Patent Grant
8247424
This application is a national stage application (under 35 U.S.C. 371) of PCT/EP03/02922 filed Mar. 20, 2003 which claims benefit to German Application Serial No. 102 48 451.1 filed Oct. 17, 2002 and German Application Serial No. 102 13 580.0 filed Mar. 26, 2002.
The invention relates to novel fredericamycin derivatives, to drugs containing said derivatives or the salts thereof, and to the use of the fredericamycin derivatives for treating diseases, particularly cancer diseases.
Fredericamycin has been isolated 1981 from Streptomyces griseus, and demonstrates anti-cancer activity.
Fredericamycin and several fredericamycin derivatives are known.
In Heterocycles 37 (1994) 1893-1912, J. Am. Chem. Soc. 116 (1994) 9921-9926, J. Am. Chem. Soc. 116 (1994) 11275-11286, J. Am. Chem. Soc. 117 (1995) 11839-11849, JP 2000-072752, and in J. Am. Chem. Soc. 123 (2001), various total syntheses of fredericamycin A have been described, some being enantio-selective.
In U.S. Pat. No. 4,673,768, alkali salts of the fredericamycin A are described. In U.S. Pat. No. 4,584,377, fredericamycin derivatives are described, particularly derivatives acylated in ring E and F. In U.S. Pat. No. 5,166,208, fredericamycin derivatives are described as well, particularly derivatives carrying thio and amino substituents in ring F. The derivatives are generated semi-synthetically or fully synthetically.
Surprisingly it was found that fredericamycin derivatives, especially those derivatized in ring A, represent potent drugs. Also, a possibility was found to introduce such residues in ring A semi-synthetically, with which the water solubility and/or the biological effect, the spectrum of action in comparison with fredericamycin, can be significantly increased. Furthermore, an alternative method was found to make fredericamycin and its derivatives water-soluble by generating cyclodextrin inclusion compounds.
The invention relates to novel fredericamycin derivatives with the general Formula Ia or Ib:
wherein in each,
and the (CH2)r-chain elongated residue (CH2)rCH═N—N—(C3NX′R211R212R213R214) (with X′═NR215, O, S, and R211, R212, R213, R214, R215 being independently H or C1-C6 alkyl), —(CH2)rCH═N—NHSO2 aryl, —(CH2)rCH═N—NHSO2 heteroaryl, with r=0, 1, 2, 3, 4, 5, preferably 0,
Preferred are compounds of Formula IIa or IIb
wherein the meaning of the residues R1-R41, X is as described above, their tautomers and their physiologically tolerable salts or inclusion compounds, wherein the residues for Formula Ia may not concomitantly adopt the following meaning, except in the case of cyclodextrin inclusion compounds: R1: H, C1-C6 alkyl, R2: C1-C6 alkyl, C2-C6 alkenyl, R3: H, R4 and R6 identical, and independently H, C1-C6 alkyl, CO—R41, with R41 being C1-C6 alkyl, aryl, and R7 being H, C1-C6 alkyl, Y: O, and for Formula Ib: R1: H, R2: pentyl, 1-pentenyl, 3-pentenyl, 1,3-pentdienyl, R3: H, R4 and R6 being H, and X—R5 being methoxy, Y: O.
The invention further relates to compounds of Formula Ia, Ib, IIa or IIb, in which the residues R, except for R2, have the above described meanings, and the water solubility of R2 is at least two times higher, preferably at least five timer higher, more preferred at least ten times higher, especially preferred at least fifty time higher, particularly one hundred times higher, or even five hundred times higher than of R2 being CH═CH—CH═CH—CH3, when all other residues are maintained. The increase in the water solubility is achieved e.g. by introduction of groups which can form additional hydrogen bonds, and/or are polar, and/or are ionic. A key intermediate are compounds with an aldehyde function in R2.
For R2 preferred is also the group of the residues CmH2m+o−pY′p (with m=1 to 6, for o=1, p=1 to 2m+o; for m=2 to 6, o=−1, p=1 to 2m+o; for m=4 to 6, o=−2, p=1 to 2m+o; Y′=independently selected from the group of halogen, OH, OR21, NH2, NHR21, NR21R22, SH, SR21), (CH2)rCH2NHCOR21, (CH2)rCH2OCOR21, (CH2)rCH2NHCSR21, (CH2)rCH2S(O)nR21, with n=0, 1, 2, (CH2)rCH2SCOR21, (CH2)rCH2OSO2—R21, (CH2)rCH(OH)R21, (CH2)rCOOH, (CH2)rCOOR21, (CH2)rCONR21R22. Still particularly preferred is the group of the aldehyde-derived residues (CH2)rCHO, (CH2)rCH═NOH, —(CH2)rCH═NOR21, (CH2)rCH═NOCOR21, (CH2)rCH═NOCH2CONR21R22, (CH2)rCH═N—NHCO—R23, (CH2)rCH═N—NHC(O)NH—R23, (CH2)1CH═N—NHC(S)NH—R23, (CH2)rCH═N—NHC(NH)NH—R23, (CH2)rCH═N—NHC(NH)—R23, (CH2)rCH═N—NHCO—CH2NHCOR21, (CH2)rCH═N—O—CH2NHCOR21, (CH2)rCH═N—NHCS—R23, (CH2)rCH═CR24R25 (trans or cis), (CH2)rCH═NR21, (CH2)rCH═N—NR21R22,
and the (CH2)r-chain elongated residue (CH2)rCH═N—N—(C3NX′R211R212R213R214) (with X′═NR215, O, S, and R211, R212, R213, R214, R215 being independently H or C1-C6 alkyl), —(CH2)rCH═N—NHSO2 aryl, (CH2)rCH═N—NHSO2 heteroaryl, (CH2)rCH═CH heteroaryl, with r=0, 1, 2, 3, 4, 5, preferably 0.
From the aldehydes and thereof derived compounds, such are preferred in which at least R1 or R3 are not H, if R4 to R7 are H or alkyl.
Preferred residues in R2 are further heteroaryl, cycloaryl, alkylcycloalkyl, heterocycloalkyl, C1-C4 alkyl heterocycloalkyl, CmH2m+o−pY′p (with m=1 to 6, for o=1, p=1 to 2m+o; for m=2 to 6, o=−1, p=1 to 2m+o; for m=4 to 6, o=−2, p=1 to 2m+o; Y′=independently selected from the group of halogen, OH, OR21, NH2, NHR21, NR21R22, SH, SR21), CH2NHCOR21, CH2NHCSR21, CH2S(O)nR21, with n=0, 1, 2, CH2SCOR21, CH2OSO2—R21, CH(OH)R21, CH═NOCOR21, —CH═NOCH2CONR21R22, —CH═NOCH(CH3)—CONR21R22, CH═NOC(CH3)2CONR11R22, CH═N—NHCO—R23, —CH═N—NHCO—CH2NHCOR21, CH═N—O—CH2NHCOR21, —CH═N—NHCS—R23, CH═CR24R25 (trans or cis), CONR21R22, —CH═NR21, —CH═N—NR21R22,
(with X′═NR215, O, S, and R211, R212, R213, R214, R215 being independently H or C1-C6 alkyl), CH═N—NHSO2 aryl, H═N—NHSO2 heteroaryl.
Furthermore, compounds as described above are preferred, in which R3 means F, Cl, Br, I, OH, OR31, NO2, NH2, NHR31, NR31R32, NHCHO, NHCOR31, NHCOCF3, CH3-mhalm (with hal=Cl, F, particularly F, and m=1, 2, 3), OCOR31, with the above described meanings for R31, R32.
Also preferred are compounds as described above, in which X means N or S, especially when R3 is H or halogen, and/or R2 is alkenyl, particularly butadienyl or 1,3-pentdienyl.
Also preferred are compounds as described above, in which X—R5 is OH, and particularly their salts, and preferred in compounds of Formula Ia or Ia, since this acidic OH group may easily be deprotonized, which increases the water solubility and/or the biological efficacy.
Furthermore preferred are still compounds as described above, wherein the residues R preferably independently adopt one or more of the following meanings:
(with X′═NR215, O, S, and R211, R212, R213, R214, R215 being independently H or C1-C6 alkyl), —CH═N—NHSO2 aryl, —CH═N—NHSO2 heteroaryl, CH═N—NHCO—R23,
For R2 also preferred is the residue —CHOHCHOHCHOHCHOHCH3.
Furthermore, the following residues are preferred for R2: —CHCH-2-methyl-4-thiazyl, particularly
wherein R″ particularly is alkyl or NHCO alkyl, CH═NOR21, with R21 being methyl, ethyl, n-propyl, isopropyl, n-butyl, n-hexyl, benzyl, halogen benzyl, particularly fluorobenzyl and chlorobenzyl, —CH2CH2 morpholinyl.
Especially preferred are the compounds, the stereo isomers, tautomers, and physiologically tolerable salts or inclusion compounds of which, selected from the group consisting of the compounds of the examples and the compounds, demonstrate combinations of the various substituents of the examples.
Particularly preferred for R3 is H, F, Cl, Br, J, particularly F, Cl, Br, J.
Particularly preferred for R2 is C1-C8 alkyl, C2-C8 alkenyl, CH═NOR1, with R21 being C1-C8 alkyl, C1-C8 alkenyl, aryl or heteroaryl, C1-C2 alkylaryl, particularly benzyl, C1-C2 alkylheteroaryl, wherein aryl or heteroaryl in particular have only one ring system which may be substituted once or twice with a substituent such as halogen, methyl, CF3, OH, OMe.
Particularly preferred are derivatives of fredericamycin A in which only the above indicated, particularly preferred meanings of R2 and/or R3 are realized.
The invention furthermore relates to drugs containing the above compounds of Formula I or II together with the usual carriers and adjuvants.
Also preferred are the above mentioned drugs in combination with other agents for cancer treatment.
These compounds according to the invention are used for preparation of drugs for treatment of cancers, particularly such that may be treated by inhibition of the topoisomerases I and/or II. Cancers that can be treated with the substances according to the invention are e.g. leukemia, lung cancer, melanomas, uterus tumors, prostate tumors and colon tumors.
Also, fredericamycin A and its derivatives act against an unknown target in the cell cycle leading to apoptosis in tumor cells. Furthermore, the compounds according to the invention, and compounds which have concomitantly adopted the following meanings in Formula Ia: R1: H, C1-C6 alkyl, R2: C1-C6 alkyl, C2-C6 alkenyl, R3: H, R4 and R6 identically and independently H, C1-C6 alkyl, CO—R41, with R41 being C1-C6 alkyl, aryl, and R7 being H, C1-C6 alkyl, and in Formula Ib: R1: H, R2: pentyl, 1-pentenyl, 3-pentenyl, 1,3-pentdienyl, R3: H, R4 and R6 being H and X—R5 being methoxy, are used for preparation of drugs for treatment of neurodermitis, parasites and for immunosuppression.
The invention also relates to a method for preparation of fredericamycin derivatives in which R2 as intermediate is —CHOHCHOHCHOHCHOHCH3. These compounds are preferably transformed into aldehydes for further derivatization.
In the description and the claims, the substituents are described by the following definitions:
The term “alkyl” by itself or as part of another substituent means a linear or branched alkyl chain radical of the respectively indicated length, in which optionally a CH2 group may be substituted by a carbonyl function. Thus, C1-4 alkyl may be methyl, ethyl, 1-propyl, 2-propyl, 2-methyl-2-propyl, 2-methyl-1-propyl, 1-butyl, 2-butyl, C1-6 alkyl, e.g. C1-4 alkyl, pentyl, 1-pentyl, 2-pentyl, 3-pentyl, 1-hexyl, 2-hexyl, 3-hexyl, 4-methyl-1-pentyl, or 3,3-dimethylbutyl.
The term “C1-C6 alkylhydroxy” by itself or as part of another substituent means a linear or branched alkyl chain radical of the respectively indicated length which may be saturated or unsaturated, and which carries an OH group, e.g. hydroxymethyl, hydroxymethyl, 1-hydroxypropyl, 2-hydroxypropyl.
The term “alkenyl” by itself or as part of another substituent means a linear or branched alkyl chain radical with one or more C═C double bonds of the respectively indicated length, several double bonds being preferably conjugated. Thus, C2-6 alkenyl may for example be ethenyl, 1-propenyl, 2-propenyl, 2-methyl-2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 1,3-butdienyl, 2,4-butdienyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 1,3-pentdienyl, 2,4-pentdienyl, 1,4-pentdienyl, 1-hexenyl, 2-hexenyl, 1,3-hediexyl, 4-methyl-1-pentenyl, or 3,3-dimethylbutenyl.
The term “alkinyl” by itself or as part of another substituent means a linear or branched alkyl chain radical with one or more C—C triple bonds of the respectively indicated length. Thus, C2-6 alkinyl may for example be ethinyl, 1-propinyl, 2-propinyl, 2-methyl-2-propinyl, 2-methyl-1-propinyl, 1-butinyl, 2-butinyl, 1,3-butdiinyl, 2,4-butdiinyl, 1-pentinyl, 2-pentinyl, 3-pentinyl, 1-hexinyl, 2-hexinyl, 4-methyl-1-pentinyl, or 3,3-dimethylbutinyl.
The term “halogen” stands for fluorine, chlorine, bromine, iodine, preferably bromine and chlorine.
The term “NR21R22” preferably stands for a dialkylamino group, wherein the two alkyl groups together with the N can form a ring with 5 or 6 members with optionally one more heteroatom N or O.
The term “cycloalkyl” by itself or as part of another Substituent comprises unsaturated (mono or poly, preferably mono) or saturated, cyclic hydrocarbon groups with 3 to 10 C atoms, preferably 3 to 8 C atoms, such as e.g. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohex-2-enyl, cyclohex-3-enyl, cyclohex-2,4-dienyl, 4-methylcyclohexyl, 3-methylcyclohexyl, cycloheptyl or cyclooctyl. Saturated cycloalkyls are preferred. The cycloalkyls may be substituted with up to 3 substituents, preferably with up to 1 substituent, wherein the substituents independently can have the meaning C1-C6 alkyl, OH, NO2, CN, CF3, OR11, SH, SR11, C1-C6 alkylhydroxy, C1-C6 alkyl-OR11, COOH, COOR11, NH2, NHR11, NR11R12, halogen, aryl, C1-C4 alkylaryl, heteroaryl, C1-C4 heteroalkylaryl, wherein the residues R11 and R12 independently can mean C1-C10 alkyl, cycloalkyl, C1-C4 alkylcycloalkyl.
The term “heterocycloalkyl” by itself or as part of another substituent includes cycloalkyl groups, wherein up to two CH2 groups may be substituted by oxygen, sulfur or nitrogen atoms, and one or two other CH2 groups may be substituted by one or two carbonyl function(s), carbothionyl function(s), or a carbonyl function and a carbothionyl function, for example pyrrolidine, piperidine, morpholine or
The heterocycloalkyls may be substituted as with the cycloalkyls.
The term “aryl” by itself or as part of another substituent includes aromatic ring systems with up to 3 rings, in which at least 1 ring system is aromatic, and those with up to 3 substituents, preferably up to 1 substituent, wherein the substituents independently can have the meaning C1-C6 alkyl, OH, NO2, CN, CF3, OR11, SH, SR11, C1-C6 alkylhydroxy, C1-C6 alkyl-OR11, COOH, COOR11, NH2, NHR11, NR11R12, halogen, wherein the residues R11 and R12 independently can mean C1-C10 alkyl, cycloalkyl, C1-C4 alkylcycloalkyl, or R11 and R12, together with the N, form a ring with 4, 5, 6, 7 or 8 members optionally containing still another heteroatom selected from the group N, O, S.
Apart from phenyl and 1-naphthyl and 2-naphthyl, preferred aryls are:
The term “heteroaryl” by itself or as part of another substituent includes aromatic ring systems with up to 3 rings and with up to 3 identical or different heteroatoms N, S, O, in which at least 1 ring system is aromatic, and those with up to 3 substituents, preferably up to 1 substituent, wherein the substituents independently can have the meaning C1-C6 alkyl, OH, NO2, CN, CF3, OR11, SH, SR11, C1-C6 alkylhydroxy, C1-C6 alkyl-OR11, COOH, COOR11, NH2, NHCOR11, NHR11, NR11R12, halogen, or phenyl, wherein the residues R11 and R12 independently can have the above indicated meanings.
Preferred heteroaryls are:
The term “ring system” generally refers to rings with 3, 4, 5, 6, 7, 8, 9, or 10 members. Preferred are rings with 5 and 6 members. Furthermore, ring systems with one or 2 annealed rings are preferred.
The compounds of Formula I may be present as such, or, if they contain acidic or basic groups, in the form of their salts with physiologically tolerable bases or acids. Examples for such acids are: hydrochloric acid, citric acid, trifluoracetic acid, tartaric acid, lactic acid, phosphoric acid, methane sulfonic acid, acetic acid, formic acid, maleic acid, fumaric acid, succinic acid, hydroxysuccinic acid, sulfuric acid, glutaric acid, aspartic acid, pyruvic acid, benzoic acid, glucuronic acid, oxalic acid, ascorbic acid, and acetylglycine. Examples for bases are alkali ions, preferably Na, K, particularly preferred the tri-potassium and tri-sodium salts, alkaline earth ions, preferably C, Mg, ammonium ions.
The compounds according to the invention may be administered orally in the usual way. The application may also be i.v., i.m., with vapors, or sprays through the nasopharynx.
The dosage depends on age, condition and weight of the patient as well as on the type of application. Usually, the daily dose of the active ingredient per person is between 0.1 μg/kg and 1 g/kg orally. This dosage may be given as 2 to 4 split dosages, or once per day as a slow release form.
The novel compounds may be used in the usual solid or liquid pharmaceutical application forms, e.g. as tablets, film tablets, capsules, powder, granules, coated tablets, solutions, or sprays. These are produced in the usual way. The agents can be processed with the usual pharmaceutical adjuvants such as tablet binders, fillers, preservatives, disintegrants, flow regulators, plasticizers, wetting agents, dispersants, emulsifiers, solvents, retardation agents, antioxidants, and/or propellants (see H. Sucker et al.: Pharmazeutische Technologie, Thieme-Verlag, Stuttgart, 1978). Usually, the so obtained application forms contain the active ingredient in amounts of 0.1 to 99 percent per weight.
Experimental Part
Fredericamycin A can be prepared by fermentation or fully synthetically according to the known methods. The reduced forms of the Formulas Ib and IIb can be obtained from the appropriate compounds of Formulas Ia and Ia using mild reducing agents.
Preparation of the Substances
For synthesis of water soluble fredericamycin derivatives, fredericamycin (1) was first hydroxylized with osmium(IV)oxide at the diene side chain. The resulting compound (2) shows significantly higher water solubility compared to the original compound fredericamycin (1). In order to further increase the water solubility, (2) was transformed into the tri-potassium salt (3) (see diagram 1).
The fredericamycin tetrol (2) serves, among others, as an important intermediate for the synthesis of other fredericamycin derivatives with increased solubility and/or better action profile. By iodate cleavage with sodium periodate or carrier-bound periodate, the tetrol side chain may be degraded with very high yields to fredericamycin aldehyde (4) (see diagram 2).
The fredericamycin aldehyde (4) can be reacted with acylhydrazones, hydroxylamine, and O-alkylhydroxylamine to the appropriate hydrazone (see diagram 3), or oxime and oximether (see diagram 4). The reaction can be performed at room temperature in solvents such as DMF or pyridine, and is finished after a few minutes to hours.
Synthesis of Hydrazones
Synthesis of Oximether
Analogously, the compounds 100-242 can be generated according to the instructions below (table 3). The hereby used hydrazines, hydrazones and hydroxylamines are available commercially, or have been produced according to instructions known from the literature.
Reduction and Oxidation of Fredericamycin Aldehyde (4)
Fredericamycin aldehyde (4) can be reacted with a common reducing agent such as sodium borohydrid in a solvent such as DMF or pyridine to hydroxymethyl fredericamycin (11). The reaction can be summarized as a single pot reaction (iodate cleavage of fredericamycin tetrol (2) to fredericamycin aldehyde (4) (see diagram 2) and reduction without isolation of the intermediates to fredericamycin alcohol (11)).
Fredericamycin aldehyde (4) can be oxidized with the oxidizing agent sodium chlorite (NaClO2), a buffer such as sodium dihydrogenphosphate in presence of an alkene such as 2,3-dimethylbutene with very good yields to fredericamycin carboxylic acid (12). The usually employed oxidation methods such as those being used in preparative chemistry for the oxidation of aldehydes to carboxylic acids (oxidation with chromium(VI) compounds, manganese(VII) compounds as well as peroxo acid) did not lead to success. Only the use of the above described oxidation method provided the desired product. The literature describes oxidations of 2-pyridone-6-aldehydes with silver ions and potassium permanganate in an alkaline medium. This method, however, is not suited for fredericamycin and its derivatives since fredericamycin (1) contains base-labile (-reactive) groups (OH groups) causing undesired side reactions.
The potassium salt of the fredericamycin acid (13) was obtained according to a common method by stoichiometric neutralization.
Substitution in the B Ring
Fredericamycin (1) can be reacted with halogenation agents such as N-bromosuccinimide (NBS) and N-iodosuccinimide (NIS) with good yields to the substituted 5-bromo or 5-iodo fredericamycin derivatives (14) and (15) (diagram 6). The fredericamycin aldehyde (4) and (36) can be transformed with elemental bromine, NBS, BrI, NIS, and NCS to the appropriate halogen-substituted fredericamycin aldehyde (37), (38) and (39).
The appropriate fluorine compound is accessible, too.
Both of the two following fredericamycin compounds (23) and (24) are also precursors. (23) is the precursor for an amino acid-linked fredericamycin derivative.
The preparation of (23) may be recognized as proof that the aldehyde (4) may be reacted with phosphorylides according to Wittig or Wittig-Horner (see diagram 7).
The compound (24) is the precursor of an N-methylated fredericamycin derivative (diagram 8).
Fredericamycin may be transformed by palladium/hydrogen almost quantatively to tetrahydro fredericamycin (25), and may be halogenated in the nucleus according to the above described methods, e.g. to the bromine compound (26) (diagram 9):
Surprisingly it has also been found that the methoxy groups in fredericamycin and the derivatives according to the invention can be exchanged under alkali or earth alkali acetate catalysis by oxygen nucleophiles such as alcohols or polyols. Thereby, the alcohols can carry a multitude of different substituents (table 4).
Exchange of the Methoxy Group at the F Ring
The exchange of the methoxy groups at the F ring of the fredericamycin and at the derivatives is possible by primary, secondary or aromatic amines. Thereby, the components are stirred with the appropriate primary or secondary amines at room temperature in DMF or in another inert solvent. With aromatic amines, a catalysis with Lewis acids such as stannous(IV)chloride, etc. is required.
Preparation of Heterocyclic Fredericamycin Derivatives
The fredericamycin aldehyde (4) can be reacted to pyridal acetone (271) according to Wittig or Wittig-Horner. Bromation with bromine in DMF yields the dibromo-derivative (272) substituted in the side chain and at the B ring. With the appropriately substituted thioamides or thioureas, the respective thiazole derivatives (273-276) are accessible.
Preparation of Thioanalogoues of Fredericamycin Derivatives
By sulfurization of fredericamycin or its derivatives with Lawesson reagent or P4S10 in pyridine, the derivatives analogous to thiopyridone are accessible (see diagram 13).
Preparation of Thioanalogoues of Fredericamycin Derivatives
By sulfurization of fredericamycin or its derivatives with Lawesson reagent or P4S10 in pyridine, The derivatives analogous to thiopyridone are accessible (see diagram 13).
Fredericamycin (1) forms inclusion compounds such as (25) with polysugars such as α-cyclodextrin, that have good water solubility compared to the original substance.
The dextrin inclusion compounds form easily if the components are mixed in the appropriate stoichiometric ratio in a suitable solvent such as DMSO (see diagram 11).
Biological Activity Against 12 Cancer Cell Lines:
LCL (H460, lung), MACL (MCF7, breast), LXFL (52L, lung), LXFA (629L, lung), MEXF (462NL, melanoma), MEXF (514L, melanoma), MAXF (401NL, breast), RXF (944L, renal), RXF (486L, renal), UXF (1138L, uterus), PRXF (PC3M, prostate), PRXF (22RV1).
Efficacy (IC70) Averaged Over all Cell Lines in μg/mL at 5 Test Concentrations
Two hundred (200) mg (0.38 mmol) fredericamycin A (1) are dissolved in 30 mL dichloromethane. After addition of 20 mL methanol and 4.4 ml water, 350 mg (2.6 mmol) N-methylmorpholine-N-oxide are added. Under vigorous stirring, 0.2 ml of a 2.5% osmium(IV)oxide solution in t-butanol is added dropwise. The reaction mixture is acidified with 2-3 drops of trifluoracetic acid. After stirring for 48 hours, the reaction is complete according to HPLC control (RP18, acetonitrile water (0.2% acetic acid)). The reaction mixture is added to 400 ml water under vigorous stirring, and the dark red crystalline solid is sucked off through a filter. Drying in HV. Yield: 195 mg (87% of the theoretical value) dark red powder. ES−: M/e=606.2 (M+−H), λmax: 504.0.
Twelve (12.0) mg (19.8 μmol) fredericamycin tetrol (2) are dissolved in 1.5 mL absolute pyridine under nitrogen atmosphere. The solution is gassed for 30 min with argon at 0° C. Under the argon atmosphere, 5.94 mL of a 0.01 N KOH solution are added at once at 0° C. The reaction solution immediately turns turquoise. The reaction mixture is stirred for another 1 hour, and subsequently is frozen and lyophilized. Yield: 13.2 mg (100% of the theoretical value); deep blue crystal mass.
1.) Fifty (50) mg (82.3 μmol) tetrahydroxy fredericamycin (tetrol (2)) are dissolved in 4 mL DMF. Under vigorous stirring, an aqueous sodium iodate solution (300 mg NaIO4 in 1 mL water) is added dropwise within one hour. After 1 h stirring at room temperature, 2 drops of trifluoracetic acid are added. After stirring for another 30 min, the reaction solution is diluted with 3 ml DMF, and 150 mg NaIO4 dissolved in 0.5 ml water are added.
After another hour, 100 mL water are added. The supernatant over the precipitate is sucked off, and dryed in HV. Dark red crystal powder. Yield: 41 mg (100% of the theoretical value). M/e=501.3, UVmax: 504.0 nm.
2.) One hundred and nine (109) mg (179 μmol) fredericamycin tetrol (2) are dissolved in 8 mL pyridine. 180 μL water are added. To the reaction mixture, 450 mg (1.08 mmol, 6 eq.) (polystryrylmethyl)trimethylammonium periodate resin are added. Then the mixture is stirred for 12 h at RT. The resin is filtered off; washing and concentrating until dry. Dark red residue.
Yield: 89.9 mg (100% of the theoretical value). M/e=501.3, UVmax: 504.0 nm.
Twenty (20) mg (39.9 μmol) fredericamycin aldehyde (4) are dissolved under argon in 1.5 mL absolute DMF. Addition of 9.1 mg (47.9 μmol, 1.2 eq.) acetylhydrazide dimethylammoniumchloride (Girard reagent D) and 20 mg polyvinylpyridine (2% DVB). The mixture is stirred for 2.5 h. Then, 27 mg (80 μmol, 2.0 eq.) aldehyde Wang resin (coating: 3.0 mmol/g) are added and stirred for another 1 h. Then, the resin is filtered, and washed 3× with DMF. Concentration in high vacuum. The residue is dissolved in 1 ml trifluoracetic acid, and concentrated after 10 min until dry.
Red solid; Yield: 28.5 mg (100%); ES+: M/e=601.3, UVmax: 504.0 nm.
Fifteen (15) mg (29.9 μmol) fredericamycin aldehyde (4) are dissolved in 3 mL DMF. At room temperature 7.5 mg (40.0 μmol) acethydrazinopyridinium chloride (Girard reagent P) dissolved in 75 μL water are added. The reaction mixture is stirred for 1.5 h at room temperature, and the course of the reaction is monitored by HPLC. When finished, acetic acid ethyl ester is added to the reaction mixture, until a precipitation occurs. After the crystallization is finished, the red solid is sucked off.
Yield: 9.1 mg (44% of the theoretical value). M/e=635.2; λmax: 486.0.
Ten (10) mg (19.4 μmol) fredericamycin aldehyde (4) are dissolved in 2 mL DMF. After addition of 3.1 mg (44.6 μmol) hydroxylammonium chloride, 3.2 μl pyridine are added. Stirring for 2 h at room temperature. The reaction mixture is added to 50 ml water and extracted 3 times with ethyl acetate. After drying and concentration, a deep red amorphous crystal powder was left (HPLC clean).
Yield: 7.4 mg (72% of the theoretical value). ES−: M/e=516.1; λmax: 500.0 nm.
Ten (10) mg (19.4 μmol) fredericamycin aldehyde (4) are dissolved in 2 mL DMF. After addition of 3.4 mg (40.7 μmol) O-methylhydroxylammonium chloride and 3.2 μl pyridine, the reaction mixture is stirred for 2 h at room temperature. Then, it is added to 100 ml water, and the supernatant is sucked off from the red precipitate (HPLC clean).
Yield: 7.6 mg (71% of the theoretical value). ES+: M/e=531.2; λmax: 500.0.
Ten (10) mg (19.4 μmol) fredericamycin aldehyde (4) are dissolved in 2 mL DMF. After addition of 6.4 mg (43.2 μmol) O-benzylhydroxylammonium chloride and 3.2 μl pyridine, the reaction mixture is stirred for 2 h at room temperature. Then, it is added to 50 ml water, and the supernatant is sucked off from the red precipitate (HPLC clean).
Yield: 6.8 mg (57% of the theoretical value). ES+: M/e=607.2; λmax: 504.0 nm.
Two (2.0) mg (4.0 μmol) fredericamycin aldehyde (4) are dissolved in 150 μL DMF, and 0.86 mg (4.4 μmol) β-aminoxy-D-glucopyranose is added. The mixture is stirred for 24 h at room temperature, and 5 mg (15.0 μmol) aldehyde Wang resin (coating: 3.0 mmol/g) is added. After stirring for another 3 h, the resin is filtered off, washed with DMF, and the filtrate is concentrated in high vacuum until dry.
Yield: 2.7 mg (99% of the theoretical value), red powder; ES−: M/e=678.1; λmax: 504.0 nm.
Thirty (30) mg (49.4 μmol) tetrahydroxy fredericamycin (2) were dissolved in 2 mL pyridine. Twenty (20) mg (93.0 μmol) sodium metaperiodate dissolved in 0.3 ml water are added. After stirring for 4 h, 10 mg (260 μmol) sodium borohydride are added. After 12 h, concentration until dry, and the residue is separated by preparative HPLC.
Yield: 2.6 mg (13% of the theoretical value) red powder. ES−: M/e=503.2; λmax: 504.0 nm.
Fifteen (15) mg (29.9 μmol) fredericamycin aldehyde (4) are dissolved in 1 mL dichloromethane and 0.5 ml t-butanol. Addition of 250 μl 2,4-dimethylbutene. Under stirring at room temperature, a solution of 6.0 mg (53.1 μmol) sodium chlorite (80%) and 5.1 mg sodium hydrogenphosphate in 250 μl water are added dropwise.
After 2.5 h, again a solution of 10.0 mg (88.5 μmol) sodium chlorite and 5 mg sodium dihydrogenphosphate in 200 μl water are added. After altogether 4 h, it is put on water, and extracted with ethyl acetate.
The raw mixture was purified by preparative HPLC (RP18, acetonitrile-water-acetic acid). Red amorphous powder.
Yield: 68.3 mg (53.5% of the theoretical value). E−: M/e=516.1; λmax: 504.0 nm.
6.9 mg (13.3 μmol) Fredericamycin carboxylic acid (12) are dissolved in 5 mL DMF under nitrogen. At room temperature and under oxygen exclusion and vigorous stirring, 1.27 mL (12.7 μmol) of an aqueous 0.01 N KOH solution is added dropwise. It is stirred for 15 minutes at room temperature, and concentrated in high vacuum until dry.
Yield: 7.40 mg (100% of the theoretical value). E−: M/e=516.1; λmax: 504.0 nm.
Twenty (20) mg (37.1 μmol) fredericamycin (1) were dissolved in 250 μl DMF, and then 6. 3 mg (35.3 μmol) N-bromosuccinimide in 250 μl DMF were added within one hour at 0° C. The reaction was stirred in a slowly thawing ice bath over night. Then, the DMF is removed in high vacuum, and the residue is purified by preparative HPLC.
Yield: 7 mg (32% of the theoretical value) red crystal mass. M/e=616.1/618.1; λmax: 486.0 nm.
Eighty four (84) mg (158 μmol) fredericamycin (1) were dissolved in 1.0 μl DMF, and then 33.0 mg (150.0 μmol) N-iodosuccinimide in 500 μl DMF were added within one hour at 0° C. The reaction was stirred in a slowly thawing ice bath over night. Then, the DMF is removed in high vacuum, and the residue (120 mg (14) with a content of 80%) is purified by preparative HPLC (gradient CH3CN 50-90% over 16 min.)
Yield: 18 mg (17% of the theoretical value) red crystal mass. M/e=665.0; λmax: 484.0 nm.
Sixty six (66) mg (200 μmol) Z-α-phosphonoglycine trimethylester are dissolved under argon in 1 mL absolute pyridine, and 25 μL 1,1,3,3-tetramethylguanidine are added at 0° C. After 40 min. 20 mg (40 μmol) fredericamycin aldehyde (4) is added at 0° C. After 15 min. 20 ml 1 M acetic acid is added, and the mixture is extracted 3× with acetic acid. The raw product is purified by preparative HPLC (RP18, acetonitrile-water).
Yield: 10.0 mg (36% of the theoretical value). M/e=706.4; λmax: 492.0 nm.
Ten (10) mg (15 μmol) fredericamycin (1) were dissolved under protective gas in 4 ml absolute DMF. At RT, 400 μl (4311 μmol) methyliodide and 81 mg powdered potassium carbonate are added. The reactions mixture is then stirred at RT for 20 h, and is then transferred onto water. Extraction with ethyl acetate, and purification of the residue by separating chromatography on chloroform/methanol 30/1.
Yield: 4 mg (37% of the theoretical value). Yellow residue. M/e=582.3; λmax: 368.0 nm.
Ten (10) mg fredericamycin (0.025 mMol) are added to a solution of 50 mg α-cyclodextrin (0.050 mMol) in 500 μl dimethylsulfoxide. The solution is then diluted with 5 ml water. A stock solution prepared in such way can be diluted as desired with water.
λmax=504.0 nm.
Five (5) mg (9.42 μmol) fredericamycin aldehyde (4) are dissolved in 500 μl DMF and 25 μl trifluoracetic acid. At room temperature, 1.30 mg (11.3 μmol) 1-amino-4-methyl-piperazine is added. After stirring for 4.5 h at room temperature, 1 equivalent each of Wang aldehyde resin and sulfonohydrazide resin is added and stirred for 2 h.
Filtration and concentration of the reaction solution at high vacuum.
Red powder. Yield: 5.4 mg (91% of the theoretical value). M/e=599 (M+H)+; λmax: 504.0 nm.
Five (5.00) mg (9.42 μmol) fredericamycin aldehyde (4) are dissolved in 500 μl DMF and 25 μl trifluoracetic acid. At room temperature, 2.05 mg (11.3 μmol) 2-hydrazino-2-imidazolin hydrobromide is added. After stirring for 4.5 h at room temperature, 1 equivalent each of Wang aldehyde resin and sulfonohydrazide resin are added and stirred for 2 h. Separation of the resin by filtration and concentration of the reaction solution at high vacuum.
Red powder. Yield: 3.9 mg (67% of the theoretical value). M/e=584 (M+H)+; λmax: 504.0 nm.
Five (5.00) mg (9.42 μmol) fredericamycin aldehyde (4) are dissolved in 500 μl DMF and 25 μl trifluoracetic acid. At room temperature, 1.67 mg (11.3 μmol) 2N-aminorhodanide are added. After stirring for 4.5 h at room temperature, 1 equivalent each of Wang aldehyde resin and sulfonohydrazide resin are added and stirred for 2 h.
Filtration and concentration of the reaction solution.
Red powder. Yield: 4.1 mg (65% of the theoretical value). M/e=599 (M+H)+; λmax: 504.0 nm.
Five (5.00) mg (9.42 μmol) fredericamycin aldehyde (4) are dissolved in 500 μl DMF and 25 μl trifluoracetic acid. At room temperature, 2.47 mg (11.3 μmol) N-(aminoxyethyl)morpholine dihydrochloride is added. After stirring for 4.5 h at room temperature, 1 equivalent of Wang aldehyde resin (3.1 mg, 9.4 μmol, coating: 3.0 mmol/g) as well as 1 equivalent sulfonohydrazide resin (6.1 mg, 9.4 mmol, 1.5 mmol) are added and stirred for 2 h.
Filtration and concentration of the reaction solution.
Red powder. Yield: 6.1 mg (98% of the theoretical value). M/e=630 (M+H)+; λmax: 504.0 nm.
Three hundred (300) mg (556.6 μmol) fredericamycin (1) are dissolved under argon in 10 μl DMF, and then 75.0 mg (556.6 μmol) N-chlorosuccinimide are added. The reaction is stirred for 5 h at 40° C. The reaction mixture is then added to 400 ml methanol/water 1:1, and the red precipitate is sucked off and dried at high vacuum.
Yield: 305 mg (96% of the theoretical value) red crystal mass. M/e=573/575; λmax: 504.0 nm.
Fifty (50) mg (92.8 μmol) fredericamycin (1) are dissolved in 5 ml DMF under argon, and then 33.0 mg (93.5 μmol) 1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate) Selectfluor (is added. The reaction is stirred for 24 h at room temperature. The reaction mixture is then added to 200 ml water, and is extracted with ethyl acetate. The concentrated raw product is purified by preparative HPLC (RP18, acetonitrile-water-acetic acid).
Yield: 7.1 mg (14% of the theoretical value) red crystal mass. M/e=557; λmax: 504.0 nm.
Hundred twenty (120) mg (209 mmol) chlorofredericamycin (34) are dissolved in 25.0 ml dichloromethane. After addition of 3.6 ml methanol and 0.8 ml water, 197 mg (1.46 mmol) N-methylmorpholine-N-oxide is added. Under vigorous stirring, 0.12 ml of a 2.5% solution of osmium(IV)oxide in t-butanol is added dropwise. After stirring for 27 hours, the reaction is complete, according to HPLC monitoring (RP18, acetonitrile-water (0.2% acetic acid)). The reaction mixture is added to 200 ml water under vigorous stirring, and the dark red solid is sucked off. Drying in HV.
Yield: 101 mg (75% of the theoretical value) dark red powder. M/e=641/643; λmax: 504.0.
Hundred (100) mg (200 μmol) fredericamycin aldehyde (4) are dissolved under argon in 5 ml DMF. Then, 200 μl of a 1M bromine solution in DMF is added. After stirring for 1.5 h at RT, another 20 μl bromine solution are added. According to HPLC monitoring, the reaction mixture is complete after 3.5 h.
Add to 150 ml water, and shake out with dichloromethane.
Yield: 96 mg (83% of the theoretical value) dark red powder. M/e=579/581; λmax: 504.0.
Eight (8.0) mg (0.0128 mmol) 1,2,3,4-tetrahydrofredericamycin (25) are dissolved in 1 ml absolute DMF under nitrogen. Then a solution of 2.3 mg (0.0128 mmol) bromine in 0.25 ml DMF is added dropwise to the solution. Stirring at room temperature over 24 h. The reaction mixture is concentrated to half volume in high vacuum, and is then transferred onto 100 ml water. The supernatant is sucked off from the precipitate and dried in a vacuum.
Red crystal powder 8.1 mg (88% of the theoretical value) m/e=621/623; λmax: 499 nm.
Twenty (20) mg (37.1 μmol) fredericamycin are dissolved in 1 ml DMF under argon, then 4.76 mg (44.50 μmol) benzylamine are added at room temperature. According to HPLC (RP18, acetonitrile/water), a homogenous new product has formed after 3 h. The reaction mixture is concentrated at high vacuum until dry.
Red crystal mass; Yield: 23 mg (100% of the theoretical value) M/e=615.3 (M+H); λmax: 492 nm.
Five (5.0) mg (8.71 μmol) 5-chlorofredericamycin are dissolved in 1 ml DMF under argon, then 1.12 mg (10.45 μmol) benzylamine are added at room temperature. After 29 h, the reaction mixture is concentrated at high vacuum until dry.
Red crystal mass; Yield: 5 mg (89% of the theoretical value) M/e=649.1 (M+H); λmax: 492 nm.
Ten (10) mg (18.6 μmol) fredericamycin are dissolved in 1 ml DMF under argon, then 1.36 mg (22.3 μmol) ethanolamine are added at room temperature. According to HPLC (RP18, acetonitrile/water), a homogenous new product has formed after 3 h. The reaction mixture is concentrated at high vacuum until dry.
Red crystal mass; Yield: 9 mg (85% of the theoretical value) M/e=569.3 (M+H); λmax: 500 nm.
Ten (10) mg (18.6 μmol) fredericamycin are dissolved in 1 ml DMF under argon, then 2.7 PI (22.3 μmol) 4-aminomethylpiperidine are added at room temperature. The reaction mixture is concentrated at high vacuum until dry after 24 h.
Red crystal mass; Yield: 11 mg (99% of the theoretical value) M/e=622.3 (M+H); λmax: 492 nm.
The compounds 100-142 can be generated analogously to examples 7, 8, 9, 10, 18, 19 and 20:
Yield: (95% of the theoretical value) MS: M/e=593.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=562.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=621.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=621.1; λmax: 500.0 nm.
Yield: (80% of the theoretical value) MS: M/e=568.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=584.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=610.0; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=635.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=558.0; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=626.0; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=673.1; lλmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=599.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=587.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=632.0; λmax; λmax:500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=583.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=635.1; λmax: 492.0 nm.
Yield: (85% of the theoretical value) MS: M/e=630.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=558.1; λmax: 500.0 nm.
Yield: (85% of the theoretical value) MS: M/e=601.1; λmax: 492.0 nm.
Yield: (85% of the theoretical value) MS: M/e=635.1; λmax: 492.0 nm.
Yield: (90% of the theoretical value) MS: M/e=531.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=607.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=517.1; λmax: 482.0 nm.
Yield: (95% of the theoretical value) MS: M/e=679.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=635.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=559.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=641.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=641.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=641.1; λmax: 492.0 nm.
Yield: (90% of the theoretical value) MS: M/e=624.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=615.1; λmax: 492.0 nm.
Yield: (50% of the theoretical value) MS: M/e=656.1; λmax: 492.0 nm.
Yield: (60% of the theoretical value) MS: M/e=643.1; λmax: 492.0 nm.
Yield: (70% of the theoretical value) MS: M/e=587.1; λmax: 492.0 nm.
Yield: (70% of the theoretical value) MS: M/e=629.1; λmax: 492.0 nm.
Yield: (90% of the theoretical value) MS: M/e=588.1; λmax: 492.0 nm.
Yield: (85% of the theoretical value) MS: M/e=753.1; λmax: 492.0 nm.
Yield: (70% of the theoretical value) MS: M/e=602.1; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=627.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=696.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=655.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=655.0; λmax: 500.0 nm.
Yield: (90% of the theoretical value) MS: M/e=602.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=618.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=644.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=669.0; λmax: 500.0 mm.
Yield: (95% of the theoretical value) MS: M/e=609.0; λmax:500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=660.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=707.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=633.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=621.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=665.3; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=617.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=669.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=664.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=592.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=635.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=669.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=565.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=641.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=551.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=713.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=669.1; λmax: 500.0 nm.
Yield: (90% of the theoretical value) MS: M/e=593.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=675.1; λmax: 500.0 nm.
Yield: (90% of the theoretical value) MS: M/e=675.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=675.0; λmax: 500.0 nm.
Yield: (90% of the theoretical value) MS: M/e=658.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=649.0; λmax: 500.0 nm.
Yield: (60% of the theoretical value) MS: M/e=690.1; λmax: 500.0 nm.
Yield: (60% of the theoretical value) MS: M/e=677.1; λmax: 500.0 nm.
Yield: (70% of the theoretical value) MS: M/e=621.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=675.1; λmax: 500.0 nm.
Yield: (60% of the theoretical value) MS: M/e=622.0; λmax: 500.0 nm.
Yield: (90% of the theoretical value) MS: M/e=787.1; λmax: 500.0 nm.
Yield: (75% of the theoretical value) MS: M/e=636.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=670.9; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=739.9; λmax: 500.0 nm.
Yield: (90% of the theoretical value) MS: M/e=699.0; λmax: 500.0 nm.
Yield: (90% of the theoretical value) MS: M/e=699.0; λmax: 500.0 nm.
Yield: (70% of the theoretical value) MS: M/e=645.9; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=662.0; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=688.9; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=713.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=653.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=704.0; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=751.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=677.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=665.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=709.9; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=661.0; λmax: 500.0 nm.
Yield: (70% of the theoretical value) MS: M/e=713.0; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=708.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=636.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=679.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=713.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=609.0; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=685.0; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=595.0; λmax: 492.0 nm.
Yield: (90% of the theoretical value) MS: M/e=757.0; λmax: 500.0 nm.
Yield: (90% of the theoretical value) MS: M/e=713.0; λmax; λmax:500.0 nm.
Yield: (90% of the theoretical value) MS: M/e=637.0; λmax: 492.0 nm.
Yield: (90% of the theoretical value) MS: M/e=719.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=718.0; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=718.9; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=702.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=693.0; λmax: 492.0 nm.
Yield: (90% of the theoretical value) MS: M/e=734.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=721.1; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=665.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=707.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=666.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=831.0; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=680.0; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=559.2; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=601.3; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=625.2; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=641.2; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=625.3; λmax: 500.0 mm.
Yield: (80% of the theoretical value) MS: M/e=593.2; λmax: 500.0 nm.
Yield: (90% of the theoretical value) MS: M/e=635.3; λmax: 500.0 nm.
Yield: (85% of the theoretical value) MS: M/e=659.3; λmax: 500.0 nm.
Yield: (80% of the theoretical value) MS: M/e=675.3; λmax: 500.0 nm.
Yield: (80% of the theoretical value) MS: M/e=659.3; λmax: 500.0 nm.
Yield: (90% of the theoretical value) MS: M/e=639.3; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=679.3; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=703.3; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=719.3; λmax: 492.0 nm.
Yield: (95% of the theoretical value) MS: M/e=705.3; λmax: 492.0 nm.
Yield: (99% of the theoretical value) MS: M/e=685.3; λmax:500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=727.4; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=751.3; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=767.3; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=751.3; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=733.3; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=756.3; λmax: 500.0 nm.
Yield: (95% of the theoretical value) MS: M/e=657.3; λmax: 492.0 nm.
Yield: (99% of the theoretical value) MS: M/e=767.3; λmax; λmax:492.0 nm.
Yield: (99% of the theoretical value) MS: M/e=879.4; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=643.3; λmax: 492.0 nm.
Yield: (99% of the theoretical value) MS: M/e=782.3; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=782.3; λmax: 500.0 nm.
Yield: (99% of the theoretical value) MS: M/e=713.3; λmax: 500.0 nm.
Five (5) mg (0.0095 mmol) fredericamycin (1) are suspended in 2.0 ml ethanol. Under N2 atmosphere, 90 mg sodium acetate are added and boiled under reflux. After a few minutes, the suspension turns into a deep blue solution. After 24 h it is cooled, transferred onto water and shaken out with ethyl acetate (0.1% CF3COOH). After drying and concentration, a chromatographically homogenous, red powder is left.
Yield: 5.0 mg (97% of the theoretical value) MS=554 (M+H)+; λmax: 504.0 nm.
Six (6) mg (0.0114 mmol) fredericamycin (1) are suspended in 3.0 ml n-butanol. Under N2 atmosphere, 50 mg potassium acetate are added and heated to 100° C. After a few minutes, the suspension turns into a deep blue solution. The solution is left for 1 h at this temperature, and is then cooled. It is transferred onto water and shaken out with ethyl acetate (0.1% CF3COOH). After drying and concentration, a chromatographically homogenous red powder is left.
Yield: 6.2 mg (96% of the theoretical value) MS=582 (M)+; λmax: 500.0 nm.
Five (5) mg (0.0095 mmol) fredericamycin (1) are suspended in 3.0 ml n-propanol. Under N2 atmosphere, 50 mg potassium acetate (anhydrous) are added and heated to 80° C. After a few minutes, the suspension turns into a deep blue solution. The solution is left for 48 h at this temperature, and is then cooled. It is transferred onto water and shaken out with ethyl acetate (0.1% CF3COOH). After drying and concentration, a chromatographically homogenous red powder is left.
Yield: 3.7 mg (70% of the theoretical value) MS=568 (M+H)+; λmax: 500.0 nm.
6.1 mg (0.01159 mmol) fredericamycin (1) are suspended in 3.5 ml N,N-Dimethylaminoethanol. Under N2 atmosphere, 52 mg anhydrous potassium acetate are added and heated to 80° C. After a few minutes, the suspension turns into a deep blue solution. The solution is left for 1.5 h at this temperature, and is then cooled. It is transferred onto water and shaken out with ethyl acetate (0.1% CF3COOH). After drying and concentration, a chromatographically homogenous red powder is left.
Yield: 2.4 mg (36% of the theoretical value); MS=597 (M+H)+; λmax: 504.0 nm.
Ten (10.0) mg (0.019 mmol) bromofredericamycin (14) are suspended in 3.0 ml ethanol. Under N2 atmosphere, 50 mg anhydrous potassium acetate are added and heated to 80° C. After a few minutes, the suspension turns into a deep blue solution. The solution is left for 48 h at this temperature, and is then cooled. It is transferred onto water and shaken out with ethyl acetate (0.1% CF3COOH). After drying and concentration, a chromatographically homogenous red powder is left.
Yield: 7.2 mg (71% of the theoretical value); MS=632/634 (M+H)+; λmax: 504.0 nm.
9.6 mg (0.01824 mmol) fredericamycin (1) are suspended in 3.0 ml allyl alcohol. Under N2 atmosphere, 58 mg anhydrous potassium acetate are added and heated to 70° C. After a few minutes, the suspension turns into a deep blue solution. The solution is left for 2.5 h at this temperature, and is then cooled. It is transferred onto water and shaken out with ethyl acetate (0.1% CF3COOH). After drying and concentration, a chromatographically homogenous red powder is left.
Yield: 9.2 mg (91% of the theoretical value); MS=566 (M+H)+; λmax: 500.0 nm.
The compounds 249, 250, 251, 252, 253, 254, 255 were generated analogously to the instructions 244-248:
Yield: 5.2 mg (52% of the theoretical value); MS=569 (M)+; λmax: 499.0 nm.
Yield: 10.2 mg (99% of the theoretical value); MS=616 (M+H)+; λmax: 504.0 nm.
Yield: 12.9 mg (99% of the theoretical value); MS=580 (M)+; λmax:500.0 nm.
Yield: 2.0 mg (20% of the theoretical value); MS=622 (M+H)+; λmax: 499.0 nm.
Yield: 12.9 mg (99% of the theoretical value); MS=669 (M)+; λmax: 500.0 mm.
Yield: 5.8 mg (48% of the theoretical value); MS=653 (M+H)+; λmax: 500.0 nm.
Yield: 5.5 mg (50% of the theoretical value); MS=594 (M+H)+; λmax: 500.0 nm.
10.6 mg (0.02014 mmol) fredericamycin (1) are suspended in 2.0 ml bromoethanol. Under N2 atmosphere, 150 mg anhydrous potassium acetate are added and heated to 120° C. After a few minutes, the suspension turns into a deep blue solution. After 12 hours, addition of another 150 mg potassium acetate. The solution is left for another 12 h at this temperature, and is then cooled. It is transferred onto water and shaken out with ethyl acetate (0.1% CF3COOH). After drying and concentration, a chromatographically homogenous red powder is left.
Yield: 11.5 mg (99% of the theoretical value); MS=632/634 (M+H)+; λmax: 499.0 nm.
Five (5.0) mg (7.5 μmol) 5-iodofredericamycin (15) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 0.64 mg (11.2 μmmol) cyclopropylamine, it is stirred at room temperature for 3 h. Excess cycloprolylamine and DMF are removed at high vacuum. After drying and concentration, a chromatographically homogenous red powder is left.
Yield: 5.1 mg (99%); MS=691.3 (M+H)+; λmax: 504.0 nm.
Five (5.0) mg (7.5 μmol) 5-iodofredericamycin (15) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 0.82 mg (11.2 μmmol) n-butylamine, it is stirred at room temperature for 20 h. Excess n-butylamine and DMF are removed at high vacuum. After drying and concentration, a chromatographically homogenous red powder is left.
Yield: 5.3 mg (99%); MS=707.3 (M+H)+; λmax: 504.0 nm.
Five (5.0) mg (8.1 μmol) 5-bromofredericamycin (15) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 0.89 mg (12.2 μmmol) n-butylamine, it is stirred at room temperature for 20 h. Excess n-butylamine and DMF are removed at high vacuum. After drying and concentration, a chromatographically homogenous red powder is left.
Yield: 5.3 mg (99%); MS=659.4/661.4 (M+H)+; λmax: 504.0 nm.
Five (5.0) mg (9.3 μmol) fredericamycin (1) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 2.12 mg (37.2 μmmol) cyclopropylamine, it is stirred at room temperature for 2 h. Excess cyclopropylamine and DMF are removed at high vacuum. After drying and concentration, a chromatographically homogenous red powder is left.
Yield: 5.1 mg (99%); MS=565.4 (M+H)+; λmax: 510.0 nm.
Five (5.0) mg (9.3 μmol) fredericamycin (1) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 3.46 mg (37.2 μmmol) aniline and 37.2 μg stannous(IV)chloride (1.0 M in CH2Cl2), it is heated to 60° C. The reaction mixture is stirred for 24 h, and then excess diethanolaminomethyl polystyrene resin is added. Stir for 1 h. Exhaust off the resin and wash with DMF. The organic phase is concentrated at high vacuum. A chromatographically homogenous red powder is left.
Yield: 5.5 mg (99%); MS=601.1 (M+H)+; λmax: 504.0 nm.
Five (5.0) mg (9.3 μmol) fredericamycin (1) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 3.16 mg (37.2 μmmol) piperidine, it is stirred for 22 h at room temperature. Excess amine and DMF are removed in high vacuum. A chromatographically homogenous red powder is left.
Yield: 5.5 mg (99%); MS=593.4 (M+H)+; λmax: 504.0 nm.
Five (5.0) mg (9.3 μmol) fredericamycin (1) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 1.67 mg (37.2 μmmol) dimethylamine (2M in MeOH), it is stirred for 4 h at room temperature. Excess amine and DMF are removed in high vacuum. A chromatographically homogenous red powder is left.
Yield: 5.5 mg (99%); MS=553.6 (M+H)+; λmax: 526.0 nm.
Five (5.0) mg (9.3 μmol) fredericamycin (1) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 2.19 mg (37.2 μmmol) isopropylamine, it is stirred for 4 h at room temperature. Excess amine and DMF are removed in high vacuum. A chromatographically homogenous red powder is left.
Yield: 5.2 mg (99%); MS=567.3 (M+H)+; λmax: 504.0 nm.
Five (5.0) mg (9.3 μmol) fredericamycin (1) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 0.34 mg (11.1 μmmol) methylamine (2M in CH3OH), it is stirred for 19 h at room temperature. Excess amine and DMF are removed in high vacuum. A chromatographically homogenous red powder is left.
Yield: 5.0 mg (99%); MS=539.2 (M+H)+; λmax: 504.0 nm.
Five (5.0) mg (7.5 μmol) 5-iodofredericamycin (1) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 0.28 mg (9.0 μmmol) methylamine (2M in CH3OH), it is stirred for 2 h at room temperature. Excess amine and DMF are removed in high vacuum. A chromatographically homogenous red powder is left.
Yield: 5.0 mg (99%); MS=665.2 (M+H)+; λmax: 492.0 nm.
Five (5.0) mg (9.3 μmol) fredericamycin (1) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 3.24 mg (37.2 μmmol) morpholine, it is stirred for 18 h at room temperature. Excess amine and DMF are removed in high vacuum. A chromatographically homogenous red powder is left.
Yield: 5.5 mg (99%); MS=595.5 (M+H)+; λmax: 518.0 nm.
Five (5.0) mg (9.3 μmol) fredericamycin (1) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 0.67 mg (37.2 μmmol) ammonia (2M in EtOH), it is stirred for 24 h at room temperature. Excess ammonia and DMF are removed in high vacuum. A chromatographically homogenous red powder is left.
Yield: 4.8 mg (99%); MS=525.4 (M+H)+; λmax: 504.0 nm.
Five (5.0) mg (9.3 μmol) fredericamycin (1) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 0.99 mg (13.9 μmmol) pyrrolidine, it is stirred for 19 h at room temperature. Excess amine and DMF are removed in high vacuum. A chromatographically homogenous red powder is left.
Yield: 5.3 mg (99%); MS=579.2 (M+H)+; λmax: 554.0 nm.
Five (5.0) mg (8.1 μmol) 5-bromofredericamycin (1) are dissolved under argon in 1.0 ml anhydrous DMF. After addition of 0.70 mg (12.2 μmmol) cyclopropylamine, it is stirred for 5 h at room temperature. Excess cyclopropylamine and DMF are removed in high vacuum. A chromatographically homogenous red powder is left.
Yield: 5.0 mg (99%); MS=643.4/645.4 (M+H)+; λmax: 492.0 nmn.
79.5 mg (479 μmol) (2-oxo-propyl)-phosphonic acid dimethylester are dissolved under argon in 8 ml absolute pyridine, and 60.2 μl (479 μmol) 1,1,3,3-tetramethylguanidine are added at 0° C. After 5 minutes, 80.0 mg (159.7 μmol) fredericamycin aldehyde (4) is added at 0° C. After 2 hours, 100 ml 1 M hydrochloric acid are added, and the supernatant is sucked off from the precipitate. Dry under high vacuum.
Yield: 60.0 mg (69% of the theoretical value); M/e=542.2; λmax: 492.0 nm.
Fifty (50.0) mg (92.4 μmol) acetyl fredericamycin are dissolved under argon in 5 ml absolute DMF, and then 36.9 mg (231.1 μmol) bromine as a 1 M bromine solution in DMF are added under exclusion of light. It is stirred for 23 h under exclusion of light, and then 100 ml water are added. The precipitate is sucked off and dried under high vacuum.
Yield: 57.0 mg (87% of the theoretical value) red powder; M/e=697.9/699.9/701.9; M+; λmax: 504.0 nm.
Twenty (20.0) mg (28.7 μmol) bromoacetyl fredericamycin (273) are dissolved under argon in 4 ml absolute DMF. At room temperature, first 3.3 mg (43.0 μmol) thiourea, and then 20 mg IR120H+ are added. After 2 hours, it is filtered off the resin, and added to 50 ml water. The precipitate is dried under high vacuum. Red powder.
Yield: 18.0 mg (93% of the theoretical value); M/e=676.1/678.1; (M+H); λmax: 492.0 nm.
Five (5.0) mg (7.2 μmol) bromoacetyl fredericamycin (273) are dissolved under argon in 1 ml absolute DMF. At room temperature, first 1.5 mg (10.8 μmol) thiobenzamide, and then 5 mg IR120H+ are added. After 3.5 h, addition of hydrazinosulfonyl resin, and stirring for 2 h. It is filtered off the resin, and added to 10 ml water. The precipitate is dried under high vacuum. Red powder.
Yield: 3.0 mg (57% of the theoretical value); M/e=737.2/739.2; (M+H); λmax: 492.0 nm.
Five (5.0) mg (7.2 μmol) bromoacetyl fredericamycin (273) are dissolved under argon in 1 ml absolute DMF. At room temperature, first 1.3 mg (10.8 μmol) acetylthiourea, and then 5 mg IR120H+ are added. After 22 h, addition of hydrazinosulfonyl resin, and stirring for 2 h. It is filtered off the resin, and added to 10 ml water. The precipitate is dried under high vacuum. Red powder.
Yield: 2.0 mg (39% of the theoretical value); M/e=718.3/720.4; (M+H); λmax: 492.0 nm.
Five (5.0) mg (7.2 μmol) bromoacetyl fredericamycin (273) are dissolved under argon in 1 ml absolute DMF. At room temperature, first 0.81 mg (10.8 μmol) thioacetamide, and then 5 mg IR120H+ are added. After 2 h, addition of hydrazinosulfonyl resin, and stirring for 2 h. It is filtered off the resin, and added to 10 ml water. The precipitate is dried at high vacuum. Red powder.
Yield: 3.0 mg (62% of the theoretical value); M/e=675.2/677.2; (M+H); λmax: 492.0 nm.
Ten (10.0) mg (18.5 μmol) fredericamycin (1) are dissolved under argon in 2 ml absolute pyridine. After addition of 20.5 mg (92.5 mmol) phosphorous-V-sulfide, it is heated for 12 h to 60° C. Addition of another 20.5 mg (92.5 mmol) phosphorous-V-sulfide. According to HPLC (acetonitrile/water CF3COOH), the reaction was complete after 1 h. It is transferred onto water and shaken out with ethyl acetate. Dry and concentrate. Purple-red powder.
Yield: 5.0 mg (49% of the theoretical value); M/e=55.7; (M+H); λmax: 504.0 nm.
The water solubility of the various fredericamycin derivatives can be determined in a 0.9% NaCl solution with a pH of 7.
The compounds (22) and (3) dissolve very well. Compound (6) dissolves well, and compounds (2), (10), and (13) are soluble. Compounds (5), (7), (11) and (12) are sufficiently and markedly better soluble than fredericamycin (compound (1)).
| Number | Date | Country | Kind |
|---|---|---|---|
| 102 13 580 | Mar 2002 | DE | national |
| 102 48 451 | Oct 2002 | DE | national |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/EP03/02922 | 3/20/2003 | WO | 00 | 9/24/2004 |
| Publishing Document | Publishing Date | Country | Kind |
|---|---|---|---|
| WO03/080582 | 10/2/2003 | WO | A |
| Number | Name | Date | Kind |
|---|---|---|---|
| 4584377 | Yokoi et al. | Apr 1986 | A |
| 4673678 | Misra | Jun 1987 | A |
| 5166208 | Kelly et al. | Nov 1992 | A |
| 20050153997 | Simon et al. | Jul 2005 | A1 |
| 20050215579 | Simon et al. | Sep 2005 | A1 |
| Number | Date | Country |
|---|---|---|
| 61 044868 | Mar 1986 | JP |
| Number | Date | Country | |
|---|---|---|---|
| 20050256066 A1 | Nov 2005 | US |
