Transesterification method for production of synthetic chlorophyll or bacteriochlorophyll derivatives

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

 

The SCOPE of the INVENTION

The present invention relates to new derivatives of chlorophyll and bacteriochlorophyll, to receive them, use in the methods of photodynamic treatment (PDT) in vivo for diagnosis and photodynamic destruction of viruses and microorganisms in vitro.

DEFINITIONS AND ABBREVIATIONS

hl = bacteriochlorophyll and (MD-containing 7,8,17,18-tetrahydropyran formula (a) in the following diagram And in which M stands for MD, and having vitilevu or geranylgeranyl group in position 173SOON3group in position 132the atom N in position 132hazelgrove in position 3 and italgroup in position 8).

Derived hl = BChl derivative with modifications in the macrocycle, the Central metal atom and/or at the periphery.

BChlide = bacteriochlorophyll and (free from a C-172carboxylic acid, obtained from BChl).

BPhe = bacteriopheophytin and (hl, in which the Central magnesium replaced by two atoms of H).

BPheid = bacteriopheophorbide and (free from a C-172carboxylic acid, obtained from BPhe).

Chl (b) = chlorophyll a (b) (MD-containing 17,18-dihydroporphyrin formula (b) in the following diagram And in which M MD means, and having failgroup in position 173SOON3group in position 132the atom N in position 132, the vinyl group at position 3, the double bond in positions 7-8 and either methyl in position 7 and ethyl at position 8 (Chl a), or formylpropyl in position 7 and ethyl at position 8 (Chl b)).

Chlide = chlorophyllide and (free from a C-172carboxylic acid, obtained from Chl).

DMF = dimethylformamide; ESI = ionization electrospray;

et = ethyl; gg = geranylgeranyl; GLC = glucose;

HPLC = liquid chromatography high pressure (ghvd);

FITC = fluoresceinisothiocyanate.

[M]-BChl = BChl derivative, in which the Central atom MD substituted by a metal M according to the below description.

me = methyl; MS = mass spectroscopy; NMR = nuclear magnetic resonance; NtBoc-ser=N-tert-butoxycarbonylmethyl; PDT=photodynamic treatment.

Phe = pheophytin a (Chl, in which the Central MD replaced by two atoms of H).

Pheid = pheophorbide and (free from a C-172carboxylic acid derived from Phe); WG = propyl; SDP = site-directed photodynamic treatment; ser = seril, serine;

tbb = para-tert-butylbenzyl; THF = tetrahydrofuran;

Ti(OEt)4=tetraethyl-ortho-titanate.

Throughout the description, use the nomenclature and numbering (bacterial plankton)chlorophyllose structures according to IUPAC (international Union of pure and applied chemistry) (see below Diagram). According to this nomenclature native (bacterial plankton)chlorophylls are two groups of ester carboxylic acids at positions C-132and C-172, esterified at positions C-133and C-17 3. In the nomenclature and abbreviations used in the examples appears first tarifitsiruemih residue in the C-133then the Central metal atom, if not the MD, and then tetrapyrrol (noun), followed by the rest of ester C-173. For example, the connection in the following example 1 is designated as a-173methyl ether 133-tert-butylbenzyl-PD-bacteriopheophorbide (abbreviated tbb-Pd-BPheid-me).

BACKGROUND of the INVENTION

Chlorophylls and bacteriochlorophylls, ubiquity pigments of photosynthesis has been studied intensively with the aim of understanding their photophysique and photochemistry (Scheer, 1991). Together with more affordable, but by a spectroscope less informative porphyrins they were also used for more General insights into phenomena of energy transfer and electron, the interaction of large aromatic molecules with a Central metal, and the Central metal with excess ligand.

Interest is the use of photosensitizers for photodynamic treatment (PDT) of tumors. In accordance with this method used a combination of non-toxic drug that absorbs light at a suitable wavelength, non-hazardous photosensitizing irradiation of the patient after administration of the drug.

It was found that porphyrins accumulate in the tumor tissue and the settlement of the E. its exposure, absorb light in situ, providing the opportunity to identify tumors by location fluorescence. As for discovery, and for the photodynamic treatment of tumors has been proposed the use of raw hematoporphyrin derivatives, known as HPD. Form HPD, which is considered to be more effective, includes a portion of the HPD, which have an aggregate weight in excess of 10 kDa and is the subject of U.S. patent No. 4649151. HPD or its active components have been described in U.S. patent No. 4753958 for the local treatment of skin diseases, and Matthews et al., 1988, for sterilization of biological samples containing infectious organisms such as bacteria and viruses. The mixture, known as hematoporphyrin derivatives (HPD), containing a large number of related complex ether oligomers of hematoporphyrin (HP), available commercially (Photofrin II, Quadra Logic Technologies Inc., Vancouver, BC, Canada).

To optimize the properties of porphyrin drugs in the treatment and diagnosis suggested several porphyrin derivatives, in which, for example, there is a Central metal atom in a complex with four pyrrole rings, and/or peripheral substituents of the rings pyrrole modified, and/or the macrocycle digidrive to derivatives of Chl (chlorins) or tetrahydrofurane to BChl derivatives (bacteriochlorin).

Chlorophyll derivatives and bacteriochlorophyll which have better properties compared to porphyrins, however, they are not as readily available and more difficult to handle. Was investigated the possibility of using chlorophyll derivatives (Spikes and Boomer, 1991) and derived bacteriochlorophyll (Beems et al., 1987; Dougherty, 1992; Fiedor et al., 1993; Kessel et al., 1993; Moser, 1998; Pandey et al., 1994; Tregub et al., 1993) for the diagnosis and treatment of cancer. Due to their intense absorption in a favorable spectral ranges (650-850 nm) and the light decay after treatment chlorophyll derivatives and bacteriochlorophyll are excellent sensitizers for tumors PDT.

Were investigated tetrapyrrole complexes with metals other than MD, the series of porphyrin and 17,18-dihydroporphyrin to understand their spectroscopic and redox properties (Hynninen, 1991). Bacteriochlorophylls have a potential advantage in comparison with chlorophyll, because they demonstrate the intense, near infra-red band, i.e. with a substantially greater wavelengths than chlorophyll derivatives.

Publication of PCT International application no WO 90/12573 name Dougherty describes derivatives of bacteriochlorophyll-a or-b corresponding bacteriochlorin that do not have a Central metal atom or in which the Central metal atom can represent separability metal selected from Mg2+Sn2+and Zn2+and C-172-carboxyl group etherification n is sasenum or unsaturated hydrocarbonyl the remainder of the 8-25S, to obtain a composition applicable in the method of destruction or deterioration of unwanted target biological substrates, including photosensitization specified substrate an effective amount of the specified derivative, followed by irradiation target substrate by radiation in the wavelength range absorbed by the specified derived over a period of time sufficient for the deterioration or destruction of the substrate. In addition, the use of these compounds is considered to be suitable for photodynamic therapy and diagnosis. It should be noted that despite the claimed complexes Sn2+and Zn2+bacteriochlorophyll-a or-b, the data derived metal has not been illustrated examples, as well as no way of obtaining them was not disclosed in the description of the specified patent application WO 90/12573.

Under normal shipping conditions, i.e. in the presence of oxygen at room temperature and under normal light conditions remains BChl labile and have a slightly lower quantum yield for obtaining the triplet state compared to, for example, a derivative of hematoporphyrin (HPD). However, their possible initiation of biological redox reactions, favorable spectral characteristics and their easy dissolution in vivo lead to potential superiority bacteriochlorin low over other connections, for example, porphyrins and chlorophylls, when the PDT treatment and diagnosis, as well as for destruction of cells, viruses and bacteria in the samples and living tissue. It is expected that chemical modification of bacteriochlorophylls will lead to further improvement of their properties, but it is very limited due to the lack of suitable methods for such modified bacteriological (Hynninen, 1991).

The application for the European patent published under No. 0584552, the same applicant as the applicant of the present application, describes the new conjugates of chlorophylls and bacteriochlorophylls in position C-173with amino acids, peptides and proteins that are applicable for the treatment of PDT and for diagnostics. Amino acid, peptide or protein residue linked directly or through a spacer with a C-172-carboxyl group of the molecule of chlorophyll or bacteriochlorophyll. These conjugates produced by the methods, is soft enough to hold colorability Central atom MD.

S-132-carbomethoxy chlorophylls and bacteriochlorophylls biosynthetic derived from the side chain of the C-13 propionic acid and the reactive part β-ketoamine system present in socilism the ring most of the chlorophylls. However, unlike the side-chain complex C-17 propionic ester there is no way for any x the economic, neither for the enzyme transesterification in position C-133. The only previously known reaction for this group is splitting, leading to the 132-dimethoxybenzoyl or prochlorophytes. In the application at the German patent No. DE 4121876 and PCT publication no WO 97/19081 owned by the applicant, the proposed derivatives of bacteriochlorophylls with modified residues of esters in position 133and 173. However, these applications are described only derivatives bacteriochlorophyll with native remnants methyl ester in position 133and not describes how to get the other esters in position 133.

There is a need to obtain new derivatives of chlorophyll and bacteriochlorophyll applicable for PDT, with the objective of maintaining or even improving the favorable optical and physiological properties of Chl and BChl, while optimizing their photosensitizing potential, as well as to improve chemical stability and optimizing their physiological validity.

BRIEF description of the INVENTION

In accordance with the present invention it was found that new diesters With-133/C-173chlorophylls and bacteriochlorophylls can be obtained by selective transesterification With-132-carpometacarpi chlorophyll derivatives and bacteriochlorin the illa as such or together with the side chain of the C-17 propionic acid anhydrous and anaerobic conditions in the presence of excess alcohol and using tetraethyl-o-titanate as a catalyst. This procedure is sufficiently soft to provide a modified, i.e. transesterification, cyclotouring pigments, such as native, MD-containing chlorophyll.

In the present invention discloses new derivatives of chlorophyll and bacteriochlorophyll General formula I:

in which

M is a Central metal atom or represents two atoms N;

each of R3and R5independently represents acetyl, vinyl, ethyl, 1-hydroxyethyl or a simple or a complex ester of the specified 1-hydroxyethylrutoside;

R4represents methyl or formyl;

the dashed line in positions 7-8 represents an optional double bond; and

R1and R2the same or different, selected from the group including:

(i) C1-C25hydrocarbonrich, which can be straight or branched, saturated or unsaturated, optionally substituted by one or more radicals selected from halogen, oxo (=O), HE, Cho, COOH or NH2or interrupted by one or more heteroatoms selected from O, S and NH, or carbocyclic or heterocyclic residues;

(ii) the balance of amino acids, oligopeptides or polypeptide containing the hydroxy-group or their derivative, selected from the group comprising esters and N-saxeseni is derived, where specified gidrauxilirovannaya amino acid or its derivative is associated with soo-the remainder derived Chl or BChl through the said hydroxy-group;

(iii) a residue of a peptide described in (ii)associated with soo-the remainder through C1-C25hydrocarbon described in (i)where the specified C1-C25saturated or unsaturated hydrocarbonyl residue, optionally substituted by one or more radicals selected from halogen, oxo (=O), HE, Cho, COOH or NH2or this balance is interrupted by one or more heteroatoms selected from O, S and NH, or phenyl ring is further substituted terminal functional group selected from HE, COOH or NH2;

(iv) the balance of cell - or tissue-specific ligand selected from oligopeptides and protein directly associated with soo-the remainder or via C1-C25hydrocarbon described in (i), wherein said C1-C25saturated or unsaturated hydrocarbonyl residue, optionally substituted by one or more radicals selected from halogen, oxo (=O), HE, Cho, COOH or NH2or this balance is interrupted by one or more heteroatoms selected from O, S and NH, or phenyl ring is further substituted terminal functional group selected from HE, COOH or NH2; and

(v) the remainder of the mono-ol is St - or polysaccharide or polyoxyethylene, directly associated with soo-the remainder or via C1-C25hydrocarbon described in (i)where the specified C1-C25saturated or unsaturated hydrocarbonyl residue, optionally substituted by one or more radicals selected from halogen, oxo (=O), HE, Cho, COOH or NH2or this balance is interrupted by one or more heteroatoms selected from O, S and NH, or phenyl ring is further substituted terminal functional group selected from HE, COOH or NH2.

The compounds of formula I in which R3is vinyl, R4is methyl or formyl, R5is ethyl, and the dotted line in positions 7-8 represents a double bond, are derivatives of chlorophyll a and b, respectively. The compounds of formula I in which R3is acetyl, R4is methyl, R5is ethyl, and the provisions 7-8 gidrirovanny, are derived bacteriochlorophyll A.

The Central metal atom M in the compound of formula I may be missing, may be native atom MD natural pigments chlorophyll and bacteriochlorophyll, or it can be a divalent metal selected from the group comprising Pd, Co, Ni, Cu, Zn, Hg, Er, It, S, Sn and MP.

The present invention relates to a new method of transesterification to obtain synthetic grass is the x derivative of chlorophyll and bacteriochlorophyll the above General formula I, includes the following stages:

(a) interaction in the anaerobic conditions of a respective derivative (bacterial plankton)chlorophyll, metal(bacterial plankton)chlorophyll or (bacterial plankton)pheophytin having a position-132a group of SOON3and in position C-172group COOR2with alcohol R1OH, where R1and R2have the above values, provided that R1is not methyl, in the presence of tetraethyl-ortho-titanate, and the interactions are carried out either in a solvent such as free from peroxide tetrahydrofuran (THF) or dimethylformamide (DMF); in this case preferably receive a W C-132-COOR1C-172-OR2or alcohol R1OH, used in large excess, serves as a solvent; in this case, get the W C-132-COOR1C-l72-COOR1; and

(b) separating the desired product from the reaction mixture.

Methods in accordance with this invention can be implemented in combination with other known methods of modification of the molecule, for example, conjugation in position C-173described in EP 0584552, modifications on the periphery of the molecule and/or Parametrierung, for example, described in WO 97/19081. Preferably demeterova or the replacement of the Central metal atom is carried out before the transesterification.

the new compounds of chlorophyll and bacteriochlorophyll, obtained by the process of transesterification of the present invention, applicable as photosensitizers as therapeutic and diagnostic agents, for example, cancer and age-related macular degeneration, destruction of cells, viruses and bacteria in the samples and living tissues, also known in the field of application of PDT and other photosensitizers.

BRIEF DESCRIPTION of DRAWINGS

Figa-1D show toxicity in the dark (black squares) and phototoxicity (white squares) with respect to melanoma cells M2R after incubation with the use tbb-Pd-BPheid-tbb (1A), tbb-Pd-BPheid-me (1B), Pd-BPheid-et (1C), control (1D). Sensitizers are added to liposomes. Cell viability was determined by introducing the DNA of [3H]-thymidine.

Figure 2 shows the toxicity in the dark (--+--) and phototoxicity Pd-BPheid-Nglc (black squares) and Pd-BPheid-ser light (black triangles), dark (white triangles) melanoma cells M2R in mice. Cell viability was determined by entering [3H] -thymidine.

DETAILED description of the INVENTION

The present invention relates to a new method of transesterification to obtain new derivatives of s-132-COOR1C-172-OOR2and C-132-COOR1C-172-COOR1compounds of chlorophyll and bacteriochlorophyll.

In accordance with one possible implementation of this and is gaining R 1and R2identical; in accordance with another way they are different.

In accordance with one embodiments of the present invention R1and R2can imagine hydrocarbonyl radical. In this description, the term "hydrocarbon" means any linear or branched, saturated or unsaturated, including aromatic, gidrolabilna radicals, preferably having 1-25 carbon atoms, such as alkyl, preferably having 1-4 carbon atoms, e.g. methyl, ethyl, propyl, butyl or alkenyl, quinil, cycloalkyl, aryl, such as phenyl, or kalkilya group, such as benzyl or substituted benzyl, for example, tert-butylbenzyl. If R1and R2various, R1preferably represents methyl, radical, present in natural compounds Chl and BChl, a R2preferably represents ethyl or a radical derived from natural compounds Chl and BChl, for example, geranylgeranyl(2,6-dimethyl-2,6-octadienal) or fail(2,6,10,14-tetramethylhexadecane-14-ene-16-yl). If R1and R2various, R1and R2can also represent a hydrocarbon chain substituted by one or more radicals selected from halogen, such as F, Br, Cl and I, or HE, oxo (=O), Cho, COOH or NH2or this optional substituted gidrolabilna chain, preran the I O S or NH, preferably O, for example, R1or R2is the rest of oligoastrocytoma having from 4 to 10 carbon atoms, preferably pentoxifyllin, or carbocyclic, for example, phenyl, or heterocyclic, for example, pyridyl, remains.

In accordance with another variant of R1and R2you can imagine the rest of the amino acid or peptide (oligo - or polypeptide)that contains the hydroxy-group, such as serine, threonine and tyrosine, or containing peptides, or a derivative of the specified amino acid or peptide, selected from esters, for example esters of alkyl and N-protected derivatives in which the N-protective group is, for example, tert-Butylochka, carbobenzoxy or trityl, as specified gidrauxilirovannaya amino acid or peptide, or a derivative associated with a group COO-derived Chl or BChl through its hydroxy-group. Examples of such derivatives of amino acids are serine methyl ester, N-tert-butyloxycarbonyl, methyl ester N-Fritillaria, methyl ester of tyrosine and methyl ester of N-tert-butoxysilane, and an example of such a peptide is the methyl ester of N-carbobenzoxy, all of the above compounds obtained in accordance with the description in EP 0584552. According to a preferred variant about svodnoe Chl or BChl atrificial L-serine or N-tert-butyloxycarbonyl.

In accordance with a further variant of R1and R2you can imagine the rest of the peptide (oligo - or polypeptide)that is associated with Chl or BChl through radical C1-C25hydrocarbide as described above; in this case, the radical hydrocarbide serves as a spacer for the specified peptide or polypeptide/protein and has a terminal functional group selected from HE, COOH and NH2through which the peptide or protein is bound ester or amide bond.

In accordance with the following option R1and R2you can imagine the rest of the cell - or tissue-specific ligand selected from peptides and proteins, examples of which include, but are not limited to, peptide hormones, for example, melanocytestimulating hormones (melanotropin), and antibodies, such as immunoglobulins and tumor-specific antibodies. In this case, the peptide or protein can also be connected with Chl or BChl through radical C1-C25hydrocarbide, as indicated above, while the radical hydrocarbide serves as a spacer for the specified peptide or polypeptide/protein and has a terminal functional group selected from HE, COOH and NH2through which the peptide or protein is bound ester or amide bond.

In accordance with another variant of R1and R2can depict ablate the residue is mono-, oligo - or polysaccharide, directly associated with soo-molecules of Chl or BChl or through radical C1-C25hydrocarbide, as described above. In accordance with the preferred monosaccharide is glucosamine.

To obtain the esters in accordance with this invention the transesterification only in position C-133preferably realized by the interaction of the C-173-133derived diapir Chl or BChl with the native carpometacarpus in position C-133the desired alcohol R1OH, in which R1is not methyl, in the presence of tetraethyl-ortho-titanate, when this reaction is carried out in an aprotic solvent such as free from peroxide tetrahydrofuran (THF) or dimethylformamide (DMF). Some esters get this way in the following description with ethanol, tert-butylbenzyl alcohol, propanol, tert-butyloxycarbonyl and serine.

In accordance with another variant of the transesterification as in position C-133and in position C-173carried out simultaneously with alcohol R1OH. After synthesis follows the above procedure, however, as the solvent used alcohol (not Applicable for NtBoc-ser, which are solid substance). In this way, in accordance snisarivgeny description get some esters, including esters tbb, Pr, NtBoc-Ser and ser. The reaction time for para-tert-butylbenzyl alcohol and n-propanol is 48 and 12 hours, respectively.

Type of alcohol and the temperature will determine whether there will be substantial stronger in position C-133or as in position C-173and-133. Large alcohols R1OH preferably atrificial position C-133, while a small alcohols atrificial as the position of the C-173and the position-133.

The preferred solvent in accordance with this invention is THF. DMF is used if the alcohol insoluble in THF. The temperature of the reaction mixture may be 75°in a few days, for example, using 1-propanol, para-tert-butylbenzyl alcohol and N-tBoc-serine.

The separation of the products from the reaction mixture is performed by conventional means, for example, by adding diethyl ether and water before phase separation, triple extraction of the aqueous phase by simple ether, drying of the combined organic phases with the use of NaCl, evaporation of the solvent in vacuum, remove excess alcohol in high vacuum (<10-3RA) and highlight the desired transesterification derived Chl or BChl as a result of application jhud or column chromatography.

Transesterification clonality Chl and BChl in accordance with this invention can be then treated with pyridine at elevated temperature with the purpose of removal of C-13 2-carbomethoxyamino and get periprostatic formulas IV and V below in Scheme C. the Pigments of the formula IV can then be transetherification, tolerowany or lidirovali in position 173.

Derivatives of Chl and BChl, obtained by both the methods themselves can be used as sensitizers in accordance with this invention, or they can serve as a bridge/spacer to link other suitable molecules with macrocycle Chl/BChl.

Upon receipt of ester when the desired peptide or protein is attached to one of the provisions, devoid of hydroxyl-containing amino acid residue, the macrocycle Chl or BChl may be initially connected to the serine or any other hydroxyl-containing residue or its derivatives by transesterification of native compounds or esterification of the corresponding free acids (Chlide or BChlide), and then the peptide or protein is associated with the macrocycle through the same amino acid residue.

To obtain the substituted metal derivatives of Chl and BChl native Central atom MD replace the desired metal M before conjugation pigment with an amino acid or a cell-specific ligand. Substitution of the Central atom MD chlorophyll and its derivatives on Pd, It, si, Ni, Zn, V, Co, Sn, Hg and other divalent metals carried out by conventional means, for example,by treatment of the corresponding pheophytin salt of the desired metal, for example, acetate Zn or si, in absolute ethanol at room temperature (Hambright, 1975; Hynninen, 1991). In the case of bacteriochlorophyll and its derivatives Central atom MD may be substituted Zn, cu or Pd in the similar procedure includes treatment with acetate Zn, cu or Pd in argon at elevated temperatures, as described in WO 97/19081.

If R1is substituted hydrocarbon, it may contain terminal functional group through which it can be attached to other desirable residues, for example, ester group obtained by interaction or the terminal carboxyl group R1with a carboxyl group of another compound, such as an amino acid or saccharide, or the terminal hydroxyl group of R1with a carboxyl group of another of the compounds; amide group is produced by interaction of the terminal carboxyl groups of R1with the amino group of another compound, such as an amino acid or the terminal amino group, R1with a carboxyl group of another compound, such as an amino acid.

New esters, obtained by the process of transesterification of the present invention, have the same characteristics of the optical absorption and photophysical characteristics as the corresponding Chl and BChl. Therefore, it is expected that after implementation in the treated fabric new words is by esters of Chl and BChl will become effective photodynamic agents. Thus, they can be used as photosensitizers as therapeutic and diagnostic agents for the destruction of cells, viruses and bacteria in the samples and living tissues, is well known in the field of HPD and other photosensitizers. Connection information is applicable, for example, sensitizing neoplastic cells or other abnormal tissue to destruction by irradiation in vivo or ex vivo using light with an appropriate wavelength. I believe that the photoactivation energy is transferred to the endogenous oxygen into singlet oxygen, which is thought to be responsible for the cytotoxic effect. In addition, photo form (bacterial plankton)chlorophylls flyuorestsiruyut that can help in the localization of tumors or other places, which is administered (bacterial plankton)chlorophylls.

Examples known in the field of indications for which can be assigned to new derivatives of (bacterial plankton)chlorophylls, obtained by the process of transesterification of the present invention include the destruction of tumor tissue in solid tumors, the dissolution of plaques in blood vessels (see, for example, U.S. patent No. 4512762); treatment of local conditions such as acne, athlete's foot, warts, papilloma and psoriasis, as well as the processing of biological products (such as blood) to destroy infecti is the R agents.

Derivatives (bacterial plankton)chlorophyll, obtained by the process of transesterification of the present invention, prepared in the form of finished pharmaceutical compositions, administered to the patient or used for in vitro using methods well known in this field, for example, described in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton Penna., the latest edition. The composition can be introduced systemically, in particular by injection, or can be a local application.

For the diagnosis of derivatives (bacterial plankton)chlorophyll can be used as such or may be labeled with a radioisotope or other means of detection known in this field.

According to practice, the number of input derivative (bacterial plankton)chlorophyll corresponds to other porphyrins used for PDT, and varies depending on the choice of a derivative used as an active ingredient, being treated condition, method of administration, the age and condition of the patient and doctor.

The wavelength of the irradiating light is preferably chosen so that it would correspond to the maximum absorption of the photosensitizer (bacterial plankton)chlorophyll. A suitable wavelength for any connection can be easily identified by its absorption spectrum.

In addition to the use of in vivo-derived bacterial plankton) chlorophyll, obtained by the process of transesterification of the present invention can be used for processing materials in vitro to destroy harmful viruses or infectious agents such as dangerous bacteria. For example, blood and blood plasma intended for transfusion, can be processed by the connection in accordance with this invention and exposed to sterilization.

Conjugation of proteins, such as hormones, growth factors or their derivatives and antibodies and nutrients to cells, for example, tyrosine, with the remnants of Chl and BChl, as implied, helps them better retention in tumors and treated areas. The increased shift in the red area provides deeper penetration, while maintaining ubiquitously natural system. Replacement MD other metals, as implied, to optimise their own and metabolic stability of the balance of Chl or BChl and intersystem crossing to an excited triplet state, and opens the possibility for new diagnostic procedures.

Tumor-specific antibodies preferentially target the remains of Chl and BChl in the tumor or on the area treated, while the hormones and nutrients to cells can also be captured by normal nontransgenic counterparts is. However, cells that are selected as targets for hormones and nutrients to cells, such as melanocytes, scattered among the other cells in normal conditions and during transformation into malignant cells together, forming a solid tumor. The result is that the concentration of the photosensitizer in malignant tissue will rise sharply relative to its concentration in normal tissue, where cells are more dispersed, providing amplification steps of PDT on tumor site. This allows you to effectively use low doses, below the damage threshold of normal tissue, thus reducing the need to use clear directional space radiation. In addition, having a very strong fluorescence, site-directed Chl or BChl can be used for fluorescent labeling of tumor plot (plots) or other targets.

New photosensitizers Chl and BChl, obtained by the process of transesterification of the present invention are suitable for treatment of tumors, melanomas for several reasons: (a) early (non-metastatic) malignant melanoma and other skin tumors readily available for PDT; (b) photodynamic treatment using green light, as well as conventional chemotherapy and radiotherapy still has not achieved success in the treatment of malignant melanoma; (C) however, the creature is no several melanoma-specific ligands, which can be targeted photosensitizing balance at the tumor site, and (d) it is expected that the use of induced long waves residues Chl and BChl to eliminate the deficiencies commonly used photosensitizers, which are due to the absorption of melanin.

Tumor-melanoma arise carcinogenic transformation (including UV-induced mutagenesis) melanocytes. Normal melanocytes include a small amount of normal cell population of human skin and are usually located in the basal cell layer between epidermis and dermis, where each of them surrounded by 30-40 keratinocytes and one cell of Langerhans. Treatment of tumors, melanomas especially difficult when using PDT as tumor cells of melanoma may contain black insoluble eumelanin (poly-5,6-indolinone)having a wide absorption band around 540 nm and therefore competing with any photosensitizer for radiation at wavelengths shorter than 650 nm. In addition, molecules of melanin can absorb the formed oxygen radicals, thereby preventing intoxication organelles of viable cells. Therefore, PDT melanholicheskij melanomas commonly used HPD is not very encouraging. However, having a maximum optical absorption at the wavelength of 650 nm, the excited Chl and BChl and their synthetic is such derivatives should not Salomatina melanin. Moreover, tumor cells of melanoma (i.e. transformed melanocytes) consume a significant amount of tyrosine during the synthesis of melanin, exhibit high affinity towards melanotropin (α-, β- and γ-melanocytestimulating hormones (MSH) of the pituitary gland) and to several well-known antibodies. Therefore, they can serve as a good target for tyrosine, melanocortin - or antibody-conjugates Chl and BChl, provided that conjugation does not greatly affect the recognition of ligands for cellular receptors. Since the concentration of melanocytes increases almost 40 times in the areas of melanoma (relative to normal skin tissue), the expected sharp increase in photodynamic action.

New derivatives of Chl and BChl, obtained by the process of transesterification of the present invention, can be used as active agents in pharmaceutical compositions for photodynamic treatment of some types of cancer, including brain cancer, ovarian cancer, breast cancer, tumors, skin cancer, lung, esophagus and bladder, as well as other hormonetestosterone tumors.

For example, the compounds may be used in photodynamic treatment of malignant melanomas. Photodynamic action of compounds observed in melanoma cells in tumor and cell cultures. Examples of derivatives which may be used for this purpose, include conjugates derived Chl and BChl C α-melanotropin associated with the remainder of the pigment or through their residues serine, tyrosine or lysine, or through a terminal amino group.

The pharmaceutical compositions can be administered to the patient in the usual methods used in PDT. The number of input connections and route of administration are determined according to the kind of tumor, stage of disease, age and health condition of the patient, however, this number is much lower than the currently used doses of Photofrin II, which constitutes about 20-40 mg HPD/kg of body weight. The preferred method of administration is intravenous route or direct injection into a solid tumor of an aqueous solution of the active compounds include conventional pharmaceutically acceptable carriers and additives, as well as local treatment of skin tumors suitable compositions for local application.

Thus, derivatives of Chl and BChl obtained by the method of this invention can be used for multiple purposes, such as photo-destruction of benign or malignant cells or tissue site-targeted photodynamic therapy (SDP). Conjugate gives the molecule Chl or BChl to the cells, grouped in tumor tissues after transformation, but well separated from each other in normal tissues (in the example, the melanocytes to melanoma). In the photodynamic action of the photosensitizer in the tumor can be several orders of magnitude higher than its action in normal tissue. Therefore, it is expected that the threshold destructive tumor lighting will be reduced to a level that is not disruptive to normal tissue. In such conditions phototoxic action will be restricted to the tumor site, even if non-specific irradiation. This application is especially important when you have tumors that are inaccessible to conventional surgery.

Another way to extend the range of sensitization to the level of near-IR, is photodynamic treatment with the use of a biphoton ways (Leupold and Freyer, 1992). High endogenous level crossing derivatives of Chl and BChl and wavelength for maximum absorption make them very good candidates for this type of PDT.

The conjugates obtained by the method of transesterification of the present invention are also applicable for photodegradation of normal or malignant cells, animals, and microorganisms in culture with or without the use of SDP, providing selective photodestruction of some types of cells in culture or infectious agents; to target the rest of the porphyrin on the selected cells by attaching, for example, conjugate Chl or BChl-serine to specifications the economic polypeptides, such as a hormone or other ligand receptors, cell - or tissue-specific antibodies or other ligands such as lectins; for fluorescent labeling molecules for analytical purposes in the laboratory, diagnostic and industrial applications; as well as for fluorescent labeling of animal cells or microorganisms, or particles for laboratory, diagnostic or industrial purposes. Due to its high attenuation coefficient and high fluorescence, they can replace some of the currently used fluorescent labels, such as fluoresceinisothiocyanate (FITC) or phycoerythrin.

For diagnostic purposes derivatives of Chl and BChl obtained by the method of this invention, can be labeled with radioactive isotopes with conventional methods, for example,67Ga111In201Tl,99MTS, after which the radioactive diagnostic agent is administered to a patient, preferably by intravenous injection. After a few hours the location of the cancer can be visualized using conventional methods.

The expected positive results of PDT using site-directed sensitizers, such as those obtained by the process of transesterification of the present invention, including a dramatic reduction in side effects and doses of the sensitizers. Not what the quiet of the particular advantages of the use of derivatives of Chl and BChl, obtained by the method of this invention for PDT are as follows:

1. Previously unavailable functional group Chl and BChl, i.e. s-133ester group, becomes available as such or in combination with C-173ester group for the transesterification. The obtained pigments retain their positive absorption and other optical properties, and properties of the excited state, resulting in better regulation of hydrophilic, hydrophobic balance and/or target.

2. These compounds exhibit a maximum optical absorbance at a wavelength at which optical absorption/attenuation human tissues/animal is significantly reduced (660-830 nm in Monomeric form and 1,000 nm dimers or higher aggregates). Together with the reduction in light scattering that provides deeper penetration or the use of less powerful and expensive light sources.

3. Their attenuation coefficient in the visible and near-IR spectrum is approximately ten times higher than the attenuation coefficient of porphyrins, currently used for PDT.

4. This procedure is gentle enough to preserve the native atom MD. However, the substitution of the Central atom MD other metals are possible and are able to increase the yield of singlet oxygen due to the higher level of the education triplet state of the photosensitizer, and also able to significantly stabilize the connection.

5. Sensitizers Chl and BChl, obtained by the process of transesterification of the present invention must have a high specificity for recognition of target cells and, consequently, cell necrosis can be achieved at lower doses. In addition, they show good photochemical properties compared with many of the fluorophore used in the present time, and, therefore, can be used for other purposes.

6. There are several works describing the high level of excretion of some derivatives of Chl from the body (Spikes and Boomer, 1991).

7. Typically, the irradiation with PDT perform laser sources such as laser, dye Ah-pumped discontinuous field that is configured for the emission at 630 nm, or vapor laser gold (pulsating), emitting at 628 nm. The high cost of such equipment limits the use of PDT in large medical centers. The use of red or near-IR absorbing photosensitizers received in accordance with the method of this invention makes it possible to use cheaper and more convenient means, such as a xenon flash lamp, a halogen lamp, diode lasers or direct solar radiation.

8. Derived Chl and hl labeled with radioactive or asset is diversified isotopes, can be used both for diagnostic and for therapeutic purposes.

Below, the invention is illustrated in the following non-limiting examples.

EXAMPLES

General methods

(a) Complex diesters, preferably modified at C-132the group of carboxylic acids (bacterial plankton)chlorophyll can be obtained in the following way:

Derived (bacterial plankton)chlorophyll (3 mg, 4 mmol) dissolved in 15 ml dry and free from peroxide tetrahydrofuran (THF) (or in dimethylformamide (DMF) using insoluble in THF alcohols). To the reaction solution was added to 500-fold amount of ethanol and 1 ál (4 μm) tetraethyl-ortho-titanate. The mixture was kept at 75°for 2, 8 or 14 days using 1-propanol, para-tert-butylbenzyl alcohol or N-tBoc-serine, respectively. The reaction mixture is usually definitively process as follows: (i) by adding diethyl ether and water before phase separation; (ii) three-time extraction of the aqueous phase by simple ether; (iii) drying the combined organic phases with the use of NaCl; (iv) evaporation of the solvent under vacuum; and (v) removing excess alcohol in high vacuum (<10-3Pa).

(b) Transesterification With-133and C-173can be carried out simultaneously in accordance with the above-described way is with the difference, as a solvent used alcohol. Reacting para-tert-butylbenzyl alcohol and n-propanol is 48 and 12 hours, respectively.

The above-described method (a) and (b) have obtained several esters using, for example, methanol, ethanol, propanol, tert-butylbenzylamine alcohol, tert-butoxycarbonylamino and serine. Examples of such esters of formulas I, II and III in accordance with the scheme below in table 1. These derivatives can be used by themselves when applying this invention, or they can serve as a bridge/spacer for connecting other suitable molecules with macrocycles Chl and BChl.

(C) Processing transesterification esters of formulas I, II and III, obtained in accordance with the above description, pyridine at high temperatures results in periprostatic formulas IV and V in the diagram, examples of which are shown below in table 2.

Example 1. 133-tert-butylbenzyl-PD-bacteriopheophorbide-a-173methyl ether (tbb-Pd-BPheid-me) (R1-tbb; R2-CH3; M-Pd)

According to the above General procedure (a) 3 mg Pd-BPhe-me subjected to interaction with 250 μl of para-tert-butylbenzyl alcohol (tbb). After 10 days, the main product tbb-Pd-BPheid-me allot with the yield amounting to 65%, after chromatography on silica the acetone/toluene 5:95 (about./vol.).

Fluids a-173-tert-butylbenzylamine ether 133-tert-butylbenzyl-Pd-bacteriopheophorbide (tbb-Pd-BPheid-tbb) was isolated as a by-product.

Analytical data for tbb-Pd-BPheid-me: tr=16,3 min in the system Ehud N: rf=0,27 on silica with acetone/toluene =5:95 (about./vol.).

UV/Vis: λmax[nm] (Arel, task) = 332 (0,67) By, 385 (0,53)x, 530 (0,19) Qx, 755 (1) Qy.

1H-NMR: δ [parts per million] (multipletness, task) = to 9.57 (s, 5-H), 8,67 (s, 10-H), 8,63 (s, 20-H), of 7.48 and 7,28 (2 d,3JA'b'=3JAB=4 Hz, o - and m-H tbbat s-133), of 6.52 (s, 132(H)of 5.53 (s, 133CH2), 4,37, 4,14, 4,11, 3,97 (4m, 7-H, 8-H, 17-h, 18-H), 3,50 (173SOON3), 3,06 (31The PINES3), to 3.41 (s, 12 CH3), 3,39 (s, 2-CH3), 2,60 for 2.01 (m, 171,2-CH2), and 1.63 (m, 7-CH3), and 0.98 (t,3JAB=7 Hz, 81-CH3), of 1.18 (s, tbb-CH3when 133).

MS (FAB): M+=860,0 (calculated 860,4 for12C

1
46
H
14
50
N
16
4
O
106
6
Pd), 669,3 (35%, M+- COO-tbb); to 713.3 (18%, M+- tbb); 877 (30%, the addition of O and N); 819 (17%, adding O and N plus the loss of NA(CH3)3).

Example 2. 133-tert-butylbenzyl-Pd-bacteriopheophytin-a-173-geranylgeranylated ether (tbb-Pd-BPhe-gg)

3 mg of Pd-BPhe-gg transesterification 250 μl of para-tert-butylanisole alcohol (tbb). After 14 days at 75°tbb-Pd-BPhe-gg highlight exit, constituting 63%, after chromatography on silica with acetone/toluene =5:95 (about./vol.).

Analytical data for tbb-Pd-BPhe-gg: tr=16,3 min ghvd (silica, gradient of 3-10% And In, with A = toluene. In: toluene/methanol/n-propanol = 100:4:0.5 to (about./about./vol.), flow rate = 1 ml/min); rf=0,63 on silica with acetone/toluene =5:95 (about./vol.).

UV/Vis: λmax[nm] (Arel, task) = 332 (0,51 By), 384 (0,42, Bx), 528 (0.45 and Qx), 756 (1, Qy).

1H-NMR: δ [parts per million] (multipletness, task) = 9,58 (s, 5-H), 8,67 (s, 10-H), 7,49 and 7,24 (2m, o - and m-H 133-tbb), 6,53 (s, 132-N, 5,54 (s, 133-CH2), 4,37, 4,14, 4,11, 3,97 (4m, 7-H, 8-H, 17-h, 18-H), is 3.08 (s, 3 PINES3), to 3.41 (s, 12 CH3), 3,39 (s, 2-CH3), 3,50 (s, 173SOON3), br4.61 (m, gg-OCH2), 5,15 (m, gg-OCH2-), and 1.63 (s, gg-CH3), 2,59-1,98 (4m, 17-CH2), of 1.18 (s, 133-tgg-C(CH3)3), 1.69 in (d3JAB=7 Hz, 18-CH3) and 0.98 (t, 3JAB=7 Hz, 81-CH3-).

MS (FAB) M+=1118,6 (calculated 1118,6 for12C

1
65
H
14
80
N
16
4
O
106
6
Pd), 927 (<5%) (M+- COO-tbb).

Example 3. 133-propyl-Pd-bacteriopheophytin-a-173-geranylgeranylated ether (pr-Pd-BPhe-gg)

6 mg of Pd-BPhe-gg transetherification 1 ml of 1-propanol (rgon) in 20 ml of THF. After 14 days at 75°pr-Pd-BPhe-gg highlight exit, constituting 60%, after chromatography on silica with acetone/toluene = 5:95 (about./vol.).

Analytical data for tbb-Pd-BPhe-gg: rf=0.44 in the system of the silica with acetone/toluene = 5:95 (about./vol.).

UV/Vis: λmax[nm] (Arel, task) = 332 (0,50, By), 384 (0,44, Inx), 528 (0,47, Qx), 756 (1, Qy).

1H-NMR: δ [parts per million] (multipletness, task) = 9,59 (s, 5-H), 8,66 (s, 10-H), 6,53 (s, 132-H), 4,38, 3,90, 3,89, 3,97 (4m, 7-H, 8-H, 17-h, 18-H), is 3.08 (31The PINES3), 3,42 (s, 12 CH3), 3,39, (s, 2-CH3 ), 4,71 (m, gg-och2when 173), 5,46 (m, gg-OCH2-when 173), of 1.62 (s, gg-CH3when 173), 2,59-1,98 (4 m, 171,2-CH2), 1,74-of 1.56 (m, propyl-when 133), or 0.62 to 0.98 (m, propyl-CH3when 133).

MS (FAB) M+=1014,4 (calculated 1014,4 for C57H72N4O

106
6
Pd), 927 (<3%) (M+- COO-pr).

Example 4. 133-tert-butyloxycarbonyl-PD-heid-a-173methyl ether (N-tBoc-ser-Pd-BPheid-me)

50 mg of tert-butyloxycarbonyl (N-tBoc-ser) are added to the pigment to a solution of Pd-BPheid-me in DMF. After 14 days at 75°N-tBoc-ser-Pd-BPheid-me allot with the yield amounting to 7%, the distribution between water and ethyl acetate and chromatography on silica with acetone/toluene =5:95 (about./vol.).

Analytical data for N-tBoc-ser-Pd-BPheid-me: rf=0.03 in the system of the silica with acetone/toluene = 10:90 (vol./vol.).

UV/Vis: (l3) λmax[nm] (Arel, task) = 332 (0,45, By), 387 (0,34, Inx), 537 (0,16), 764 (1, Qy).

MS (ESI) M+=901,2 (calculated = 901,4 for12C

1
43
H 14
49
N
16
5
O
106
10
Pd), 845,4 (40%Appendix H and the loss of C(CH3)3); 801/5 (10%Appendix H plus loss NtBoc); 669 (11%, loss NtBoc-ser).

Example 5. 133-O-seryl-PD-bacteriopheophorbide-a-173methyl ether (O-ser-Pd-BPheid-me)

The protective group of the compound of example 4 otscheplaut by adding 2 ml triperoxonane acid to obtain dry N-tBoc-ser-Pd-BPheid-me. Triperoxonane acid is removed within 15 minutes by a stream of argon and the residue is gently shaken out three times with ethyl acetate and water, receiving ser-Pd-BPheid-me (<5%) of Pd-BPheid-me. The pigment is purified on silica with acetone/toluene = 40:60 (vol./about.) (further purification to interact with in order to obtain ser-Pd-Bpheid-me can not hold).

Analytical data for ser-Pd-BPheid-me: rf=0,65 C18silica reversed-phase with methanol/toluene =5:95 (about./vol.).

UV/Vis: (l3) λmax[nm] (Arel, task) = 334 (0,36, By), 387 (0,29, Bx), 534 (0,09, Qx), 765 (1, Qy).

MS (ESI) M+=801,2 (calculated 801,3 for12C

1
38
H
14
41
N
16
5
O
106
8
Pd); 698,3 (10%Appendix H and the loss of serine).

Example 6. 133-methyl-Pd-bacteriopheophorbide-a-173-n-propyl ether (me-Pd-BPheid-pr)

Applying a 7% n-propanol in THF, during the synthesis of pr-Pd-BPheid-pr allocate a by-product pr-Pd-BPheid-me with the yield amounting to 5%, after chromatography on silica with acetone/toluene = 5:95 (about./vol.).

Analytical data for me-Pd-BPheid-pr: rf=0,42 on silica with acetone/toluene = 10:90 (vol./vol.).

UV/Vis: (DE) λmax[nm] (Arel, task) = 332 (0,50, By), 385 (0,38, Bx), 528 (0,15, Qx), 755 (1, Qy).

1H-NMR: δ [parts per million] = to 9.57 (s, 5-H), of 8.95 (s, 10-H), 6,50 (s, 132-H), 4,32, 4,24, 3,88 (3 m, 7-H, 8-H, 17-h, 18-H), a 3.87 (m, propyl-och2when 133), of 3.07 (s, 31The PINES3), 3,43 (s, 12 CH3), 3,38, (s, 2-CH3), 2,62-of 2.09 (m, 171,2-CH2), 1,72-of 1.56 (m, propyl-och2CH2when 173-CH2), by 1.68 (m, 7-CH3), or 0.63 to 0.98 (m, propyl-SN 3when 173).

MS (ESI): M+=756,6 (calculated 756,3 for12C

1
38
H
14
42
N
16
4
O
106
6
Pd); 697,5 (27%, M+SOON3).

Example 7. 133-n-propyl-bacteriochlorophyll-173-n-propyl ether (pr-BChlide-pr) (Central MD metal instead of Pd)

According to the above General method (b) and using as a starting material hl with a 100-fold excess of n-propanol, 3 days to receive the product pr-BChlide-pr with access constituting 40%, after chromatography on C18silica reversed-phase gradient components 25-10% (phase A: HEPES/KOH (20 mm; pH 7.5), phase: acetone).

Analytical data for pr-BChlide-pr: rf=0,73 C18silica reversed-phase with HEPES/KOH (20 mM, pH 7.5)/acetone =15:85 (about./vol.).

UV/Vis: (DE) λmax[nm] (Arel, task) = 357 (0,78, By), 392 (0,52, Bx), 574 (0,23, Qx), 771 (1, Qy).

1H-NMR: δ [parts per million] (multipletness, task) = 9,51 (s, 5-H), 8,65 (s, 10-H), 8,54 (s, 20-H), to 6.57 (s, 132-H), 4,35 (m, 7-H, 8-H, 17-h, 18-H), 3,99 (propyl-och2the 173and C-133), 3,11 (31The PINES3), 3,57 (s, 12 CH3), of 3.46 (s, 2-CH3), 2,62-of 2.09 (m, 171,2-CH2), and 1.63 (m, 7-CH3), 0,81 (t3JAB=7 Hz, 81-CH3)

MS (ESI): M+=702,4 (calculated 702,4 for12C

1
40
H
14
46
N
16
4
O
24
6
Mg); 616,4 (Appendix H and loss SOOS3H7).

Example 8. 133-tert-butylbenzyl-PD-bacteriopheophorbide-a-173-tert-butylbenzylamine ether (tbb-Pd-BPheid-tbb)

In the result of the interaction of Pd-BPheid-me in para-tert-butylbenzyl alcohol within 48 hours receive tbb-Pd-BPheid-tbb (50%) after chromatography on silica with acetone/toluene =5:95 (about./vol.).

Analytical data for tbb-Pd-BPheid-tbb: tr=10,8 min ghvd (silica, gradient 2-10% And In, etc is this A = toluene, In: toluene/methanol/n-propanol = 100:4:0.5 to (about./about./vol.); rf=0,50 on silica with acetone/toluene = 5:95 (about./vol.), flow rate = 1 ml/min).

UV/Vis: (DE) λmax[nm] (Arel, task) = 332 (0,49, By), 385 (0,36, Inx), 528 (0,15, Qx), 755 (1, Qy).

1H-NMR: δ [parts per million] (multipletness, task) = 9,58 (s, 5-H), is 8.75 (s, 10-H), 8,63 (s, 20-H), 7,50 and 7,26 (2 m, o - and m-benzyl-N C-133), 6,53 (s, 132-N), of 5.53 (s, CH2group 133), 5,21, 5,16, 5,13, 5,03 (4 and s, 173-CH2), 4,47, 4,22, 4,15, 3,97 (4 m, 7-H, 8-H, 17-h, 18-H), 3,06 (s, 31The PINES3), to 3.41 (s, 12 CH3), to 3.38 (s, 2-CH3), a 2.36 (m, 171,2-CH2), and 1.63 (m, 7-CH3), of 0.95 (t,3JAB=7 Hz, 81-CH3), 1,65 (d3JAB=7 Hz, 18-CH3), of 1.2 and 1.8 (tbb-C(CH3)3).

MS (ESI): M+=992,3 (calculated 992,5 for12C

1
56
H
14
62
N
16
4
O
106
6
Pd); 1015 (20%, M++Na), 801,4 (24%, M+- COO-tbb).

Example 9. P is torching 13 3-n-propyl-PD-bacteriopheophorbide-a-173-n-propyl ether (pr-Pd-BPheid-pr)

The transesterification start with Pd-BPheid-gg in propanol, following the General methodology. After 12 hours of receiving the product pr-Pd-BPheid-pr with output, constituting 71%, after chromatography on silica with acetone/toluene = 5:95 (about./vol.).

Analytical data for pr-Pd-BPhe-pr: rf=0,54 on silica with acetone/toluene = 10:90 (vol./vol.).

UV/Vis: (DE) (Areltask) = 332 (0,48, By), 385 (0,41, Inx), 527 (0,15, Qx), 755 (1, Qy).

1H-NMR: δ [parts per million] = 9,58 (s, 5-H), is 8.75 (s, 10-H), 8,65 (s, 20-H), 6,50 (s, 132-H), 4,38, 3,97, 3,89, 3,80 (4 m, 7-H, 8-H, 17-h, 18-H), of 3.07 (s, 31The PINES3), 3,89-a 3.83 (m, 2H, propyl-och2when 173), 3,42 (s, 12 CH3), 3,39 (s, 2-CH3), 2,80-2,00 (m, 171,2-CH2), 1,74-of 1.56 (m, N propyl-och2when 173and 133), and 1.63 (m, 7-CH3), to 1.70 (s, 18-CH3), 0.98 and of 0.62 (t,3JAB=7 Hz, propyl-CH3when 173and 133).

MS (ESI): M+=784,7 (calculated 784,4 for12

1
40
H
14
46
N
16
4
O
106
6
Pd); 697,5 (17%, M+- SOOS3H7).

Example 10. Probationaries-a-173-n-propyl ether (pyro-BChlide-pr) (formula V in the diagram, the Central metal of the MD instead of Pd)

After 6 days of pyro-BChlide-gg get pyro-BChlide-pr (30%) after chromatography on silica with acetone/toluene = 5:95 (about./vol.).

Analytical data for pyro-BChlide-pr: rf=0,76 C18silica reversed-phase with HEPES/KOH (20 mm, pH 7.5)/acetone = 15:85 (about./vol.).

UV/Vis: (DE) λmax[nm] (Arel, task) - 357 (0,75, By), 391 (0,52, Bx), 575 (0,21, Qx), 771 (1, Qy).

1H-NMR: δ [parts per million] (multipletness, task) = 9,48 (s, 5-H), 8,65 (s, 10-H), 4,50-4,00 (4 m, 7-H, 8-H, 17-h, 18-H), 3,11 (31The PINES3), 3,57 (s, 12 CH3), of 3.46 (s, 2-CH3), 2,73-of 2.09 (m, 17-CH2group), and 1.63 (m, 7-CH3), 0,81 (s, 81-CH3), to 1.70 (s, 18-CH3), a 1.75 (m, H, propyl-och2when 173).

MS (ESI): M+=616,5 (calculated 616,3 for12

1
36
H
14
40
N
16
4
O
24
4
Mg)

Example 11. Pyro-PD-bacteriopheophorbide-a-173-tert-butylbenzylamine ether (pyro-Pd-BPheid-tbb) (formula V in the diagram)

(a) the Pyro-Pd-BPheid-me expose interaction within 48 hours with para-tert-butylbenzyl alcohol to pyro-Pd-BPheid-tbb. (b) Using as a starting material pyro-Pd-BPhe-gg, get the same product under other equal conditions, the output of which is 70% after chromatography on silica with acetone/toluene =5:95 (about./vol.).

Analytical data for pyro-Pd-BPheid-tbb: rf=0,25 on silica with acetone/toluene = 5:95 (about./vol.).

UV/Vis: (DE) λmax[nm] (Arel, task) = 332 (0,50, By), 384 (0,37, Bx), 530 (0,15, Qx), 755 (1, Qy)

1H-NMR: δ [parts per million] (multipletness, task) = 9,63 (s, 5-H), 8,73 (s, 10-H), 7,31 (s, o - and m-benzyl-N C-133), 5,09 and is 5.18 (dd3JAA'=12 Hz, 132-H), 5,12 and 5.17 (dd,3JAA'=6 Hz, CH2C-173), 4,8, 4,36, 4,25, 4,02 (4 m, 7-H, 8-H, 17-h, 18-H), of 3.07 (s, 31The PINES3), 3,48 (s, 12 CH3), 3,40 (s, 2-CH3), 2,78 is 2.33 (m, 171,2-CH2), was 1.58 (m, 7-CH3), 0,99 (t3JAB=8 Hz, 8-CH3), 1,68 (d3JAB=7 Hz, 18-CH3), 1,17 (ttbb-C (CH3 )3).

MS (FAB): M+=the 802.1 (calculated 802,4 for12C

1
44
H
14
48
N
16
4
O
106
4
Pd).

Example 12. Getting Pd-bacteriopheophorbide a-173-N glucosamine (Pd-BPheid-Nglc) (formula VI in scheme (C)

Another reaction mechanism results in derivatives, in which the ester group in position C-173replaced by the more stable amide bond (see scheme).

In carefully dried the device dissolve 60 mg (88 µmol) free from acid Pd-BPheid in 20 ml of dry DMF. After cooling the flask to 0°add 70 mg (324 mmol) of glucosamine hydrochloride. The pH value was adjusted to 8-9 using 31,2 ál (317 mmol) diisopropylethylamine. To determine the pH drop of the reaction mixture and a drop of water mix on a strip of pH indicator paper. Add 30 mg (91 mmol) TBTU (2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium of tetrafluoroborate) and the flask was incubated for 16 cha is s at room temperature. During the night the flask and allow to warm to room temperature. The reaction mixture is partitioned between chloroform and water. Any formed precipitate is removed by filtration. The chloroform phase was removed and the pigment is dried toluene in a rotary evaporator. After chromatography in the system SII receive the product from the market, constituting 20%.

Analytical data for Pd-BPheid-Nglc: rf=0,75 C18silica reversed-phase with methanol.

UV/Vis: (l3) λmax[nm] (Arel, task) = 334 (0,32, By), 388 (0,27, Bx), 537 (0,11, Qx), 763 (1, Qy).

1H-NMR: δ [parts per million] (multipletness, task) = 9,58 (s, 5-H), of 8.95 (s, 10-H), 8,46 (s, 20-H), 6,37 (s, 132-H), 4,51, to 4.41, 3,83 (3 m, 7-H, 8-H, 17-h, 18-H), of 3.07 (s, 31The PINES3), 3,43 (s, 12 CH3), 3,37 (s, 2-CH3), by 1.68 (m, 7-CH3), 3,85 (s, 132SOON3), 2,8-2,0 (m, 171,2-CH2).

MS(ESI): M+=875,1 m/z (calculated 875,3 for12C

1
4l
H
14
47
N
16
5
O
106
10
Pd); 898,1 (M++Na+).

Example 13. Phototoxicity derivatives M-BChl in relation to melanoma cells in cultures

Obtaining liposomes

Liposomes L-α-dipalmitoylphosphatidylcholine (DPPC) as a carrier for pigments, insoluble in water, get in line with the way Toledano, 1998, and by the way Cuomo et al., 1990. 1,4×10-8mol. (≅ 130 µg) of photosensitizer and 5 mg DPPC dissolved in 400 μl of chloroform. Placed on top of 250 μl of N2O and 250 μl of phosphate buffer (pH 7,2; 1.5 mm KN2RHO4; 7.6 mm Na2HPO4; 0.15 M NaCl). The chloroform is removed within 5 minutes rapid stream of argon, while the mixture is treated with ultrasound and maintained at a temperature of 45°C. the sonication continued for another 20 minutes and liposomes loaded with pigment, get in the supernatant after centrifugation (16000×g, 10 min). The concentration of liposomes determined photometrically at 750 nm (in accordance with the method Grossweiner and Grossweiner, 1982).

Photodynamic activity in single-layer cell culture

Melanoma cells (mice) M2R is cultivated as monolayers in 96-well tablets for titration in DMEM (modified by way of Dulbecco Wednesday Needle)/F121/1 (vol./about.) at 37°C in a humid atmosphere containing the th 8% CO 2. The medium (pH 7.4) supplemented with HEPES buffer (25 mm), fetal bovine serum (FBS) (10%), glutamine (2 mm), penicillin (0.06 mg/ml) and streptomycin (0.1 mg/ml). Within 24 hours the density of cells increases from 1×104up to 2×104cells/100 μl. To cells add an increasing number of liposomal preparation containing BChl derivative. (Pd-BPheid-ser or Pd-BPheid-Nglc added as ethanol solution (10-4M) so that the maximum ethanol concentration was <1%). Initially, cells are kept in the dark for 4 hours, washed with 100 μl of medium, treated with 100 μl of fresh medium and then irradiated from below through the bottom of the tablets of the Russian lamp BS LS3-PDT, equipped with a filter (600-1300 nm). Within 10 minutes the cells are irradiated with a dose of light, part 10 mW × s × cm-2. After another 24 hours in the dark at 37°cell viability is determined by the microscope (the size of the cells and form) and by introducing the DNA of [3H]-thymidine (Chen et al. 1988). For this, cells incubated in the end of the experiment for 2 hours at 37°With 1 MX/ml [3H]-thymidine (in water). Then twice washed with phosphate buffer, incubated with 7.5% cold trichloroacetic acid for 30 minutes at 4°With, again washed with 95% ethanol and finally treated with 200 μl of 1N NaOH for 10 minutes at 37°C. 100 µl of the prepared suspension is NaOH removed neutralize 100 μl of 1N HCl, mixed with 4 ml of scintillation fluid (20,8 (about./about.) Xylen: mix scintillator Lumax) and 5 ml imidazoline buffer (0.1 M).

The results are presented in figure 1(A-D) and 2. Three of the sensitizers represented in figure 1 (A-D) and 2, were any phototoxic against melanoma cells of mice M2R (Pd-BPheid-et (LD90=0.02 mm) and tbb-Pd-BPheid-me (LD90=1.1 µm)), Pd-BPheid-ser ((LD90=0.1 ám). tbb-Pd-BPheid-tbb and Pd-BPheid-NgIc ineffective in these conditions, because they form aggregates in liposomes, are not effective for PDT.

Scheme And

Table 1.

[M]-BPhe Transetherification selectively in position C-133or both provisions-133and C-173
ConnectionFormula (schema)R2R1R1M
Me-BChl-ggIggme-Mg
Me-Pd-BPhe-meImeme-Pd
Me-Pd-BPhe-ggIggme-Pd
Tbb-Pd-BPhe-tbbIIItbb-tbb Pd
Tbb-Pd-BPhe-meIIme-tbbPd
Tbb-Pd-BPhe-ggIIgg-tbbPd
NtBoc-ser-Pd-BPhe-meIIme-serPd
Pr-Pd-Bphe-prIIIpr-prPd
Me-Pd-Bphe-prIIpr-mePd
Pr-Pd-Bphe-ggIIgg~prPd
Pr-BChl-prIIIpr-prMg

Table 2.

Transesterification products pyro[M]BPhe
Connection:Formula (schema)R1R1M
Pyro-BCh1-ggIVggMg
Pyro-Pd-BPhe-ggIVggPd
Pyro-Pd-BPhe-tbbV-tbbPd
Pyro-Pd-BPhe-prV -prPd
Pyro-BChl-prV-prMg

Scheme C. the Reaction scheme for the derivative bacteriochlorophyll, transesterification in position C-133and/or C-173

VI-R4is glucosamine residue

Scheme C. the Combination of glucosamine with C-173Pd-Bpheid

LINKS

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L. Chen, S. Gross, V. Rosenbach-Belkin, A. Brandis, Y. Salomon and A. Scherz (1988) Microbial photoinactivation by serine and IgG conjugates of chlorophyll and bacteriochlorophyll. In: Proceedings of the 7th Biennial Congress., International Photodynamic Association (ed. T. Patrice).

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Dougherty, T.J. (1992) Bacteriochlorophyll a Derivatives Useful in Photodynamic Therapy., Pat No. 5171741, USA. Int. Class A 61 K 31/40, Appl. No. 341,591.

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Fiedor, L., Rosenbach-Belkin, V., M. Sai and A. Scherz (1996) Preparation of tetrapyrrole-amino acid covalent complexes. Plant Physiol. Biochem. 34, 393-398.

S.Gross, A.Brandis, L.Chen, V.Rosenbach-Belkin, S.Roehrs, A. Scherz and Y. Salomon (1997) Protein A mediated Bacteriochlorophyll-IgG targeting to Staphylococcus aureus:

A model for enhanced site-specific photocytotoxicity. Photochem. Photobiol 66, 872-878.

Grossweiner, L.I.P.A.S. and Grossweiner, J.B. (1982). Photobiol. 36: 159-167.

Hunt, J.E., & Michalski, T.J. (1991). Chlorophylls, H.Scheer. (ed.) Boca Raton: CRC Press.

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S. Katz, Y.Vakrat, V.Brumfeld, L.Weiner, E.Gabelman, A. Brandis, A. Paul, M. Hild, R.Lendt, D.Leopold, J.R. Norris, Y.Salomon and A.Scherz (1998), Bacteriochlorophyll-serine generates only hydroxy radicals under near infra-red illumination In: Proceedings of the 7th Biennial Congress., International Photodynamic Association (ed. T. Patrice).

D.K. Kelleher, O.Thews, J.Rzeznik, A.Scherz, Y.Salomon and P.Vaupel (1999) Water-filtered infrared-A radiation: a novel technique for localized hyperthermia in combination with bacteriochlorophyll-based photodynamic therapy, Int.J. of Hyperthermia (In press).

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V. Rosenbach-Belkin, L.Chen, L.Fiedor, Y.Salomon and A. Scherz (1998) Chlorophyll and bacteriochlorophyll derivatives as photodynamic agents. In: Photodynamic Tumor Therapy; 2nd and 3rd Generation Photosensitizers (ed. J.G. Moser), Harwood Academic Publishers, Australia, 117-125.

Scheer, H. (ed.) (1991), Chlorophylls, CRC Press, Boca Raton.

A.Scherz, S.Katz, Y.Vakrat, V.Brumfeld, E.Gabellman, J.Zilberstein, D.Leopold, J.R. Norris, H.Scheer, and Y.Salomon (1998) Bacteriochlorophyll-serine based photochemotherapy; type III PDT? In: "Photosynthesis: Mechanisms and Effects (Ed. G. Garab), Kluwer Acad. Publishers, Dordrecht, the Nederlands, V.V pp.4207-4212.

Schneider, C.H. & Eberle, A.N. (1992). Peptides. ESCOM Science Publishers B.V.

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Spikes, J.D. & Boomer, J.C. (1991) in 'Chlorophylls' (Scheer, H., ed.), pp 1181-1204, CRC Press, Boca Raton.

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Toledano, F.; Edrei, R. and Kimel, S. (1998) J. Photochem. Photobiol. B: Biol.; 42:20-27.

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J.Zilberstein, A.Bromberg, A. Franz, V. Rosenbach-Belkin, A.Kritzmann, R.Pfeferman, Y.Salomon and A.Scherz (1997) Light dependent oxygen consumption in bacteriochlorophyll-serine treated melanoma tumors: on-line determination using a tissue inserted oxygen microsensor. Photochem. Photobiol 65, 1012-1020.

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1. The method of transesterification to obtain a synthetic derivative of chlorophyll or bacteriochlorophyll General formula I

where X=O

M is a Central metal atom selected from the group including MD, Pd, Co, Ni, Si, Zn, Nd, Er, It, S, Sn and mn, or represents two atoms N;

each of R3and R5independently represents acetyl, vinyl, ethyl, 1-hydroxyethyl or a simple or a complex ester of the specified 1-hydroxyethylrutoside;

R4represents methyl or formyl;

the dashed line in positions 7-8 represents an optional double bond; and

R1and R2are different and are selected from the group comprising: (i) C1-C25hydrocarbonrich, which can be straight or branched, saturated or unsaturated, optionally substituted by one or more radicals selected from halogen, oxo (=O), HE, Cho, COOH or NH2or interrupted by one or more heteroatoms selected from O, S and NH, or carbocyclic or heterocyclic residues;

(ii) the balance of amino acids, oligopeptides or polypeptide containing the hydroxy-group or their derivative, selected from the group comprising esters and N-protected derivatives, in which the specified gidrauxilirovannaya amino acid or its derivative is associated with soo-the remainder derived Chl or BChl through the said hydroxy-group;

(iii) a residue of a peptide described in (ii)associated with soo-the remainder through C1-C25hydrocarbon described in (i)where the specified C1- 25saturated or unsaturated hydrocarbonyl residue, optionally substituted by one or more radicals selected from halogen, oxo (=O), HE, Cho, COOH or NH2or this balance is interrupted by one or more heteroatoms selected from O, S and NH, or phenyl ring is further substituted terminal functional group selected from HE, COOH or NH2;

(iv) the balance of cell - or tissue-specific ligand selected from oligopeptides and protein directly associated with soo-the remainder or via C1-C25hydrocarbon described in (i)where the specified C1-C25saturated or unsaturated hydrocarbonyl residue, optionally substituted by one or more radicals selected from halogen, oxo (=O), HE, Cho, COOH or NH2or this balance is interrupted by one or more heteroatoms selected from O, S and NH, or phenyl ring is further substituted terminal functional group selected from HE, COOH or NH2; and

(v) the residue of a mono-, oligo - or polysaccharide or polyoxyethylene associated with soo-the remainder either directly or through C1-C25hydrocarbon described in (i)where the specified C1-C25saturated or unsaturated hydrocarbonyl residue, optionally substituted by one or more of the radicals is AMI, selected from halogen, oxo (=O), HE, Cho, COOH or NH2or this balance is interrupted by one or more heteroatoms selected from O, S and NH, or phenyl ring is further substituted terminal functional group selected from HE, COOH or NH2;

including interaction under anaerobic conditions derived (bacterial plankton)chlorophyll, metal(bacterial plankton)chlorophyll or (bacterial plankton)pheophytin having a position-132a group of SOON3and in position C-172group COOR2with alcohol R1OH, provided that R1is not methyl, in the presence of tetraethyl-ortho-titanate, and the interaction is carried out in an aprotic solvent such as free from peroxide tetrahydrofuran (THF) or dimethylformamide (DMF), resulting in the desired W C-132-COOR1C-172-COOR2which is then separated from the reaction mixture.

2. The method of transesterification to obtain a synthetic derivative of chlorophyll or bacteriochlorophyll General formula I

where X=O

M is a Central metal atom selected from the group including MD, Pd, Co, Ni, cu, Zn, Nd, Er, It, S, Sn and mn, or represents two atoms N;

each of R3and R5independently represents acetyl, vinyl, ethyl, 1-hydroxyethyl or question is Oh or an ester of the specified 1-hydroxyethylrutoside;

R4represents methyl or formyl;

the dashed line in positions 7-8 represents an optional double bond; and

R1and R2are identical and are selected from the group including:

(i) C1-C25hydrocarbonrich, which can be straight or branched, saturated or unsaturated, optionally substituted by one or more radicals selected from halogen, oxo (=O), HE, Cho, COOH or NH2or interrupted by one or more heteroatoms selected from O, S and NH, or carbocyclic or heterocyclic residues;

(ii) the balance of amino acids, oligopeptides or polypeptide containing the hydroxy-group or their derivative, selected from the group comprising esters and N-protected derivatives, in which the specified gidrauxilirovannaya amino acid or its derivative is associated with soo-the remainder derived Chl or BChl through the said hydroxy-group;

(iii) a residue of a peptide described in (ii)associated with soo-the remainder through C1-C25hydrocarbon described in (i)where the specified C1-C25saturated or unsaturated hydrocarbonyl residue, optionally substituted by one or more radicals selected from halogen, oxo (=O), HE, Cho, COOH or NH2or this balance is interrupted by one or more heteroatoms, selected from O, S and NH, or phenyl ring is further substituted terminal functional group selected from HE, COOH or NH2;

(iv) the balance of cell - or tissue-specific ligand selected from oligopeptides and protein directly associated with soo-the remainder or via C1-C25hydrocarbon described in (i)where the specified C1-C25saturated or unsaturated hydrocarbonyl residue, optionally substituted by one or more radicals selected from halogen, oxo (=O), HE, Cho, COOH or NH2or this balance is interrupted by one or more heteroatoms selected from O, S and NH, or phenyl ring is further substituted terminal functional group selected from HE, COOH or NH2; and

(v) the residue of a mono-, oligo - or polysaccharide or polyoxyethylene associated with soo-the remainder either directly or through C1-C25hydrocarbon described in (i)where the specified C1-C25saturated or unsaturated hydrocarbonyl residue, optionally substituted by one or more radicals selected from halogen, oxo (=O), HE, Cho, COOH or NH2or this balance is interrupted by one or more heteroatoms selected from O, S and NH, or phenyl ring is further substituted terminal functional group selected from HE, COOH NH 2;

including interaction under anaerobic conditions derived (bacterial plankton)chlorophyll, metal(bacterial plankton)chlorophyll or (bacterial plankton)pheophytin having a position-132a group of SOON3and in position C-172group COOR2with alcohol R1OH, (provided that R1does not represent methyl)in the presence of tetraethyl-ortho-titanate, resulting in the desired W-132-COOR1C-172-COOR2and where in the solvent used R1OH.

3. The method according to claim 1 or 2, where M represents a divalent metal selected from the group including MD, Pd, Co, Ni, cu, Zn, Nd, Er, It, S, Sn and MP.

4. The method for obtaining compounds of chlorophyll according to claim 1 or 2, where R3is vinyl, R4is methyl or formyl, R5is ethyl, and the dotted line in positions 7-8 represents a double bond.

5. The method for obtaining compounds of bacteriochlorophyll according to claim 1 or 2, where R3is acetyl, R4is methyl, R5is ethyl, and the provisions 7-8 gidrirovanny.

6. The method according to claim 5, where M is Pd, R1represents tert-butylbenzyl, a R2represents methyl.

7. The method according to claim 5, where M is Pd, R1represents tert-butylbenzyl, and R2is geranylgeranyl.

8. The method according to claim 5, where M is Pd, R1 is propyl, a R2is geranylgeranyl.

9. The method according to claim 5, where M is Pd, R1is N-tert-butyloxycarbonyl, and R2represents methyl.

10. The method according to claim 5, where M is Pd, R1is 0-seril, a R2represents methyl.

11. The method according to claim 5, where M is the MD, a R1and R2are propyl.

12. The method according to claim 5, where M is Pd, a R1and R2represent tert-butylbenzyl.

13. The method according to claim 5, where M is Pd, a R1and R2are propyl.



 

Same patents:

The invention relates to biotechnology, in particular to the production of antibiotics

The invention relates to pharmaceutically active bicyclic heterocyclic amines (XXX) and can be used as pharmaceuticals for the treatment of diseases and injuries

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 chemistry of biologically active compounds, and specifically to an improved process for the preparation of a derivative of hematoporphyrin, which is used as a photosensitizer for photodynamic therapy (PDT) of malignant tumors

FIELD: organic chemistry and medicine.

SUBSTANCE: invention relates to new imidazole derivatives of general formula I useful as adenosine A3-receptor modulators, as well as to method for cancer treatment and detection of tumor cells using claimed derivatives.

EFFECT: compounds for treatment and diagnosis of improved activity.

13 cl, 4 dwg, 19 ex

FIELD: medicine; veterinary medicine.

SUBSTANCE: method involves using biologically active substances obtainable by means of bacteriological synthesis. Individual avermectines or their complexes are applied in vitro in concentrations of 0.01-10 ng/ml and in vivo at a dose of 0.01-1 mg/kg.

EFFECT: enhanced effectiveness of treatment; reduced toxic effect.

4cl, 2 tbl

FIELD: pharmaceutical chemistry.

SUBSTANCE: invention relates to treatment of patients suffering from diseases associated with pathologic activity of matrix proteases. Treatment involves administration of compounds depicted by general formula (I).

EFFECT: increased treatment efficiency.

136 cl, 448 ex

FIELD: organic chemistry, pharmaceutical compositions.

SUBSTANCE: invention relates to novel pyrasolbenzodiazepines of formula I 1 (in formula R1 is hydrogen, -NO2, -CN, halogen, -OR5, -COOR7, -CONR8R9, -NR10R11, NHCOR12, NHSO2R13; each R2 and R4 independently of one another are hydrogen, halogen, -NO2, -CF3; R3 is hydpegen, C3-C8-cycloalkyl, aryl, in particular C6-C10-aromatic group having 1 or 2 rings, 5-10-membered heteroaryl, having 1 or 2 rings and1-3 heteroatoms, selected from N, O, and S, -COOR7, CN, C2-C6-alkenyl, -CONR8R9 or C1-C6-alkyl optionally substituted with OR9-group, F or aryl as mentioned above; R5 is C1-C6-alkyl; R7 is hydrogen or C1-C6-alkyl; each independently of one another are hydrogen or C1-C6-alkyl optionally substituted with hydroxyl or NH2, or alternatively R8 and R9 together form morpholino group; each R10,R11 and R12 independently of one another are hydrogen or C1-C6-alkyl; R13 is C1-C6-alkyl optionally substituted with halogen or -NR14R15; each R14 and R15 independently of one another are hydrogen or C1-C6-alkyl optionally substituted with halogen; or alternatively -NR14R15 is morpholino group) or pharmaceutically acceptable salts thereof, as well as to certain pyrasolbenzodiazepine derivatives, thiolactam intermediates for production of compound (I) and pharmaceutical compositions containing the same. Compound and pharmaceutical composition of present invention are cycline-dependent kinase (CDK2) inhibitors and antiproliferation agents used in treatment or controlling disorders associated with cell proliferation, in particular breast, colon, lung and/or prostate tumors.

EFFECT: new antiproliferation agents.

20 cl, 12 tbl, 8 ex

FIELD: organic chemistry, biochemistry, pharmacy.

SUBSTANCE: invention relates to new compounds of the formula (I):

eliciting inhibitory activity with respect to metalloproteinases and wherein R1 means phenoxy-group wherein phenyl residue can be substituted with one or some halogen atoms, hydroxy-, (C1-C6)-alkoxy-group, (C1-C6)-alkyl, cyano- or nitro-group; R2 means pyrimidine, pyrazine or its N-oxide or phenyl substituted with -SO2NR3R4 wherein R3 and R4 can be similar or different and mean hydrogen atom, direct-chain or branch-chain (C1-C6)-alkyl that can be substituted once or some times with the group OH, N(CH3)2, or it can be broken by oxygen atom, or it represents COR5 wherein R5 means (C1-C)-alkyl group that can be substituted with NH2. Also, invention relates to a pharmaceutical composition comprising above said compounds.

EFFECT: valuable biochemical properties of compounds and composition.

5 cl, 1 sch, 1 tbl, 10 ex

FIELD: organic chemistry, heterocyclic compounds, medicine, pharmacy.

SUBSTANCE: invention relates to nitrogen-containing heterocyclic derivatives of the formula (I): A-B-D-E (I) wherein A means 5- or 6-membered heteroaryl comprising one or two nitrogen atoms in ring; B means ethenylene; D mean phenylene; E means group -N(COR)-SO2-G wherein G means phenyl; R means 5- or 6-membered heteroaryl or heteroarylmethyl comprising one or two nitrogen atoms in ring, or group -(CH2)n-N(R5)R6 wherein n means a whole number from 1 to 5; R5 and R6 are similar or different and mean: hydrogen atom, (C1-C6)-alkyl, hydroxyalkyl, aminoalkyl; or R5 and R6 in common with nitrogen atom can form 5-7-membered cyclic amino-group -N(R5)R6 that can comprise, except for nitrogen atom, also oxygen, sulfur or nitrogen atom as a component forming the ring, or their N-oxides. Compounds of the formula (I) elicit anticancer activity and can be used in medicine.

EFFECT: valuable medicinal properties of compounds.

10 cl, 1 tbl, 24 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to new derivatives of benzene of the formula (I): wherein A represents a group taking among the following groups: -C≡C-, -CH=CH-, -CH2-CH2; n = 1 or 2; X represents hydrogen, chlorine or fluorine atom or methyl or methoxy-group; Y represents hydrogen, chlorine or fluorine atom; R1 represents cyclohexyl group monosubstituted, disubstituted, trisubstituted or tetrasubstituted with methyl group, phenyl group monosubstituted or disubstituted with fluorine or chlorine atom or methoxy-group, cycloheptyl, tert.-butyl, dicyclopropylmethyl, 4-tetrahydropyranyl or 1- or 2-adamantyl, or adamantine-2-ol group; or R1 represents phenyl group and in this case X and Y both represents chlorine atom; R2 represents hydrogen atom or (C1-C4)-alkyl group; R3 represents (C5-C7)-cycloalkyl, and salts of these compounds formed by addition of pharmaceutically acceptable acids, and their solvates and hydrates also. Also, invention relates to methods for preparing compounds of the formula (I) and to pharmaceutical composition able to interact with receptors sigma-2 based on these compounds. Invention provides preparing new compounds and medicinal agents based on thereof for treatment of autoimmune states, disturbance on heart contraction frequency and control against proliferation of tumor cells.

EFFECT: improved preparing methods, valuable medicinal properties of compositions.

18 cl, 14 tbl, 78 ex

FIELD: medicine, oncology.

SUBSTANCE: the present innovation deals with systemic chemotherapy in case of different oncological diseases. At first, one should carry out 4 courses of systemic chemotherapy at 2-mo-long interval between each other, then, since 10-11th mo against the onset of chemotherapeutic treatment one should additionally conduct 6 courses more at 4-mo-long interval between them. The innovation enables to considerably prolong the period of remissions and that of patient's life duration due to prolonging the terms of therapy by carrying out chemotherapeutic courses before the highest lethal risk.

EFFECT: higher efficiency of therapy.

4 dwg, 1 ex

FIELD: organic chemistry, medicine, pharmacy.

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

wherein R represents the group of the general formulae -COOR1, -CONR2R3, -CONR4CONR2R5 or -CH2OR6 wherein R1 represents (C2-C6)-alkyl group, (C1-C6)-alkyl group substituted with hydroxyl group or 5-8-membered saturated nitrogen-containing heterocyclic group (it can comprise oxygen atom in addition to nitrogen atom) or 5-6-membered nitrogen-containing aromatic heterocyclic group or (C3-C6)-cycloalkyl group; R2 and R3 are similar or different and represent independently hydrogen atom or (C1-C6)-alkyl group that can be substituted optionally with hydroxyl, (C2-C5)-alkoxycarbonyl or 5-8-membered saturated nitrogen-containing heterocyclic group (it can comprises oxygen atom in addition to nitrogen atom) or 5-6-membered aromatic homocyclic group or aromatic heterocyclic group comprising oxygen and/or nitrogen atom, 5-6-membered cycloalkyl or heteroaryl group; R4 and R5 are similar or different and represent independently hydrogen atom or (C3-C6)-cycloalkyl group; R6 represents hydrogen atom; also, invention relates to tautomers, solvates of these compounds, their mixtures and acid-additive salts. Also, invention relates to pharmaceutical compositions comprising compounds of the general formula (I) as an active component. Angiogenesis inhibitors of the present invention inhibit formation of new vessels in tissues of live organisms and can be used for prophylaxis and inhibition of the angiogenesis process arising in the tumor proliferation, and for prophylaxis of formation of tumor metastasis. Invention provides preparing new derivatives of borrelidin eliciting the value physiological effect.

EFFECT: valuable medicinal properties of compounds.

8 cl, 15 ex

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