BQC-G, a tumor-selective anti-cancer prodrug
The invention relates to the synthesis of a second-generation camptothecin glucuronide prodrug (BQC-G) of a potent anticancer camptothecin derivative 5,6-dihydro-4H-benzo[de]quinoline-camptothecin (BQC). BQC-G was over 4000 times more water soluble than BQC, displayed good stability in human plasma and was an excellent substrate for enzymatic hydrolysis by bacterial and human β-glucuronidases. BQC-G was about 30 times less toxic than BQC, but was as toxic as BQC after hydrolysis of the glucuronide moiety by β-glucuronidase. In the presence of human serum albumin, BQC-G displayed lower cytotoxicity (IC 50 =1080 nM) but could be activated by β-glucuronidase to display potent activity (IC 50 =13.3 nM).
1. A compound according to the formula I, below:
wherein R is a glucuronide linked via an immolative spacer, or a pharmaceutically acceptable salt thereof.
2. A compound according to claim 1 wherein R is
3. The compound according to claim 1 wherein the glucuronide moiety is hydrolyzed by β-glucuronidases.
4. The compound according to claim 1 wherein the solubility of the compound in water based buffers is at least approximately 775 micrograms per milliliter.
5. The compound according to claim 1 wherein the compound is stable in human serum.
6. The compound according to claim 3 wherein the compound is cytotoxic to human cancer cells.
7. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to claim 1 and a pharmaceutically acceptable carrier.
8. A method of treating a cancer patient which comprises administering to the cancer patient an effective amount of a pharmaceutical composition containing a therapeutically effective amount of a compound of formula (I) and a pharmaceutically acceptable carrier.
9. A method for synthesizing the compound of formula I comprising:
(a) contacting D(+)-glucurono-3,6-lactone and sodium methoxide to give methyl 1,2,3,4-tetra-O-acetyl-β-D-glycopyranuronate;
(b) reacting methyl 1,2,3,4-tetra-O-acetyl-β-D-glycopyranuronate and TiBr 4 in CH 2 Cl 2 to give methyl 1-bromo-1-deoxy-2,3,4-tri-O-acetyl-α-D-glycopyranuronate;
(c) contacting methyl 1-bromo-1-deoxy-2,3,4-tri-O-acetyl-α-D-glycopyranuronate, 4-hydroxy-3-nitrobenaldehyde, and Ag 2 O to give methyl 1-O-(2-nitro-4-formylphenyl)-2,3,4-tri-O-acetyl-β-D-glycopyranuronate;
(d) contacting methyl 1-O-(2-nitro-4-formylphenyl)-2,3,4-tri-O-acetyl-β-D-glycopyranuronate, NaBH 4 and silica gel in i-PrOH/CHCl 3 to give methyl 1-O-[2-nitro-4-(hydroxymethyl)phenyl]-2,3,4-tri-O-acetyl-β-D-glycopyranuronate;
(e) reacting methyl 1-O-[2-nitro-4-(hydroxymethyl)phenyl]-2,3,4-tri-O-acetyl-β-D-glycopyranuronate with methanesulfonyl chloride and triethylamine to give methyl 1-O-[4-(methanesulfonyloxymethyl)-2-nitrophenyl]-2,3,4-tri-O-acetyl-β-D-glycopyranuronate;
(f) reacting methyl 1-O-[4-(methanesulfonyloxymethyl)-2-nitrophenyl]-2,3,4-tri-O-acetyl-β-D-glycopyranuronate with BQC and Cs 2 CO 2 to give 10-[4-O-(methyl-2,3,4-tri-O-acetyl-β-D-glycopyranuronate)-3-nitrobenzyloxy]5,6-dihydro-4H-benzo[de]quinoline-camptothecin;
(g) reacting 10-[4-O-(methyl-2,3,4-tri-O-acetyl-β-D-glycopyranuronate)-3-nitrobenzyloxyl]5,6-dihydro-4H-benzo[de]quinoline-camptothecin with sodium hydroxide to produce 10-[4-O-(methyl-β-D-glycopyranuronate)-3-nitrobenzyloxy]5,6-dihydro-4H-benzo[de]quinoline-camptothecin; and
(h) purifying 10-[4-O-(methyl-β-D-glycopyranuronate)-3-nitrobenzyloxy]5,6-dihydro-4H-benzo[de]quinoline-camptothecin to give 10-[4-O-β-D-glucopyranuronate)-3-nitrobenzyloxy]camptothecin.
10. A method for synthesizing the compound of formula I comprising:
(a) mixing D(+)-glucurono-3,6-lactone and sodium methoxide in methanol to give methyl 1,2,3,4-tetra-O-acetyl-β-D-glycopyranuronate;
(b) reacting a solution of methyl 1,2,3,4-tetra-O-acetyl-β-D-glycopyranuronate and TiBr 4 in CH 2 Cl 2 to give methyl 1-bromo-1-deoxy-2,3,4-tri-O-acetyl-α-D-glycopyranuronate;
(c) mixing a suspension of methyl 1-bromo-1-deoxy-2,3,4-tri-O-acetyl-α-D-glycopyranuronate, 4-hydroxy-3-nitrobenaldehyde, and Ag 2 O to give methyl 1-O-(2-nitro-4-formylphenyl)-2,3,4-tri-O-acetyl-β-D-glycopyranuronate;
(d) mixing methyl 1-O-(2-nitro-4-formylphenyl)-2,3,4-tri-O-acetyl-β-D-glycopyranuronate, NaBH 4 and silica gel in i-PrOH/CHCl 3 to give methyl 1-O-[2-nitro-4-(hydroxymethyl)phenyl]-2,3,4-tri-O-acetyl-β-D-glycopyranuronate;
(e) reacting a solution of methyl 1-O-[2-nitro-4-(hydroxymethyl)phenyl]-2,3,4-tri-O-acetyl-β-D-glycopyranuronate in CH 2 Cl 2 with methanesulfonyl chloride and triethylamine to give methyl 1-O-[4-(methanesulfonyloxymethyl)-2-nitrophenyl]-2,3,4-tri-O-acetyl-β-D-glycopyranuronate;
(f) reacting methyl 1-O-[4-(methanesulfonyloxymethyl)-2-nitrophenyl]-2,3,4-tri-O-acetyl-β-D-glycopyranuronate with a suspension of BQC and Cs 2 CO 2 in anhydrous DMF and purified on silica gel to give 10-[4-O-(methyl-2,3,4-tri-O-acetyl-β-D-glycopyranuronate)-3-nitrobenzyloxy]5,6-dihydro-4H-benzo[de]quinoline-camptothecin;
(g) reacting a suspension of 10-[4-O-(methyl-2,3,4-tri-O-acetyl-β-D-glycopyranuronate)-3-nitrobenzyloxy]5,6-dihydro-4H-benzo[de]quinoline-camptothecin with sodium hydroxide in anhydrous MeOH to produce 10-[4-O-(methyl-β-D-glycopyranuronate)-3-nitrobenzyloxy]5,6-dihydro-4H-benzo[de]quinoline-camptothecin; and
(h) purifying 10-[4-O-(methyl-β-D-glycopyranuronate)-3-nitrobenzyloxy]5,6-dihydro-4H-benzo[de]quinoline-camptothecin to give 10-[4-O-(β-D-glucopyranuronate)-3-nitrobenzyloxy]camptothecin.