1,7-DIARYL-1,6-HEPTADIENE-3,5-DIONE DERIVATIVES, METHODS FOR THE PRODUCTION AND USE THEREOF
20220054671 · 2022-02-24
Inventors
- Andreas Späth (Regensburg, DE)
- Kristjan PLÄTZER (Salzburg, AT)
- Tim Maisch (Nürnberg, DE)
- Anja EICHNER (Regensburg, DE)
Cpc classification
C07C217/08
CHEMISTRY; METALLURGY
C07C217/20
CHEMISTRY; METALLURGY
C07C279/08
CHEMISTRY; METALLURGY
A01N2300/00
HUMAN NECESSITIES
A01N35/02
HUMAN NECESSITIES
A61P43/00
HUMAN NECESSITIES
A01N2300/00
HUMAN NECESSITIES
C07C217/24
CHEMISTRY; METALLURGY
C07C217/18
CHEMISTRY; METALLURGY
A01N57/34
HUMAN NECESSITIES
A01N35/02
HUMAN NECESSITIES
C07C217/22
CHEMISTRY; METALLURGY
International classification
A01N35/02
HUMAN NECESSITIES
A01N57/34
HUMAN NECESSITIES
C07C217/08
CHEMISTRY; METALLURGY
C07C217/18
CHEMISTRY; METALLURGY
C07C217/20
CHEMISTRY; METALLURGY
C07C217/22
CHEMISTRY; METALLURGY
C07C217/24
CHEMISTRY; METALLURGY
C07C279/08
CHEMISTRY; METALLURGY
Abstract
1,7-diaryl-1,6-heptadiene-3,5-dione derivatives, methods for the production and use thereof.
Claims
1. A method for the inactivation of microorganisms, wherein the method comprises the following steps: (A) bringing the microorganisms into contact with at least one photosensitizer, wherein the photosensitizer is at least one 1,7-diaryl-1,6-heptadiene-3,5-dione derivative with formula (100): ##STR00197## and/or with formula (101): ##STR00198## or respectively a pharmacologically acceptable salt and/or ester and/or complex thereof, wherein the residues Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, represent one substituted or unsubstituted, monocyclic or polycyclic aromatic residue or one substituted or unsubstituted, monocyclic or polycyclic heteroaromatic residue, wherein K represents hydrogen or a cation, and wherein M.sup.z+ represents a cation of a metal or boron, wherein z is the formal oxidation number of the metal M or boron and z represents a whole number from 1 to 7, and wherein (a1) at least one of the residues Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, is an unsubstituted, monocyclic or polycyclic heteroaromatic residue, which has at least 5 ring atoms, wherein the ring atoms contain at least one carbon atom and at least one nitrogen atom which optionally can be protonated, or (a2) at least one of the residues Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, is substituted with at least one organic residue W1, wherein the at least one organic residue W1 has the general formula (4), (5), (6), (7), (8), or (9):
-(C(D)(E)).sub.h-X, (4)
-A-(C(D)(E)).sub.h-X, (5)
-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (6)
-A-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (7)
-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (8)
-A-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (9), wherein h represents a whole number from 1 to 20, wherein k represents a whole number from 0 to 10, wherein l represents a whole number from 0 to 10, and wherein m, n and p, respectively independently of each other, represent a whole number from 1 to 6, and wherein A, respectively independently of each other, represents oxygen or sulphur, wherein D and E, respectively independently of each other, represent hydrogen, halogen, G-R.sup.(I), or G-C(=G)-R.sup.(II), wherein G, respectively independently of each other, represents oxygen or sulphur, and wherein the residues R.sup.(I) and R.sup.(II), respectively independently of each other, represent hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl, phenyl or benzyl, wherein phenyl and benzyl may be unsubstituted or substituted, wherein aryl represents a substituted or unsubstituted aromatic group or a substituted or unsubstituted heteroaromatic group which does not contain a nitrogen atom, wherein X, respectively independently of each other, is an organic residue which (i) contains at least one neutral nitrogen atom which can be protonated, or (ii) contains at least one positively charged nitrogen atom, or (iii) contains at least one positively charged phosphorus atom, and wherein the residues R1, R2, R3, R4 and R5, respectively independently of each other, represent hydrogen, halogen, alkyl containing 1 to 12 C atoms, cycloalkyl containing 1 to 12 C atoms, alkylaryl containing 1 to 12 C atoms, aryl containing 5 to 20 C atoms, ether containing 2 to 12 C atoms or glycol containing 2 to 12 C atoms, or wherein (b) the residue R3 is an organic residue W2, wherein the one organic residue W2 has the general formula (4), (5), (6), (7), (8), (9), or (10):
-(C(D)(E)).sub.h-X, (4)
-A-(C(D)(E)).sub.h-X, (5)
-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (6)
-A-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (7)
-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (8)
-A-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (9),
-(-C(D)=C(E)-).sub.r-X, (10), and wherein, optionally, at least one of the residues Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, is substituted with at least one organic residue W1 which has the general formula (4), (5), (6), (7), (8), or (9), wherein h represents a whole number from 1 to 20, wherein k represents a whole number from 0 to 10, wherein l represents a whole number from 0 to 10, and wherein m, n, p, and r, respectively independently of each other, represent a whole number from 1 to 6, and wherein A, respectively independently of each other, represents oxygen or sulphur, wherein D and E, respectively independently of each other, represent hydrogen, halogen, G-R.sup.(I), or G-C(=G)-R.sup.(II), wherein G, respectively independently of each other, represents oxygen or sulphur, and wherein the residues R.sup.(I) and R.sup.(II), respectively independently of each other, represent hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl, phenyl or benzyl, wherein phenyl and benzyl may be unsubstituted or substituted, wherein aryl represents a substituted or unsubstituted aromatic group or a substituted or unsubstituted heteroaromatic group which does not contain a nitrogen atom, wherein X, respectively independently of each other, is an organic residue which (i) contains at least one neutral nitrogen atom which can be protonated, (ii) contains at least one positively charged nitrogen atom, or (iii) contains at least one positively charged phosphorus atom, and wherein the residues R1, R2, R4 and R5, respectively independently of each other, represent hydrogen, halogen, alkyl containing 1 to 12 C atoms, alkylaryl containing 1 to 12 C atoms, aryl containing 5 to 20 C atoms, ether containing 2 to 12 C atoms or glycol containing 2 to 12 C atoms, and (B) irradiating the microorganisms and the at least one photosensitizer with electromagnetic radiation of a suitable wavelength and energy density.
2. The method as claimed in claim 1, wherein, in the compound with formula (100), K is a cation M.sup.z+ of a metal M or boron, wherein z is the formal oxidation number of the metal M or boron and wherein z represents a whole number from 1 to 7, and wherein the compound has the formula (102): ##STR00199## wherein L.sup.1 and L.sup.2, respectively independently of each other, represent water, fluoride, chloride, bromide, iodide, phosphate, hydrogen phosphate, dihydrogen phosphate, sulphate, hydrogen sulphate, tosylate, mesylate or at least one carboxylation of a carboxylic acid containing 1 to 15 carbon atoms and/or mixtures thereof.
3. The method as claimed in claim 1, wherein the irradiation of the microorganisms and of the at least one photosensitizer with electromagnetic radiation of a suitable wavelength and energy density is carried out in the presence of at least one oxygen-donating compound and/or at least one oxygen-containing gas.
4. The method as claimed in claim 1, wherein the at least one photosensitizer is a compound with formula (1): ##STR00200## wherein the residues Q.sup.1 and Q.sup.2, respectively independently of each other, represent one substituted or unsubstituted, monocyclic or polycyclic aromatic residue, wherein the compound with formula (1) does not contain an OH group which is bonded directly to the organic residue Q.sup.1 or Q.sup.2, and wherein K represents hydrogen or a cation, and wherein (a) at least one of the residues Q.sup.1 and Q.sup.2, respectively independently of each other, is substituted with at least one organic residue W1a, wherein the at least one organic residue W1a has the general formula (5a), (6a), (7a), (8a), or (9a):
-A-(C(D)(E)).sub.h-X.sup.a, (5a)
-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X.sup.a, (6a)
-A-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X.sup.a, (7a)
-((C(D)(E)).sub.mA).sub.p-(C(D)(E)).sub.n-X.sup.a, (8a)
-A-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X.sup.a, (9a) wherein h represents a whole number from 1 to 20, wherein k represents a whole number from 0 to 10, wherein l represents a whole number from 0 to 10, and wherein m, n and p, respectively independently of each other, represent a whole number from 1 to 6, and wherein A, respectively independently of each other, represents oxygen or sulphur, wherein D and E, respectively independently of each other, represent hydrogen, halogen, G-R.sup.(I), or G-C(=G)-R.sup.(II), wherein G, respectively independently of each other, represents oxygen or sulphur, and wherein the residues R.sup.(I) and R.sup.(II), respectively independently of each other, represent hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl, phenyl or benzyl, wherein phenyl and benzyl may be unsubstituted or substituted, wherein aryl represents a substituted or unsubstituted aromatic group or a substituted or unsubstituted heteroaromatic group which does not contain a nitrogen atom, wherein X.sup.a, respectively independently of each other, represents a residue with formula (20c), (20d), or (21): ##STR00201## wherein each of the residues R.sup.(VII), R.sup.(VIII), and R.sup.(IX), respectively independently of each other, represents hydrogen, an aryl residue containing 5 to 12 C atoms, an alkylaryl residue containing 5 to 12 C atoms, an alkyl residue, which may be linear or branched, containing 1 to 8 C atoms, or an ether residue, which may be linear or branched, containing 1 to 8 C atoms, and wherein the residues R1, R2, R3, R4 and R5, respectively independently of each other, represent hydrogen, halogen, alkyl containing 1 to 12 C atoms, cycloalkyl containing 1 to 12 C atoms, alkylaryl containing 1 to 12 C atoms, aryl containing 5 to 20 C atoms, ether containing 2 to 12 C atoms or glycol containing 2 to 12 C atoms, or wherein (b) the residue R.sup.3 is an organic residue W2a which contains at least one neutral nitrogen atom which can be protonated, and/or at least one positively charged nitrogen atom and/or at least one positively charged phosphorus atom, wherein the one organic residue W2a has the general formula (4b), (5b), (6b), (7b), (8b), or (9b):
-(C(D)(E)).sub.h-X.sup.b, (4b)
-A-(C(D)(E)).sub.h-X.sup.b, (5b)
-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X.sup.b, (6b)
-A-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X.sup.b, (7b)
-[(C(D)(E)).sub.mA].sub.p-(C(D)(E)).sub.n-X.sup.b, (8b)
-A-[(C(D)(E)).sub.m-A].sub.p-(C(D)(E)).sub.n-X.sup.b, (9b) and wherein, optionally, at least one of the residues Q.sup.1 and Q.sup.2, respectively independently of each other, is substituted with at least one organic residue W1 b which has the general formula (4b), (5b), (6b), (7b), (8b), or (9b), wherein h represents a whole number from 1 to 20, wherein k represents a whole number from 0 to 10, wherein l represents a whole number from 0 to 10, and wherein m, n and p, respectively independently of each other, represent a whole number from 1 to 6, and wherein A, respectively independently of each other, represents oxygen or sulphur, wherein D and E, respectively independently of each other, represent hydrogen, halogen, G-R.sup.(I), or G-C(=G)-R.sup.(II), wherein G, respectively independently of each other, represents oxygen or sulphur, and wherein the residues R.sup.(I) and R.sup.(II), respectively independently of each other, represent hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl, phenyl or benzyl, wherein phenyl and benzyl may be unsubstituted or substituted, wherein aryl represents a substituted or unsubstituted aromatic group or a substituted or unsubstituted heteroaromatic group which does not contain a nitrogen atom, wherein X.sup.b, respectively independently of each other, is an organic residue which (i) contains at least one neutral nitrogen atom which can be protonated, or (ii) contains at least one positively charged nitrogen atom, or (iii) contains at least one positively charged phosphorus atom, and wherein the residues R1, R2, R4 and R5, respectively independently of each other, represent hydrogen, halogen, alkyl containing 1 to 12 C atoms, alkylaryl containing 1 to 12 C atoms, aryl containing 5 to 20 C atoms, ether containing 2 to 12 C atoms or glycol containing 2 to 12 C atoms, and/or a pharmacologically acceptable salt and/or ester and/or complex thereof.
5. A method for photodynamic inactivation of microorganisms on or within a patient, wherein the method comprises the following steps: (A) bringing microorganisms on or within a patient into contact with at least one photosensitizer, wherein the photosensitizer is at least one 1,7-diaryl-1,6-heptadiene-3,5-dione derivative with formula (100) ##STR00202## and/or with formula (101): ##STR00203## or respectively a pharmacologically acceptable salt and/or ester and/or complex thereof, wherein Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, represent one substituted or unsubstituted, monocyclic or polycyclic aromatic residue or one substituted or unsubstituted, monocyclic or polycyclic heteroaromatic residue, wherein K represents hydrogen or a cation, and wherein M.sup.z+ represents a cation of a metal or boron, wherein z is the formal oxidation number of the metal M or boron and z represents a whole number from 1 to 7, and wherein (a1) at least one of the residues Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, is an unsubstituted, monocyclic or polycyclic heteroaromatic residue, which has at least 5 ring atoms, wherein the ring atoms contain at least one carbon atom and at least one nitrogen atom which optionally can be protonated, or (a2) at least one of the residues Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, is substituted with at least one organic residue W1, wherein the at least one organic residue W1 has the general formula (4), (5), (6), (7), (8):
-(C(D)(E)).sub.h-X, (4)
-A-(C(D)(E)).sub.h-X, (5)
-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (6)
-A-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (7)
-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (8)
-A-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (9), wherein h represents a whole number from 1 to 20, wherein k and l, respectively independently of each other, represents a whole number from 0 to 10, and wherein m, n and p, respectively independently of each other, represent a whole number from 1 to 6 and wherein A, respectively independently of each other, represents oxygen or sulphur, wherein D and E, respectively independently of each other, represent hydrogen, halogen, G-R.sup.(I), or G-C(=G)-R.sup.(II), wherein G, respectively independently of each other, represents oxygen or sulphur, and wherein the residues R.sup.(I) and R.sup.(II), respectively independently of each other, represent hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl, phenyl or benzyl, wherein phenyl and benzyl may be unsubstituted or substituted, wherein aryl represents a substituted or unsubstituted aromatic group or a substituted or unsubstituted heteroaromatic group which does not contain a nitrogen atom, wherein X, respectively independently of each other, is an organic residue which (i) contains at least one neutral nitrogen atom which can be protonated, or (ii) at least one positively charged nitrogen atom or (iii) at least one positively charged phosphorus atom, and wherein the residues R1, R2, R3, R4 and R5, respectively independently of each other, represent hydrogen, halogen, alkyl containing 1 to 12 C atoms, cycloalkyl containing 1 to 12 C atoms, alkylaryl containing 1 to 12 C atoms, aryl containing 5 to 20 C atoms, ether containing 2 to 12 C atoms or glycol containing 2 to 12 C atoms, or wherein (b) the residue R3 is an organic residue W2, wherein the one organic residue W2 has the general formula (4), (5), (6), (7), (8), (9), or (10):
-(C(D)(E)).sub.h-X, (4)
-A-(C(D)(E)).sub.h-X, (5)
-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (6)
-A-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (7)
-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (8)
-A-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (9),
-(-C(D)=C(E)-).sub.r-X, (10), and wherein, optionally, at least one of the residues Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, is substituted with at least one organic residue W1 which has the general formula (4), (5), (6), (7), (8), or (9), wherein h represents a whole number from 1 to 20, wherein k and l, respectively independently of each other, represents a whole number from 0 to 10 and wherein m, n, p, and r, respectively independently of each other, represent a whole number from 1 to 6, and wherein A, respectively independently of each other, represents oxygen or sulphur, wherein D and E, respectively independently of each other, represent hydrogen, halogen, G-R.sup.(I), or G-C(=G)-R.sup.(II), wherein G, respectively independently of each other, represents oxygen or sulphur, and wherein the residues R.sup.(I) and R.sup.(II), respectively independently of each other, represent hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl, phenyl or benzyl, wherein phenyl and benzyl may be unsubstituted or substituted, wherein aryl represents a substituted or unsubstituted aromatic group or a substituted or unsubstituted heteroaromatic group which does not contain a nitrogen atom, wherein X, respectively independently of each other, is an organic residue which (i) contains at least one neutral nitrogen atom which can be protonated, (ii) contains at least one positively charged nitrogen atom or (iii) contains at least one positively charged phosphorus atom, and wherein the residues R1, R2, R4 and R5, respectively independently of each other, represent hydrogen, halogen, alkyl containing 1 to 12 C atoms, alkylaryl containing 1 to 12 C atoms, aryl containing 5 to 20 C atoms, ether containing 2 to 12 C atoms or glycol containing 2 to 12 C atoms; and (B) irradiating the microorganisms and the at least one photosensitizer with electromagnetic radiation of a suitable wavelength and energy density to inactivate said microorganisms.
6. The method according to claim 5, wherein said microorganisms are selected from the group consisting of viruses, archaea, bacteria, bacterial spores, fungi, fungal spores, protozoa, algae and blood-transmissible parasites.
7. The method according to claim 5, wherein the microorganisms on or within the patient are brought into contact with the at least one photosensitizer in at least one selected from the group consisting of cleaning of teeth, dental prostheses and dental braces, treatment of a disease of the dental tissue and of the periodontium.
8. The method according to claim 5, wherein the microorganisms on or within the patient are brought into contact with the at least one photosensitizer for treatment of an infectious skin disease.
9. The method according to claim 5, wherein the microorganisms on or within the patient are brought into contact with the at least one photosensitizer for reduction of the bacterial count in a wound.
10. A pharmaceutical composition comprising: (A) at least one 1,7-diaryl-1,6-heptadiene-3,5-dione derivative with formula (100) ##STR00204## and/or with formula (101) ##STR00205## or respectively a pharmacologically acceptable salt and/or ester and/or complex thereof, wherein the residues Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, represent one substituted or unsubstituted, monocyclic or polycyclic aromatic residue or one substituted or unsubstituted, monocyclic or polycyclic heteroaromatic residue, wherein K represents hydrogen or a cation, and wherein M.sup.z+ represents a cation of a metal or boron, wherein z is the formal oxidation number of the metal M or boron and z represents a whole number from 1 to 7, and wherein (a1) at least one of the residues Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, is an unsubstituted, monocyclic or polycyclic heteroaromatic residue, which has at least 5 ring atoms, wherein the ring atoms contain at least one carbon atom and at least one nitrogen atom which optionally can be protonated, or (a2) at least one of the residues Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, is substituted with at least one organic residue W1, wherein the at least one organic residue W1 has the general formula (4), (5), (6), (7), (8), or (9):
-(C(D)(E)).sub.h-X, (4)
-A-(C(D)(E)).sub.h-X, (5)
-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (6)
-A-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (7)
-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (8)
-A-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (9), wherein h represents a whole number from 1 to 20, wherein k represents a whole number from 0 to 10, wherein l represents a whole number from 0 to 10, and wherein m, n and p, respectively independently of each other, represent a whole number from 1 to 6, and wherein A, respectively independently of each other, represents oxygen or sulphur, wherein D and E, respectively independently of each other, represent hydrogen, halogen, G-R.sup.(I), or G-C(=G)-R.sup.(II), wherein G, respectively independently of each other, represents oxygen or sulphur, and wherein the residues R.sup.(I) and R.sup.(II), respectively independently of each other, represent hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl, phenyl or benzyl, wherein phenyl and benzyl may be unsubstituted or substituted, wherein aryl represents a substituted or unsubstituted aromatic group or a substituted or unsubstituted heteroaromatic group which does not contain a nitrogen atom, wherein X, respectively independently of each other, is an organic residue which (i) contains at least one neutral nitrogen atom which can be protonated, or (ii) contains at least one positively charged nitrogen atom, or (iii) contains at least one positively charged phosphorus atom, and wherein the residues R1, R2, R3, R4 and R5, respectively independently of each other, represent hydrogen, halogen, alkyl containing 1 to 12 C atoms, cycloalkyl containing 1 to 12 C atoms, alkylaryl containing 1 to 12 C atoms, aryl containing 5 to 20 C atoms, ether containing 2 to 12 C atoms or glycol containing 2 to 12 C atoms, or wherein (b) the residue R.sup.3 is an organic residue W2, wherein the one organic residue W2 has the general formula (4), (5), (6), (7), (8), (9), or (10):
-(C(D)(E)).sub.h-X, (4)
-A-(C(D)(E)).sub.h-X, (5)
-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (6)
-A-(C(D)(E)).sub.k-aryl-(C(D)(E)).sub.l-X, (7)
-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (8)
-A-((C(D)(E)).sub.m-A).sub.p-(C(D)(E)).sub.n-X, (9),
-(-C(D)=C(E)-).sub.r-X, (10), and wherein, optionally, at least one of the residues Q.sup.3, Q.sup.3a, Q.sup.4 and Q.sup.4a, respectively independently of each other, is substituted with at least one organic residue W1 which has the general formula (4), (5), (6), (7), (8), or (9), wherein h represents a whole number from 1 to 20, wherein k represents a whole number from 0 to 10, wherein l represents a whole number from 0 to 10, and wherein m, n, p, and r, respectively independently of each other, represent a whole number from 1 to 6, and wherein A, respectively independently of each other, represents oxygen or sulphur, wherein D and E, respectively independently of each other, represent hydrogen, halogen, G-R.sup.(I), or G-C(=G)-R.sup.(II), wherein G, respectively independently of each other, represents oxygen or sulphur, and wherein the residues R.sup.(I) and R.sup.(II), respectively independently of each other, represent hydrogen, methyl, ethyl, n-propyl, n-butyl, n-pentyl, phenyl or benzyl, wherein phenyl and benzyl may be unsubstituted or substituted, wherein aryl represents a substituted or unsubstituted aromatic group or a substituted or unsubstituted heteroaromatic group which does not contain a nitrogen atom, wherein X, respectively independently of each other, is an organic residue which (i) contains at least one neutral nitrogen atom which can be protonated, (ii) contains at least one positively charged nitrogen atom, or (iii) contains at least one positively charged phosphorus atom, and wherein the residues R1, R2, R4 and R5, respectively independently of each other, represent hydrogen, halogen, alkyl containing 1 to 12 C atoms, alkylaryl containing 1 to 12 C atoms, aryl containing 5 to 20 C atoms, ether containing 2 to 12 C atoms or glycol containing 2 to 12 C atoms; and (B) one or more physiologically acceptable excipients.
11. The pharmaceutical composition as claimed in claim 10, wherein, in the compound with formula (100), K is a cation M.sup.2+ of a metal M or boron, wherein z is the formal oxidation number of the metal M or boron and wherein z represents a whole number from 1 to 7, and wherein the compound has the formula (102): ##STR00206## wherein L.sup.1 and L.sup.2, respectively independently of each other, represent water, fluoride, chloride, bromide, iodide, phosphate, hydrogen phosphate, dihydrogen phosphate, sulphate, hydrogen sulphate, tosylate, mesylate or at least one carboxylation of a carboxylic acid containing 1 to 15 carbon atoms and/or mixtures thereof.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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EXAMPLES
Example 1) Production of Various 1,7-diaryl-1,6-heptadiene-3,5-dione Derivatives
Overview of the Syntheses
[0619] All of the chemicals were purchased from conventional suppliers (TCI, ABCR, Acros, Merck and Fluka) and used without further purification. The solvents were distilled before use and if required, were dried in the normal manner. Dry DMF was purchased from Fluka (Taufkirchen, DE).
[0620] Thin film chromatography was carried out on thin film aluminium foils coated with silica gel 60 F254, from Merck (Darmstadt, DE). Preparative thin film chromatography was carried out on commercially available glass plates coated with silica gel 60 (20 cm×20 cm, Carl Roth GmbH & Co. KG, Karlsruhe, DE). The compounds were detected with UV light (λ=254 nm, 333 nm) and some detected with the naked eye or stained with ninhydrin. The chromatography was carried out with silica gel (0.060-0.200) from Acros (Waltham, US). NMR spectra were recorded on a Bruker Avance 300 spectrometer (300 MHz [.sup.1H-NMR], 75 MHz [.sup.13C-NMR]) (Bruker Corporation, Billerica, US).
[0621] All of the chemical displacements are given in δ [ppm] relative to an external standard (tetramethylsilane, TMS). The coupling constants are respectively given in Hz; Characterization of the signals: s=singlet, d=doublet, t=triplet, m=multiplet, dd=doublet of doublets, br=broad. Integration determined the relative number of atoms. The definitive identification of the signals in the carbon spectra was carried out using the DEPT method (pulse angle: 135°). Error limits: 0.01 ppm for .sup.1H-NMR, 0.1 ppm for .sup.13C-NMR and 0.1 Hz for coupling constants. The solvent used is noted for each spectrum.
[0622] The IR spectra were recorded on a Biorad Excalibur FTS 3000 spectrometer (Bio-Rad Laboratories GmbH, Munich, DE).
[0623] ES-MS was measured using a ThermoQuest Finnigan TSQ 7000 spectrometer, all of the HR-MS were determined on a ThermoQuest Finnigan MAT 95 (respectively Thermo Fisher Scientific Inc, Waltham, US) spectrometer; argon was used as the ionization gas for FAB.
[0624] The melting points were determined with the aid of the Büchi SMP-20 melting point instrument (Büchi Labortechnik GmbH, Essen, DE) using a glass capillary.
[0625] All of the UV/VIS spectra were recorded using a Varian Cary 50 Bio UV/VIS spectrometer; the fluorescence spectra were recorded with a Varian Cary Eclipse spectrometer.
[0626] The solvents for absorption and emission measurements were purchased in special spectroscopic purity grade from Acros or Baker, or Uvasol from Merck. Millipore water (18 MO, Milli Q.sub.plus) was used for all of the measurements.
1. Synthesis of Aldehyde Components
[0627] The educts and alkylation agents listed in Table 1 were obtained commercially.
[0628] The alkylation agents used were the corresponding alkyl bromides or alkyl tosylates.
##STR00095##
Overview 1: Synthesis of synthesis components: Conditions: (a) K.sub.2CO.sub.3, KI, DMF or DMSO, T=60-80° C.
TABLE-US-00001 TABLE 1 Synthesis of aldehyde components: Conditions: (a) K.sub.2CO.sub.3, KI, DMF or DMSO, T = 60-80° C. Educt Alkylation agent Product Example R.sup.6a= R.sup.7a= R.sup.9a= w v Y R.sup.6a= R.sup.7a= R.sup.9a= w v Yield E-1 H H H 1 0 Br H H H 1 0 76% E-2 H Me H 1 0 Br H Me H 1 0 73% E-3 H OMe H 1 0 Br H OMe H 1 0 79% E-4 OMe H H 1 0 Br OMe H H 1 0 71% E-5 H OMe I 1 0 Br H OMe I 1 0 66% E-6 H OMe H 1 1 TsO H OMe H 1 1 72% E-7 H OBz H 2 0 Br H OBz H 2 0 82% E-8 H OH H 1 0 Br H OH H 1 0 17% E-9 H OH H 1 0 Br H OC.sub.8H.sub.17 H 1 0 39% (2 steps) E-10 H OH H 1 0 Br H O(CH.sub.2CH.sub.2).sub.3OH H 1 0 33% (2 steps) E-11 H OH H 1 0 Br H OCH.sub.2CH.sub.2NHBoc H 1 0 69% E-12 H OH OH 1 0 Br H OCH.sub.2CH.sub.2NHBoc OCH.sub.2CH.sub.2NHBoc 1 0 61%
##STR00096##
Overview 2: Synthesis of further aldehyde components: Conditions: (a) K.sub.2CO.sub.3, KI, DMF or DMSO, T=60-80° C.
1.1 Substituted Vanillins
1.1.1 General Procedure:
[0629] 10 mmol of the appropriate educt was placed under nitrogen in 20 mL of DMF: [0630] (I) 4-hydroxybenzaldehyde (1.22 g, 10 mmol) [0631] (II) vanillin (1.52 g, 10 mmol) [0632] (III) benzyloxyvanillin (2.28 g, 10 mmol) [0633] (IV) 6-hydroxy-2-naphthaldehyde (1.72 g, 10 mmol) [0634] (V) 4-hydroxy-3-methoxy-acetophenone (1.66 g, 10 mmol)
[0635] The corresponding alkylation reagent (12-15 mmol, 1.2-1.5 eq) in DMF (10 mL) and K.sub.2CO.sub.3 (2.76 g, 20 mmol) were added one after the other. The formulation was stirred under a nitrogen atmosphere for 24 h at 80° C. After cooling to room temperature, the reaction mixture was diluted with 60 mL ethyl acetate and the organic phase was shaken three times, each time with 40 mL of saturated aqueous sodium chloride solution, and once with 50 mL of water. The organic phase was separated, dried over MgSO.sub.4 and rotary evaporated.
[0636] In order to purify the product, column chromatography was carried out on silica gel.
E-1: 4-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde
[0637] ##STR00097##
[0638] Educt: 4-hydroxybenzaldehyde.
[0639] The alkylation reagent used was 2-(tert-butoxycarbonylamino)ethyl bromide (3.34 g, 15 mmol).
[0640] Yield: 76% of theory, 2.5 g of pale yellow solid
[0641] Molecular mass=265.30 g/mol; Empirical formula=C.sub.14H.sub.19NO.sub.4
[0642] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=9.87 (s, 1H), 7.82 (d, J=8.8 Hz, 2H), 6.98 (d, J=8.7 Hz, 2H), 5.05 (s, 1H), 4.09 (t, J=5.2 Hz, 2H), 3.55 (dd, J=10.4, 5.1 Hz, 2H), 1.44 (s, 9H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 288.1 (17%, MNa.sup.+), 265.1 (1%, MH.sup.+), 210.1 (100%, MH.sup.+—C.sub.4H.sub.9), 166.1 (19%, MH.sup.+-boc)
E-3: 3-methoxy-4-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde
[0643] ##STR00098##
[0644] Educt: vanillin (4-hydroxy-3-methoxybenzaldehyde).
[0645] The alkylation reagent used was 2-(tert-butoxycarbonylamino)ethylbromide (3.34 g, 15 mmol).
[0646] Yield: 79% of theory, 2.54 g of pale yellow solid
[0647] Molecular mass=295.34 g/mol; Empirical formula=C.sub.15H.sub.21NO.sub.5
[0648] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=9.85 (s, 1H), 7.43 (d, J=1.8 Hz, 1H), 7.42 (m, 1H), 6.98 (d, J=8.0 Hz, 1H), 5.10 (s, 1H), 4.15 (t, J=5.1 Hz, 2H), 3.92 (s, 3H), 3.60 (dd, J=10.6, 5.3 Hz, 2H), 1.44 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 318.1 (13%, MNa.sup.+), 296.1 (100%, MH.sup.+), 240.1 (45%, MH.sup.+—C.sub.4H.sub.9), 196.1 (9%, MH.sup.+-boc).
E-15: 3-methoxy-4-(2-N-tert-butyloxycarbonyl-aminoethoxy)acetophenone
[0649] ##STR00099##
[0650] Production as described in A.1).
[0651] Educt: 4-hydroxy-3-methoxyacetophenone (acetovanillone).
[0652] The alkylation reagent used was 2-(tert-butoxycarbonylamino)ethyl bromide (3.34 g, 15 mmol).
[0653] Yield: 77% of theory, 2.75 g of pale yellow solid
[0654] Molecular mass=309.36 g/mol; Empirical formula=C.sub.16H.sub.23NO.sub.5
[0655] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=7.51 (d, J=1.9 Hz, 1H), 7.49 (s, 1H), 6.94 (d, J=7.9 Hz, 1H), 5.19 (s, 1H), 4.12 (t, J=5.1 Hz, 2H), 3.89 (s, 3H), 3.58 (q, 5.3 Hz, 2H), 2.52 (s, 3H), 1.43 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 332.1 (29%, MNa.sup.+), 309.1 (2%, MH.sup.+), 254.1 (33%, MH.sup.+—C.sub.4H.sub.9), 210.1 (100%, MH.sup.+-boc).
E-14: 6-(2-N-tert-butyloxycarbonyl-aminoethoxy)naphthalene-2-carbaldehyde
[0656] ##STR00100##
[0657] Educt: 6-hydroxy-2-naphthaldehyde.
[0658] The alkylation reagent used was 2-(tert-butoxycarbonylamino)ethylbromide (3.34 g, 15 mmol).
[0659] Yield: 71% of theory, 2.30 g of pale yellow solid
[0660] Molecular mass=315.37 g/mol; Empirical formula=C.sub.18H.sub.21NO.sub.4
[0661] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=10.09 (s, 1H), 8.25 (s, 1H), 7.94-7.85 (m, 2H), 7.79 (d, J=8.5 Hz, 1H), 7.23 (dd, J=8.9, 2.5 Hz, 1H), 7.17 (d, J=2.3 Hz, 1H), 5.05 (s, 1H), 4.18 (t, J=5.1 Hz, 2H), 3.62 (dd, J=10.5, 5.3 Hz, 2H), 1.46 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 338.2 (11%, MNa.sup.+), 316.2 (12%, MH.sup.+), 260.1 (100%, MH.sup.+—C.sub.4H.sub.9), 216.1 (14%, MH.sup.+-boc)
E-7: 3-benzyloxy-4-(2-N-tert-butyloxycarbonyl-aminopropyloxy)benzaldehyde
[0662] ##STR00101##
[0663] Educt: benzyloxyvanillin (4-benzyloxy-3-methoxybenzaldehyde).
[0664] The alkylation reagent used was 2-(tert-butoxycarbonylamino)propyl bromide (3.34 g, 15 mmol).
[0665] Yield: 82% of theory, 2.64 g of pale yellow solid
[0666] Molecular mass=385.45 g/mol; Empirical formula=C.sub.22H.sub.27NO.sub.5
[0667] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=9.74 (s, 1H), 7.44-7.16 (m, 7H), 6.91 (dd, J=8.1, 1.4 Hz, 1H), 5.47 (s, 1H), 5.21 (m, 2H), 4.08 (td, J=5.6, 2.2 Hz, 2H), 3.33 (d, J=5.2 Hz, 2H), 2.03-1.92 (m, 2H), 1.36 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 408.2 (37%, MNa.sup.+), 386.1 (2%, MH.sup.+), 330.1 (63%, MH.sup.+—C.sub.4H.sub.9), 286.1 (100%, MH.sup.+-boc).
E-6: 3-methoxy-4-(2-N-tert-butyloxycarbonyl-aminoethoxy-ethoxy)benzaldehyde
[0668] ##STR00102##
[0669] Educt: vanillin (4-hydroxy-3-methoxybenzaldehyde).
[0670] The alkylation reagent used was 2-[2-(tert-butoxycarbonylamino)ethoxy]ethyl-4-methyl benzenesulphonate (4.26 g, 12 mmol).
[0671] Yield: 72% of theory; 2.78 g of pale yellow solid
[0672] Molecular mass=339.39 g/mol; Empirical formula=C.sub.17H.sub.25NO.sub.6
[0673] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=9.76 (s, 1H), 7.42-7.26 (m, 2H), 6.92 (d, J=8.1 Hz, 1H), 5.08 (s, 1H), 4.21-4.12 (m, 2H), 3.86-3.78 (m, 2H), 3.84 (s, 3H), 3.54 (dd, J=7.4, 2.6 Hz, 2H), 3.26 (m, 2H), 1.35 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 356.9 (100%, MNH.sub.4.sup.+), 339.9 (41%, MH.sup.+), 239.9 (6%, MH.sup.+-boc).
1.1.2 General Procedure:
[0674] 15 mmol of the appropriate educt was placed under nitrogen in 10 mL of DMF: [0675] (VI) 4-hydroxy-3-methyl-benzaldehyde (2.04 g, 15 mmol) [0676] (VII) 4-hydroxy-2-methoxy-benzaldehyde (2.28 g, 15 mmol)
[0677] The corresponding alkylation reagent (12-15 mmol, 1.2-1.5 eq) in DMF (20 mL) and K.sub.2CO.sub.3 (4.14 g, 30 mmol) were added one after the other. The formulation was stirred under a nitrogen atmosphere for 24 h at 80° C. After cooling to room temperature, the reaction mixture was diluted with 120 mL ethyl acetate and the organic phase was shaken three times, each time with 40 mL of saturated aqueous sodium chloride solution, and once with 40 mL of water. The organic phase was separated, dried over MgSO.sub.4 and rotary evaporated. In order to purify the product, column chromatography was carried out on silica gel.
E-2: 3-methyl-4-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde
[0678] ##STR00103##
[0679] Educt: 4-hydroxy-3-methyl-benzaldehyde.
[0680] The alkylation reagent used was 2-(tert-butoxycarbonylamino)ethyl bromide (5.02 g, 22.5 mmol).
[0681] Yield: 73% of theory, 3.44 g yellowish solid
[0682] Molecular mass=279.34 g/mol; Empirical formula=C.sub.15H.sub.21NO.sub.4
[0683] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=9.85 (s, 1H), 7.75-7.66 (m, 2H), 6.90 (d, J=8.9 Hz, 1H), 4.95 (s, 1H), 4.11 (t, J=5.2 Hz, 2H), 3.59 (m, 2H), 2.27 (s, 3H), 1.45 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 302.1 (11%, MNa.sup.+), 280.1 (9%, MH.sup.+), 224.1 (100%, MH.sup.+—C.sub.4H.sub.9), 180.1 (4%, MH.sup.+-boc).
E-4: 2-methoxy-4-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde
[0684] ##STR00104##
[0685] Educt: 4-hydroxy-2-methoxy-benzaldehyde.
[0686] The alkylation reagent used was 2-(tert-butoxycarbonylamino)ethyl bromide (5.02 g, 22.5 mmol).
[0687] Yield: 71% of theory, 3.94 g of pale yellow solid
[0688] Molecular mass=295.34 g/mol; Empirical formula=C.sub.15H.sub.21NO.sub.5
[0689] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=10.25 (d, J=1.2 Hz, 1H), 7.76 (dd, J=8.6, 1.5 Hz, 1H), 6.58-6.46 (m, 1H), 6.44 (s, 1H), 5.06 (s, 1H), 4.07 (t, J=5.0 Hz, 2H), 3.87 (s, 3H), 3.53 (m, 2H), 1.43 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 318.1 (8%, MNa.sup.+), 296.1 (100%, MH.sup.+), 240.1 (87%, MH.sup.+—C.sub.4H.sub.9), 196.1 (3%, MH.sup.+-boc).
E-5: 3-methoxy-4-(2-N-tert-butyloxycarbonyl-aminoethoxy)-5-iodo-benzaldehyde
[0690] ##STR00105##
[0691] 5-lodovanillin (4.17 g, 15 mmol) was placed under nitrogen in 10 mL of DMF.
[0692] The alkylation reagent used was 2-(tert-butoxycarbonylamino)ethyl bromide (5.02 g, 22.5 mmol) dissolved in 20 mL of DMF. The alkylation reagent and K.sub.2CO.sub.3 (4.14 g, 30 mmol) were added one after the other. The formulation was stirred under a nitrogen atmosphere for 24 h at 80° C. After cooling to room temperature, the reaction mixture was diluted with 120 mL ethyl acetate and the organic phase was shaken three times, each time with 40 mL of saturated aqueous sodium chloride solution, and once with 40 mL of water. The organic phase was separated, dried over MgSO.sub.4 and rotary evaporated. In order to purify the product, column chromatography was carried out on silica gel.
[0693] Yield: 66% of theory, 3.07 g of yellow solid
[0694] Molecular mass=421.23 g/mol; Empirical formula=C.sub.15H.sub.20INO.sub.5
[0695] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=9.82 (s, 1H), 7.84 (d, J=1.7 Hz, 1H), 7.39 (d, J=1.8 Hz, 1H), 5.40 (s, 1H), 4.17 (t, J=4.9 Hz, 2H), 3.91 (s, 3H), 3.52 (dd, J=10.3, 5.3 Hz, 2H), 1.45 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 444.1 (6%, MNa.sup.+), 422.1 (3%, MH.sup.+), 366.0 (63%, MH.sup.+—C.sub.4H.sub.9), 322.0 (100%, MH.sup.+-boc).
1.2 Multi-Substituted Aldehydes
[0696] The corresponding educt was placed under nitrogen in 10 mL of DMF: [0697] (VIII) 3,4-dihydroxybenzaldehyde (3.45 g, 25 mmol) [0698] (IX) 3,4,5-trihydroxybenzaldehyde (2.31 g, 15 mmol)
[0699] The alkylation reagent used was 2-(tert-butoxycarbonylamino)ethyl bromide (11.2 g, 50 mmol, 2 eq) dissolved in 30 mL of DMF. The alkylation reagent and K.sub.2CO.sub.3 (13.8 g, 100 mmol) were added one after the other. The formulation was stirred under a nitrogen atmosphere for 16 h at 80° C. A second portion of the alkylation reagent (50 mmol, 2 eq) in 20 mL of DMF was added and the formulation was stirred for a further 24 h at 80° C. After cooling to room temperature, the reaction mixture was diluted with 250 mL ethyl acetate and the organic phase was shaken three times, each time with 150 mL of saturated aqueous sodium chloride solution, and once with 100 mL of water. The organic phase was separated, dried over MgSO.sub.4 and rotary evaporated. In order to purify the product, column chromatography was carried out on silica gel (Eluent: initially PE:EE=3:1, then PE:EE=3:2).
E-11: 3,4-bis-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde
[0700] ##STR00106##
[0701] Educt: 3,4-dihydroxybenzaldehyde
[0702] Yield: 8.1 g of pale yellow glass, 69% of theory
[0703] Molecular mass=424.50 g/mol; Empirical formula=C.sub.21H.sub.32N.sub.2O.sub.7
[0704] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=9.83 (s, 1H), 7.52-7.37 (m, 2H), 6.99 (d, J=8.2 Hz, 1H), 5.20 (s, 2H), 4.17-4.06 (m, 4H), 3.56 (m, 4H), 1.44 (s, 9H), 1.43 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 447.2 (100%, MNa.sup.+), 425.2 (70%, MH.sup.+), 369.2 (49%, MH.sup.+—C.sub.4H.sub.9), 269.1 (78%, MH.sup.+-boc).
E-12: 3,4,5-tris-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde
[0705] ##STR00107##
[0706] Educt: 3,4,5-Trihydroxybenzaldehyde
[0707] Yield: 6.22 g pale yellow glass, 61% of theory
[0708] Molecular mass=583.68 g/mol; Empirical formula=C.sub.28H.sub.45N.sub.3O.sub.10
[0709] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=9.82 (s, 1H), 7.11 (s, 2H), 5.71 (s, 1H), 5.21 (s, 2H), 4.12 (t, J=4.6 Hz, 6H), 3.62-3.51 (m, 4H), 3.42 (d, J=4.9 Hz, 2H), 1.44 (d, J=8.7 Hz, 27H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 606.3 (47%, MNa.sup.+), 584.3 (100%, MH.sup.+), 528.3 (4%, MH.sup.+—C.sub.4H.sub.9), 484.3 (23%, MH.sup.+-boc).
1.3 Mixed Substituted Benzaldehydes
Step 1:
[0710] 3,4-dihydroxybenzaldehyde (3.45 g, 25 mmol) was placed under nitrogen in 20 mL of DMF. 2-(tert-butoxycarbonylamino)ethyl bromide (6.72 g, 30 mmol) in 30 mL of DMF and K.sub.2CO.sub.3 (4.6 g, 33 mmol) were added one after the other. The formulation was stirred under a nitrogen atmosphere for 24 h at 80° C. After cooling to room temperature, the reaction mixture was diluted with 150 mL ethyl acetate and the organic phase was shaken three times, each time with 100 mL of saturated aqueous sodium chloride solution, and once with 100 mL of water. The organic phase was separated, dried over MgSO.sub.4 and rotary evaporated. In order to purify the products, column chromatography was carried out on silica gel.
E-8: 3-hydroxy-4-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde
[0711] ##STR00108##
[0712] Yield: 1.48 g of pale yellow glass, 17% of theory
[0713] Molecular mass=281.31 g/mol; Empirical formula=C.sub.14H.sub.19NO.sub.5
[0714] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=9.78 (s, 1H), 7.42 (d, J=1.9 Hz, 1H), 7.36 (dd, J=8.2, 1.9 Hz, 1H), 7.14 (s, 1H), 6.89 (d, J=8.2 Hz, 1H), 5.53 (s, 1H), 4.13 (t, J=4.9 Hz, 2H), 3.58 (dd, J=10.6, 5.2 Hz, 2H), 1.42 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 304.1 (19%, MNa.sup.+), 282.1 (2%, MH.sup.+), 226.1 (100%, MH.sup.+—C.sub.4H.sub.9), 182.1 (42%, MH.sup.+-boc).
E-8a: 4-hydroxy-3-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde
[0715] ##STR00109##
[0716] Yield: 2.60 g of pale yellow glass, 37% of theory
[0717] Molecular mass=281.31 g/mol; —Empirical formula=C.sub.14H.sub.19NO.sub.5
E-11: 3,4-bis-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde
[0718] ##STR00110##
[0719] Yield: 1.38 g of pale yellow glass, 13% of theory
[0720] Molecular mass=424.50 g/mol; Empirical formula=C.sub.21H.sub.32N.sub.2O.sub.7
[0721] .sup.1H-NMR and MS as above.
Step 2:
[0722] 1.41 g (5 mmol) of 3-hydroxy-4-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde (E-8) was placed under nitrogen in 10 mL of DMF. The alkylation reagent, each time dissolved in 10 mL of DMF, namely triethylene glycol monotosylate or 1-bromooctanol, as well as K.sub.2CO.sub.3 (2.07 g, 15 mmol) were added one after the other. The formulation was stirred under a nitrogen atmosphere for 24 h at 80° C. After cooling to room temperature, the reaction mixture was diluted with 80 mL ethyl acetate and the organic phase was shaken three times, each time with 50 mL of saturated aqueous sodium chloride solution, and once with 50 mL of water. The organic phase was separated, dried over MgSO.sub.4 and rotary evaporated. In order to purify the product, column chromatography was carried out on silica gel.
E-10: 3-(2-(2-(2-hydroxy-ethoxy)ethoxy)ethoxy)-4-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde
[0723] ##STR00111##
[0724] The alkylation reagent used was triethylene glycol monotosylate (3.04 g, 10 mmol). Column chromatography with EE.fwdarw.EE/EtOH=3:1
[0725] Yield: 1.53 g of pale yellow glass, 74% of theory
[0726] Molecular mass=413.47 g/mol; Empirical formula=C.sub.20H.sub.31NO.sub.8
[0727] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=9.83 (s, 1H), 7.48-7.38 (m, 2H), 6.96 (d, J=8.1 Hz, 1H), 4.24 (dd, J=5.3, 3.4 Hz, 2H), 4.12 (t, J=4.8 Hz, 2H), 3.92 (dd, J=5.3, 3.4 Hz, 2H), 3.74 (t, J=4.6 Hz, 6H), 3.66-3.61 (m, 2H), 3.58 (t, J=4.7 Hz, 2H), 1.43 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 436.2 (35%, MNa.sup.+), 414.2 (33%, MH.sup.+), 370.2 (5%, MH.sup.+—C.sub.4H.sub.9), 314.2 (100%, MH.sup.+-boc).
E-9: 3-octyloxy-4-(2-N-tert-butyloxycarbonyl-aminoethoxy)benzaldehyde
[0728] ##STR00112##
[0729] The alkylation reagent used was 1-bromooctanol (1.93 g, 10 mmol).
[0730] Column chromatography with PE:EE=3:1
[0731] Yield: 1.53 g of pale yellow glass, 78% of theory
[0732] Molecular mass=393.53 g/mol; Empirical formula=C.sub.22H.sub.35NO.sub.5
[0733] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=9.81 (s, 1H), 7.43-7.32 (m, 2H), 6.96 (d, J=8.4 Hz, 1H), 4.11 (t, J=5.3 Hz, 2H), 4.02 (t, J=6.7 Hz, 2H), 3.55 (dd, J=10.2, 5.0 Hz, 2H), 1.91-1.76 (m, 2H), 1.47-1.39 (m, 2H), 1.42 (s, 9H), 1.36-1.16 (m, 8H), 0.90-0.80 (m, 3H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 416.2 (51%, MNa.sup.+), 396.2 (4%, MH.sup.+), 338.2 (7%, MH.sup.+-C.sub.4H.sub.9), 294.2 (100%, MH.sup.+-boc).
1.4 Substituted naphthaldehyde
E-13: 4-(2-N-tert-butyloxycarbonyl-aminoethoxy)naphthalene-1-carbaldehyde
[0734] ##STR00113##
[0735] 4-hydroxynaphth-1-aldehyde (1.74 g, 10 mmol) was placed under nitrogen in 10 mL of DMF. 2-(tert-butoxycarbonylamino)ethyl bromide (4.4 g, 20 mmol) in 10 mL of DMF, potassium iodide (10 mmol, 1.66 g) and Cs.sub.2CO.sub.3 (6.5 g, 20 mmol) were added one after the other. The formulation was stirred under a nitrogen atmosphere for 24 h at 80° C. After cooling to room temperature, the reaction mixture was diluted with 80 mL ethyl acetate and the organic phase was shaken three times, each time with 50 mL of saturated aqueous sodium chloride solution, and once with 100 mL of water. The organic phase was separated, dried over MgSO.sub.4 and rotary evaporated. The impure product was washed with EE/PE 1:19 in order to remove the excess reagent. Next, the orange solid was extracted with EE/PE 1:2 (3×, each 100 mL) and the precipitate was filtered out. The solution was rotary evaporated and the residue was dissolved in 250 mL of diethylether. The organic phase was shaken twice, each time with 50 mL of 3 M NaOH, and once with 100 mL of water. The organic phase was separated, dried over MgSO.sub.4 and rotary evaporated. In order to further purify the product, column chromatography was carried out on silica gel. Yield: 2.08 g of pale yellow solid, 47% of theory Molecular mass=315.37 g/mol; Empirical formula=C.sub.18H.sub.21NO.sub.4
[0736] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=10.19 (s, 1H), 9.30 (d, J=8.5 Hz, 1H), 8.33 (d, J=8.4 Hz, 1H), 7.94-7.86 (m, 1H), 7.76-7.64 (m, 1H), 7.62-7.51 (m, 1H), 6.94-6.82 (m, 1H), 5.07 (s, 1H), 4.29 (dd, J=6.9, 3.2 Hz, 2H), 3.73 (m, 2H), 1.46 (s, 9H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 338.2 (16%, MNa.sup.+), 316.2 (19%, MH.sup.+), 260.1 (100%, MH.sup.+—C.sub.4H.sub.9), 216.1 (4%, MH.sup.+-boc).
2. Synthesis of Substituted Curcumins
[0737] The corresponding curcumins were synthesized from the preliminary steps described above as curcumins protected with tert-butyloxycarbonyl (boc). The boc protective group was then removed.
2.1 Synthesis of Symmetrically Substituted Curcumins
[0738] Overview 3: Synthesis of various symmetrically substituted curcumins: Conditions: (a) acetyl acetone, B.sub.2O.sub.3, B(OBu).sub.3, n-butylamine, ethyl acetate, 80° C., 6 h, then hydrolysis with HOAc 40% overnight, RT; (b) acetyl acetone, B.sub.2O.sub.3, B(OBu).sub.3, n-butylamine, DMF, 80° C., 6 h, then hydrolysis with HOAc 40% overnight, RT; (c) 3,5-heptanedione, B.sub.2O.sub.3, B(OBu).sub.3, n-butylamine, DMF, 80° C., 6 h, then hydrolysis with HOAc 40% overnight, RT; (d) DCM, TFA, RT, 5 h; then Amberlite IRA-958 ion exchange resin, water.
##STR00114## ##STR00115## ##STR00116## ##STR00117## ##STR00118## ##STR00119##
Overview 4: Synthesis of symmetrically substituted curcumins starting from E-3: Conditions: (a) B.sub.2O.sub.3, B(OBu).sub.3, n-butylamine, DMF or ethyl acetate, 80° C., 6 h, then hydrolysis with HOAc 40% overnight, RT; (b) DCM, TFA, RT, 5 h; (c) 1,3-di-boc-2-(trifluoromethyl-sulphonyl)guanidine, DCM, triethylamine, 0° C..fwdarw.RT, 4 h; (d) DCM, TFA, RT, 5 h; then Amberlite IRA-958 ion exchange resin, water
2.1.1 General Procedure:
[0739] Acetyl acetone (0.1 g, 1 mmol) and boron oxide B.sub.2O.sub.3 (0.07 g, 1 mmol) were dissolved in ethyl acetate (2 mL) and stirred for 30 minutes at 50° C. The substituted benzaldehyde (20 mmol) in ethyl acetate (3 mL) along with tributylborate (0.7 g, 3 mmol) were added one after the other and the formulation was stirred for a further half an hour. Next, n-butylamine (0.1 mL in 1 mL EE) was added dropwise over 5 minutes. After stirring for a further five hours at 50° C., the reaction was left to stand overnight. The solution was rotary evaporated and the residue was quickly dried under high vacuum. 10 mL of ethyl acetate was added per 1 g of impure product and the impure product was dissolved. For the subsequent hydrolysis of the boron complex, double the volume of 50% acetic acid was added (20 mL per 1 g of impure product). After stirring for 24 h at room temperature protected from light, the mixture of solvents was withdrawn under reduced pressure. The residue was extracted three times with EE (3×20 mL) and the insoluble salt was filtered off. The combined organic phases were washed with water (30 mL), dried over MgSO.sub.4 and finally, the solvent was withdrawn under reduced pressure. Purification was carried out using column chromatography on silica gel with acetone/PE. The purified fraction of the curcumin which was obtained was then dissolved in as little ethyl acetate as possible. By dripping this solution into an excess of petroleum ether, the product was precipitated out as a fine yellow-orange powder.
1,7-bis(3,4-dimethoxyphenyl)hepta-1,6-diene-3,5-dione (tetramethoxycurcumin)
[0740] ##STR00120##
[0741] Quantity of corresponding aldehyde used: 334 mg=2 mmol
[0742] Column chromatography on silica gel with acetone/PE=2:5.fwdarw.1:2
[0743] Yield: 289 mg, 73% of theory, orange, viscous solid or orangey-yellow powder. Molecular
[0744] mass=396.44 g/mol; Empirical formula=C.sub.23H.sub.24O.sub.6
C-1: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-phenoxy)-ethyl]-carbamic acid tert-butyl ester
[0745] ##STR00121##
[0746] Quantity of corresponding aldehyde E-1 used: 530 mg=2 mmol
[0747] Column chromatography on silica gel with acetone/PE=1:3.fwdarw.2:3
[0748] Yield: 363 mg, 67% of theory, yellow solid or yellow powder
[0749] Molecular mass=594.71 g/mol; Empirical formula=C.sub.33H.sub.42N.sub.2O.sub.8
[0750] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=7.61 (d, J=15.8 Hz, 2H), 7.50 (d, J=8.8 Hz, 4H), 6.89 (d, J=8.7 Hz, 4H), 6.50 (d, J=15.8 Hz, 2H), 5.02 (s, 2H), 4.05 (t, J=5.1 Hz, 4H), 3.60-3.48 (m, 4H), 1.45 (s, 18H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 595.3 (100%, MH.sup.+), 539.2 (18%, MH.sup.+—C.sub.4H.sub.9).
C-5: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-3-methyl-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-2-methyl-phenoxy)-ethyl]-carbamic acid tert-butylester
[0751] ##STR00122##
[0752] Quantity of corresponding aldehyde E-2 used: 559 mg=2 mmol
[0753] Column chromatography on silica gel with acetone/PE=1:3.fwdarw.2:3
[0754] Yield: 442 mg, 59% of theory, orangey-yellow solid or orangey-yellow powder. Molecular
[0755] mass=622.77 g/mol; Empirical formula=C.sub.35H.sub.46N.sub.2O.sub.8
[0756] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=7.59 (d, J=15.8 Hz, 2H), 7.36 (d, J=10.8 Hz, 4H), 6.80 (d, J=8.3 Hz, 2H), 6.49 (d, J=15.8 Hz, 2H), 4.94 (s, 2H), 4.06 (t, J=5.0 Hz, 4H), 3.58 (m, 4H), 2.25 (s, 6H), 1.46 (s, 18H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 623.3 (100%, MH.sup.+), 567.3 (21%, MH.sup.+—C.sub.4H.sub.9).
C-3: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-2-octyloxy-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-3-octyloxy-phenoxy)-ethyl]-carbamic acid tert-butylester
[0757] ##STR00123##
[0758] Quantity of corresponding aldehyde E-4 used: 591 mg=2 mmol
[0759] Column chromatography on silica gel with acetone/PE=1:3.fwdarw.2:3
[0760] Yield: 393 mg, 58% of theory, orange, viscous solid or orangey-yellow powder. Molecular
[0761] mass=654.76 g/mol; Empirical formula=C.sub.35H.sub.46N.sub.2O.sub.10
[0762] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=7.89 (d, J=16.0 Hz, 2H), 7.47 (d, J=8.5 Hz, 2H), 6.62 (d, J=16.0 Hz, 2H), 6.52-6.39 (m, 4H), 4.99 (s, 2H), 4.06 (t, J=5.1 Hz, 4H), 3.88 (s, 6H), 3.55 (m, 4H), 1.45 (s, 18H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 655.3 (100%, MH.sup.+).
C-4: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-3-methoxy-5-iodo-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-2-octyloxy-6-iodo-phenoxy)-ethyl]-carbamic acid tert-butylester
[0763] ##STR00124##
[0764] Quantity of corresponding aldehyde E-5 used: 842 mg=2 mmol
[0765] Column chromatography on silica gel with acetone/PE=1:4.fwdarw.1:2
[0766] Yield: 580 mg, 64% of theory, orange, viscous solid or orangey-yellow powder. Molecular
[0767] mass=906.56 g/mol; Empirical formula=C.sub.35H.sub.4412N.sub.2O.sub.10
[0768] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=7.57 (d, J=1.5 Hz, 2H), 7.50 (d, J=15.8 Hz, 2H), 7.02 (d, J=1.5 Hz, 2H), 6.51 (d, J=15.8 Hz, 2H), 5.47 (s, 2H), 4.11 (t, J=4.7 Hz, 4H), 3.90 (s, 6H), 3.52 (m, 4H), 1.46 (s, 18H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 929.1 (100%, MNa.sup.+), 907.1 (100%, MH.sup.+), 807.1 (42%, MH.sup.+-boc).
C-2: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-3-methoxy-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-2-methoxy-phenoxy)-ethyl]-carbamic acid tert-butylester
[0769] ##STR00125##
[0770] Quantity of corresponding aldehyde E-15 used: 590 mg=2 mmol
[0771] Column chromatography on silica gel with acetone/PE=1:3.fwdarw.2:3
[0772] Yield: 354 mg, 54% of theory, orange, viscous solid or orangey-yellow powder. Molecular
[0773] mass=654.76 g/mol, Empirical formula=C.sub.35H.sub.46N.sub.2O.sub.10
[0774] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=7.58 (d, J=15.7 Hz, 2H), 7.14-6.99 (m, 4H), 6.87 (d, J=8.3 Hz, 2H), 6.48 (d, J=15.8 Hz, 2H), 5.16 (s, 2H), 4.09 (t, J=4.9 Hz, 4H), 3.90 (s, 6H), 3.56 (m, 4H), 1.44 (s, 18H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 655.3 (100%, MH.sup.+), 555.3 (26%, MH.sup.+-boc).
C-6: (2-{2-[4-(7-{4-[2-(2-tert-butoxycarbonylamino-ethoxy)-ethoxy]-3-methoxy-phenyl}-3,5-dioxo-hepta-1,6-dienyl)-2-methoxy-phenoxy]-ethoxy}-ethyl)-carbamic acid tert-butyl ester
[0775] ##STR00126##
[0776] Quantity of corresponding aldehyde E-7 used: 679 mg=2 mmol
[0777] Yield: 349 mg, 47% of theory, orange, viscous solid or orange powder
[0778] Column chromatography on silica gel with acetone/PE=1:2.fwdarw.1:1
[0779] Molecular mass=742.87 g/mol; Empirical formula=C.sub.39H.sub.54N.sub.2O.sub.12
[0780] .sup.1H NMR: (300 MHz, CDCl.sub.3), δ=7.59 (d, J=15.8 Hz, 2H), 7.18-7.03 (m, 4H), 6.90 (d, J=8.3 Hz, 2H), 6.49 (d, J=15.8 Hz, 2H), 5.07 (s, 2H), 4.24-4.15 (m, 4H), 3.91 (s, 6H), 3.88-3.83 (m, 4H), 3.61 (t, J=5.1 Hz, 4H), 3.33 (m, 4H), 1.43 (s, 18H). MS: (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 671.3 (51%, MNa.sup.+), 655.3 (86%, MH.sup.+), 555.3 (100%, MH.sup.+-boc).
C-7: {2-[4-{7-[3,4-bis-(2-tert-butoxycarbonylamino-ethoxy)-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-2-(2-tert-butoxycarbonylamino-ethoxy)-phenoxy]-ethyl}-carbamic acid tert-butyl ester
[0781] ##STR00127##
[0782] Quantity of corresponding aldehyde E-11 used: 850 mg=2 mmol
[0783] Yield: 374 mg, 41% of theory, orange, very viscous solid or orangey-red powder
[0784] Column chromatography on silica gel with acetone/PE=1:3.fwdarw.1:2
[0785] Molecular mass=913.08 g/mol; Empirical formula=C.sub.47H.sub.68N.sub.4O.sub.14
[0786] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.57 (d, J=15.8 Hz, 2H), 7.19-7.12 (m, 4H), 6.98-6.86 (m, 2H), 6.50 (d, J=15.8 Hz, 2H), 5.28 (s, 4H), 4.10 (t, J=4.4 Hz, 8H), 3.55 (m, 8H), 1.46 (s, 18H), 1.45 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 913.5 (100%, MH.sup.+), 457.3 (7%, (M+2H.sup.+).sup.2+):
C-8: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-3-benzyloxy-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-2-benzyloxy-phenoxy)-ethyl]-carbamic acid tert-butyl ester
[0787] ##STR00128##
[0788] Quantity of corresponding aldehyde E-7 used: 738 mg=2 mmol
[0789] Yield: 518 mg, 62% of theory, orange solid or orange powder
[0790] Column chromatography on silica gel with acetone/PE=1:3.fwdarw.2:3
[0791] Molecular mass=835.02 g/mol; Empirical formula=C.sub.49H.sub.58N.sub.2O.sub.10
[0792] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.55 (d, J=15.8 Hz, 2H), 7.45-7.31 (m, 10H), 7.14-7.04 (m, 4H), 6.86 (d, J=8.3 Hz, 2H), 6.46 (d, J=15.8 Hz, 2H), 5.39 (s, 2H), 5.22 (s, 4H), 4.14 (t, J=5.9 Hz, 4H), 3.39 (m, 4H), 2.09-2.00 (m, 4H), 1.41 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 857.4 (100%, MNa.sup.+), 835.4 (76%, MH.sup.+), 735.4 (53%, MH.sup.+-boc).
C-9: {2-[4-{7-[3,4,5-tris-(2-tert-butoxycarbonylamino-ethoxy)-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-2,6-bis-(2-tert-butoxycarbonylamino-ethoxy)-phenoxy]-ethyl}-carbamic acid tert-butyl ester
[0793] ##STR00129##
[0794] Quantity of corresponding aldehyde E-11 used: 1166 mg=2 mmol
[0795] Yield: 554 mg, 45% of theory, orange, very viscous solid or orangey-red powder. Column chromatography on silica gel with acetone/PE=1:4.fwdarw.1:2. Molecular mass=1231.46 g/mol;
[0796] Empirical formula=C.sub.61H.sub.94N.sub.6O.sub.20
[0797] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.52 (d, J=15.1 Hz, 2H), 6.79 (s, 4H), 6.51 (d, J=15.1 Hz, 2H), 5.75 (s, 2H), 5.25 (s, 3H), 4.10 (m, 12H), 3.57 (m, 8H), 3.41 (m, 4H), 1.47 (s, 18H), 1.47 (s, 36H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 1253.6 (100%, MNa.sup.+), 1231.6 (49%, MH.sup.+), 1131.3 (51%, MH.sup.+-boc).
C-10: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-3-octyloxy-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-2-octyloxy-phenoxy)-ethyl]-carbamic acid tert-butyl ester
[0798] ##STR00130##
[0799] Quantity of corresponding aldehyde E-10 used: 787 mg=2 mmol
[0800] Column chromatography on silica gel with acetone/PE=1:4.fwdarw.1:2
[0801] Yield: 400 mg, 47% of theory, orange, viscous solid or orangey-yellow powder. Molecular
[0802] mass=851.14 g/mol; Empirical formula=C.sub.49H.sub.74N.sub.2O.sub.10
[0803] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.58 (d, J=15.7 Hz, 2H), 7.15-7.05 (m, 4H), 6.89 (d, J=8.2 Hz, 2H), 6.48 (d, J=15.8 Hz, 2H), 5.17 (s, 2H), 4.09 (t, J=5.0 Hz, 4H), 4.03 (t, J=6.7 Hz, 4H), 3.54 (m, 4H), 1.91-1.80 (m, 4H), 1.41 (s, 18H), 1.51-1.26 (m, 20H), 0.89 (t, 4.5 Hz, 6H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 873.5 (100%, MNa.sup.+), 851.4 (13%, MH.sup.+), 751.5 (38%, MH.sup.+-boc).
2.1.2 General Procedure:
[0804] The reaction was carried out under an atmosphere of nitrogen and protected from light. The beta-diketone used was: [0805] (X) acetyl acetone (0.1 g, 1 mmol) [0806] (XI) 3-methyl-2,4-pentanedione (0.12 g, 1 mmol) [0807] (XII) 3,5-heptanedione (0.13 g, 0.137 mL, 1 mmol), or [0808] (XIII) 2-acetylcyclohexanone (0.14 g, 1 mmol).
[0809] The respective beta-diketone and boron oxide B.sub.2O.sub.3 (0.05 g, 0.7 mmol) were dissolved in dry DMF (2 mL) and stirred for 30 minutes at 70° C. The corresponding substituted aldehyde (2 mmol) along with tributylborate (0.46 g, 2 mmol) were dissolved in dry DMF (5 mL). This solution was added to the formulation and it was stirred for a further half hour at 85° C. Next, n-butylamine (0.1 mL in 1 mL of DMF) was added dropwise and the formulation was stirred at 70° C. for 4 hours. After cooling to room temperature, all of the volatile components in the nitrogen stream were driven off overnight (outlet, pipe behind baffle wall). 10 mL of ethyl acetate was added per 1 g of impure product and the impure product was dissolved. For the subsequent hydrolysis of the boron complex, double the volume of 50% acetic acid was added (20 mL per 1 g of impure product). After stirring for 24 h at room temperature protected from light, the mixture of solvents was withdrawn under reduced pressure (max. water bath temperature 50° C.). The residue was extracted three times with EE (3×50 mL) and the insoluble salt was filtered off. The combined organic phases were washed with water (50 mL), dried over MgSO.sub.4 and finally, the solvent was withdrawn under reduced pressure. Purification was carried out using column chromatography on silica gel with acetone/PE. The pure fraction of the curcumin was then dissolved in as little ethyl acetate as possible. By dripping this solution into an excess of petroleum ether, the product was precipitated out as a fine yellow-orange powder.
C-26: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-3-hydroxy-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-2-hvdroxv-phenoxy)-ethyl]-carbamic acid tert-butyl ester
[0810] ##STR00131##
[0811] Beta-diketone: acetyl acetone
[0812] Quantity of corresponding aldehyde E-10 used: 560 mg=2 mmol
[0813] Column chromatography on silica gel with acetone/PE=1:2.fwdarw.2:3
[0814] Yield: 28% of theory, orange, viscous solid (180 mg).
[0815] Molecular mass=626.71 g/mol; Empirical formula=C.sub.33H.sub.42N.sub.2O.sub.10
[0816] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.57 (d, J=15.8 Hz, 2H), 7.34 (m, H), 7.19-7.08 (m, 2H), 6.85 (d, J=8.4 Hz, 2H), 6.64 (d, J=15.8 Hz, 2H), 5.17 (s, 2H), 4.07 (t, J=5.0 Hz, 4H), 3.58-3.44 (m, 4H), 1.43 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 627.2 (100%, MH.sup.+), 527.2 (37%, MH.sup.+-boc).
C-18: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-napthyl]-3,5-dioxo-hepta-1,6-dienyl}-naphthoxy)-ethyl]-carbamic acid tert-butyl ester
[0817] ##STR00132##
[0818] Beta-diketone: acetyl acetone
[0819] Quantity of corresponding aldehyde E-13 used: 630 mg=2 mmol
[0820] Column chromatography on silica gel with acetone/PE=2:7.fwdarw.1:2
[0821] Yield: 62% of theory, orange, viscous solid (430 mg).
[0822] Molecular mass=694.83 g/mol; Empirical formula=C.sub.41H.sub.46N.sub.2O.sub.8
[0823] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=8.48 (d, J=15.5 Hz, 2H), 8.33 (d, J=8.1 Hz, 2H), 8.25 (d, J=8.0 Hz, 2H), 7.80 (d, J=8.1 Hz, 2H), 7.63 (t, J=6.9 Hz, 2H), 7.55 (t, J=6.8 Hz, 2H), 6.86 (d, J=8.2 Hz, 2H), 6.68 (d, J=15.5 Hz, 2H), 5.04 (s, 2H), 4.26 (t, J=5.0 Hz, 4H), 3.72 (q, J=5.0 Hz, 5H), 1.47 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 717.3 (100%, MNa.sup.+), 695.3 (56%, MH.sup.+), 639.3 (73%, MH.sup.+—C.sub.4H.sub.9).
C-19: [2-(6-{7-[6-(2-tert-butoxycarbonylamino-ethoxy)-napthyl]-3,5-dioxo-hepta-1,6-dienyl}-naphthoxy)-ethyl]-carbamic acid tert-butyl ester
[0824] ##STR00133##
[0825] Beta-diketone: acetyl acetone
[0826] Quantity of corresponding aldehyde E-19 used: 1.26 g=4 mmol
[0827] Yield: 820 mg, 59% of theory, reddish, viscous solid or orange powder.
[0828] Molecular mass=694.83 g/mol; Empirical formula=C.sub.41H.sub.46N.sub.2O.sub.8
[0829] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.85 (d, J=15.8 Hz, 2H), 7.79-7.63 (m, 8H), 7.20-7.08 (m, 4H), 6.71 (d, J=15.8 Hz, 2H), 5.06 (s, 2H), 4.15 (t, J=5.0 Hz, 4H), 3.61 (m, 4H), 1.46 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 717.3 (58%, MNa.sup.+), 695.3 (100%, MH.sup.+), 639.3 (97%, MH.sup.+—C.sub.4H.sub.9).
C-13: tert-butyl-N-[2-[4-[(E)-3-[(3E)-3-[[4-(2-(tert-butoxycarbonylamino)ethoxy]-3-methoxy-phenylmethylene]-2-oxo-cyclohexyl]-3-oxo-prop-1-enyl]-2-methoxy-phenoxy]ethyl]carbamate
[0830] ##STR00134##
[0831] Beta-diketone: 2-acetylcyclohexanone
[0832] Quantity of corresponding aldehyde used: 580 mg=2 mmol
[0833] Flash chromatography on silica gel with acetone/PE=2:5.fwdarw.1:2
[0834] The fractions containing product were rotary evaporated and the impure product was recrystallized from acetone/PE 1:3. After cooling slowly to RT, it was first cooled slowly overnight in a refrigerator, then cooled for 3 h in the freezer. The solid was extracted several times with a little ice-cold acetone/PE 1:3 and finally washed with PE. The product was dried in the air. A further, small crystalline fraction was obtained by concentrating the mother liquor.
[0835] Yield: 68% of theory, orange, matted crystal needles (472 mg).
[0836] Molecular mass=694.83 g/mol; Empirical formula=C.sub.38H.sub.50N.sub.2O.sub.10
[0837] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.76-7.62 (m, 2H), 7.17 (dd, J=8.3, 1.5 Hz, 1H), 7.09 (d, J=1.7 Hz, 1H), 7.06-6.85 (m, 5H), 5.22-5.08 (m, 2H), 4.11 (t, J=4.5 Hz, 4H), 3.92 (s, 3H), 3.89 (s, 3H), 3.57 (m, 4H), 2.83-2.73 (m, 2H), 2.71-2.64 (m, 2H), 1.86-1.77 (m, 2H), 1.45 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 717.4 (24%, MNa.sup.+), 695.4 (100%, MH.sup.+), 595.3 (30%, MH.sup.+-boc).
C-9: {2-[4-{7-[3,4,5-tris-(2-tert-butoxycarbonylamino-ethoxy)-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-2,6-bis-(2-tert-butoxycarbonylamino-ethoxy)-phenoxy]-ethyl}-carbamic acid tert-butyl ester
[0838] ##STR00135##
[0839] Beta-diketone: acetyl acetone
[0840] Quantity of corresponding aldehyde E-12 used: 1160 mg=2 mmol
[0841] Yield: 66% of theory, orange, viscous solid or orangey-red powder (810 mg).
[0842] Column chromatography on silica gel with acetone/PE=1:4.fwdarw.2:3.
[0843] Molecular mass=1231.46 g/mol; Empirical formula=C.sub.61H.sub.94N.sub.6O.sub.20
[0844] .sup.1H-NMR and MS as above.
C-11: tert-butyl-N-[2-[4-[(1E,6E)-7-[4-[2-(tert-butoxycarbonylamino)ethoxy]-3-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]phenyl]-3,5-dioxo-hepta-1,6-dienyl]-2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]phenoxy]ethyl]carbamate
[0845] ##STR00136##
[0846] Beta-diketone: acetyl acetone
[0847] Quantity of corresponding aldehyde E-10 used: 830 mg=2 mmol
[0848] Yield: 37% of theory, orange, very viscous solid (330 mg)
[0849] Column chromatography on silica gel, acetone/PE=1:2.fwdarw.1:1; prep. TLC, acetone/petroleum ether (PE)=1:1. Molecular mass=891.03 g/mol; Empirical formula=C.sub.45H.sub.66N.sub.2O.sub.16
[0850] .sup.1H NMR (600 MHz, CDCl.sub.3), δ=7.58 (d, J=15.7 Hz, 2H), 7.20-7.06 (m, 4H), 6.88 (d, J=8.1 Hz, 2H), 6.48 (d, J=15.8 Hz, 2H), 5.91 (s, 2H), 4.25-4.17 (m, 4H), 4.12-4.04 (m, 4H), 3.93-3.86 (m, 4H), 3.83-3.46 (m, 20H), 1.44 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 891.7 (92%, MH.sup.+), 446.3 (100%, (M+2H.sup.+).sup.2+).
C-23: tert-butyl-N-[2-[4-[(1E,6E)-7-[4-[2-(tert-butoxycarbonylamino)ethoxy]-3-methoxy-phenyl]-2,6-dimethyl-3,5-dioxo-hepta-1,6-dienyl]-2-methoxy-phenoxy]ethyl]carbamate
[0851] ##STR00137##
[0852] Beta-diketone: 3,5-heptanedione.
[0853] Quantity of corresponding acetophenone E-3 used: 608 mg=2 mmol
[0854] Yield: 72% of theory, orange solid (490 mg).
[0855] Column chromatography on silica gel, acetone/PE=2:5.fwdarw.1:2; prep. TLC, acetone/PE=1:2. Molecular mass=682.82 g/mol; Empirical formula=C.sub.37H.sub.50N.sub.2O.sub.10
[0856] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.55 (s, 2H), 7.07-6.88 (m, 6H), 6.31 (s, 1H), 5.16 (s, 2H), 4.12 (t, J=5.0 Hz, 4H), 3.89 (s, 6H), 3.57 (q, J=5.1 Hz, 4H), 2.18 (s, 6H), 1.45 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 705.2 (MNa.sup.+, 67%), 683.4 (MH.sup.+, 100%), 627.3 (MH.sup.+-C.sub.4H.sub.9, 4%), 583.3 (MH.sup.+-boc, 63%), 527.2 (MH.sup.+-boc-C.sub.4H.sub.9, 27%).
C-24: tert-butyl-N-[2-[4-[(1E,6E)-7-[4-[2-(tert-butoxycarbonylamino)ethoxy]-3-methoxy-phenyl]-1,2,6-trimethyl-3,5-dioxo-octa-1,6-dienyl]-2-methoxy-phenoxy]ethyl]carbamate
[0857] ##STR00138##
[0858] Beta-diketone: 3,5-heptanedione
[0859] Quantity of corresponding acetophenone E-15 used: 608 mg=2 mmol
[0860] Yield: 19% of theory, orange, very viscous solid (137 mg)
[0861] Column chromatography on silica gel, acetone/PE=2:5.fwdarw.1:2; prep. TLC, acetone/PE=1:2.
[0862] Molecular mass=710.87 g/mol; Empirical formula=C.sub.39H.sub.54N.sub.2O.sub.10
[0863] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.12-6.91 (m, 6H), 6.26 (s, 1H), 5.14 (s, 2H), 4.13 (t, J=5.3 Hz, 4H), 3.91 (s, 6H), 3.56 (q, J=5.2 Hz, 4H), 2.17 (s, 6H), 2.12 (s, 6H), 1.44 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 732.4 (MNa.sup.+, 88%), 710.4 (MH.sup.+, 100%), 654.4 (8%, MH.sup.+—C.sub.4H.sub.9), 610.4 (53%, MH.sup.+-boc).
C-12: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-3-methoxy-phenyl]-3,5-dioxo-hepta-4-methyl-1,6-dienyl}-2-methoxy-phenoxy)-ethyl]-carbamic acid tert-butyl ester
[0864] ##STR00139##
[0865] Beta-diketone: 3-methyl-2,4-pentanedione
[0866] Quantity of corresponding aldehyde E-3 used: 2.94 g=0.01 mol
[0867] Column chromatography on silica gel with acetone/PE=1:3.fwdarw.2:3
[0868] Yield: 59% of theory, orange, viscous solid or orangey-yellow powder (403 mg).
[0869] Molecular mass=682.82 g/mol; Empirical formula=C.sub.36H.sub.48N.sub.2O.sub.10
[0870] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.74-7.56 (m, 2H), 7.20-6.82 (m, 7H), 6.70 (d, J=15.9 Hz, 1H), 5.13 (s, 2H), 4.10 (dd, J=10.7, 5.3 Hz, 4H), 3.92 (s, 3H), 3.88 (s, 3H), 3.56 (m, 4H), 2.17 (m, 3H), 1.44 (s, 9H), 1.43 (s, 9H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 691.3 (43%, MNa.sup.+), 669.3 (100%, MH.sup.+), 569.3 (17%, MH.sup.+-boc).
2.2 Modification with Guanidine
C-14: tert-butyl-(2,2′-(4,4′-((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene))bis(oxy)bis(ethane-2,1-diyl))bis(azanediyl)bis((tert-butoxycarbonylamino methan-1-yl-1-ylidene dicarbamate
[0871] ##STR00140##
[0872] N,N′-di-boc-N″-triflylguanidine was produced in a manner analogous to that described in Organic Syntheses, Coll. Vol. 10, p. 266 (2004); Vol. 78, p. 91 (2002.
[0873] Curcumin C-2 was precipitated in dichloromethane (DCM) at room temperature (RT) for 5 h with trifluoroacetic acid (TFA). The trifluoroacetate salt obtained was centrifuged off. Triethylamine (0.51 g, 0.66 mL, 5 mmol) was slowly added dropwise to N,N′-di-boc-N″-triflylguanidine (0.82 g, 2 mmol) in dichloromethane (10 mL) using a syringe at 2-5° C. Curcumin C-2 trifluoroacetate (550 mg, 0.8 mmol) was added. After stirring for 5 h at room temperature, it was diluted with dichloromethane (30 mL) and the organic phase was washed with aqueous potassium hydrogen sulphate (3%, 20 mL) and water (20 mL). After drying over MgSO.sub.4, the solution was filtered and rotary evaporated. The impure material was purified using column chromatography on silica gel with acetone/petroleum ether (PE) (acetone/PE=1:3.fwdarw.1:2). Yield: 47% of theory, orange, viscous solid or orangey-yellow powder (437 mg).
[0874] Molecular mass=930.08 g/mol; Empirical formula=C.sub.47H.sub.66N.sub.6O.sub.14
[0875] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=11.47 (s, 2H), 8.79 (s, 2H), 7.59 (d, J=15.8 Hz, 2H), 7.16-7.05 (m, 4H), 7.01 (d, J=8.3 Hz, 2H), 6.50 (d, J=15.8 Hz, 2H), 4.20 (t, J=5.3 Hz, 4H), 3.92 (s, 6H), 3.89-3.81 (m, 4H), 1.51 (s, 18H), 1.49 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 939.6 (9%, MH.sup.+), 470.4 (100%, (M+2H.sup.+).sup.2+).
2.3 Synthesis of Substituted Curcumins Using the Mitsunobu Reaction
[0876] The synthesis was carried out in a manner analogous to the methods described in Lepore, S. D. and He, Y.: (“Use of Sonication for the Coupling of Sterically Hindered Substrates in the phenolic Mitsunobu Reaction”; J. Org. Chem. 68, 2003, pages 8261 to 8263).
##STR00141##
Overview 5: Synthesis of substituted curcumins by the Mitsunobu reaction starting from curcumin: Conditions: (a) 2-N-tert-butyloxycarbonyl-aminoethanol, DEAD, PPh.sub.3, DMF or THE or DCM, 0° C..fwdarw.RT; (b) 2-(2-N-tert-butoxycarbonyl-aminoethoxy)ethanol, DEAD, PPh.sub.3, DMF or THE or DCM, 0° C..fwdarw.RT; (c) 3-bromo-propan-1-ol, DEAD, PPh.sub.3, THE or DCM, 0° C..fwdarw.RT;
2.3.1 General Procedure:
[0877] Curcumin (0.36 g, 1 mmol) was provided together with triphenylphosphine (1.04 g, 4 mmol) and the corresponding boc-protected aminoalcohol (1 mmol or 3 mmol) in dry THE (4 mL). Diethylazodicarboxylate (DEAD) (0.7 g, 4 mmol, 40% in toluene) in dry THE (6 mL) was added dropwise over 20 minutes at approximately 2° C. to 5° C. and the formulation was then stirred for 4 h at RT in the dark. The formulation was diluted with 40 mL of acetic acid ethyl ester (EE) and the organic solution was shaken three times, each time with 20 mL of water. The organic phase was separated, dried over MgSO.sub.4 and rotary evaporated. The residue was purified by CC on silica gel with acetone/PE (dryload). For further purification, the product was dissolved in as little EE as possible and precipitated by adding a 10-fold quantity of petroleum ether.
[2-(4-{7-[4-hydroxy-3-methoxy-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-2-methoxy-phenoxy)-ethyl]-carbamic acid tert-butyl ester
[0878] ##STR00142##
[0879] Quantity of corresponding boc-protected aminoalcohol tert-butyl-N-(2-hydroxyethyl)carbamate used: 161 mg=1 mmol.
[0880] Column chromatography on silica gel with acetone/petroleum ether (PE)=1:2;
[0881] Preparative thin layer chromatography with acetone/petroleum ether (PE)=2:3.
[0882] Yield: 41% of theory, orange, viscous solid or orangey-yellow powder (210 mg).
[0883] Molecular mass=511.58 g/mol; Empirical formula=C.sub.28H.sub.33NO.sub.8
C-2: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-3-methoxy-phenyl]-3,5-dioxo-hepta-1,6-dienyl}-2-methoxy-phenoxy)-ethyl]-carbamic acid tert-butyl ester
[0884] ##STR00143##
[0885] Quantity of corresponding boc-protected aminoalcohol tert-butyl-N-(2-hydroxyethyl)carbamate used: 483 mg=3 mmol.
[0886] Column chromatography on silica gel with acetone/petroleum ether (PE)=1:3.fwdarw.1:2. Yield: 71% of theory, orange, viscous solid (465 mg).
[0887] Molecular mass=654.76 g/mol; Empirical formula=C.sub.35H.sub.46N.sub.2O.sub.10
[0888] .sup.1H-NMR and MS as above.
C-6: (2-{2-[4-(7-{4-[2-(2-tert-butoxycarbonylamino-ethoxy)-ethoxy]-3-methoxy-phenyl}-3,5-dioxo-hepta-1,6-dienyl)-2-methoxy-phenoxyl]-ethoxy}-ethyl)-carbamic acid tert-butyl ester
[0889] ##STR00144##
[0890] Quantity of corresponding boc-protected aminoalcohol 2-(2-N-tert-butoxycarbonyl-aminoethoxy)ethanol used: 715 mg=3 mmol.
[0891] Yield: 52% of theory, orange, viscous solid or orange powder (386 mg)
[0892] Column chromatography on silica gel with acetone/PE=1:2.fwdarw.2:3
[0893] Molecular mass=742.87 g/mol; Empirical formula=C.sub.39H.sub.54N.sub.2O.sub.12
[0894] .sup.1H-NMR and MS as above.
C-25: (1E,6E)-1,7-bis[4-(3-bromopropoxy)-3-methoxy-phenyl]hepta-1,6-diene-3,5-dione
[0895] ##STR00145##
[0896] Curcumin (2.00 g, 5.4 mmol) was placed together with triphenylphosphine (5.26 g, 20 mmol) and 3-bromo-propan-1-ol (2.25 g, 1.51 mL, 16.2 mmol) in dry THE (40 mL) and degassed at 0° C. Diethylazodicarboxylate (DEAD) (7 mL, 40% in toluene, 20 mmol) was added dropwise over 20 minutes and the formulation was stirred overnight in a thawing ice bath in the dark and with the exclusion of moisture. The formulation was poured into a three-fold quantity of diethylether. After the precipitate had settled out, the supernatant solution was carefully decanted off.
[0897] The residue was extracted twice, each time with 50 mL of diethylether. The combined organic solutions were shaken with 100 mL of water. The organic phase was separated, dried over MgSO.sub.4 and rotary evaporated. The residue was suspended in acetone/PE 1:2 and the yellow-orange solution was filtered off from the colourless crystals. The filter cake was washed several times with small portions of the cold solvent mixture and the filtrate was rotary evaporated. A preliminary purification of the residue was carried out by plug filtration over silica gel with acetone/PE 1:2. After withdrawing the solvent mixture, the remaining solid was suspended in ethanol in an ultrasound bath (30 mL), centrifuged and the supernatant solution was poured off. This washing step was repeated a total of three times, and afterwards, the product was air dried.
[0898] Yield: 2.08 g of orange powder, 63% of theory. Molecular mass=610.34 g/mol; Empirical formula=C.sub.27H.sub.30Br.sub.2O.sub.6
[0899] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.61 (d, J=15.8 Hz, 2H), 7.16-7.04 (m, 4H), 6.92 (d, J=8.3 Hz, 2H), 6.50 (d, J=15.8 Hz, 2H), 4.20 (t, J=6.0 Hz, 4H), 3.91 (s, 6H), 3.64 (t, J=6.4 Hz, 4H), 2.39 (p, J=6.1 Hz, 4H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 609.1 (57%, MH.sup.+), 611.0 (100%, MH.sup.+).
[0900] The curcumins C-27 to C-30 were produced from bis-(3-bromo-propoxy)curcumin C-25:
##STR00146##
Overview 6: Synthesis of symmetrically substituted curcumins starting from C-25: Conditions: (a) pyridine, DMF, 50° C., overnight; (b) trimethylamine in ethanol, DMF, 50° C., overnight; (c) trimethylphosphine in toluene, DMF, argon, 50° C., overnight; (d) triphenylphosphine, DMF, 50° C., overnight;
2.4 Curcumins with Quaternary Charges
[0901] The bis-(3-bromo-propoxy)curcumin (61 mg, 0.1 mmol) C-25 was placed in dry DMF (3 mL). Trimethylamine (2 mL, 5.6 M in ethanol, 11 mmol) or pyridine (790 mg, 0.8 mL, 10 mmol) in DMF (2 mL) was added dropwise through a septum using a syringe over 5 mins and the formulation was stirred overnight in the dark at 50° C. with the exclusion of moisture.
[0902] The formulation was poured into a five-fold quantity of diethylether. The precipitate was allowed to settle out and the supernatant solution was carefully decanted off. The residue was washed several times with diethylether and then suspended in 15 mL of chloroform/diethylether 1:1. It was allowed to settle out completely, the supernatant solution was poured away and the precipitate was dried using a high vacuum pump. The product was purified by HPLC.
Ion Exchange Chromatography
[0903] A column was loaded with Amberlite 954 and the ion exchange resin was conditioned with 0.1M HCl. After washing with water, it was reconditioned to water/MeOH/MeCN 3:1:1. Next, the TFA salt in a little of the solvent mixture was slowly eluted over the resin and then rinsed with a little solvent. After withdrawing the solvent under reduced pressure, the remaining aqueous solution was freeze-dried.
C-28: 3,3′-(4,4′-((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene))bis(oxy)bis(N,N,N-trimethylpropan-1-aminium) chloride
[0904] ##STR00147##
[0905] Yield: 30 mg orange solid, 45% of theory
[0906] Molecular mass: 568.76+2x 35.45=639.66 g/mol; Empirical formula: C.sub.33H.sub.48N.sub.2O.sub.6Cl.sub.2
[0907] .sup.1H-NMR and MS: see below (SA-CUR-10a).
C-27: 1,1′-(3,3′-(4,4′-((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene))bis(oxy)bis(propane-3,1-diyl))dipyridinium chloride
[0908] ##STR00148##
[0909] Yield: 36 mg orange solid, 78% of theory
[0910] Molecular mass: 608.74+2x 35.45=679.64 g/mol; Empirical formula: C.sub.37H.sub.40N.sub.2O.sub.6Cl.sub.2
[0911] .sup.1H-NMR and MS: see below (SA-CUR-10c).
2.5 Curcumins with Phosphonium Groups
[0912] The bis-(3-bromo-propoxy)curcumin C-25 (122 mg, 0.2 mmol) was placed in dry dichloromethane (DCM) (10 mL) and stirred under nitrogen. The phosphine used, in toluene (2 mL, 1 M, 2 mmol), was added dropwise over 5 minutes through a septum using a syringe. The formulation was stirred overnight in a Schlenk tube at 50° C. with the exclusion of moisture, in the dark and in a protective gas atmosphere. All of the volatile components were withdrawn under reduced pressure and the residue was suspended in 30 mL of diethylether with the aid of an ultrasound bath. The precipitate was allowed to settle out and the supernatant solution was carefully decanted. The residue was washed several times with diethylether. After settling of the precipitate, the supernatant solution was poured off and the precipitate was dried using a high vacuum pump.
Ion Exchange Chromatography
[0913] A short column was loaded with Amberlite 954 and the ion exchange resin was conditioned with 0.1M HCl. After washing with water, it was reconditioned to water/MeOH/MeCN 3:1:1. Next, the TFA salt in a little of the solvent mixture was slowly eluted over the resin and then rinsed with a little solvent. After withdrawing the solvent under reduced pressure, the remaining aqueous solution was freeze-dried. Quantitative yield.
C-29: 3,3′-(4,4′-((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene))bis(oxy)bis(trimethylpropan-1-phosphonium) chloride
[0914] ##STR00149##
[0915] Phosphine used: trimethylphosphine
[0916] Yield: 58 mg orange solid, 21% of theory
[0917] Molecular mass: 602.69+2x 35.45=673.59 g/mol; Empirical formula: C.sub.33H.sub.48P.sub.2O.sub.6Cl.sub.2
[0918] .sup.1H-NMR and MS: see below SA-CUR-15a.
C-30: 3,3′-(4,4′-((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene))bis(oxy)bis(triphenylpropan-1-phosphonium) chloride
[0919] ##STR00150##
Phosphine used: triphenylphosphine
Yield: 107 mg orange solid, 56% of theory
Molecular mass: 975.12+2×35.45=1046.02 g/mol; Empirical formula: C.sub.63H.sub.60P.sub.2O.sub.6Cl.sub.2
[0920] .sup.1H-NMR and MS: see below SA-CUR-15b.
2.6 Synthesis of Substituted Curcumins by Alkylation
[0921] ##STR00151##
Overview 7: Synthesis of substituted curcumins by alkylation: Conditions: (a) 2-N-tert-butyloxycarbonyl-aminobromide, DBU, toluene or DCM or THF, 0° C..fwdarw.RT.fwdarw.60° C.; (b) DCM, TFA, RT, 5 h; then Amberlite IRA-958 ion exchange resin, water
[0922] Tetramethoxycurcumin (0.4 g, 1 mmol) or curcumin C-2 (0.6 g, 1 mmol) and 2-N-tert-butoxycarbonylaminoethyl bromide (0.34 g, 1.5 mmol) was placed in toluene (4 mL). 1.8-diazabicyclo[5.4.0]undec-7-ene (DBU) (0.15 g, 1 mmol) was added and the mixture was stirred for 15 h at room temperature. The solution was diluted with ethyl acetate (30 mL), and washed with sodium chloride solution (30 mL), potassium hydrogen sulphate solution (5%, 30 mL) and water (30 mL). After drying over MgSO.sub.4, it was rotary evaporated and the residue was purified by column chromatography on silica gel with ethyl acetate/petroleum ether and then by preparative thin layer chromatography.
C-15: 1,7-bis-(3,4-dimethoxyphenyl)-hepta-4-(2-tert-butoxycarbonylamino-ethyl)-1,6-dien-3,5-dione
[0923] ##STR00152##
[0924] Column chromatography on silica gel with acetone/PE=1:1
[0925] Yield: 14% of theory, orange solid or orangey-yellow powder (76 mg)
[0926] Molecular mass=539.63 g/mol; Empirical formula=C.sub.30H.sub.37NO.sub.8
[0927] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.72-7.53 (m, 2H), 7.22-7.04 (m, 4H), 6.86 (d, J=8.1 Hz, 2H), 6.54 (d, J=15.7 Hz, 2H), 5.11 (s, 1H), 3.88-3.72 (m, 2H), 3.93 (s, 12H), 2.81-2.70 (m, 2H), 1.43 (s, 9H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 562.3 (MNa.sup.+, 13%), 540.3 (MH.sup.+, 100%), 484.2 (2%, MH.sup.+—C.sub.4H.sub.9), 440.3 (61%, MH.sup.+-boc).
C-20: [2-(4-{7-[4-(2-tert-butoxycarbonylamino-ethoxy)-3-methoxy-phenyl]-3,5-dioxo-hepta-4-(2-tert-butoxycarbonylamino-ethyl)-1,6-dienyl}-2-methoxy-phenoxy)-ethyl]-carbamic acid tert-butyl ester
[0928] ##STR00153##
[0929] Column chromatography on silica gel with acetone/PE=1:3.fwdarw.1:1
[0930] Yield: 12% of theory, orange, orangey-yellow powder (96 mg).
[0931] Molecular mass=797.95 g/mol; Empirical formula=C.sub.42H.sub.59N.sub.3O.sub.12
[0932] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.64 (d, J=15.7 Hz, 2H), 7.24-7.01 (m, 6H), 6.78 (d, J=15.8 Hz, 2H), 5.16 (s, 2H), 4.12 (dd, J=10.6, 5.4 Hz, 4H), 3.93 (s, 3H), 3.90 (s, 3H), 3.86-3.75 (m, 2H), 3.54 (m, 4H), 2.82-2.71 (m, 2H), 1.45 (s, 9H), 1.44 (s, 9H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 820.4 (MNa.sup.+, 69%), 798.4 (MH.sup.+, 100%), 742.4 (6%, MH.sup.+—C.sub.4H.sub.9), 698.3 (36%, MH.sup.+-boc).
Synthesis of Unsymmetrically Substituted Curcumins
General Procedure:
Step 1:
[0933] The beta-diketones used were acetyl acetone (1.5 g, 15 mmol) or 3-methyl-2,4-pentanedione (3.42 g, 30 mmol). The corresponding beta-diketone and boron oxide B.sub.2O.sub.3 (1.5 g, 21 mmol) were suspended in ethyl acetate (20 mL) and stirred for 60 minutes at 70° C. The substituted benzaldehyde E-3 (1.02 g, 3.5 mmol) in ethyl acetate (5 mL) and tributylborate (1.68 g, 7 mmol) was added and the formulation was stirred for half an hour at 85° C. Next, n-butylamine (0.5 mL in 3 mL ethyl acetate) was added dropwise over 10 minutes. After stirring for a further three hours at 80° C., it was cooled to 50° C. and in order to hydrolyse the boron complex, 100 mL of 50% acetic acid was added. After stirring overnight at room temperature, the mixture of solvents, protected from light, was removed and the residue was extracted three times with EE (30 mL each time). The combined organic phases were washed twice with water (50 mL each time), dried over MgSO.sub.4 and finally, the solvent was withdrawn under reduced pressure.
##STR00154##
Overview 7: Synthesis of unsymmetrically substituted curcumins via the corresponding intermediates (E-16 and E-17): Conditions: (a) acetyl acetone, B.sub.2O.sub.3, B(OBu).sub.3, n-butylamine, DMF or ethyl acetate, 60-80° C., 6 h, then hydrolysis with HOAc 40% overnight; (b) 3-methyl-pentane-2,4-dione, B.sub.2O.sub.3, B(OBu).sub.3, n-butylamine, ethyl acetate, 80° C., 6 h, then hydrolysis with HOAc 40% overnight; (c) 3,4-dimethoxybenzaldehyde, B.sub.2O.sub.3, B(OBu).sub.3, n-butylamine, DMF or ethyl acetate, 70° C., 6 h, then hydrolysis with HOAc 40% overnight; (d) E-11, B.sub.2O.sub.3, B(OBu).sub.3, n-butylamine, DMF or ethyl acetate, 80° C., 6 h, then hydrolysis with HOAc 40% overnight; (e) DCM, TFA, RT, 5 h; then Amberlite IRA-958 ion exchange resin, water
E-16: tert-butyl 2-(4-(3,5-dioxohex-1-enyl)-2-methoxyphenoxy)ethyl carbamate
[0934] ##STR00155##
[0935] Acetyl acetone was used as the beta-diketone. Purification was carried out using column chromatography on silica gel with acetone/PE=2:5.fwdarw.1:2. The corresponding symmetrically substituted curcumin has poorer solubility in EtOH than the product.
[0936] Yield: 687 mg, 52% of theory, yellow solid.
[0937] Molecular mass=377.44 g/mol; Empirical formula=C.sub.20H.sub.27NO.sub.6
[0938] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.53 (d, J=15.8 Hz, 1H), 7.12-7.00 (m, 2H), 6.88 (d, J=8.3 Hz, 1H), 6.34 (d, J=15.8 Hz, 1H), 5.16 (s, 1H), 4.10 (t, J=5.1 Hz, 2H), 3.90 (s, 3H), 3.56 (d, J=5.3 Hz, 2H), 2.16 (s, 3H), 1.44 (s, 9H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 777.4 (46%, 2MNa.sup.+), 400.2 (38%, MNa.sup.+), 378.2 (12%, MH.sup.+), 322.1 (100%, MH.sup.+—C.sub.4H.sub.9), 278.1 (47%, MH.sup.+−boc).
E-17: tert-butyl 2-(4-(3,5-dioxo-4-methyl-hex-1-enyl)-2-methoxyphenoxy)-ethyl carbamate
[0939] ##STR00156##
[0940] 3-methyl-2,4-pentanedione was used as the beta-diketone. Purification was carried out using column chromatography on silica gel with acetone/PE=1:3 and preparative TLC with acetone/PE=1:2.
[0941] Yield: 644 mg, 47% of theory, yellow solid.
[0942] Molecular mass=391.47 g/mol; Empirical formula=C.sub.21H.sub.29NO.sub.6
[0943] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.58 (dd, J=15.7, 6.5 Hz, 1H), 7.13 (d, J=8.3 Hz, 1H), 7.05 (dd, J=4.9, 1.8 Hz, 1H), 6.89 (d, J=8.3 Hz, 1H), 6.74 (dd, J=37.9, 15.7 Hz, 1H), 5.11 (s, 1H), 4.10 (t, J=5.1 Hz, 2H), 3.91 (s, 3H), 3.56 (m, 2H), 2.25+2.18 (s, 3H), 2.02 (s, 3H), 1.44 (s, 10H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 805.5 (24%, 2MNa.sup.+), 414.3 (53%, MNa.sup.+), 392.3 (19%, MH.sup.+), 336.2 (100%, MH.sup.+—C.sub.4H.sub.9), 292.2 (34%, MH.sup.+−boc).
Step 2:
[0944] Tert-butyl-2-(4-(3,5-dioxohex-1-enyl)-2-methoxyphenoxy)ethyl carbamate (185 mg, 0.5 mmol) or tert-butyl-2-(4-(3,5-dioxo-4-methyl-hex-1-enyl)-2-methoxyphenoxy)ethyl carbamate (191 mg, 0.5 mmol) and boron oxide B.sub.2O.sub.3 (0.07 g, 1 mmol) were suspended in ethyl acetate (3 mL) and stirred for 60 minutes at 80° C. The substituted benzaldehyde (0.6 mmol) in ethyl acetate (3 mL) along with tributylborate (0.24 g, 1 mmol) were added one after the other and the formulation was stirred for half an hour at 80° C. Next, n-butylamine (0.1 mL in 1 mL EE) was added dropwise over 5 minutes. After stirring for a further three hours at 80° C., the slightly cooled solution, still at approximately 50° C., was poured into 40 mL of 50% acetic acid. After stirring overnight at room temperature protected from light, the mixture of solvents was withdrawn under reduced pressure and the residue was extracted three times with EE (20 mL each time). The combined organic phases were washed twice with water (20 mL each time), dried over MgSO.sub.4 and finally, the solvent was withdrawn under reduced pressure. Purification was carried out using column chromatography on silica gel.
C-16: tert-butyl 2-(4-((1E,6E)-7-(3,4-dimethoxyphenyl)-3,5-dioxohepta-1,6-dienyl)-2-methoxyphenoxy)ethyl carbamate
[0945] ##STR00157##
[0946] Quantity of corresponding aldehyde E-16 used: 100 mg=0.6 mmol
[0947] Column chromatography with acetone/PE=1:2.fwdarw.2:3
[0948] Yield: 121 mg, 46% of theory, orange, viscous solid or orangey-yellow powder.
[0949] Molecular mass=525.60 g/mol; Empirical formula=C.sub.29H.sub.35NO.sub.8
[0950] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.60 (dd, J=15.7, 4.5 Hz, 2H), 7.15-7.06 (m, 4H), 6.89 (dd, J=8.3, 3.3 Hz, 2H), 6.50 (d, J=15.8 Hz, 2H), 5.13 (s, 1H), 4.11 (t, J=5.0 Hz, 2H), 3.94 (s, 3H), 3.93 (s, 3H), 3.92 (s, 3H), 3.62-3.52 (m, 2H), 1.45 (s, 9H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 1073.5 (19%, 2M+Na.sup.+), 526.2 (100%, MH.sup.+), 470.2 (21%, MH.sup.+—C.sub.4H.sub.9).
C-21: 2-(4-((1E,6E)-7-(3,4-bis(2-(tert-butoxycarbonylamino)ethoxy)phenyl)-3,5-dioxohepta-1,6-dienyl)-2-methoxyphenoxy)ethyl carbamate
[0951] ##STR00158##
[0952] Quantity of corresponding aldehyde E-16 used: 255 mg=0.6 mmol
[0953] Column chromatography with acetone/PE=2:5.fwdarw.1:2
[0954] Yield: 172 mg, 44% of theory, orange, very viscous solid.
[0955] Molecular mass=783.92 g/mol; Empirical formula=C.sub.41H.sub.57N.sub.3O.sub.12
[0956] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.57 (dd, J=15.7, 9.3 Hz, 2H), 7.17-7.05 (m, 4H), 6.90 (dd, J=8.2, 6.6 Hz, 2H), 6.49 (d, J=15.7 Hz, 2H), 5.27 (s, 2H), 5.14 (s, 1H), 4.10 (m, 6H), 3.91 (s, 3H), 3.55 (m, 6H), 1.46 (s, 9H), 1.44 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 804.6 (73%, MNa.sup.+), 784.4 (100%, MH.sup.+), 684.3 (71%, MH.sup.+-boc).
C-17: tert-butyl-2-(4-((1E,6E)-7-(3,4-dimethoxyphenyl)-3,5-dioxo-4-methyl-hepta-1,6-dienyl)-2-methoxyphenoxy)ethyl carbamate
[0957] ##STR00159##
[0958] Quantity of corresponding aldehyde E-17 used: 100 mg=0.6 mmol
[0959] Column chromatography with acetone/PE=1:2.fwdarw.2:3
[0960] Yield: 113 mg, 42% of theory, orange, viscous solid
[0961] Molecular mass=539.63 g/mol; Empirical formula=C.sub.30H.sub.37NO.sub.8
[0962] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.75-7.56 (m, 2H), 7.20-6.82 (m, 7H), 6.70 (d, J=15.9 Hz, 1H), 5.15 (s, 1H), 4.15-4.06 (m, 2H), 3.95 (s, 3H), 3.91 (s, 3H), 3.89 (s, 3H), 3.57 (m, 2H), 2.18 (s, 3H), 1.44 (s, 9H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 562.2 (MNa.sup.+, 41%), 540.3 (MH.sup.+, 100%), 484.2 (MH.sup.+—C.sub.4H.sub.9, 46%), 440.3 (MH.sup.+-boc, 3%).
C-22: 2-(4-((1E,6E)-7-(3,4-bis(2-(tert-butoxycarbonylamino)ethoxy)phenyl)-3,5-dioxo-4-methyl-hepta-1,6-dienyl)-2-methoxyphenoxy)ethyl carbamic acid
[0963] ##STR00160##
[0964] Quantity of corresponding aldehyde E-17 used: 255 mg=0.6 mmol
[0965] Column chromatography with acetone/PE=2:5.fwdarw.1:2
[0966] Yield: 160 mg, 40% of theory, orange, very viscous solid.
[0967] Molecular mass=797.95 g/mol; Empirical formula=C.sub.42H.sub.59N.sub.3O.sub.12
[0968] .sup.1H NMR (300 MHz, CDCl.sub.3), δ=7.74-7.52 (m, 2H), 7.21-6.88 (m, 7H), 6.70 (d, J=15.9 Hz, 1H), 5.29 (s, 2H), 5.16 (s, 1H), 4.10 (t, J=4.8 Hz, 6H), 3.90 (m, 3H), 3.56 (m, 6H), 2.17 (m, 3H), 1.46 (s, 9H), 1.45 (s, 18H). MS (ESI, CH.sub.2Cl.sub.2/MeOH+10 mmol NH.sub.4OAc): 820.4 (100%, MNa.sup.+), 798.3 (43%, MH.sup.+), 698.4 (67%, MH.sup.+-boc).
3. Boc Deprotection of the Curcumins and Ion Exchange Chromatography
[0969] The corresponding boc-protected curcumin (0.2 mmol, 120-160 mg) was dissolved in DCM (6 mL). 4 mL of a 10% solution of TFA in DCM (containing 6% TIS) was slowly added dropwise, with stirring. After stirring for 5 h at RT protected from light, the product was precipitated by adding diethylether (10 mL). The precipitate was centrifuged off and then the supernatant was discarded. The solid was suspended in diethylether (30 mL) and centrifuged off once again. The supernatant was also discarded. This washing step was repeated once more and then the product was air dried in darkness.
Ion Exchange Chromatography
[0970] A column was loaded with Amberlite 954 and the ion exchange resin was conditioned with 0.1M HCl. After washing with water to a very weak acidic reaction, reconditioning to a water/MeOH/MeCN mixture was carried out as necessary. Next, the TFA salt in as little of the solvent mixture as possible was slowly eluted over the resin and then rinsed several times with a little solvent. After withdrawing the solvent under reduced pressure, the remaining aqueous solution was freeze-dried. Quantitative yield.
Eluent:
[0971] Curcumins 12a and 12b: water/MeOH/MeCN 1:1:1
Curcumins 04, 07, 09b, 11 b/c and 14b: water/MeOH/MeCN 9:4:2
All other curcumins: pure water
4. Curcumin Complexes
[0972]
TABLE-US-00002 Curcumin 01a BF- complex (BF-SA- CUR-1a)
4.a) SA-CUR-01a TEA salt (68 mg, 0.1 mmol) was placed in dry DCM (10 mL). Boron trifluoride etherate (20 μL, 0.12 mmol) was added dropwise and the formulation was stirred overnight. The solution was diluted with diethylether (20 mL), distributed onto two Blue Caps and centrifuged. The pellet was washed several times with diethylether and air dried.
[0973] Red powder (TEA salt), quantitative yield.
Ion Exchange Chromatography
[0974] A short column was loaded with Amberlite 954 and the ion exchange resin was conditioned with 0.1M HCl. After washing with water to a very weak acidic reaction, the TFA salt was washed in as little water as possible, eluted slowly over the resin and rinsed several times with a little solvent. The aqueous solution was freeze-dried. Quantitative yield.
4.b) SA-CUR-01a chloride (108 mg, 0.2 mmol) and boron trioxide (0.05 mmol) were stirred overnight in aqueous HCl (1 M, 2 mL) at room temperature. The solvent was extracted in a stream of nitrogen and the residue was dried.
[0975] Red powder, quantitative yield.
4.c) SA-CUR-01a chloride (54 mg, 0.1 mmol) and zinc acetate dihydrate (0.05 mmol) were refluxed in ethanol/acetic acid/water 3:2:1 (3 mL) for 2 days. The solvent was extracted in a stream of nitrogen and the residue was dried.
[0976] Orange powder, quantitative yield.
[0977] A summary of the compounds produced is shown in Overview 8.
TABLE-US-00003 Description/ serial number Structure Analysis Curcumin 0 hydrochloride (SA-CUR-0) compound (40)
Example 2) Phototoxicity Experiments
a) Production of Growing Cultures of Bacterial Strains
[0978] All of the experiments were carried out under sterile conditions in a safety cabinet (Biosafe 4-130, Ehret, Emmedingen, Germany). After adding the photoactive substances, the work was carried out entirely in darkness.
[0979] A sample of the bacterial cell Staphylococcus. aureus (ATCC number: 25923) or Escherichia. coli (ATCC number: 25922) was removed from a cryo-freeze culture and cultured under aerobic conditions at 37° C. and 175 rpm overnight in an orbital shaker (MAXQ4000, Thermo Scientific, Dubuque, Iowa, USA). Growth was carried out in 20 mL Todd-Hewitt broth (Carl Roth, Karlsruhe, Germany) supplemented with 0.3% of yeast extract (AppliChem, Darmstadt, Germany).
[0980] Alternatively, Müller-Hinton media were used for culture:
[0981] Müller-Hinton liquid medium (Merck KGaA, Darmstadt, Germany)
[0982] 2.0 g/L meat extract, 17.5 g/L casein hydrolysate, 1.5 g/L starch, pH: 7.4+0.2.
[0983] Müller Hinton agar (Merck KGaA, Darmstadt, Germany)
[0984] 2.0 g/L meat extract, 17.5 g/L casein hydrolysate, 1.5 g/L starch, 15 g/L Agar, pH: 7.4+0.2.
b) Production of a Culture in the Exponential Growth Phase
[0985] Subsequently, dilutions (0, 5, 10, 25 and 20% v/v of the overnight culture) and their absorptions were measured at 600 nm in triplicate for 100 μL (Infinite 200 M Pro, Tecan, Männedorf, Switzerland). A calibration graph (Microsoft Excel) was used to calculate the volume which was required to produce 20 mL of a culture with an absorption of 0.05 at 600 nm. The volume calculated for the overnight culture was topped up with Todd-Hewitt broth (composition as above) to 20 mL and incubated for two hours at 37° C. with constant movement (175 rpm, MAXQ4000). The cultures were then in the exponential growth phase; the absorption at 600 nm was between 0.3 and 0.45.
[0986] The subsequent incubation with the photoactive substances as well as irradiation with electromagnetic radiation and determination of the phototoxicity was carried out with two different methods.
c.1) Incubation, Irradiation and Determination of Phototoxicity
[0987] A 2-hour culture was divided into 1800 μL aliquots. After centrifuging at 20° C., 830 rcf, 5 min (5417R centrifuge, Eppendorf, Hamburg, Germany), the pellets were re-suspended in phosphate buffer (Dulbeccos' Modified Phosphate Saline, DPBS, Sigma-Aldrich) with either 10 or 50 μM of the corresponding photosensitizer, wherein the final volume was kept at 1800 μL. The solutions obtained were immediately incubated on the orbital shaker (see above for parameters) for 5 or 25 minutes.
[0988] Three controls were run at the same time for each photosensitizer. The “light only” control contained DPBS without photosensitizer. The “photosensitizer only” (“PS only”) control was incubated like the PDI samples, but not irradiated and kept strictly in darkness. A further control (double negative, “Co−/−”), received neither light nor photosensitizer.
[0989] After the incubation, duplicates of the samples (500 μL each) were transferred into a 24-well microtitre plate (Cellstar, Greiner Bio-One, Frickenhausen, Germany). The “PS only” and “Co−/−” samples were placed in their own microtitre plate which was packed in aluminium foil so as to be lightproof.
[0990] The irradiation was carried out under constant shaking (MTS4, IKA, Staufen, Germany, ˜175 rpm) from below on a LED array with maximum homogeneity of the lighting surface. All of the controls were shaken in the same manner. The technical data for the light source are shown in Table 2; the total dose of light applied was 33.8 J/cm.sup.2.
TABLE-US-00004 TABLE 2 Technical data for LED Arrays: Diode manufacturer Roithner Lasertechnik, Wien, Austria Description of diodes LED 435-12-30 Dominant wavelengths 430 nm-435 nm Number of diodes in array 432 Intensity 9.4 mW/cm.sup.2
[0991] The determination of the colony forming units (CFU) was carried out in accordance with the method published by Miles and Misra (Miles, A A; Misra, S S, Irwin, J O (1938 November). “The estimation of the bactericidal power of the blood” The Journal of hygiene 38 (6): 732-49). In this regard, serial dilutions (1:10) of the corresponding bacterial suspension were produced in DPBS. 5×10 μL of each bacterial dilution was then dripped onto Todd-Hewitt plates (with broth, additional 1.5% agar (Agar-Agar, Kobe I, Roth, Karlsruhe, Germany) and incubated at 37° C. for 24 h. Next, the number of surviving colony forming units was determined. All of the tests were carried out four times.
c.2) Incubation, Irradiation and Determination of Phototoxicity
[0992] In a second experiment, the photosensitizers (PS) used were dissolved in Millipore water and adjusted to various concentrations. 25 μL of a bacterial suspension grown overnight (˜108/mL) was incubated with 25 μL of photosensitizer solution of the various concentrations at room temperature for 10 seconds in darkness in a 96-well plate.
[0993] Next, the suspension was irradiated for 5-20 minutes. For the irradiation, the light source BlueV from Waldmann (Villingen-Schwenningen, Germany) was used, which emits light from 380 to 480 nm (emission maximum at approximately 420 nm). The applied energy density was 17.5 mW/cm.sup.2.
[0994] Each experiment was accompanied by three controls in order to exclude side effect of the irradiation/photosensitizer (PS) on the survival of the bacteria: (i) no PS, only light (=light control), (ii) no light, only PS (=dark control) and (iii) neither light nor PS (=reference control). The determination of the colony forming units (CFU) per mL was also carried out in accordance with the method published by Miles, Misra and Irwin described above in section c.1). All of the tests were carried out four times.
d) Result of Phototoxicity Experiments
[0995] The results of the phototoxicity experiments described above in section c.1) are shown in
[0996] The results of the phototoxicity experiments described in section c.2) are shown in
[0997]
[0998]
[0999]
[1000]
[1001]
[1002]
[1003]
[1004]
[1005]
[1006]
[1007]
[1008]
[1009]
[1010] As can be seen from
[1011] Table 3 shows the effect of the tested substances against E. coli ATCC 25922 with an applied light dose of 33.8 J/cm.sup.2 for an emission maximum at approximately 435 nm (60 minutes illumination with an intensity of 9.4 mW/cm.sup.2). The average reference control (arithmetic mean with standard deviation) (no light, no PS) was 3.6×10.sup.8/mL. The logarithmic reduction after illumination is shown with respect to the reference control. The respective upper value, indicated by a *, refers to an incubation period of 5 min and the respectively lower value, indicated by a #, refers to an incubation period of 25 min.
TABLE-US-00005 TABLE 3 Phototoxicity test for substances against E. coli ATCC 25922. Effectiveness against E. Coli Reduction in CFU in log.sub.10 at concentration of Description 10 μM 50 μM 100 μM 250 μM Curcumin 0 hydrochloride >4* >6* n.d. n.d. (SA-CUR-0) >5.sup.# >6.sup.# Curcumin 01a hydrochloride >1* >5* n.d. n.d. (SA-CUR-1a) >2.sup.# >5.sup.# Curcumin 01b hydrochloride >2* >5* n.d. n.d. (SA-CUR-1b), >3.sup.# >7.sup.# Curcumin 01e hydrochloride >5* >4* n.d. n.d. (SA-CUR-1e), >5.sup.# >7.sup.# Curcumin 01c hydrochloride >2* >3* n.d. n.d. (SA-CUR-1c >2.sup.# >4.sup.# Curcumin 02 hydrochloride >2* >6* n.d. n.d. (SA-CUR-2) >4.sup.# >7.sup.# Curcumin 08 hydrochloride —* ~1* n.d. n.d. (SA-CUR-8) —.sup.# >1.sup.# Curcumin 04 hydrochloride >1* >3* n.d. n.d. (SA-CUR-4) >3.sup.# >4.sup.# Curcumin 03 hydrochloride >7* >7* n.d. n.d. (SA-CUR-3) >7.sup.# >7.sup.# Curcumin 05 hydrochloride >4* >7* n.d. n.d. (SA-CUR-5) >4.sup.# >6.sup.# Roseo-curcumin 01a hydrochloride >5* >7* n.d. n.d. (RO-SA-CUR-1a) >5.sup.# >6.sup.# Curcumin 01a zinc complex >3* >5* n.d. n.d. (Zn-SA-CUR-1a) >2.sup.# >5.sup.# Curcumin 09b hydrochloride —* >1* n.d. n.d. (cyclo-SA-CUR-9b) —.sup.# >2.sup.# Curcumin 10a hydrochloride >1* >5* n.d. n.d. (SA-CUR-10a) >1.sup.# >5.sup.# Curcumin 10b hydrochloride >5* >7* n.d. n.d. (GUA-SA-CUR-10b) >4.sup.# >6.sup.# Curcumin 10c hydrochloride >4* >7* n.d. n.d. (SA-CUR-10c) >5.sup.# >7.sup.# Curcumin 11b hydrochloride ~1* ~3* n.d. n.d. (SA-CUR-11b) >1.sup.# >3.sup.# Curcumin 12a hydrochloride >1* >6* n.d. n.d. (SA-CUR-12a) >4.sup.# >6.sup.# Curcumin 13b hydrochloride >7* >7* n.d. n.d. (SA-CUR-13b) >7.sup.# >7.sup.# Curcumin 14b hydrochloride —* —* n.d. n.d. (SA-CUR-14b) —.sup.# >1.sup.#
[1012] Table 4 shows the action of the tested substances against E. coli ATCC 25922 for an applied light dose of 15.7 J/cm.sup.2 with an emission maximum at approximately 420 nm (15 min illumination at an intensity of 17.5 mW/cm.sup.2). The average reference control (arithmetic mean with standard deviation) (no light, no PS) was 3.6×10.sup.8/mL for E. coli. The logarithmic reduction after illumination is shown with respect to the reference control.
TABLE-US-00006 TABLE 4 Phototoxicity test for substances against E. coli ATCC 25922 Effectiveness against E. Coli Reduction in CFU in log.sub.10 at concentration of Description/serial number 10 μM 50 μM 100 μM 250 μM Curcumin 01a hydrochloride >4 >5 n.d. n.d. (SA-CUR-1a) Curcumin 07 hydrochloride >1 >4 n.d. n.d. (SA-CUR-07) Curcumin 03 hydrochloride >4 >5 n.d. n.d. (SA-CUR-3)
[1013] Table 5 shows the action of the tested substances against S. aureus ATCC 25923. The irradiation period was 5 minutes (emission maximum at approximately 420 nm) for an applied intensity of 17.5 mW/cm.sup.2, i.e. an applied light energy (dosage) of 5.3 J/cm.sup.2. The average reference control (arithmetic mean with standard deviation) (no light, no PS) was 3.9×10$/mL. The logarithmic reduction after illumination is shown with respect to the reference control.
TABLE-US-00007 TABLE 5 Phototoxicity test against S. aureus ATCC 25923. Effectiveness against S. aureus Reduction in CFU in log.sub.10 at concentration of Description/serial number 10 μM 50 μM 100 μM 250 μM Curcumin 01a hydrochloride >3 >5 n.d. n.d. (SA-CUR-1a) Curcumin 07 hydrochloride >2 >5 n.d. n.d. (SA-CUR-07) Curcumin 03 hydrochloride >4 >5 n.d. n.d. (SA-CUR-3) Curcumin 01a BF-complex >5 >5 n.d. n.d. (BF-SA-CUR-1a) Curcumin 09a hydrochloride >2 >5 n.d. n.d. (Me-SA-CUR-9a) Curcumin 11a hydrochloride — >1 >2 >5 (SA-CUR-11a) Curcumin 11c hydrochloride — >1 >2 >5 (SA-CUR-11c) Curcumin 12b hydrochloride — >1 ~2 >3 (SA-CUR-12b) Curcumin 13a hydrochloride >1 >5 n.d. n.d. (SA-CUR-13a) Curcumin 13c hydrochloride >1 >5 n.d. n.d. (SA-CUR-13c) Curcumin 14a hydrochloride — >1 >2 >3 (SA-CUR-14a) Curcumin 15a hydrochloride ~1 >5 n.d. n.d. (SA-CUR-15a) Curcumin 15b hydrochloride >4 >5 n.d. n.d. (SA-CUR-15b)
[1014] As can clearly be seen from
Comparative Example 3
[1015] In a further test, the stability and phototoxicity of the following compounds were tested.
##STR00196##
[1016] The compound CRANAD-2 is a good fluorophore which has a fluorescence quantum yield which is comparable with rhodamine or Cy5 dyes.
[1017] Incubation with CRANAD-2 and the bacterial strains S. aureus ATCC 25923 and E. coli ATCC 25922 employed, as well as irradiation with electromagnetic radiation and determination of the phototoxicity were carried out as described in sections c.1) and c.2) above. An inactivation of the tested bacterial strains S. aureus ATCC 25923 and E. coli ATCC 25922 upon illumination in the presence of CRANAD-2 could not be ascertained with either of the methods.
[1018] Because of the amine substituents directly on the aromatic ring, which can act as electron pair donors, the photophysics are displaced into a singlet process and substantially less energy is available for triplet processes. A good transfer of energy into the triplet level should be the prerequisite for a photodynamic effect which, however, was not observed for the compound CRANAD-2.
[1019] Incubation with the compound (71) (SACUR-01d) and the bacterial strains S. aureus ATCC 25923 and E. coli ATCC 25922 employed as well as irradiation with electromagnetic radiation and determination of the phototoxicity were carried out as described in sections c.1) and c.2) above. Hardly any inactivation of the tested bacterial strains S. aureus ATCC 25923 and E. coli ATCC 25922 was observed with either of the methods upon illumination in the presence of compound (71) (SACUR-01d), because the decomposition of the compound in the measurement solution meant that no reliable values were obtained. Compound (71) (SACUR-01d) exhibited a low stability in aqueous solution which was comparable with the natural basic substance curcumin. The free OH groups contribute greatly to this photo-instability because due to them, the compound can readily transform into the quinoid mesomer, and thus cleavage of one half of the molecule with the formation of ferulic acids or substituted vanillins is facilitated.
[1020] The molar extinction coefficient for compound (71) (SACUR-01d) at 420 nm in the aqueous solutions A-C given below varied between 8000 and 16000 M.sup.−1 cm.sup.−1 and thus was significantly smaller than the value known for curcumin (ε.sub.420, H2O=23800 M.sup.−1 cm.sup.−1) (see Arnaut LG, Formosinho S J. J. Photochem. Photobiol. A: Chem. 75, 1993, pages 1 to 20). These results show that the compound (71) does not have sufficient stability in aqueous solution to establish a photodynamic effect after irradiation.
Aqueous solution A: distilled water.
Aqueous solution B: isotonic sodium chloride solution (0.9% by weight NaCl).
Aqueous solution C: PBS buffer, pH 7.4 (composition: see Sambrook, J.; Maniatis, T.; Russel, D. W.: Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press; 3rd edition (2001)).