| 摘要 |
Conjugate (A) consisting of an oligo-nucleotide (I) covalently bound to at least one cationic metallo-porphyrin deriv. (II) is new. MORE SPECIFICALLYU} (A) are of formula (I): @GRAPHIC = O:/EDNA/IMAGES/9429/FR/2697254.A1/D1.STR,367,134,0 B' = natural or modified nucleic acid base; L = O, S or NH; J = H or OH; n' = 2-50; X1 = O<->, S<->, alkyl, alkoxy, aryloxy or alkyl substd. by N-contg. heterocycle, aminoalkyl, aminoalkoxy, thioalkyl or -Y1-Z1; R'1, R'2 = -Y1-Z1 or -Y2-Z2; one of them may also be H; Y1, Y2 = divalent hydrocarbyl; Z1, Z2 = radical of intercalating or cleaving agent carrying at least one functional gp. reacting (in)directly with the nucleotide chain; at least one of them has formula (II). A, B = R-(CH2)n-phenyl or a gp. (b); @GRAPHIC = O:/EDNA/IMAGES/9429/FR/2697254.A1/D2.STR,432,287,0 @GRAPHIC = O:/EDNA/IMAGES/9429/FR/2697254.A1/D3.STR,176,78,0 Z = N<+>R1 or C-N<+>R1R2R3; R1 = 1-10C aliphatic gp.; R2, R3 = R1 or H; n = 0-10, opt. making a branched alkylene; M = transition metal; X<-> = anion of pharmaceutically acceptable carboxylic acid; Y = O, CO or CONH. USEU} (A) are able to cleave complementary nucleic acid sequences, i.e. they are artificial, sequence-specific nucleases which can be used as antiviral, antibacterial, antiparasitic or antitumour agents by selective destruction of appropriate exogenous or endogenous genes (or related RNA). They can also be used to produce nucleic acid fragments or as reagents for analysing the sec. and tert. structures of RNA and DNA. ADVANTAGEU} (II) are known for cleavage of nucleic acids but by attaching them to (I) selective cleavage of targetted sequences becomes possible. (A) can cleave RNA for which no sequence specific nucleases are known. PREFERRED MATERIALSU} (I) is bound to (II) at the 3'- or 5'-end; R1 = Me or Et; R2 = R3 = H; M = Mn or Fe; Z = N<+>R1; R'1 = Z1-Y1 and R'2 = H; Z1 = trimethylpyridinium-porphyrin -Y-(CH2)n-; Y1 = CONH-alkylene-NHCO; L = O; X1 is different from Y1-Z1. PREPARATIONU} (A) are made by coupling a functionalised (I) with (II), e.g. by peptide bond formation. Partic. (I) is functionalised by attachment of an alkylene diamine. EXAMPLEU} The cpd. MePy3(H2-PP)(PhO(CH2)4COOH) (Py = pyridinium) was metallated with Mn, then 2.9 mg of the product activated with carbonyldiimidazole and hydroxybenzotriazole. After 30 min., the activated deriv. was added to 13.4 nmole of the oligonucleotide 5'-NH2-GGCTCCAT3CTTGCTCTC (complementary to the initiation zone of the TAT gene of HIV-1) and incubated for 20 hr. at 37 deg.C in Hepes buffer. Work up on Bio-Gel P.2(F) gave a 25% yield of the conjugate. When tested against a complementary 35-mer (positions 5360-5394 of the HIV-1 genome), the conjugate had nuclease activity at a concn. above 10 nM. The same activity required 1 mu M of the free metallated porphyrin (not bound to oligonucleotide).
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