Sensitizing agent for photodynamic disruption of malignant neoplasm cells and pharmaceutical composition for its using

FIELD: organic chemistry, medicine, oncology.

SUBSTANCE: sensitizing agent for photodynamic disruption of malignant neoplasm cells represents a new compound - 18-hydroxy-substituted cycloimide of chlorine p6 or 7,8-dihydroxybacteriochlorine of the formula (I):

. Compounds of the formula (I) are used as an agent in treatment of malignant neoplasm. Invention expands assortment of sensitizing agents used in carrying out the photodynamic therapy.

EFFECT: valuable medicinal properties of agent.

6 cl, 1 dwg

 

The invention relates to the chemistry of biologically active compounds in the field of photodynamic therapy, in particular sensitizers for photodynamic destruction of malignant tumors.

Method of photodynamic therapy based on the use of substances - sensitizers, which when introduced into the body mainly localized in the tumor, and when light, in particular laser, the excitation wavelength produce cytotoxic substances, especially singlet oxygen. Currently known sensitizers based on complex mixtures hematoporphyrin derivative, for example Fotofrin-II (E.Sternberg, D.Dolphin, C.Bruckner. Tetrahedron, 54, 4151 (1998)).

The lack of sensitizers based on hematoporphyrin derivatives is that they have intensive absorption band in the ultraviolet and visible (blue, green) regions of the spectrum, and at the same time, in most long-wavelength absorption band (630+10 nm) extinction coefficient is relatively low. Consequently, for effective photodynamic therapy is a necessary introduction into the patient, significant concentrations of the drug. In addition, when entering the patient under direct solar radiation, with significant spectral density in the purple-green region of the spectrum accumulated in the skin prep the rat exhibits significant toxicity. Strong absorption desensibilisation tissues in the spectral range of 620-640 nm leads to a significant loss of radiation outside the tumor tissue, leads to the small penetration depth of the radiation in the tissue, which complicates the treatment of tumors of large size. All this leads to the necessity to use high power and at the same time is the cause of not enough high therapeutic efficiency of photodynamic therapy.

The development of photodynamic therapy of malignant tumors stimulated the search for new low-sensitizers, which have optical properties, the ability to be localized in the tumor cells and the efficiency should be higher than those used so far hematoporphyrin derivative. In this regard, as promising objects are compounds belonging to the class of tetrapyrroles with a modification of the conjugated system and has an intense absorption bands in the red and near IR spectral regions: phthalocyanines, naphthalocyanine, chlorins, a purpurin, a bacteriochlorin, and others.

Thus, the disadvantages of hematoporphyrin derivative partially addressed by the use of photodynamic therapy as a sensitizer aluminum complex of sulfonated phthalocyanine ("Photosense"), popadayuschego spectral range 660-680 nm. The extinction coefficient "Photosense" at the maximum of the absorption band is more than 105l·mol-1·cm-1the absorption desensibilisation tissues in this range decreases, therefore increasing the depth of the photodynamic effects on tissues, which improves therapeutic efficacy of photodynamic therapy (ARV and other, Questions of Oncology, 41, 134 (1995)). However, "photosense" has still some drawbacks, the main of which is a very high value of the used dose (0.5-0.8 mg per kg of body weight), resulting in the presence of his side dermal toxicity.

In recent years, in connection with the search for new effective sensitizers wide range of actions are being actively studied distribution derivative of chlorin p6. These connections are available, almost completely harmless, have good spectral and photophysical properties, as well as the ability to a variety of modifications.

The objective of the invention is the development of new sensitizers for photodynamic therapy based on the distribution of derivative of chlorin p6.

To improve the efficiency of photodynamic destruction of malignant neoplasms as a sensitizer offers new connection - 18-hydroxy-substituted cyclone the s chlorin p 6or 7,8-dihydroxyphenylalanine. Describes the method of production thereof and pharmaceutical composition with a new sensitizer. New connections - 18-hydroxy-substituted cycloimide chlorin p6or 7,8-dihydroxyphenylalanine intended for the treatment of malignant tumors.

According to the invention synthesized a number of new 18-hydroxy-substituted derivatives of chlorin p6and 7,8-dihydroxyphenylalanine with additional emenim eksotika, paired with the main the chlorin macrocycle containing different substituents at the nitrogen atom amidnogo of eksotika, preferably hydroxy-, acetoxy -, and 3-hydroxyproline deputies.

18-hydroxy-substituted cycloimide chlorin p6or 7,8-dihydroxyphenylalanine are compounds of General formula I.

where R1denotes-N-CH3, -(CH2)6HE

R2represents-CH=CH2,,

R3=R4and represents HE or R3and R4together form a chemical bond,

R5denotes-H, -OH, -och3,, -(CH2)2HE, -(CH2)3HE, -(CH2)4OH, -(CH2)3Other6, where

R6the same or different and represent-H,.

Preferred compounds of formula I, where

R1denotes-H or-CH3,

R2represents-CH=CH2,

R3and R4together form a chemical bond,

R5means-HE, -(CH2)3HE.

These compounds are selected from the group comprising (17S, 18R)-18-hydroxy-13,15-N-(3-hydroxypropyl)cycloimide chlorin p6, 175-methyl-(17S, 18R)-18-hydroxy-13,15-N-(3-hydroxypropyl)cycloimide chlorin p6, 175-methyl-(17S, 18R)-18-hydroxy-13,15-N-hydroxycinnamic chlorin p6, 175-methyl-(17S, 18R)-18-hydroxy-13,15-N-acetoxysilane chlorin p6.

The most preferred compounds of formula I, where

R1denotes-H or-CH3,

R2represents-CH=CH2,

R3and R4together form a chemical bond,

R5represents -(CH2)3HE.

These compounds represent (17S, 18R)-18-hydroxy-13,15-N-(3-hydroxypropyl)cycloimide chlorin p6or 175-methyl-(17S, 18R)-18-hydroxy-13,15-N-(3-hydroxypropyl)cycloimide chlorin p6.

As is well known, the introduction of hydroxyl groups in the molecule of chlorine can significantly change its amphiphilicity. This hydroxyl gr is PPI may be located directly in the macrocycle (K.R.Adams, M.C.Berenbaum, R.Bonnett, A.N.Nizhnik, A.Salgado, M.A.Valles. J.Chem. Soc., Perkin Trans. 1, 1465-1470 (1992)), and as part of side substituents (R.Bonnett, M.F.Grahn, A.Salgado, M.Turkish, M.A.Valles, N.S.Williams. Photodynamic therapy and biomedical lasers. Amsterdam:Elsevier, 866-869 (1992)). As practice shows, amphiphilic molecules better accumulate in tumors. Of great interest are compounds in which hydrophobic and hydrophilic substituents are on opposite sides of chlorin molecules. Such substances, usually, easier to penetrate into the cell and then concentrated in its vital compartments. New 18-hydroxy-substituted cycloimide chlorin p6synthesized Regio - and stereoselective method for hydroxylation of natural chlorins and their derivatives. The method is based on intramolecular cyclization residue of propionic acid under the action of 2,3-dichloro-5,6-dicyan-1,4-benzoquinone (DDQ) in an adjacent position 18 education δ-lactone and the disclosure of the latter in an acid or alkaline medium with the formation of 18-hydroxychlorides. In accordance with the scheme of distribution derivative of chlorin p6were synthesized by the reaction of purpurine 18 (1) with hydroxylamine hydrochloride and 3-aminopropanol respectively. Processing of N-acetoxysilane chlorin p6(2) DDQ leads to δ-lactone corresponding chlorin p6(4) with the release of 54%. Similarly, the compound (3) is kislitsa education δ -lactone N-(3-hydroxypropyl)cycloimide chlorin p6(5). Mass spectra of 5-lactones of cycloimide (4) and (5) contain intense peaks of molecular ions with m/z 620.3 (M++N) (4) and 619.6 (M+) (5). In the electronic spectra intensive maximum of the main absorption band Q compounds (4) and (5) is at 707-708 nm, and the band Sora - at 418-419 nm, which means a significant bathochromic shift compared to δ-lactone of purpurine 18 (699 and 411 nm, respectively).

Structure δ-lactones (4) and (5) was also confirmed by the spectra of1H-NMR. Spectra of compounds (4) and (5) a similar range δ-lactone of purpurine 18 and differ from the latter only by the additional signals of the substituents located at the nitrogen atom of cycloimide. Unlike the original cycloimide (2) and (3), in the spectra of compounds (4) and (5) no quartets 18-H protons in the field 4.32-4.33 ppm, and signals 18-CH3shifted in a weak field with 1.70-1.72 ppm up to 2.15-2.18 ppm, and manifest themselves in the form of a singlet and not a doublet, as the starting compounds. Protons 17-N are located in the area of 5.57-5.69 ppm

The study of spectra1H-NMR of compound (5) using the method of NOESY revealed the presence of strong interactions between 17-h, 18-CH3that indicates that the data substituents are located on one side from the plane of the macrocycle and, therefore, formed lacto is Noah ring is located above the plane of chlorine. Subsequent analysis of the NOESY spectra allowed us to do the assignment for protons of two methylene groups in δ-lactone cycle, and Deputy under imenom nitrogen. The protons of the N-hydroxypropyl Deputy shall have the following meanings chem. shifts: the triplet in the region of 4.67 ppm corresponds CH2group on the nitrogen atom, multiplet in the region 3.81 ppm - 134-CH2and the triplet in the region 2.24 ppm - methylene group of the hydroxyl. Similarly, there was made an assignment of all the main signals in the compound (4). These results are in good agreement with data obtained for purpurine 18 and chlorin p6under which additional δ-lactone cycle is located above the main plane of the macrocycle (A.F.Mironov, A.V.Efremov, O.A.Efremova, R.Bonnett, G.Martinez. J.Chem. Soc., Perkin Trans. 1, 3601-3608 (1998), Afronova, Averichev. Bioorg. chem., 27, 141-144 (2001)).

We have shown that 18-hydroxy-substituted N-acetoxysilane (6) is formed with the best output when opening δ-lactone cycle in methanol in the presence of triperoxonane acid. Conversely, to obtain the corresponding derivative of N-(3-hydroxypropyl)cycloimide (7a) it is better to use alkaline hydrolysis. As a result of processing of the lactone (5) 50% aqueous alkali solution in tetrahydrofuran 18-hydroxycinnamic (7a) is obtained with a yield of 41%. In acidic conditions δ-lactone N-(3-hydroxyprop the sludge)of cycloimide (5) mainly undergoes dehydration with the formation of the porphyrin.

To obtain 18-hydroxy-N-hydroxychloride (9) a convenient source is the 18-hydroxypurine 18 (8). Replacement of the N-hydroxycinnamic (9) synthesized by the interaction of 18-hydroxypurine 18 (8) with hydroxylamine hydrochloride in pyridine to yield 58%.

In the mass spectra of compounds (6), (7a) and (9) there are peaks of molecular ions with m/z 652.1, 637.4 and 610.5. Processing 18-hydroxychloride (7a) diazomethane leads to the corresponding methyl ether (7b). This group of 18 HE and 136-OH not affected. In the spectra of1H-NMR 18-gidroksietilimino (6), (7b) and (9) of a methyl group at position 18 is manifested in the form of a singlet and not a doublet, characteristic of natural chlorins. Method NOESY shown that 18-CH3and 17-N are located on one side of the chlorin macrocycle and, consequently, the resulting 18-Oh group is above the plane of the macrocycle. Electronic spectra of chlorins (6), (7a, b), and (9) do not differ from the spectra of their distribution predecessors, not containing in position 18 of the macrocycle hydroxyl group, and have the maximum of the main absorption bands in the region 710-719 nm.

The following examples serve to illustrate various aspects of the present invention.

Examples of the synthesis methods.

Below are the conditions for the synthesis of target compounds for each specific compounds given weekend Yes the data and results of analyses. The purity was controlled by thin-layer chromatography on plates HPTLC-Kieselgel 60 with concentrating zone (Merck, Germany). For preparative thin-layer chromatography was used Kieselgel 60 H (Merck, Germany) (a) and silica gel CHEMAPOL L 5/40 (Czech Republic) (B) on the plates 20×20 cm layer thickness of 1 mm Electronic spectra of compounds recorded on spectrophotometer Jasco 7800" in the region of 400-800 nm in chloroform. Spectra1H-NMR in deuterium chloroform were recorded on a spectrometer "Bruker MSL 200 and Bruker AMX-600".

Mass spectra were recorded on the instrument Micromass Autospec (EI, 70 eV, 200°C). Used in the synthesis of purpurine 18 (1) made from blue-green algae Spirulina platensis in methodology (A.S.Brandis, A.N.Kozyrev, A.F.Mironov. Tetrahedron, 48, 6491 - 6499 (1992)).

(17S, 18R)-17418-lactone 13,15-N-acetoxysilane chlorin p6(4). To a solution of 10.4 mg (0.017 mmol) 13,15-N-acetoxysilane (2) in 5 ml of chloroform added 6 mg (0.033 mmol) of DDQ and stirred for 20 min at 25°C. the Reaction mass is washed with water (5×50 ml), the organic layer is separated, dried with sodium sulfate, the solvent is distilled off in vacuum. The remainder chromatographic on the plate (A) in the system chloroform : acetone (5:1) and recrystallized from a mixture of chloroform-hexane. Received 5.7 mg (54.8%) of the desired product. Electronic spectrum, λmaxnm (ε/103): 418 (110.0), 483 (4.6), 513 (5.6), 552 (23.7), 652 (7.5), 708 (42.1). Range1H-NMR (δ, m is.): 9.68 and 9.67 (1H, 10-H), 9.49 (1H, 5-H), 8.77 and 8.76 (1H, 20-H), 7.92 DD (1H, J 18 and 12 Hz, 31-CH), 6.31 DD (1H, J 18 and 1 Hz, 32-CH2), 6.22 DD (1H, J 12 and 1 Hz, 32-CH2), 5.66 DD and 5.57 DD (1H, J 7 and 5 Hz, 17-H), 3.84 and 3.82 (3H, 12-CH3), 3.67 kV (2H, J 8 Hz, 81-CH2), 3.38 (3H, 2-CH3), 3.20 (3H, 7-CH3), 2.97 m (1H, 171-CH2), 2.65 and 2.64 (3H, 135-CH3), 2.46 m (1H, 172-CH2), 2.28 m (1H, 171-CH2), 2.16 and 2.15 (3H, 18-CH3), 1.69 t (3H, J 8 Hz, 82-CH3), 1.63 m (1H, 172-CH2), 0.5 s and 0.4 s (1H, NH), 0.12 and 0.10 s (1H, NH). The IR spectrum νmaxcm-1: 3431 Wed, 2961 SL, 2923 SL, 2867 SL, 2847 SL, 1728 SL, 1681 SL, 1640, 1601 Wed, 1545 SR 1526 SR 1383 SL, SL 1307, 1168 CL, CL 1062, 987 Wed, 898 SL. Mass spectrum, m/z: 620.3 (M++H).

(17S, 18R)-17418-lactone 13,15-N-(3-hydroxypropyl)cycloimide chlorin p6(5). Similarly, δ-lactone (4) of 9.8 mg (0.016 mmol) 13,15-N-(3-hydroxypropyl) cycloimide chlorin p6(3) in 5 ml of chloroform and 7.1 mg (0.031 mmol) of DDQ after chromatography was carried out on the plate (A) in the system chloroform : acetone (5:1) and recrystallization from a mixture of chloroform-hexane received 5.0 mg (51.5%) of the desired product. Electronic spectrum, λmaxnm (ε/103): 419 (99.7), 483 (1.5), 513 (2.4), 552 (17.2), 651 (3.55), 707 (33.9). Range1H-NMR (δ, ppm): 9.70 (1H, 10-H), 9.52 (1H, 5-H), 8.87 (1H, 20-H), 7.92 DD (1H, J 18 and 12 Hz, 31-CH), 6.28 DD (1H, J 18 and 1 Hz, 32-CH2), 6.21 DD (1H, J 12 and 1 Hz, 32-CH ), 5.69 DD (1H, J 7 and 5 Hz, 17-H), 4.67 t (2N, J 6 Hz, 133-CH2), 3.83 (3H, 12-CH3), 3.81 m (2N, 134-CH2), 3.66 kV (2H, J 8 Hz, 81-CH2), 3.40 (3H, 2-CH3), 3.19 (3H, 7-CH3), 2.97 m (1H, 171-CH2), 2.48 m (1H, 172-CH2), 2.29 (1H, 171-CH2), 2.24 t (2N, J 6 Hz, 135-CH2), 2.18 (3H, 18-CH3), 1.66 (3H, J 8 Hz, 82-CH3), 1.64 m (1H, 172-CH2), -0.18, -0.33 (2H, NH). The IR spectrum νmaxcm-1: 3380 SL, 3331 SL, 2956 Wed, 2919, 2851 Wed, 1758 Wed, 1738 Wed, 1683, 1644, 1603 SL, 1586 SL, 1545, 1526, 1454 SL, 1431 SL, 1397 Wed, 1384 Wed, 1349 SL, SL 1331, 1313 SL, 1168 Wed, 1084 SL, 988, 962 SL, cf 910, 802 Wed, 789, 702, 674 C. Mass spectrum, m/z: 619.6 (M+).

Methyl ether (17S, 18R)-18-hydroxy-13,15-N-acetoxysilane chlorin p6(6). A solution of 17.0 mg (0.026 mmol) of cycloimide (4) in a mixture of 1 ml of TFA and 10 ml Meon stirred for 2 h under argon at 25°C, the reaction mass is diluted with 40 ml of chloroform and washed with water (5×150 ml). The organic layer is separated, dried with Na2SO4and evaporated in vacuum. The remainder chromatographic on the plate in the system chloroform:Meon (10:0.75). After recrystallization from a mixture of chloroform-hexane the product (4) with the release of 6.9 mg (40%). Electronic spectrum, λmaxnm (ε/103): 425 (47.1), 517 (3.9), 559 (13.6), 663 (5.2), 719 (19.5). Range1H-NMR (5, ppm): 9.60 (1H, 10-H), 9.34 (1H, 5-H), 8.79 (1H, 20-H), 7.86 DD (1H, J 18 and 12 Hz, 31-CH), 6.28 the d (1H, J 18 and 1 Hz, 32-CH2), 6.16 DD (1H, J 12 and 1 Hz, 32-CH2), And 5.36 5.44 DD (1H, J 7 Hz, 17-H), 3.79 (3H, CH3), 3.64 kV (2H, J 8 Hz, 81-CH2), 3.34 (3H, 2-CH3), 3.23 and 3.20 (3H, 175-CH3), 3.16 (3H, 7-CH3), 2.62 and 2.60 (135-CH3), 2.57 m (1H, 171-CH2), 2.45 m (1H, 172-CH2), 2.34 m (1H, 171-CH2), 2.15 and 2.12 m (1H, 172-CH2), 1.54 (3H, 18-CH3), 1.62 t (3H, J 8 Hz, 82-CH3), 0.30 and 0.12 (2H, NH). Mass spectrum, m/z: 652.1 (M+).

(17S, 18R)-18-hydroxy-13,15-N-(3-hydroxypropyl)cycloimide chlorin p6(7a). To a solution of 27.0 mg (0.044 mmol) of cycloimide (5) in 5 ml of tetrahydrofuran, add 5 ml of 50% aqueous sodium hydroxide solution and stirred for 4 h at 25°C. the Reaction mass was diluted with 30 ml of water and neutralized with acetic acid. The product is extracted with chloroform (3×50 ml), the organic layer washed with water (3×100 ml), dried with sodium sulfate and evaporated in vacuum. The remainder chromatographic on the plate (A) in the system chloroform : methanol (10:2). After recrystallization from a mixture of chloroform-hexane obtained product (7a) with the release of 11.3 mg (41%). Electronic spectrum, λmaxnm (ε/103): 420 (78.0), 484 (4.4), 514 (4.9), 554 (16.9), 654 (6.1), 711 (29.6). Mass spectrum, m/z: 637.4 (M+).

Methyl ether (17S, 18R)-18-hydroxy-13,15-N-(3-hydroxypropyl)cycloimide chlorin p6(7b). To a solution of 11.3 mg (0.018 mmol who) chlorin (7a) in 5 ml of chloroform added excess ethereal solution diazomethane, stirred for 5 min at 25°and evaporated in vacuum. The remainder chromatographic on the plate (A) in the system chloroform : methanol (10:0.5). The selected substance is recrystallized from a mixture of chloroform-hexane. The output of 10.5 mg (91%). Electronic spectrum, λmaxnm (relative intensity): 420, 484, 514, 554, 654, 711 (2.64:0.15:0.17:0.57:0.21:1.00). Range1H-NMR (δ, ppm): 9.63 (1H, 10-H), 9.36 (1H, 5-H), 8.80 (1H, 20-H), 7.84 DD (1H, J 18 and 12 Hz, 31-CH), 6.25 DD (1H, J 18 and 1 Hz, 32-CH2), 6.14 DD (1H, J 12 and 1 Hz, 32-CH2), 5.45 (1H, J 7 Hz, 17-H), 4.64 t (2N, J 6 Hz, 133-CH2), 3.81 (3H, 12-CH3), 3.78 m (2H, 134-CH2), 3.64 kV (2H, J 8 Hz, 81-CH2), 3.47 (3H, 175-CH3), 3.24 (3H, 7-CH3), 3.16 (3H, 2-CH3), 2.22 m (2H, 171-CH2) and (2H, 172-CH2), 2.04 m (2N, 135-CH2), 1.54 (3H, 18-CH3), 1.66 (3H, J 8 Hz, 82-CH3), -0.07 and-0.06 (2H, NH).

Methyl ether (17S, 18R)-18-hydroxy-13,15-N-hydroxychloride chlorin p6(9). A mixture of 8.0 mg (0.014 mmol) of purpurine 18 (8) [4] in 3 ml of Ru and 9.4 mg (0.14 mmol) of hydroxylamine hydrochloride is stirred for 12 h at 25°C. the Reaction mass was diluted with chloroform, washed with 0.1 N HCl solution (3×50 ml) and then water. The organic layer is separated, dried with Na2SO4and the solvent evaporated in vacuum. The remainder chromatographic on the plate in the system chloroform : Meon (80:5). After the recristallisation from a mixture of chloroform-hexane obtained 4.7 mg (58%) of the product (9). Electronic spectrum, λmax, nm. (ε/103): 419 (82.8), 483 (5.4), 513 (5.8), 553 (19.0), 596 (3.0), 653 (6.8), 710 (31.8). Range1H-NMR (δ, ppm): 8.49 (1H, 10-H), 8.42 (1H, 5-H), 8.23 (1H, 20-H), 7.77 DD (1H, J 18 and 12 Hz, 31-CH), 6.20 DD (1H, J 18 and 1 Hz, 32-CH2), 6.10 DD (1H, J 12 and 1 Hz, 32-CH2), 5.30 (1H, J 7 Hz, 17-H), 4.71 (1H, 18-OH), 3.70 kV (2H, J 8 Hz, 81-CH2), 3.47 (3H, 12-CH3), 3.31 (3H, 2-CH3), 3.23 (3H, 175-CH3), 3.09 (3H, 7-CH3), 2.57 m (1H, 171-CH2), 2.35-2.00 m (3H), 171,172-CH2), 1.79 (3H, 18-CH3), 1.61 t (3H, J 8 Hz, 82-CH3), 0.43 and 0.31 (2H, NH). Mass spectrum, m/z: 610.5 (M+).

The subject of the invention is also a pharmaceutical composition for treatment of cancer comprising as active ingredient a sensitizer, a 18-hydroxy-substituted cycloimide chlorin p6General formula I, in which the symbols have the above significance, in an effective amount and a pharmaceutically acceptable carrier.

Typically, the pharmaceutical composition is supplied by injection. A sample of the pharmaceutical composition is a sterile aqueous solution. Aqueous solution, addition of the sensitizer of the formula I, contains, perhaps, physiological salt and if necessary - propylene glycol.

Describes solutions you can apply neposredno is but after their production or they can be stored in sterile containers. The pharmaceutical composition according to the invention can be used as a sensitizer in photodynamic therapy of tumors.

The new compounds of formula I were investigated in cell lines.

Absorption spectrum of the investigated derivative (7a) in 1% cremophor (45 mm Tris-HCl pH 7.4) is characterized by highs: λ1=367 nm (ε=34600), λ2=419 nm (ε=80800), λ3=483 nm (ε=2300), λ4=513 nm (ε=3300), λ5=553 nm (ε=15730), λ6=647 nm (ε=5200), λ7=710 nm (ε=30300 M-1·cm-1). Stock solution (0.5 mm) (7a) in 10% cremophor characterized by high stability when stored at +4°S: within 2 weeks from the spectra of absorption has not been found either sedimentation or aggregation or changes in the absorption spectrum, pointing to the modification of the original connection.

The compound (7a) can easily penetrate into the cells of the human lung adenocarcinoma A and accumulates in the cytoplasm in Monomeric form, is associated with the cellular membrane structures. High intracellular penetration of the compound (7a) can be achieved by ensuring the stabilization of the Monomeric form of the compound (7a) in biological environments using 0.002-0.01% Cremophor emulsion. When exposed to light of membrane-bound compound (7a) does not form hydroxyl radicals, but is characterized by high is Kim quantum yield of generation of singlet oxygen, equal to 0.67±0.03. Efficient generation of singlet oxygen under the action of the exciting light is not accompanied photoinduced degradation of the photosensitizer, which indicates a fairly high photostability.

To test the activity of compounds (7a) on the cell culture used the original solution of the compound with a concentration of 500 μm in a 5% solution of Cremophor EL (Sigma) in distilled water. Studies were performed on cell cultures of human lung adenocarcinoma A. Cells were cultured in the medium NEEDLE MEME, with the addition of 2 mm L-glutamine and 8% fetal calf serum, at 37°C in a humid atmosphere with 5% CO2. Cells subcultured in 96-well plates in a concentration of 60 tuscl/ml After 24 hours, in the early logarithmic phase of growth were made photosensitizers. The concentration of the photosensitizer was varied from 0.5 μm to 15 nm. Cells were incubated with the photosensitizer within 2 hours, and then irradiated with red light in the presence of a photosensitizer. Irradiation was performed using a halogen lamp 500 W, equipped with a broadband filter KS-13 (λ≥640 nm) and a water filter with thickness of 5 cm, the power Density was 13.6-15.0 mW/cm2. Estimated light dose of 10 j/cm2. To assess cytotoxic the STI cells were incubated in the presence of compound (7a) in the dark (CO 2-incubator). All manipulations with cells was performed in low light conditions.

Estimates of survival cells was visually estimating using light microscopy morphological changes of the cells and by the colorimetric method using the MTT-test (J.Carmichael, W.G.DeGraff, A.F.Gazador, J.D.Minna, J.B.Mitchell. Cancer Res., 47,936-942(1987)).

The level of inhibition of cell growth was calculated by the formula:

ER (%) = [(ODto-ODo)/ODto]×100%,

where: IL is the level of inhibition of cell growth in culture; ODo- the optical density in the experience; ODto- the optical density in control, λmax=550 nm.

Expected concentration of compound at which there is 50% and 90% inhibition of growth of the culture.

The result shows that the compound (7a) has the phototoxicity against cell culture A. IR50was 80 nm. Cytotoxicity in the absence of irradiation in the range of effective concentrations were absent. Photoinduced activity of compound (7a) in vitro exceeds the activity of chlorin p6(IR50=3.4 μm), and used in the clinic photosensitizers Injected (IR50=8.8 µm) and Photosense (IR50=6.5 μm).

Thus, in vitro, the compound (7a) is characterized by high photoinduced activity and low dark toxicity.

The set ek is received pilot photophysical parameters and results of these calculations suggests, the new distribution derivative of chlorin p6can be considered as an effective and promising sensitizers agents of a new generation for photodynamic therapy.

The scheme for synthesis of 18-replacement of cycloimide chlorin p6.

A - NH2OH·HCl, Ru; AU2O; C - NH2(CH2)3HE (1), Ac2O (2), NaHCO3(3), (D) DDQ, CHCl3; E - TFA/MeOH, F - NaOH/THF.

1. The sensitizer for photodynamic destruction of cells of malignant tumors, characterized by the fact that he is an 18-hydroxy-substituted cycloimide chlorin p6or 7,8-dihydroxyphenylalanine General formula I

where R1denotes-H, -CH3,

,

-(CH2)6HE

R2represents-CH=CH2,

R3=R4and represents HE or R3and R4together form a chemical bond;

R5denotes-H, -OH, -och3,

,

-(CH2)2(HE), -(CH2)3OH, -(CH2)4OH, -(CH2)3Other6;

,

where

< num="84"> R6the same or different and represent-H,

2. The sensitizer according to claim 1, which represents a compound of formula I, where

R1denotes-H or-CH3,

R2represents-CH=CH2,

R3and R4together form a chemical bond,

R5means-HE-(CH2)3HE.

3. The sensitizer according to claim 2, selected from the group comprising (17S, 18R)-18-hydroxy-13,15-N-(3-hydroxypropyl)cycloimide chlorin p6, 175-methyl-(17S,18R)-18-hydroxy-13,15-N-(3-hydroxypropyl)cycloimide chlorin p6, 175-methyl(17S,18R)-18-hydroxy-13,15-N-hydroxycinnamic chlorin p6, 175-methyl(17S,18R)-18-hydroxy-13,15-N-acetoxysilane chlorin p6.

4. The sensitizer according to claim 2, representing a compound of formula I, where

R1denotes-H or-CH3,

R2represents-CH=CH2,

R3and R4together form a chemical bond,

R5represents -(CH2)3HE.

5. The sensitizer according to claim 4, representing (17S,18R)-18-hydroxy-13,15-N-(3-hydroxypropyl)cycloimide chlorin p6or 175-methyl-(17S,18R)-18-hydroxy-13,15-N-(3-hydroxypropyl)cycloimide chlorin p6.

6. Pharmaceutical composition for the treatment of malignant newly formed the th, comprising as active ingredient a sensitizer according to any one of claims 1 to 5 in an effective amount and a pharmaceutically acceptable carrier.



 

Same patents:

FIELD: chemistry of metalloorganic compounds.

SUBSTANCE: invention relates to new derivatives of metalloporphyrazines of the general formula (I):

wherein M means Cu, Co. These compounds can be used as dyes, catalysts of different processes and material for sensitive members of gas pickups.

EFFECT: valuable properties of complexes.

4 fig, 1 dwg, 5 ex

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

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

FIELD: medicine, oncourology.

SUBSTANCE: the present innovation deals with conservative treatment of patients with malignant prostatic tumor at different stages. The method includes testicular enucleation, introduction of anti-tumor chemopreparations and radiation therapy. Moreover, in the onset of radiation therapy one should introduce 25 mg Cisplatin incubated with 10 ml patient's plasma into both prostatic lobes and paraprostatic fiber from the right and from the left. At achieving a focal dosage of 20 Gy one should repeat introduction of chemopreparation in similar dosage, and radiation therapy should be continued up to total focal dosage of 40 Gy. The innovation enables to decrease tumor sizes, side manifestations of radiation therapy at decreasing radiation loading and improve patient's life quality due to mitigating the urination.

EFFECT: higher efficiency of therapy.

1 ex

FIELD: veterinary science.

SUBSTANCE: the present innovation deals with treating tumors of external reproductive organs in dogs: in preoperational period for 3 d one should introduce 20%-hypertonic solution of ACD-2 fraction upon 0.5%-novocaine solution at 0.1-0.3 ml working solution/cu. cm tumor into tumor basis, at not more than 0.4 ml pure ACD-2/10 kg canine body weight. One should dissect the tumor within the limits of healthy tissue, close vessels and wound defect after removing the tumor due to electrocoagulation of submucous layer at scab's development.

EFFECT: higher efficiency of therapy.

1 ex

FIELD: oncological diseases.

SUBSTANCE: invention is designed for use in complex treatment of urinary bladder cancer in case of propagation of tumor to sub-epithelium connective tissue. Method comprises transurethral resection and introduction of immune preparations and furacillin solution. Once transurethral resection completed, three-way Foli catheter is transurethrally introduced to patient, through which mixture of standard furacillin solution with 10·106 ME interferon α-2β is injected from the first postoperative day and over following 3-5 postoperative days bringing summary dose of interferon to 50·106 ME.

EFFECT: prevented traumatism of urinary bladder due to single introduction of catheter and assured continuous action of drugs without development of immune complications.

FIELD: applied immunology.

SUBSTANCE: composition contains, wt parts: borax decahydrate1-25, sodium thiosulfate pentahydrate 10-5-10-4, potassium carbonate 30-150, refined sugar 30-200, and water 100-200 per 100 wt parts of sodium metasilicate pentahydrate. In addition to its capability of improving resistance to diseases, body weight increase, productivity of agricultural plants, quality of crop, and ripening term (harvest time), composition according to invention possesses nonspecific immunostimulating activity, including production of antibodies and enhancement of immunity through activation of immunocytes thereby maximally strengthening vaccination effect regarding diseases caused by malignant neoplasm viruses.

EFFECT: increased assortment of immunostimulating agents.

10 cl, 11 dwg, 12 ex

FIELD: medicine, oncology.

SUBSTANCE: method involves carrying out chemotherapy. Firstly, dose of nicotinic acid is administrated that is sufficient to initiate hyperemia of skin integuments and in appearance of first symptoms of hyperemia cytostatics are administrated by parenteral route. Method provides increasing duration of contact of cytostatics with tumor cells and micrometastasis due to inclusion of blood volumes depots in the parent state in vascular skin networks to volume of circulating blood. Invention can be used in treatment of malignant neoplasms with cutaneous and subcutaneous localization.

EFFECT: improved treatment method.

2 ex

FIELD: organic chemistry, medicine.

SUBSTANCE: invention relates to a photosensibilizing compound representing phenylthio-substituted derivative of phthalocyanine of the general formula (I): wherein R means hydrogen atom (H), t-C4H9; M means HH, AlOH, Zn. Also, invention relates to its liposomal formulation representing the composition of mixture of lipids (lecithin, cholesterol, cardiolipin) and a photosensibilizing agent, and to a method for carrying out the photodynamic therapy using this formulation. Photosensibilizing agents show high elimination rate from normal tissues, provide the deep therapeutic effect on tumor tissues, non-toxic properties and perspective for their using in oncology and other branches of medicine.

EFFECT: valuable medicinal properties of compound.

4 cl, 3 dwg, 9 ex

FIELD: medicine, oncology, pharmacy.

SUBSTANCE: invention relates to an anti-tumor agent representing benzoic acid amide (benzamide). Benzamide is used in the range of doses from 25 to 100 mg/kg.

EFFECT: expanded assortment of agents of indicated designation.

2 cl, 1 tbl

FIELD: organic chemistry, medicine, oncology, pharmacy.

SUBSTANCE: invention relates to a new pentacyclic compound derivative of taxane represented by the formula:

wherein R1 represents dimethylaminomethyl group or morpholinomethyl group; R2 represents halogen atom or alkoxy-group comprising from 1 to 6 carbon atoms, or its salt eliciting an antitumor effect, and to a medicine agent based on its. Invention provides preparing new derivatives of taxane eliciting the valuable biological effect.

EFFECT: valuable medicinal properties of compound.

13 cl, 1 dwg, 4 tbl, 16 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to new derivatives of cyclic amide of the formula (I)

or its salt, or hydrate, or solvate wherein X represents (C1-C6)-alkyl, (C1-C6)-alkyl substituted with phenyl, (C2-C6)-alkenyl substituted with phenyl or halogenphenyl, (C2-C6)-alkynyl substituted with phenyl, phenyl that can be substituted with (C1-C6)-alkyl; one or more halogen atom, nitro-group, phenyl, (C1-C6)-alkoxy-group, halogen-(C1-C6)-alkyl, halogen-(C1-C6)-alkoxy-group, phenyl-(C1-C6)-alkyl, (C1-C6)-alkoxyphenyl-(C1-C6)-alkyl, amino-group, optionally substituted with (C1-C6)-alkyl, acetyl, (C1-C6)-alkoxy-group, substituted with phenyl, phenylcarbonyl, furanyl; 1- or 2-naphthyl, monocyclic (C3-C8)-cycloalkyl, amino-group substituted with one or more substitutes taken among phenyl, halogenphenyl, (C1-C6)-alkoxyphenyl, (C1-C6)-alkyl, halogen-(C1-C6)-alkyl, phenyl-(C1-C6)-alkyl; 5- or 6-membered monocyclic heterocyclic group comprising 1 or 2 heteroatoms, such as nitrogen (N), oxygen (O), sulfur (S) atom optionally substituted with halogenphenyl, halogen atom, benzyl, (C1-C6)-alkyl, phenyl; 8-10-membered bicyclic heteroaryl group comprising 1 or 2 heteroatoms taken among N, O and optionally substituted with halogen atom; 8-10-membered polycyclic cycloalkyl group; Q means -CH2-, -CO-, -O-, -S-, -CH(OR7)- or -C(=NR8)- wherein R7 means hydrogen atom (H), (C1-C6)-alkyl; R8 means OH, (C1-C)-alkoxy-group, acylamino-group, (C1-C6)-alkoxycarbonylamino-group, phenyl-(C1-C6)-alkoxy-group; n = 0-5; B represents group or wherein each among R3, R4, R5 and R6 represents independently substitute taken among group consisting of hydrogen atom (H), halogen atom, NO2 (nitro-group), (C1-C6)-alkoxy-group, CN (cyano-group); m = 1 or 2; ring represents 5- or 6-membered aromatic heterocyclic ring comprising one or two heteroatoms taken among O, S, N. Compound of the formula (I) elicit activity inhibiting binding sigma-receptors that allows their using as component of medicinal agent.

EFFECT: valuable medicinal properties of compounds.

21 cl, 2 sch, 4 tbl, 183 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to new derivatives of indol-3-yl of the formula (I):

wherein each A and B represents independently of one another oxygen atom (O), NH, CONH, NHCO or a direct bond; X means (C1-C2)-alkylene or a direct bond; R1 means hydrogen atom (H); R2 means hydrogen atom (H); R3 means NHR6, -NR6-C(=NR6)-NHR6, -C(=NR6)-NHR6, -NR6-C(=NR9)-NHR6, -C(=NR9)-NHR6 or Het1; each R4 and R5 represents independently of one another hydrogen atom (H); R7 means -(CH2)o-Ar, Het, OR6; R6 means hydrogen atom (H); R7 means (C1-C10)-alkyl, (C3-C10)-cycloalkyl; R8 means Hal, NO2 (nitro-group), CN (cyano-group), Z, -(CH2)o-Ar, COOR1, OR1, CF3, OCF3, NHR1; R9 means CN or NO2; Z means (C1-C6)-alkyl; Ar means aryl that can represent unsubstituted, monosubstituted, or polysubstituted R8; Hal means F, Cl, Br, J; Het means saturated, partially or completely saturated monocyclic or bicyclic heterocyclic radical comprising from 5 to 10 ring members wherein 1 or 2 nitrogen atom (N) and/or 1 or two sulfur atom (S) present, and heterocyclic radical can be monosubstituted with phenyl; Het1 means saturated, partially or completely unsaturated monocyclic or bicyclic heterocyclic radical comprising from 5 to 10 ring members and from 1 to 4 nitrogen atoms (N) that can be unsubstituted or monosubstituted NHX, or oxo-group; n = 0, 1 or 2; m = 0, 1, 2, 3, 4, 5 or 6; o means 0, 1 or 2; and their physiologically acceptable salts and solvates. Compounds of the formula (I) elicit intergin-inhibitory effect that allows their using as components of pharmaceutical composition. Also, invention describes intermediate compounds.

EFFECT: valuable medicinal properties of compounds.

11 cl, 4 sch, 1 tbl, 34 ex

FIELD: organic chemistry, medicine, psychiatry, pharmacy.

SUBSTANCE: invention relates to medicinal agents used for prophylaxis and treatment of schizophrenia by inhibition or suppression of neurodegenerative disease caused by hypofunction of glutamic acid receptors. As an active component agents comprise derivative of 5-substituted 3-oxadiazolyl-1,6-naphthiridine-2(1H)-one of the formula (I):

wherein Het represents oxadiazolyl group; R1 represents hydrogen atom, lower alkyl group, lower cycloalkyl group, trifluoromethyl group, lower alkenyl group, lower alkynyl group, lower alkoxyl group, lower alkoxy-(lower)-alkyl group, lower hydroxyalkyl group, substituted or unsubstituted aryl group, substituted or unsubstituted heteroaryl group; R2 represents hydrogen atom, lower alkyl group, lower cycloalkyl group, lower cycloalkylmethyl group, lower alkenyl group, lower cycloalkenyl group, lower alkynyl group, substituted or unsubstituted aryl group and substituted or unsubstituted heteroaryl group wherein indicated groups represent phenyl or naphthyl and indicated heteroaryl groups represents furyl, thienyl or pyridyl, or their physiologically acceptable acid-additive salts.

EFFECT: valuable medicinal properties of agents.

10 cl, 1 tbl

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