Method for production of geminipeptides as nucleotic agents

FIELD: microbiology.

SUBSTANCE: invention relates to method for production of porphyrinopeptides satisfying the formula I , wherein R1 and R2 independently from one another represent amino acids or peptides comprising 2-15 of amino acid residues, wherein α-carboxylic groups of amino acids or peptides may be modified by C1-C8-alkyl ester and side functional groups of amino acids or peptides may be protected; in particular R1 is ArgOMe; R2 is -OH (III); R1 is LeuHisOMe; R2 is -OH (IV); R1 is LeuLeuValPheOMe; R2 is -OH (V); porphyrin carboxylic group may be modified by methyl or other C1-C9-ester or pharmaceutically acceptable salt; Y- represents Cl-; Me represents Zn, Cu, Fe, Mn. Claimed method includes activation of porphyrin carboxylic group with N-oxy-5-norbornene-2,3-dicarboxyimede in molar ratio of 1:1 in presence of N,N'-dicyclohexylcarbodiinide; or with diphenylphosphorylazide (DPPA) in equimolar ratio of porphyrin/DPPA in presence of base. Then porphyrin with activated carboxylic group is brought into reaction with amino component (amino acid or peptide) in form of mineral acid salt, which is neutralized with base. Also disclosed are methods for application of compounds (I) as nucleotic agents.

EFFECT: new nucleotic agents.

4 cl, 7 ex, 1 tbl

 

The invention relates to chemistry and medicine, in particular to a new method of synthesis of conjugates of porphyrins, in particular hemin oligopeptides and amino acids, and their use as nucleophilicity agents of General formula I:

where R1and R2- substituents, which may be an amino acid or a peptide consisting of 2-15 amino acid residues, with α -carboxyl group of amino acids or peptides can be modified by C1-C8alkilany ether, and the side functions of amino acids or peptides can be protected, and it is possible that R1=R2or R1R2in particular, R1=-ArgOMe, R2=-OH(III); R1=-LeuHisOMe, R2=-OH(IV); R1=-LeuLeuValPheOMe, R2=-OH(V); carboxyl group of the porphyrin can be modified methyl or other C1-C8ester or physiologically acceptable salt;

Y-represents CL-;

Me represents Zn, Cu, Fe, Mn.

It is known that one of the mechanisms of action of antiviral drugs is their ability to destroy DNA. In this regard, at present, a search for agents capable of effectively and selectively restrukturirovany DNA and RNA. Connection with such activity is detected, for example, among the peptides [Pierre J., Laval J. // J.Biol. Chem. 981. V.256. R-10220] and their conjugates with oligonucleotides. Geminus has some antiviral activity [Levere R.D., Gong Y.-F., Bucher D.J., Wormser C.P., Abraham N.G. // Proc. Natl. Acad. Sci. USA. 1991. V.88. P.1756-1759] and in special conditions can cause destruction of sites of DNA [Aft R.L., Mueller G.C. The hemin - mediated DNA strend scission. // J.Biol. 1983. V.258. P.19069-19079].

Given the above, Geminate can be a better reagent for cleavage of DNA (RNA).

Porphyrinoid, specifically Geminate corresponding to the formula (I), namely, (IV), (V), previously obtained with the methods of classical synthesis in solution and on solid phase. In both cases, as an activated derivative of hemin was used N-oxysuccinimide ether [Radygin VA, Philipovich H., Evstigneeva R.P. // Ukr. General. chemistry. 1980. T. S-678.; Evstigneeva R.P., Luvsandorj L.K., Zheltukhina GA // Bioorgan. chemistry. 1993. T. No. 6. S-669.; Evstigneeva R.P., Zheltukhina GA, Zarubina T.V., Nebolsin V.E., Nose, D.N., Spout I.I.// Copyright patent application No. 2002111028, filing date 25.04.2002].

Solid-phase synthesis method of gemination allows purposefully with high yield monopeptide hemin derivatives and has advantages over the classical synthesis in solution when receiving gemination containing relatively long peptides with complex amino acid sequence is. However, the solid-phase synthesis method gemination containing residues of amino acids or short peptides, unreasonably complicated and time-consuming compared with the synthesis in solution.

Known aminoaniline and peptide hemin derivatives corresponding to formula I, namely (III-V)were previously obtained by interaction aminoboronic ester of the amino acid or peptide with N-oxysuccinimide ether hemin [Evstigneeva R.P., Rozhkova EA, Nemykin V.N., Voronov S.A., Zheltukhina G.A., " DOKL. 1997. T. S-644.; Evstigneeva R.P., Zheltukhina GA, Zarubina T.V., Nebolsin V.E., Nose, D.N., Spout N.N.// Original patent application No. 2002111028, filing date 25.04.2002].

The last is a mixture of mono - and bis-esters with Eminem at a ratio of approximately 2:1:1, which is introduced into the reaction aminocompounds without selection. As a consequence, to highlight mono-aminoaniline and mono-peptide derivatives of hemin was necessary to use time-consuming two-step purification, involving the use of column and preparative thin-layer chromatography. As a result, the outputs of gemination (III-V) were, respectively, 17, 47 and 41%.

These shortcomings make known methods are unsuitable for preparative synthesis of low molecular weight gemination corresponding to the General formula I.

Task claimed and the possession is the simplification of the process, increase the yield and quality of the target product.

The problem is solved by the claimed method, according to which the synthesis of porphyrinoids, in particular gemination formula I in solution is carried out by acylation of the amino group of the amino acid or peptide N-hydroxy-5-norbornene-2,3-dicarboximide ether hemin or Eminem, carboxyl group which is activated by the action of DPPA [herskowitz AV, Kibirev VK Chemical synthesis of peptides. // Kiev: Naukova Dumka. 1992. 359 S.].

Activation of the carboxyl group of hemin is carried out by conversion to N-hydroxy-5-norbornene-2,3-dicarboximide ether DCC method (scheme 1) with effect concentration dilution. The latter is a slow addition of 5 to 6 hours solution of 1 EQ. DCC in dimethylformamide solution protohemin IX and N-hydroxy-5-norbornene-2,3-dicarboximide (1:1) at 0° and under vigorous stirring. As a result, the ratio of geminus:mono-activated ester:bis-activated ester in the reaction mixture is about 2:4:1, respectively. Thus, the number of mono-activated ester in the reaction mixture is doubled, and the number of bis-activated ester is decreased compared to a similar reaction with N-oxysuccinimide.

Scheme 1

Synthesis of 6(7)-mono-N-hydroxy-5-norbornene-2,-dicarboximide ether protohemin IX

The obtained mono-ONb-ether hemin (II) no selection enter into reaction with aminocompounds H-ArgOMe or H-Leu-HisOMe or H-Leu-Leu-Val-PheOMe. The reactions take place at a higher speed compared with similar reactions involving N-oxysuccinimide ether hemin. Thus, there is a complete conversion of mono-N-hydroxy-5-norbornene-2,3-dicarboximide ether and aminocompounds with predominant formation of mono-aminoacylase or monopeptide hemin derivatives; the ratio of the formed mono - and bis-aminoacyl or peptide derivatives of hemin in the reaction mixture is about 4:1.

The above factors and a significant difference in Rf mono - and bis-N-hydroxy-5-norbornene-2,3-dicarboximide esters of hemin, and between N-hydroxy-5-norbornene-2,3-dicarboximide esters and aminoaniline and peptide derivatives of hemin simplify the allocation process and to increase the yields of target mono-peptide derivatives of hemin (III, IV, V). To clean these gemination was sufficient single column chromatography on silica gel, while using N-oxysuccinimide ether of hemin to highlight the corresponding gemination (III, IV, V) was required rather complicated purification: column chromatography in combination with preparative TLC or dvuhkomnatnaya [Evstigneeva R.P., Rozhkova EA, Nemykin V.N., Voronov S.A., Zheltukhina G.A., " DOKL. 1997. T. S-644], which complicates the whole process and reduces the yield of products.

The outputs of gemination (III, IV, V), obtained using N-hydroxy-5-norbornene-2,3-dicarboximide esters of hemin were slightly higher than in the case of N-oxysuccinimide esters of hemin, and are, respectively, 36, 60 and 51%.

In addition, an objective of the claimed invention to increase the yield and quality mono-derivatives protohemin IX and facilitate the selection can be solved in another alternative method using diphenylphosphinite (DPPA) for activation of the COOH-group of hemin.

Activation of the COOH-group of hemin effect DPPA carried out generally in accordance with the methodology used in peptide synthesis for the activation of amino acids [herskowitz AV, Kibirev VK Chemical synthesis of peptides. // Kiev: Naukova Dumka. 1992. 359 S.], by successively added to a solution of hemin in cooling the base, DPPA and aminocompounds. The advantage of the proposed method, as in the case of ONb-esters of hemin are almost complete conversion aminocompounds, the predominant formation of mono-peptide derivative (the ratio of generated mono - and bis-peptide derivatives of hemin is approximately 4:1).

The above factors and the absence of the sustainable activated derivatives of hemin after completion of the reaction, you can simplify the allocation process and to increase the yields of target mono-derivatives of hemin. To clean them turned out to be a single column chromatography on silica gel and outputs aminoacylase derived hemin and gemination were, respectively, 63, 66, and 62%.

Previously when receiving gemination (IV, V) were used terminal of amine component, obtained by neutralization of the corresponding trifurcation by triethylamine. It is known that triperoxonane acid in the presence of a base and an activated carboxylic component in anhydrous environment during the formation of the various methods of the peptide bond between the activated carboxyl and aminocompounds can serve as a source of adverse reactions [gross, E., Myencoder N. The peptides. The main methods of formation of the peptide bond. // Moscow. World. 1983].

In addition, triptorelin can act as a counterion for Jonah metal instead of CL-in the composition of the porphyrin residue.

The above may change and affect the quality and output of gemination, including their stability during storage.

So, Geminate (IV, V), obtained with the use of triperoxonane acid is unstable in solutions of DMSO, which is confirmed by TLC, electronic and mass spectrometry, which may be the cause of poor reproducibility of biological and biochemical tests performed on solutions containing DMSO.

the left in the application for the invention is the goal of improving quality and yield of gemination solved by replacing triperoxonane acid on mineral, for example, hydrochloric, when the removal of the N-protection, such as Vos, from N-protected peptide aminocompounds in the synthesis of gemination formula I. thus Obtained Geminate stable in solutions containing DMSO.

Conclusion

Thus, the activation of the COOH-groups of hemin with the use of mono-N-hydroxy-5-norbornene-2,3-dicarboximide ether and DCC or DPPA in the presence of a base and using aminocompounds - peptides in the form of salts with a mineral acid, such as clorhidrato, followed by neutralization with base ensures simplification of the process, increase output and quality of gemination.

List of abbreviations

Vos - tert-butyloxycarbonyl

DCC is N,N’-dicyclohexylcarbodiimide

DCU - N,N’-dicyclohexylmethane

DMF - N,N’-dimethylformamide

DMSO - dimethyl sulfoxide

DPPA - diphenylphosphinite

Meon - methyl alcohol

OMe methyl ether

ONb - norbornenedicarboxylic

Tris-Hcl - (hydroxymethyl)aminomethane hydrochloride

TLC - chromatography in thin layer

EDTA - ethylenediaminetetraacetate

Experimental part

Synthesis of porphyrinoids reflected in examples 2-7.

The proposed porphyrinoid get with high yields and high purity and characterized by UV, IR spectroscopy, mass spectrometry, TLC.

Individuality polucen the x connection is checked by TLC on plates Kieselgel 60 F 254(company Merck, Germany) in the system chloroform - methanol 9:1 (1), chloroform - methanol - acetic acid 5:3:1 (2), chloroform - methanol 4:1 (3), on plates Alufol (neutrons., firm Merck, Germany) in methanol (4)in the system chloroform - methanol 9:1 (5).

Chromatogram showing the ninhydrin, chlorine-toleenum reagent and glow under UV light, imidazolidinone derivative reagent Pauli.

Mass spectra of high resolution receive on time-of-flight mass spectrometer REFLEX™ III (BRUKER, Germany) by the method of matrix usernamescorpionbay ionization, as the matrix used was 2,5-dihydroxybenzoic acid.

IR spectra were recorded on the instrument: Perkin ELMER FT-IR 1725X Spectrometer (Switzerland) and Magna 750 (Nicolet, USA).

Electronic spectra shoot at the instrument Jasco model UV/VS 7800 Spectrophotometer (Japan).

Example 1. Synthesis of 6(7)-mono-N-hydroxy-5-norbornene-2,3-dicarboximide ether protohemin IX (II)

To a solution of 0.7 g (1.08 mmol) protohemin IX in 7 ml DMF and 0.20 g (1.08 mmol) of N-hydroxy-5-norbornene-2,3-dicarboximide when cooled to -10° and With stirring is added dropwise 0.22 g (1.08 mmol) of DCC in 3 ml of DMF for 6 hours the Solution is kept for 48 h at 0° C. DCU separated. Additional cleaning from DCU spend presidenial from DMF-ether. In the sediment obtain a mixture of compounds with Rfcore.(II)0.27(1), Rfminor.0.5(1), Rfhemin0.15 (1).

Example 2. Nα -[6(7)-(�protohemin IX)-yl]-Arg-OMe (III)

To a solution of 0.16 ml (1.2 mmol) Et3N in 2 ml of DMF was added 0.16 g (0.6 mmol) of methyl ester hydrochloride arginine, leave for 15 min at 0° C. To the solution obtained aminocompounds add a solution of 20.8 ml (0.6 mmol) 6(7)-mono-N-hydroxy-5-norbornene-2,3-dicarboximide ether protohemin (IX) in DMF obtained by the above method. The reaction mixture is stirred for 5 h until the complete disappearance of the mono-activated ester of hemin Rf 0.27(1). The solution was concentrated in vacuo to 2 ml and add 6 ml of ether. The precipitate separated from the solution by decanting and dried over anhydrous CaCl2. The substance is purified on a column (24× 1.2 cm) with Kieselgel 60, the target product elute with a mixture of chloroform-methanol (7:3). The fractions containing the substance with Rf 0.78(2), unite. The solvent is removed in vacuum. The substance is dissolved in 8 ml of butanol and washed with water (3× 3 ml). The solvent of the organic layer is removed in vacuum. For the introduction of the counterion CL-Geminate (III) in 10 ml of a mixture of chloroform-methanol (9:1) shake with 6 ml of 0.5 n hydrochloric acid, 6 ml of 2% aqueous sodium hydrogen carbonate solution and water until neutral. The organic layer is dried over anhydrous Na2SO4the solvent is removed in vacuum and the residue is dried over anhydrous CaCl2in a vacuum. Yield 0.14 g (36%). Rf 0.78 (2). Electronic spectrum, λmaxnm, l3-Meon (1:1), (ε· 10-3): 397.4 (101.5), 500.80 (4.49), 540.40 (2.63). 626.0 (1.14). The IR spectrum ν cm-1, liquid paraffin: 1745 (WITH slap.), 1650 (amide I), 1514 (amide II). Mass spectrum, m/z: [M]+786.41.

Example 3. Nα -[6(7)-(Protohemin IX)-yl]-Leu-His-OMe (IV)

A solution of 0.14 g (0.36 mmol) of Boc-Leu-His-OMe 3.25 n Hcl in a mixture of dioxane and methanol incubated 25 min at 20° C. the Acid and the solvent is removed in vacuo, the residue triturated with ether. The precipitate was separated by centrifugation and dried in vacuum over anhydrous NaOH. To a solution of dichlorhydrate methyl ester lacilities in 3.0 ml of DMF under stirring and cooling to 0° add 0.1 ml (0.72 mmol) Et3N. Leave for 15 min at 0° C. To the solution obtained aminocompounds add a solution of 8 ml (0.36 mmol) 6(7)-mono-N-hydroxy-5-norbornene-2,3-dicarboximide ether protohemin IX (I)obtained previously. The reaction mixture was stirred for 4 h until the disappearance aminocompounds. The solution was concentrated in vacuo to 1.5 ml and add 6 ml of ether. The precipitate was separated from the solvent by decantation and dried over anhydrous CaCl2. The substance is purified on a column (19× 3.5 cm) with Kieselgel 60, the target product elute with a mixture of chloroform-methanol (7:3). The fraction containing a mixture of mono - and bis-(methyl ester Nα -lacilities)-protohemin IX with Rf 0.4 (3) and Rf 0.38 (3), respectively, and the fraction containing individual product with Rf 0.38 (3), 0 (4) is collected about the individual. The solvent is removed in vacuum. The fraction with Rf 0.4 (3) optionally purified column chromatography, elute with a mixture of chloroform-methanol (9:1), selected fractions separately with Rf 0.38 respectively (3), 0 (4) (compound IV), 0.4 (3), 0.8 (4) (bis-derived). The solvent is removed in vacuum. The target substance (IV) perioada ether of chloroform. For the introduction of the counterion CL-Geminate (IV) treated in accordance with the procedure described for compound (III). Yield 0.19 g (60%). Rf 0.38 (3), Rf 0 (4), Electronic spectrum, λmaxnm, l3-Meon (8:2), (ε · 10-3): 401.00 (102), 497.40 (10.6), 575.80 (3.87), 630.20 (2.27). IR spectrum, KBr, ν cm-1: 3274 (NH, Sh.S.), 1744 (WITH Slater), 1663 (amide I), 1552 (amide II). Mass spectrum, m/z: [M]+880.8.

Example 4. Nα -[6(7)-(Protohemin IX)-yl]-Leu-Leu-Val-Phe-OMe (V).

The compound (V) are obtained similarly to the compound (IV) of 0.105 g (0.18 mmol) of Boc-Leu-Leu-Val-Phe-OMe and 4.5 ml (0.18 mmol) 6(7)-mono-N-hydroxy-5-norbornene-2,3-dicarboximide ether protohemin IX (II)obtained previously. The target substance can be distinguished in column (35× 1.2 cm) with Kieselgel 60, elwira mixture of solvents chloroform-methanol (9:1). For the introduction of the counterion CL-Geminate (V) is treated in accordance with the procedure described for compound (III). Yield 0.190 g (51%). Rf 0.5 (1), Rf 0 (5). Amino acid analysis: Leu 1.9 (2), Val 1.07 (1), Phe 1.02 (1). The IR spectrum ν cm-1, KBr: 3248 (NH), 1730 (CO slap.), 1640 (amide I), 1540 (amid). Mass spectrum, m/z: [M+] 1102.8.

Example 5. Nα -[6(7)-(Protohemin IX)-yl]-Arg-OMe (IIIa)

To a solution of 0.05 g (0.20 mmol) of H-ArgOMex2HCl and 0.13 g (0.20 mmol) protohemin IX in 2 ml of DMF under cooling to -10° and With stirring was added 0.044 ml (0.20 mmol) of DPPA and 0.084 ml (0.6 mmol) Et3N. the Reaction mixture was stirred for 2 h at -10° and 15 h at 20° C. the Solution was concentrated in vacuo to 1.5 ml and add 6 ml of ether. The precipitate was separated from the solvent by decantation and dried over anhydrous l2. Connection purified on a column of Kieselgel 60 F254(12× 31.5 cm) (Merck). The target substance with Rf 0.78 (2) elute with a mixture of chloroform-methanol (7:3). The solvent is removed in vacuum. For the introduction of the counterion CL-Geminate (IIIa) is treated in accordance with the procedure described for compound (III). The output of 0.100 g (63%). Rf 0.78 (2). Electronic spectrum, λmaxnm, l3-Meon (1:1), (ε · 10-3): 398.4 (97.5). 495.80 (5.8), 542.40 (2.54), 630.0 (1.1). The IR spectrum ν cm-1, liquid paraffin: 1745 (WITH slap.), 1635 (amide I), 1560 (amide II). Mass spectrum, m/z: [M]+786.0.

Example 6. Nα -[6(7)-(Protohemin IX)-yl]-Leu-His-OMe (IVa)

A solution of 0.10 g (0.26 mmol) of Boc-Leu-His-OMe 3.25 n Hcl in a mixture of dioxane and methanol incubated 25 min at 20° C. the Acid and the solvent is removed in vacuo, the residue triturated with ether. The precipitate was separated by centrifugation and dried in vacuum over anhydrous NaOH. To a solution of the obtained aminocompounds and 0.17 g (0.26 mmol) protohemin IX in 3 ml of DMF, when cooled to -10° and With stirring was added 0.056 ml (0.26 mmol) of DPPA and 0.11 ml (0.8 mmol) Et3N. the Reaction mixture was stirred for 2 h at -10 C and 15 h at 20° C. the Solution was concentrated in vacuo to 1.5 ml and add 6 ml of ether. The precipitate was separated from the solvent by decantation and dried over anhydrous CaCl2. The compound (IVa) is purified on a column of Kieselgel 60 F254(35× 1.2 cm) (Merck). The solvent is removed in vacuum. For the introduction of the counterion CL-Geminate (IVa) is treated in accordance with the procedure described for compound (III). Output 0.156 g (65.6%). Rf 0.38 (3), Rf 0 (4). Amino acid analysis: Leu 1.12 (1), His 1.07 (1). The IR spectrum ν cm-1, KBr: 3400 (NH-), 1741 (WITH slap.), 1660 (amide I), 1534 (amide II). Mass spectrum, m/z: [M]+880.2

Example 7. Nα -[6(7)-(Protohemin IX)-yl]-Leu-Leu-Val-Phe-OMe (Va)

Release aminocompounds and synthesis of Geminate (Va) is carried out in accordance with the conditions described for compound (IVa), on the basis of 0.075 g (0.12 mmol) of Boc-Leu-Leu-Val-Phe-OMe, 0.078 g (0.12 mmol) protohemin IX in 3 ml of DMF, 0.03 ml (0.12 mmol) of DPPA and 0.033 ml (0.24 mmol) Et3N. the Target substance can be distinguished in column (40× 1.2 cm) with Kieselgel 60, elwira mixture of solvents chloroform-methanol (9:0.5). For the introduction of the counterion CL-Geminate (Va) process in accordance with the procedure described for compound (III). Output 0.084 g (62%). Rf 0.5 (1), Rf 0 (5). Amino acid analysis: Leu 2.01 (2), Vl 1.06 (1), Phe 0.9 (1). The IR spectrum ν cm-1, KBr: 3250 (NH), 1726 (CO slap.), 1642 (amide I), 1542 (amide II). Mass spectrum, m/z: [M]+1103.0

The study nucleotidase activity gemination

Another object of the invention is the use of gemination General formula I as nucleophilicity agents.

The study nucleotidase activity gemination (III, IV, V) conduct by a typical method [Maniatis T., Fritish E., Sambrook J. Molecular Cloning (A Laboratory Manual), Cold Spring Harbor; N.-Y.: Cold Spring Harbor Cloning (A Laboratory Manual), Cold Spring Harbor; N.-Y.: Cold Spring Harbor Laboratory Press, 1982] with the use of DNA plasmids pGem, and DNA plasmids pUKB244 carrying a full-sized viral gene Gag-P24 (HIV), isolated from E. coli. Geminate at a concentration of 10-4-10-8M incubated with 0.5 μg of plasmid DNA for 1 h at 37° With 10 ál of the reaction mixture. The cleavage products analyzed by electrophoresis in 1% agarose gel in TAE buffer (0.04 M Tris-acetate (pH 8.0), 0.02 M EDTA) at a voltage of 10 V/see the reaction Products produce images by painting them fluorescent dye bromide by ethidium. The DNA bands detected by using a source of UV light wavelength region with λ 300 nm and photographed with a digital camera company Canon.

The activity of the compounds tested at acidic and neutral pH values in the following buffer solutions:

buffer I:100 mm AcONH4(pH 4.5), 10 mm Mnl 2;

buffer II: 100 mm AcONH4(pH 4.5), 20 mm ZnOAC2;

buffer III: 100 mm AcONH4(pH 4.5), 10 MM MgCl2;

buffer IV: 20 mm Tris-HCl (pH 7.5), 20 mm nl2;

buffer V: 20 mm Tris-HCl (pH 7.5), 20 mm ZnOAC2;

buffer VI: 100 mm Tris-HCl (pH 7.5), 10 mm MgCl2;

the buffer VII: 20 mm Tris-HCl (pH 7.5), 2 mm FeSO4;

the buffer VIII: 10 mm Tris-HCl (pH 8.5), 10 mm MgCl2, 100 mm KCl, 0.1 mg/ml BSA, 200 mm EDTA;

the IX buffer: 10 mm Tris-HCl (pH 7.5), 10 mm MgCl2, 0.1 mg/ml BSA, 200 mm EDTA.

Geminate added to the reaction mixture in the form of a solution in DMSO, and the concentration of DMSO in the final sample is 10%.

The disappearance of bands corresponding to different forms of DNA, indicate the cleavage of DNA by oxidative type.

Results

Introduction in the reaction environment gemination causes the disappearance of the bands (splitting) DNA under different conditions (pH, metal ions and other) (see table).

tr>
Table 1

Nucleotidase activity gemination
No. of connectionsFormulaBuffer (), DNA cleavageBuffer (), no DNA cleavage
IIIHeminArgOMe(I-III, VIII, IX)

+
(IV-VII)

-
IVHeminLeuHisOMe(I, II)

+
(III-IX)

-
VHeminLeuLeuValPheOMe (I, IV)

-
+ DNA cleavage

- the absence of DNA cleavage

Conclusion

Porphyrinoid received the claimed method can be used as nucleophilicity agents, such as biomedical research, or as potential drugs for the chemotherapy of viral diseases and cancer.

1. Ways of getting porphyrinates General formula I or their pharmaceutically acceptable salts

(I)

where R1and R2- substituents, which may be an amino acid or a peptide consisting of 2÷ 15 amino acid residues, with α -carboxyl group of amino acids or peptides can be modified With the1-C8alkilany ether, and the side functions of amino acids or peptides can be protected, and it is possible that R1=R2or R1R2in particular, R1=-ArgOMe, R2=-OH (III); R1=-LeuHisOMe, R2=-OH (IV); R1=-LeuLeuValPheOMe, R2=-OH (V); carboxyl group of the porphyrin can be modified methyl or other1-C8ester or physiologically acceptable salt;

Y-represents Cl- ;

Me represents Zn, Cu, Fe, Mn;

including the activation of carboxyl groups of the porphyrin and the subsequent interaction of the activated derivative of porphyrin with an amino acid or a peptide, characterized in that the carboxyl group of the porphyrin activate the action of N-hydroxy-5-norbornene-2,3-dicarboximide at a molar ratio of initial reagents 1:1 and the process is conducted in the presence of DCC, or the action of the DPPA when equimolecular ratio porphyrin: DPPA in the presence of a base, and then activated at the carboxyl group of the porphyrin enter into reaction with aminocompounds - amino acid or a peptide, which is in the form of a salt with a mineral acid, which is neutralized by the base.

2. Ways of getting porphyrinates according to claim 1, characterized in that the compounds of General formulaIthe rest of the porphyrin is a gemin; amino acid residues or peptide represent-Arg-OMe, -Leu-HisOMe, -Leu-Leu-Val-PheOMe and the basis for the activation of the COOH group of hemin effect DPPA and neutralization aminocompounds, in the form of a hydrochloric salt, is a triethylamine.

3. The use of porphyrinoids General formulaIas nucleophilicity agents.

4. The use of porphyrinoids according to claim 3, characterized in that compounds of General formulaIarenucleotides aktivnosti in relation to viral DNA.



 

Same patents:

FIELD: biochemistry.

SUBSTANCE: invention relates to method for production of synthetic chlorophyll (Chl) or bacteriochlorophyll (Bchl) derivatives of general formula I , wherein X is O;. Claimed method includes interaction under anaerobic conditions of Chl, Bchl derivatives containing COOCH3-group in C-132-position and COOR3-group in C-172-position in presence of tetraethyl orthotitanate. Further compounds of formula I wherein R1 and R2 are different radicals are obtained in aproton solvent such as peroxide-free tetrahydrofurane and dimethyl formamide, and compounds of formula I wherein R1 and R2 are the same ones are produced by using R1OH as a solvent. Derivatives of present invention are useful as stabilizers, linkage/spacer for binding another acceptable molecules to Chl/Bchl macrocycle.

EFFECT: simplified method for production of various chlorophyll or bacteriochlorophyll derivatives.

13 cl, 3 ex, 2 tbl, 8 dwg

FIELD: organic chemistry.

SUBSTANCE: invention relates to new derivatives of metalloporphyrazine of the general formula (I): wherein M means Cu, Co. These compounds can be used as dyes, catalysts in different processes and materials of sensitive members of gas sensor.

EFFECT: valuable properties of compounds.

2 cl, 6 sch, 1 dwg, 5 ex

The invention relates to new derivatives of metalloporphyrins that can primeneniia as pigments, catalysts, materials sensitive elements gases

The invention relates to the preparation of metal complexes of meso-tetraallylsilane of tetrabenzoporphyrin that can be used as a fat-soluble dyes, and also as a source of compounds for obtaining water-soluble dyes

The invention relates to a derivative of hemin or their pharmaceutically acceptable salts and inhibitors of proteolytic enzymes, which are the compounds of General formula (I)

where R1and R2- substituents, which may represent amino acids, derivatives of amino acids, peptides, consisting of 1-15 amino acid residues, derived peptides consisting of 1-15 amino acid residues, and-carboxyl group of amino acids or peptides and side groups of amino acids or peptides can be modified, and it is possible that R1=R2or R1R2=OH; carboxyl group of the porphyrin can be modified methyl or other C2-C8-ester or a physiologically acceptable salt; Y-represents Cl-CH3SOO-; Me represents Fe, with the exception of compounds where

Me=Fe3+, Y-=Cl-,

R1=-LeuLeuValPheOMe, R2=-OH; R1=-ValPheOMe, R2=-OH; R1=-LeuHisOMe,

R2=-OH; R1=-LeuHisAlaOMe, R2=-OH; R1=-LeuHisNHC10H20COOMe, R22=-LeuHisNHC10H20COOMe; R1=-Lys(Tfa)AlaAlaOMe, R2=-OH;

R1=-ValPheOMe, R2=-LeuHisOMe; R1=-LeuLeuValPheOMe, R2=-LeuHisOMe;

R1=-LeuLys(Tfa)LeuOMe, R2=-OH; R1=-LeuLys(Tfa)LeuOMe, R2=-LeuHisOMe;

R1=-Lys(Tfa)AlaAlaOMe, R2=-AlaHisLys(Cbz)LeuOMe; R1=-GlyOBzl,

R2=-GlyOBzl; R1=-HisOMe, R2=-HisOMe; R1=-LeuHisOMe, R2=-LeuHisOMe;

R1=-LeuHisLeuGlyCys(Bzl)OBzl, R2=-LeuHisLeuGlyCys(Bzl)OBzl;

R1=-LeuHisOMe, R2=-OEt; R1=-LeuHisLeuGlyCys(Bzl)OBzl, R2=-OEt; R1=-OBzl,

R2=-OBzl; R1=-OBzl, R2=-OH; R1=-AlaOMe, R2=-OBzl; R1=-HisOMe, R2=-OBzl;

R1=-LeuHisOMe, R2=-OBzl; R1=-LeuHisLeuGlyCys(Bzl)OBzl, R2=-OBzl;

R1=-LeuHisAlaLys(Cbz)GlyCys(Bzl)OBzl, R2=-OBzl; R1=-LeuHisLys(Cbz)OMe,

R2=-OH; R1=-LeuHis(Bzl)Lys(Cbz)OMe, R2=-OH; R1=-LeuHisOMe, R2=-OMe;

R1=-LeuHis(Bzl)Lys(Cbz)OMe, R2=-OMe; R1=-AlaLeuAlaPheAlaCys(Bzl)OMe,

R2=-LeuHis(Bzl)Lys(Cbz)OMe; R1=-AlaLeuAlaPheAlaCys(Bzl)OBzl,

R2=-LeuHis(Bzl)Lys(Cbz)OMe; R1=-LeuHisAlaLys(Cbz)Cys(Bzl)OBzl,

R2=-LeuHis(Bzl)Lys(Cbz)OMe; R1=-LeuHisOMe, R2=-OMe;

R1=-GlyProArgGlyGlyOMe, R2=-OH;

R1=-ArgProProGlyPheSer(Bzl)PheArgGlyGlyOMe, R2=-OH,

two ways to get hemin derivatives of General formula I, hemin derivatives of the formula I, formerly known above, as inhibitors of proteolytic enzymes: the HIV protease, pepsin, trypsin, chymotrypsin

The invention relates to new compounds, metalcomplexes of carboranylporphyrins General formula I,

with antitumor activity and low toxicity, which can be used in boron neutron capture therapy had cancer

The invention relates to palletization derived bacteriochlorophyll formula I, I' or I"

where a is a HE, OR1, -O-(CH2)n-Y, -S-(CH2)n-Y, -NH-(CH2)n-Y, -O-(CH2)2-OH, -NH-(CH2)2-NH-BOC, or-N(CH2-CH=CH2)2where R1represents Na+, K+, (CA2+)0,5, (Mg2+)0,5Li+, NH+4,+NH3-C(CH2OH)3,+NH3-CH2-(CHOH)4-CH2OH,+NH2(CH3)-CH2(SNON)4-CH2OH, or+N(Cn'H2n'+1)4; R2represents N or C1-C12the alkyl for the compounds of formula I', and R2represents H, HE or COOR4for the compounds of formula I, where R4represents C1-C12alkyl or C3-C12cycloalkyl; R3represents N or C1-C12the alkyl for the compounds of formula I', and R3represents H, HE, or C1-C12alkyl or alkoxy with the R'3X-where R'1, R'2and R'3each independently represents-CH3or-C2H5; X represents F, Cl, Br or I, n is 1, 2, 3 or 4, and where * denotes an asymmetric carbon atom and --- represents a single saturated bond or an unsaturated double bond pharmaceutical composition having a capacity of detection or treatment of tumors containing at least one compound of formula I, I' or I", three methods for obtaining the compounds of formula I

The invention relates to new substituted the phthalocyanine, which may find application as a dye, catalyst for various redox processes

The invention relates to a phthalocyanine of the formula (I) used as a means for marking liquids, such as mineral oils

The invention relates to inhibitors tyrosinekinase type bis-indolylmaleimide compounds of the formula I

< / BR>
where Z denotes a group of General formula II

< / BR>
where A, B, X, Z, R1-R10have the meanings indicated in the claims, as well as the way they are received and drug based on these compounds

The invention relates to the field of medicine and organic chemistry and relates to a therapeutic agent against hepatitis C, containing the compounds of formula I, farbkomposition containing these compounds, and the polymerase inhibitor of hepatitis C virus

The invention relates to medicine and relates to antiviral agents for external use, with anti-herpes activity, wound healing and antiviral action

The invention relates to methods for treating diseases caused by hepatitis B virus (also known as HBV and Epstein-Barr (also known as EBV, which include the introduction of an effective amount of one or more of the active compounds disclosed here, or farmatsevticheskii acceptable derivatives or prodrugs of one of these compounds

The invention relates to medicine

The invention relates to biologically compatible carriers of oxygen for administration to patients as a Supplement or partial substitution of whole blood

The invention relates to medicine and relates to a substitute for red blood cells, and method of its production

The invention relates to medicine, namely to the production of blood preparations

The invention relates to medicine, namely to a method of chromatographic selection of hemoglobin

The invention relates to the production of polyhemoglobin, which may represent the basis of blood solutions with the function of oxygen transport

Antianemic agent // 2108102
The invention relates to the production of therapeutic and preventive drugs for diseases associated with hematopoiesis, can be used in food industry, medicine and veterinary

The invention relates to the technology of extraction of biologically active compounds, namely the allocation of hemoglobin from the blood of farm animals
Up!