.00 (q, lH, J5~ 6' = 6-3 Hz~ H-5')~ 4-93 (s, lH, H-10), 5.11 (m, lH, H-7), 5.41 (bs, lH, H-l'), 7.0 (d, lH, Jd~ = 2.3 Hz, H-d), 7.08 (dt, lH) and 7.17 (dt~ lH) (Ja,f = Jf,g = J8,h = 7 Hz~ J~,e; = J~ h = 1 Hz~ H--f and H-g), 7.32 (dd, lH, Jl3 = 1 Hz, J2 3 = 8.4 Hz, H-3), 7.33 (d, lH, Je ~ = 8.0 Hz, H-e), 7.54 (d, lH, Jg h = 7.7 Hz, H-h), 7.72 (t, lH, Jl 2 = J~ 3 = 8 Hz, H-2), 7.88 (dd, lH~ Jl,2 = 7-5 Hz~ Jl,3 = 1 Hz, H-l), 8.29 (bs, lH, CO-NH).
Cytotoxicitv of com~ound~ of the formula I for L1210 mouse leukemia cells in vitro Procedure for determining the formation of colonie6 of L1210 leukemia cell6 in soft agar 500 leukemia cells per plate were incubated with various concentrations of the test substance at 37C for 1 hour.
The cells were then washed twice with McCoy5A medium and finally, after addition of 0.3~ agar, poured into Petri dishes. Controls were incubated only with fre~h medium.
In some cases, in place of the incubation for one hour, various concentrations and test substances were mixed with the upper agar layer in order thus to achieve continuous exposure of the cells throughout the incuba-tion time. After the agar had solidified, the plate6 were incubated in an incubator at 37C for 7 days (5% by vol.
CO2, 95~ relative humidity). The number of resulting colonies with a diameter of more than 60 ~m was then counted. The results have been reported as the number of colonies in treated agar plates as a percentage of the ~''~' "' ~' ' '' , . . .
~.` " ' '. ' ' . . ,, , ' .
- - 14 - 2~ 398 untreated control. The IC50 was determined as a measure of the activity of the substance from the dose-effect plot obtained in this way. The results for the compounds described here are compiled in Table 1, comparing with adriamycin.
- 20~5398 Table 1: Cytotoxicity of the prepared compound~ of the formula I on L1210 leukemia cells in vi~ro Substance Continuou~ 1 h No. incubation incubation (Example) Rl R2 IC50 (~g/ml) IC50 (~g/ml) 1 . R NH-NH2 0.32 0,21 b c 2 H NH-CH2-CH2-NH2 0~35 1.0 b c 3 H NH-CH2-CH2-N(CH3)2 0. 12 o . 2 6 b c d e 4 H NH-CH2-CH2-CH2-CH2-NH2 0~35 1~3 b c ~
H NH CH2 CH2 ~ ~ 0.31 1~0 b c ~
ethan~l (20/1) (RF 0.32).
Yield 70 mg (0.114 mmol) = 60%
MS-FAB (M+Ht) m/e = 616 `-lH NMR (300 NHz, CDC13) ~ 1.12 (t, 3H, Jl314 = 7.4 Hz, Me-14), 1.16 (t, 3H, Jbc = 7.1 Hz, Me-c), 1.40 (d, 3H, J5 6~
= 6.8 Hz, Me-6'), 1.7-19 (m, 4H, CH2-2' and CH2-13), 2.11 (dd, lH, J7~, = 4 Hz, J8.8b = 15 Hz, H-8a), 2.26 (d, lH, J8. 6b = 15 Hz, H-8b), 2.38 (8~ 3H, N-CH3), 2.72 (d, lH, J1o ~-1o = 4 Hz, OH-10), 2.82 (m, lH, H-3'), 3.14 (bs, lH, OH-4'), 3.31 (d, lH, J,,. = 17 Hz, H-a), 3.33 (d, lH, J~
= 17 Hz, H-a'), 3.65 (bs, lH, H-4'), 4.03 (s, lH, OH-9), 4.08 (m, 3H, CH2-b and H-5'), 4-92 (d, lH~ J1oo~-1o = 4 Hz~
H-10), 5.15 (m, lH, H-7), 5.51 (bd, lH~ J1 2~ = 2-5 Hz, H-1'), 7.33 (dd, lH, J13 = 1 Hz, J23 = 8.4 Hz, H-3), 7.73 (t~ lH~ J12 = J23 = 8-4 Hz, H-2), 7.90 (dd, lH, J12 = 8.4 Hz, J13 = 1 Hz, H-l), 12.16 (bs, lH, OH-4), 12.84 (bs, lH, OH-6), 13.63 (bs, lH, OH-ll).
~,, 2 ~
, Example 1:
Preparation of a compound of the formula I in which R1 i8 H and R2 is NH-NH2 7-0-(3'-N-Hydrazinocarbonylmethyl-3'-N-methyl-~-L-daunos-S aminyl)-~-rhodomycinone (compound 1) A solution of7-0-(3~-N-ethoxycarbonylmethyl-3~-N-methyl-~-L-daunosaminyl)-~-rhodomycinone (154 mg = 0.25 mmol) in ethanol (38 ml) was mixed with 100% hydrazine hydrate (17.5 ~1 = 18 mg = O.36 mmol = 1.44 equiv.) and stirred at room temperature in the dark for 60 h. The reaction mixture was then evaporated to dryness in a rotary evaporator and chromatographed on 16 g of ~ilica gel in solvent mixture A ( RF . 26). Water wa~ added to the combined fractions to ~eparate the phases, the pH was ad~usted to 7 with 10 percent (w/v) sodium hydroxide solution and then ad~usted to pH 8 by addition of satura-ted aqueous sodium bicarbonate solution. The phases were then separated in a separating funnel, the aqueous phase was extracted several times with chloroform, and the combined organic phases were evaporated to dryness in a rotary evaporator.
Yield 51 mg (0.085 mmol ) = 34%
MS-FAB (M~H+) m/e = 602 lH-NMR (300 MHz, CDC13)~D6-DMSO (6/1)) ~ 1-10 (t, 3H~ Jl3 14 = 7.4 Hz, Me-14), 1.35 (d, 3H, J5~ 6~ = 6.5 Hz, Me-6'), 1.6-2.0 (m, 4H, CH2-2'and CH2-13), 2.~ (bs, 2H, CH2-8), 2.26 (s, 3H, N-CH3), 2.52 (bd, lH, J2~ 3. = 10-5 Hz, H-3')~
3.06 (d, lH, J~ A~ = 16.6 Hz, H-a), 3.19 (d, lH, JA ~
16.6 Hz, H-a'), 3.70 (bs, lH, H-4'), 3.90 (bs, lH, OH-9), 4.06 (q, lH, J5~6~ = 6.6 Hz, H-5'), 4.84 (8, lH, H-10), 5.12 (m, lH, H-7), 5.48 (bd, lH, J1' 2. = 3.2 Hz, H-1'), 7.31 (dd, lH, Jl3 = 1 Hz, J2 3 = 8-4 Hz, H-3), 7-72 (t~
lH~ Jl2 = J23 = 8-3 Hz, H-2), 7.87 (dd, lH, Jl 2 = 7.7 Hz, J13 = 1 Hz, H-l), 9.03 (8~ lH, hydrazide-NH).
Exam~le 2:
Preparation of the compound of the formula I in which R1 is H and R2 is NH-(CHæl7-NH~:
7-0-(3'-N-Ethylenediaminocarbonvlmethyl-3'-N-methyl-~-L-daunosaminvl~-~-rhodomvcinone (compound 2) A solution of7-0-(3'-N-ethoxycarbonylmethyl-3'-N-methyl-~-L-daunosaminyl)-~-rhodomycinone (150 mg = 0.24 mmol) in dimethylformamide (20 ml) was mixed with 1 ml of ethyl-enediamine and stirred at room temperature in the dark for 3 h. The reaction mixture was then evaporated to dryness in a rotary evaporator and chromatographed on 15 g of silica gel in solvent mixture A (Rp 0.15), and the fractions were worked up in analogy to Example 1.
Yield: 120 mg ~0.19 mmol) = 79%
MS-FAB (M+H~) m/e = 630 1H-NMR (300 MHz, CDCl3/D6-DMSO) (4/1)) ~ 1.09 (t, 3H, Jl3 14 = 7.4 Hz, Me-14), 1.30 (d, 3H, J5 6. = 6.5 Hz, ~e-6'), 1.6-1.9 (m, 4H, CH2-2' and CH2-13), 2.19 (bs, 2H, CH2-8), 2.28 (s, 3H, N-CH3), 2.55 (m, lH, H-3'), 2.84 (m, 2H, CH2-c), 3.01 (d, lH, J~ = 18 Hz, H-a), 3.15 (d, lH, J,~.
= 18 Hz, H-a'), 3.25 (m, lH, H-b), 3.38 (m, lH, H-b'), 3.62 (bs, lH, H-4'), 4.05 (q, lH, J56 = 6.6 Hz, H-5'), 4.82 (s, lH, H-10), 5.12 (m, lH, H-7), 5.48 (bd, lH, Jl-2 = 3 Hz~ H-1'), 7-32 (dd, lH, J13 = 1 Hz, J23 = 8-4 Hz, H-3), 7.73 (t, lH, J12 s J23 = 8 Hz, H-2), 7.87 (dd, lH, Jl2 = 7.5 Hz, J~3 = l Hz, H-l), 8.06 (t, lH, J~ b = 6 Hz, -CO-NH-).
ExamDle 3:
Preparation of a compound of the formula I in which Rl is H and R2 is NH-(CH2~2-N(CH?)2 7-0-(3'-N-(Dimethylaminoethylaminocarbonyl)-3'-N-methyl-~-L-daunosaminyl~-~-rhodomycinone (com~ound 3) A solution of 7-0-(3'-N-ethoxycarbonylmethyl-3'-N-methyl-~-L-daunosaminyl)-~-rhodomycinone (16 mg = 0.026 mmol) in methanol (8 ml) was mixed with 100 ~1 (81 mg = 0.92 mmol) of N,N-dimethylaminoethylamine and 6tirred at room temperature in the dark for 16 h. The mixture was then evaporated to dryness in a rotary evaporator and dried under high vacuum. The reaction mixture was chromato-graphed on 4 g of silica gel in solvent mixture A (Rp 0.14), and the fractions were worked up in analogy to Example 1.
Yield 13 mg (0.020 mmol) = 77%
H-NNR (300 MHz, CDCl3) ~ 1-12 (t, 3H, Jl314 = 7-4 Hz, Me-14), 1.30 (d, 3H, J5.6. = 6.4 Hz, Me-6'), 1.7-2.05 (m, 4H, CH2-2' and CH2-13), 2.11 (dd, lH, J78~ = 4 Hz, J~b = 15 Hz, H-8a), 2.25 (d, lH, J~ ~b = 15 Hz, H-8b), 2.28 (8, 6H, N(CH3)2), 2.31 (~l 3H, N-CH3), 2.51 (m, 3H, CH2-c and H-3~)l 2.98 (d, lH, J~a = 18 Hz, H-a), 3.19 (m, lH, H-b), 3.20 (d, lH, J~ n~ = 18 Hz, H-a'), 3.68 (m, lHI H-b'), 5.04 (q, lH J5.6 = 6.5 Hz, H-5~), 4.10 (8l lH, OH-9), 4.91 (8l lH, H-10)~ 5.15 (m, lH~ H-7), 5.52 (bd, lH, J1.2. = 3 Hz, H-1~), 7-33 (d~ lH~ J23 = 8 Hz, H-3)~ 7.72 (t, lH, Jl2 =
J23 = 8 Hz, H-2), 7-89 (d, lH, Jl2 = 7 Hz, H-1), 8.14 (m, lH, NH).
Example 4:
Pre~aration of a com~ound of the formula I in which Rl is H and R2 i8 NH-(CH?)~
7-0-(3'-N-(4-Aminobutylaminocarbonyl)-3 ~ -N-methyl-~-L-daunosaminvl)--~-rhodomycinone (com~ound 4):
A solution of7-0-(3'-N-ethoxycarbonylmethyl-3'-N-methyl-~-L-daunosaminyl)-~-rhodomycinone (16 mg = 0.026 mmol) in acetonitrile (3 ml) was mixed with 200 ~1 of 1,4-diamino-butane and stirred at room temperature in the dark for 3 h. The reaction mixture was then evaporated to dryness in a rotary evaporator, and chromatographed on 3 g of silica gel in solvent mixture C (RF 0.16), and the fractions were worked up in analogy to Example 1.
Yield: 11 mg (0.017 mmol) = 65%
MS-FAB (M+H+) m/e = 658 - 11 - 2~398 lH-NMR (300 MHz, CDCl3/D6-DMSO (4/1)) ~ 1.10 (~, 3H, J13,14 = 7.4 Hz, Me-14), 1.32 (d, 3H, J5~ 6~ = 6.5 Hz, Me-6'), 1.4-1.6 (m, 4H, CH2-c and CH2-d), 1.6-2.0 (m, 4H, CH2-2' and CH2-13), 2.17 (bs, 2H, CH2-8), 2.29 (s, 3H, N-CH3), 2.55 (m, lH, H-3'), 2.69 (bt, 2H, Jd u = 6.5 Hz, CH2-e)~
3.06 (d, lH, J~a~ = 17 Hz, H-a), 3-08 (d, lH, Ja a' = 17 Hz, H-a'), 3.20 (bt, 2H, Jb c = 6.5 Hz, CH2-b), 3.71 (bs, lH, H-4'), 4.06 (q, lH, J5~ 6~ = 6.6 Hz, H-5'), 4.82 (s, lH, H-10), 5.12 (m, lH, H-7), 5.48 (bd, lH, Jl' 2~ = 3 Hz~
H-l'), 7-29 (dd, lH, Jl 3 = 1 Hz, J23 = 8.4 Hz, H-3), 7.70 (t~ lH~ Jl2 = J23 = 8 Hz, H-2), 7.85 (dd, lH, Jl2 = 7.5 Hz, H-l, Jl3 = 1 Hz, H-l).
Example 5:
7-0-(3'-N-Hi6taminocarbonylmethyl-3'-N-methvl-~-L-daunos-aminyl)-~-rhodomycinone (compound 5) A solution of 7-0-(3~-N-ethoxycarbonylmethyl-3'-N-methyl-~-L-daunosaminyl)-~-rhodomycinone (25 mg = 0.04 mmol) in dimethylformamide (7 ml) was mixed with 20 mg (0.18 mmol) of histamine and stirred at 50C in the dark for 30 h.
The reaction mixture was then evaporated to dryness in a rotary evaporator and chromatographed in solvent mixture C (RF 0.14), and the fractions were worked up in analogy to Example 1.
Yield: 21 mg (0.03 mmol) = 75 NS-FAB (M+Ht) m/e = 681 H-NMR (300 MHz, CDCl3) ~, 1.13 (t, 3H, Jl31~ = 7.4 Hz, Me-14), 1.40 (d, 3H, J5. 6' = 6.5 Hz, Me-6'), 1.6-2.0 (m, 4H, CHz-2' and CH2-13), 2-11 (dd, lH, J7Ba = 4 Hz, J8~Bb = 15 Hz, H-8a), 2.26 (d, lH, JôuBb = 15 Hz, H-8b), 2.29 (s, 3H, N-CH3), 2.30 (bd, lH, J2.3. = 11 Hz, H-3'), 2.78 (bt, 2H, Jbc = 5.6 Hz, CH2-c), 2.96 ~d, lH, Ja- = 17 Hz, H-a), 3.14 (d, lH, J,~. = 17 Hz, H-a'), 3.49 (m, 2H, CH2-b), 3.73 (8~ lH, H-4'), 4.10 (q, lH, J5 6~ = 6-3 Hz, H-5')~
4.92 (8~ lH, H-~0), 5.16 (m, lH, H-7), 5-51 (bd, lH, Jl.2.
= 3.5 Hz, H-l'), 6.81 (8, lH, H-d), 7.33 (dd, lH, Jl 3 =
1 Hz, J23 = 8.4 Hz, H-3), 7.56 (s, lH, H-e), 7.73 (t, lH, I
` - 12 - 2~ 98 Jl,2 = J2,3 = 8 Hz, H-2), 7.89 (dd, lH, Jl 2 = 7.5 Hz, Jl 3 =
1 Hz, H-1), 8.57 (bs, lH, NH).
Example 6:
7-0-(3'-N-Methvl-3'-N-tyraminocarbonylmethyl-~-~-daunos-aminvl)-B-rhodomycinone (compound 6) The reaction was carried out in analogy to Example 5 with 20 mg (0.032 mmol) of startin~ compound and 20 mg (0.15 mmol) of tyramine in dimethylformamide (3 ml), but with stirring at 50-60C for 50 h. Purification was carried out in solvent mixture C ( RF . 5)-Yield 12 mg (0.017 mmol) = 53%
MS-FAB (M+H~) m/e = 707 lH-NMR (300 NHz, CDCl3/D6-DMSO (4/1)) ~ 1.10 (t, 3H, Jl314 = 7.4 Hz, Me-14), 1.32 (d, 3H, J5. ~. = 6.5 Hz, Me-6'), 1.6-2.0 (m, 4H, CH2-2' and CH2-13), 2.19 (bs, 2H, CH2-8), 2.20 (s, 3H, N-CH3), 2.54 (bd, lH, J2' 3~ = 11 Hz~ H-3')~
2.69 (bt, 2H, Jb c = 7 Hz, CH2-c), 3.03 (d~ lH~ J~ a~ = 17 H~, H-a), 3.05 (d, lH, J~ a~ = 17 Hz, H-a'), 3.40 (m, 2H, CH2-b), 3.65 (bs, lH, H-4'), 3.89 (bs, lH, OH-9), 4.05 (q, lH, J5~ 6~ = 6.7 Hz, H-5'), 4.85 (s, lH, H-10), 5.12 (m, lH, H-7), 5.47 (bd, lH, Jl' 2~ = 3 Hz, H-l'), 6.74 (d, 2H, Jd ~ = 8.5 Hz, CH2-d), 6.97 (d, 2H, Jd ~ = 8.5 Hz, CH2-e), 7.32 (dd, lH, Jl 3 = 1 Hz, J2 3 = 8-4 Hz~ H-3)~ 7-49 (bt, lH, J~,b = 6 Hz, NH), 7-72 t (lH, Jl,2 = J2.3 = 8 Hz~
H-2), 7.87 (dd, lH, Jl 2 = 7-5 Hz, Jl 3 = 1 Hz~ H-l)-Example 7:
7-0-(3'-N-Methyl-3'-N-tryptaminocarbonylmethyl-~-L-daunosaminyl~-g-rhodomycinone (compound 7) The reaction was carried out in analogy to Example 5 with ~ --20 mg (0.032 mmol) of starting compound and 20 mg (0.12 mmol) of tryptamine in dimethylformamide (3 ml);
but with stirring at 50-60~C for 50 h. Purification was ~ '':'' ~,-,~, . .. . . .
13 2~ ~3~8 carried out in solvent mixture C (RF 0.46).
Yield 17 mg (O.023 mmol) = 72%
Ms-FAs (M+H+) m/e = 730 lH-NMR (300 MHz, CDCl3) ~, 1.13 (t, 3H, Jl314 = 7-4 Hz, Me S 14), 1.23 (d, 3H, J5~ 6' = 6.5 Hz, Me-6~), 1.5-2.0 ~m, 4H, CH2-2' and CH2-13), 2.12 (dd, lH, J7,8n = 4 Hz, J8~,~b = 15 Hz, H-8a), 2.17 (8, 3H, N-CH3), 2.20 (d, lH, J8~,8b = 15 Hz, H-8a~), 2.48 (m, lH, H-3~), 2.96 (bt, 2H, Jb c = 6.5 Hz, CH2-c), 3.46 (bs, lH, H-4~), 3.59 (m, 2H, CH2-b), 3.96 (s~ lH, OH-9), 4.00 (q, lH, J5~ 6' = 6-3 Hz~ H-5')~ 4-93 (s, lH, H-10), 5.11 (m, lH, H-7), 5.41 (bs, lH, H-l'), 7.0 (d, lH, Jd~ = 2.3 Hz, H-d), 7.08 (dt, lH) and 7.17 (dt~ lH) (Ja,f = Jf,g = J8,h = 7 Hz~ J~,e; = J~ h = 1 Hz~ H--f and H-g), 7.32 (dd, lH, Jl3 = 1 Hz, J2 3 = 8.4 Hz, H-3), 7.33 (d, lH, Je ~ = 8.0 Hz, H-e), 7.54 (d, lH, Jg h = 7.7 Hz, H-h), 7.72 (t, lH, Jl 2 = J~ 3 = 8 Hz, H-2), 7.88 (dd, lH~ Jl,2 = 7-5 Hz~ Jl,3 = 1 Hz, H-l), 8.29 (bs, lH, CO-NH).
Cytotoxicitv of com~ound~ of the formula I for L1210 mouse leukemia cells in vitro Procedure for determining the formation of colonie6 of L1210 leukemia cell6 in soft agar 500 leukemia cells per plate were incubated with various concentrations of the test substance at 37C for 1 hour.
The cells were then washed twice with McCoy5A medium and finally, after addition of 0.3~ agar, poured into Petri dishes. Controls were incubated only with fre~h medium.
In some cases, in place of the incubation for one hour, various concentrations and test substances were mixed with the upper agar layer in order thus to achieve continuous exposure of the cells throughout the incuba-tion time. After the agar had solidified, the plate6 were incubated in an incubator at 37C for 7 days (5% by vol.
CO2, 95~ relative humidity). The number of resulting colonies with a diameter of more than 60 ~m was then counted. The results have been reported as the number of colonies in treated agar plates as a percentage of the ~''~' "' ~' ' '' , . . .
~.` " ' '. ' ' . . ,, , ' .
- - 14 - 2~ 398 untreated control. The IC50 was determined as a measure of the activity of the substance from the dose-effect plot obtained in this way. The results for the compounds described here are compiled in Table 1, comparing with adriamycin.
- 20~5398 Table 1: Cytotoxicity of the prepared compound~ of the formula I on L1210 leukemia cells in vi~ro Substance Continuou~ 1 h No. incubation incubation (Example) Rl R2 IC50 (~g/ml) IC50 (~g/ml) 1 . R NH-NH2 0.32 0,21 b c 2 H NH-CH2-CH2-NH2 0~35 1.0 b c 3 H NH-CH2-CH2-N(CH3)2 0. 12 o . 2 6 b c d e 4 H NH-CH2-CH2-CH2-CH2-NH2 0~35 1~3 b c ~
H NH CH2 CH2 ~ ~ 0.31 1~0 b c ~
6 H NH CH2 2~H 0.41 1,3 7 H NH-CH2-CH2 ~ 0.36 1, 2 .~ . ,~,.:
~, . .. . . .
~, . .. . . .
Claims (9)
1. New anthracycline derivatives with cytostatic activ-ity and the formula I
I
and the physiologically acceptable salts thereof, where R1 is hydrogen or a hydroxyl group and R2 is NRa-(CH2)n-NRaRb with n = 0-12, preferably with n = 0 or 2-5, where Ra and Rb can each be hydrogen, C1-C4-alkyl or benzyl, or R2 is NRa -CHR3-CH2-R4, where R has the said meaning, and R3 can be hydrogen or a carboxyl group and R4 can be the residue of an L-amino acid or of a biogenic amine.
I
and the physiologically acceptable salts thereof, where R1 is hydrogen or a hydroxyl group and R2 is NRa-(CH2)n-NRaRb with n = 0-12, preferably with n = 0 or 2-5, where Ra and Rb can each be hydrogen, C1-C4-alkyl or benzyl, or R2 is NRa -CHR3-CH2-R4, where R has the said meaning, and R3 can be hydrogen or a carboxyl group and R4 can be the residue of an L-amino acid or of a biogenic amine.
2. Anthracycline derivative as claimed in claim 1, wherein R2 is NRa-(CH2)n-RaRb.
3. Anthracycline derivative as claimed in claim 2, wherein R2 is NRa-NRaRb, NRa-(CH2)2-NRaRb or NRa-(CH2)4-NRaRb, where derivatives with Ra= Rb=
hydrogen are preferred.
hydrogen are preferred.
4. Anthracycline derivative as claimed in claim 1, wherein R2 is NRa-CHR3-CH2-R4, where the compounds in which R3 is hydrogen are preferred.
5. Anthracycline derivatives as claimed in claim 4, wherein R4 is the residue of an aromatic L-amino acid or of an aromatic biogenic amine.
6. Anthracycline derivatives as claimed in claim 5, wherein R4 is the residue of the biogenic amine tyramine, tryptamine, histamine, dopamine or 5-hydroxydopamine.
7. A process for the preparation of anthracycline derivatives as claimed in claim 1, which comprises reacting a compound of the formula I in which R1 has the said meaning, and R2 is C1-C4-alkoxy, branched or unbranched, or benzyloxy, with a compound of the formula HRaN-(CH2)n-NRaRb in which n, Ra and Rb have the said meaning, or with a biogenic amine or an amino acid in a suitable solvent, preferably dimethylform-amide or ethanol, at a temperature between 0°C and the boiling point of the solvent, and isolating and purifying the compound of the formula I obtained in this way, in which R1 has the said meaning, and R2 is NRa-(CH2)n-NRaRb or NRa-CHR3-CH2-R4, where n, Ra, Rb, R3 and R4 have the said meaning.
8. An anthracycline derivative as claimed in claim 1 as pharmaceutical.
9. The anthracycline derivatives as claimed in claim 1 and substantially as described herein.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE3913742A DE3913742A1 (en) | 1989-04-26 | 1989-04-26 | CYTOSTATICALLY EFFECTIVE RHODOMYCINE DERIVATIVES |
| DEP3913742.2 | 1989-04-26 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2015398A1 true CA2015398A1 (en) | 1990-10-26 |
Family
ID=6379500
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002015398A Abandoned CA2015398A1 (en) | 1989-04-26 | 1990-04-25 | Rhodomycin derivatives with cytostatic activity |
Country Status (7)
| Country | Link |
|---|---|
| EP (1) | EP0394982A3 (en) |
| JP (1) | JPH02304099A (en) |
| KR (1) | KR900016246A (en) |
| AU (1) | AU5380490A (en) |
| CA (1) | CA2015398A1 (en) |
| DE (1) | DE3913742A1 (en) |
| PT (1) | PT93863A (en) |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2591599B1 (en) * | 1985-12-17 | 1988-08-05 | Hoechst Lab | NEW ANTHRACYCLINES AND MEDICINES CONTAINING THEM |
| JPS63227599A (en) * | 1987-03-14 | 1988-09-21 | Kirin Brewery Co Ltd | Anthracycline compound and use thereof |
| DE3819092A1 (en) * | 1988-06-04 | 1989-12-14 | Behringwerke Ag | CYTOSTATICALLY EFFECTIVE ANTHRACYCLINE DERIVATIVES |
| DE3913759A1 (en) * | 1989-04-26 | 1990-10-31 | Behringwerke Ag | New dimeric anthracycline derivs. - useful as antitumour agents |
-
1989
- 1989-04-26 DE DE3913742A patent/DE3913742A1/en not_active Withdrawn
-
1990
- 1990-04-23 AU AU53804/90A patent/AU5380490A/en not_active Abandoned
- 1990-04-24 PT PT93863A patent/PT93863A/en not_active Application Discontinuation
- 1990-04-24 KR KR1019900005720A patent/KR900016246A/en not_active Application Discontinuation
- 1990-04-25 JP JP2107720A patent/JPH02304099A/en active Pending
- 1990-04-25 CA CA002015398A patent/CA2015398A1/en not_active Abandoned
- 1990-04-25 EP EP19900107812 patent/EP0394982A3/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| PT93863A (en) | 1990-11-20 |
| DE3913742A1 (en) | 1990-10-31 |
| KR900016246A (en) | 1990-11-13 |
| EP0394982A2 (en) | 1990-10-31 |
| JPH02304099A (en) | 1990-12-17 |
| AU5380490A (en) | 1990-11-01 |
| EP0394982A3 (en) | 1991-09-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| RU2116087C1 (en) | Binding agent for bioactive preparations, method of its synthesis and pharmaceutical compositions | |
| CN106749478B (en) | 1,4 sensitive pH Di-substituted phthalocyanine Zn complexes and preparation method thereof and in application pharmaceutically | |
| EP1399411B1 (en) | Aloe-emodin derivatives and their use in the treatment of neoplastic pathologies | |
| IE904543A1 (en) | Morpholinyl derivatives of of doxorubicin and process for¹their preparation | |
| IL111725A (en) | 3'-Aziridino- anthracycline glycoside derivatives their preparation and pharmaceutical compositions containing them | |
| US4191755A (en) | Novel daunomycin derivatives, their aglycones and the use thereof | |
| CA2015398A1 (en) | Rhodomycin derivatives with cytostatic activity | |
| EP0381989B1 (en) | New 4'-epi-4'-amino anthracyclines | |
| Cogan et al. | Doxsaliform: A novel N-Mannich base prodrug of a doxorubicin formaldehyde conjugate | |
| EP0317956A2 (en) | Anti-tumor prodrugs | |
| CA2015395A1 (en) | Rhodomycin derivatives with cytostatic activity | |
| CN110183471B (en) | Piperazine derivative, preparation method and application | |
| WO1992004362A1 (en) | 2-hydroxy- and 2-acyloxy-4-morpholinyl anthracyclines | |
| EP0490878A1 (en) | New 3'-(4-Morpholinyl)- anthracycline derivatives | |
| EP0683787B1 (en) | 4'-o-sulfonyl-anthracycline derivatives | |
| EP0989989B1 (en) | 13-dihydro-3'aziridino anthracyclines | |
| KR102150418B1 (en) | Sitosterol-doxorubicin derivatives for radiotherapy amplification, and composition for preventing or treating cancer diseases comprising the same | |
| RU2081878C1 (en) | Anthracycline glycoside and method of its synthesis | |
| US10806788B2 (en) | Chlorin-vitamin conjugates | |
| GB2287463A (en) | Bis-anthracycline derivatives | |
| EP1395599B1 (en) | Anti-tumour anthracycline glycoside 14-sulfonic acid derivatives | |
| IE902703A1 (en) | Anthracycline derivatives | |
| JPH03193798A (en) | Anthracyclines, preparation thereof and drug containing same | |
| CN101161290A (en) | Beta-elemi alkene polyglycol amine derivant as well as its synthesis method and purpose | |
| GB2118932A (en) | A daunorubicin derivative |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Dead |