Antagonists cdk2 as antagonists of short form of c-maf transcription factor for glaucoma treatment
SUBSTANCE: invention relates to medicine, namely to experimental ophthalmology, and can be applied for suppression of positive regulation of short form of c-Maf transcription factor in processed with steroids or transforming growth factor β2 (TGF β2) cells of trabecular net. For this purpose introduced is antagonist c-Maf, also possessing inhibiting activity with respect to cyclin-dependent kinase cdk2, for instance purvalanol A.
EFFECT: invention explains mechanism and possible new approach to treatment of primary open-angle and steroid glaucoma.
4 cl, 1 tbl, 7 ex, 3 dwg
The technical FIELD TO WHICH the INVENTION RELATES.
The present invention relates to a prophylactic and therapeutic agents for the treatment of glaucoma, including primary open-angle glaucoma and steroid glaucoma.
The LEVEL of TECHNOLOGY
Trabecular network (TM) is a combined tissue, including endothelial cells, connective tissue and extracellular matrix, located at the angle between the cornea and the iris, which provides normal resistance required to maintain the intraocular pressure (IOP). Adequate intraocular pressure necessary to maintain the shape of the eye and to ensure that the pressure gradient for flow of intraocular fluid lacking vessels of the cornea and the crystalline lens. Increased IOP associated with glaucoma, has a devastating effect on the optic nerve, which leads to loss of cells and axons of retinal ganglion and resulting in progressive loss of vision and blindness in the absence of treatment. Glaucoma is one of the main causes of blindness worldwide.
Primary glaucoma occurs due to flow of intraocular fluid, which have anatomical or physiological basis. Secondary glaucoma occurs as the result of damage is, or trauma of the eye or as a result of the existing disease. In ninety percent of cases of primary glaucoma it is primary open-angle glaucoma (POAG), also called or chronic simple glaucoma. POAG is destruction of the trabecular network, leading to pathologically high resistance to the flow of fluid from the eye. The consequence of this resistance is increased IOP, which is necessary to ensure a flow of fluid produced in the normal eye, this increased resistance.
It is known that some drugs, such as prednisone, dexamethasone and hydrocortisone, induce glaucoma, increasing IOP. In addition, IOP effect, apparently, the route of administration. For example, the introduction of dexamethasone in the eye leads to a higher increase in IOP compared with systemic administration. Glaucoma, which occurs as a result of taking steroid medicines called steroid glaucoma.
Existing methods of treatment of glaucoma include lowering IOP using the suppressor education intraocular fluid or tools that enhance uweoscleeralny leakage, laser trabeculoplasty (laser photocoagulation of the trabecular network of the eye) or trabeculectomy, which is a filtration surgery to improve drainage. Pharmaceutical approaches for the treatment of glaucoma found various who's unwanted side effects. For example, mitotic agents such as pilocarpine can cause blurred vision and other adverse visual effects. Systemically administered carbonic anhydrase inhibitors may also cause nausea, dyspepsia, fatigue and metabolic acidosis. In addition, some beta-blockers are increasingly cause serious pulmonary side effects attributed to their effect on beta-2 receptors in the lung tissue. Sympathomimetic means cause tachycardia, arrhythmia and hypertension. These negative side effects can lead to poor compliance by the patient mode and regimens of the drug or to discontinuation of treatment.
More important is that the existing methods of treatment of glaucoma is not aimed directly at the pathological disturbance in trabecular network, optic nerve and loss of cells and axons of the retinal ganglion, which continues unabated. Because of the importance of glaucoma and failure of previous treatments, it would be desirable to have an improved method of treating glaucoma, which would be directed at the causes underlying the progression of glaucoma.
The present invention relates to a method for the treatment of primary open-angle glaucoma or steroid glaucoma in patients at risk of developing primary open-angle glaucoma is or steroid glaucoma or who have their symptoms. The method involves the administration to a patient an effective amount of a composition comprising antagonist a short form of the transcription factor c-Maf and an acceptable carrier.
In accordance with the invention is identified that the short form of the transcription factor c-Maf positively regulated treated with steroids and transforming growth factor β2 (TGFβ2) cells of the trabecular network (TM) and is present in elevated levels in glaucomatous cells compared to normal cells of the tissue of the disc of the optic nerve, and in elevated levels in glaucomatous tissue TM compared to normal TM cells. Expression of the short form of the transcription factor c-Maf in these conditions indicates a causal or effector role of this factor in the pathogenesis of primary open-angle and steroid glaucoma. The methods of the invention include antagonism against transcription, expression and/or activity of a shortened form of the transcription factor c-Maf in trabecular network or other ocular tissue, such as tissue of the optic nerve, for inhibiting or weakening of the pathogenesis of glaucoma.
The antagonist of the present invention inhibits the transcription or expression of a short form of the transcription factor c-Maf. In one of the embodiments, the antagonist of a short form of the transcription factor c-Maf is going to win a purine analog, possessing inhibitory activity against cyclin-dependent kinase cdk2. This antagonist can include, for example, purvalanol And purvalanol, aminophenol, olomoucine, N9-isopropylammonium, roscovitine, methoxyresorufin, combinations thereof, or their salts.
According to another variant implementation, the antagonist possessing inhibitory activity against cyclin-dependent kinase cdk2, is administered nonpurine connection, such as, for example, indirubin, oxindol, interoperator, pyridopyrimidines, anilinoquinazoline, aminothiazol, flavopiridol, staurosporin, Paulson, geniality, combinations thereof, and their salts.
The use of antagonists of expression or activity of a short form c-Maf as therapeutic agents for the protection or treatment of patients damage caused by the pathogenesis of glaucoma, aimed at the development of the disease, along with signs and symptoms of the disease, i.e. as a result of this treatment changes the pathological process. Antagonists of expression or activity of a short form c-Maf can be used for the treatment of POAG and steroid glaucoma. Identification of a short form of the transcription factor c-Maf as a participant in the pathogenesis of glaucoma and application presented here inhibitors of the expression or activity has not been described previously.
BRIEF DESCRIPTION of DRAWINGS
Figure 1 - QPCR-EN is Liz expression of short form c-Maf in the merged cells SGTM2697 demonstrates the expression of TGFβ2-induced gene increased 16-fold compared to control.
Figure 2 - QPCR analysis of the expression of the short form c-Maf in cells TMA demonstrates the expression of dexamethasone-induced gene, increased 2.5 times in the first day and 3.2-fold at day 14, compared to control.
Figure 3 - QPCR analysis of the expression of the short form c-Maf in cells SGTM2697 (P6) in the presence and in the absence of purvalanol And for basal and TGFβ2-induced cells.
DETAILED description of the INVENTION
The present invention relates to the use of agents for antagonistic action on the expression and/or activity of a short form of the transcription factor c-Maf for the treatment of glaucoma. Microemissive human genome hybridized with normal and glaucomatous RNA gene and the short form of the transcription factor c-Maf positively regulated in cells of glaucoma compared to normal cells.
Maf-dependent genes were identified as important participants in the development of the crystalline lens and anterior segment (Yoshida, et al. (1997), Invest Ophtalmol Vis Sci 38(12): 2679-83; Ogino et al. (1998), Science 280(5360): 115-8; Kawauchi, et al. (1999), J Biol Chem. 274(27): 19254-60; Kim, et al. (1999), Proc Natl Acad Sci USA 96(7); 3781-5; Ring et al. (2000), Development 127(2): 307-17; Ishibashi et al. (2001), Mech Dev 101(1-2): 155-66; Jamieson, et al. (2002), Hum Mol Genet 11(1): 33-42; Reza, et al. (2002), Mech Dev 116(1-2): 61-73). It was shown that c-Maf activates expression of the gene of the lens when activated by the gene product of glaucoma RAH (Sakai, et al. (2001), Nucleic Acids Res29(5): 1228-37; Yoshida, et al, (2001) Curr Eye Res 23(2): 116-9) and, possibly, self-adjusting its own gene product (Kim, et al. (1999), Proc Natl Acad Sci USA 96(7): 3781-5).
c-Maf is a transcription factor latinboy main zipper area (bZIP). Members of the Maf family have ≤40% homology in their primary domain bZIP motifs. There are short form c-Maf, with one exon, (373 amino acids) and the long form of c-Maf, with two exons, (403 amino acids), but their functional difference remains unknown. Short form C-Maf ends with a methionine at the C-end. Additional carboxy-terminal amino acid sequence of the long form is a ITEPTRKLEPSVGYATFWKPQHRVLTSVFTK, SEQ ID NO:4. As used here, the term "short form of the transcription factor c-Maf" refers to a gene that encodes a short form of the transcription factor c-Maf or protein product, consisting of 373 amino acid protein sequence deposited under access number Gen Bank Accession no. AF55376.
In U.S. patent No. 6274339 issued Glimcher et al., described here as a reference in full, describes the sequence of the nucleic acid sequence of the protein c-Maf person, as well as antisense molecules and anti-c-Maf antibodies. The sequence of c-Maf U.S. patent 6274338 placed in GenPept under access number No. UAE. This sequence corresponds to the long form of c-Maf, except a few missing amino acids, includes a deletion of 3 amino acids at positions 241 to 243, compared to the sequence of the protein contained in GenBank under the numbers AF055376 (sequence short form) and AF055377 (sequence of the long form).
Antagonists of a short form of the transcription factor c-Maf
Antagonists of a short form of the transcription factor c-Maf include agents that, for example, reduce transcription of the gene short forms, inhibit the expression of short form or inhibit the activity of a short form. In particular, it was found that inhibitors of the cyclin-dependent kinase cdk2, in particular analogs of purines, negatively regulate the transcription of a short form of the transcription factor c-Maf. Table 1 presents the list of antagonists of a short form of the transcription factor c-Maf, possessing inhibitory activity against cdk2.
Additional inhibiting cdk2 agents described in U.S. patent No. 6573044, issued to Gray et al., Rosania et al., Exp. Opin. Ther. Patents (2000) 10(2): 215-230, in particular, in section 3, a relatively low molecular weight inhibitors, Fischer, P.M., Celltransmissions 19:1, pg 3-9, March, 2003. The person skilled in the art in light of the present invention will be clear that such agents can be racemic mixture, or a diastereoisomer, or enantiomer depending on the substituents.
Despite their chemical diversity, many of the compounds in Table 1 are competing with ATP for the binding site in the complex of cyclin/dk2. For example, the results of structural analysis showed that the purine part of many purine inhibitors is associated with adminisuative pocket cdk2, preventing its binding to the true ligand. Planar system heterocyclic ring is, apparently, a structural characteristic common to many of cdk2 inhibitors.
Analysis antagonist short form of the transcription factor c-Maf provides for the integration of the proposed antagonist gene of the transcription factor c-Maf in the environment in which it is possible transcription and expression. The amount present of the transcription factor c-Maf or the level of activity, less than this amount and the level of activity in the absence of the alleged antagonist, suggests that this is likely the antagonist is really an antagonist of c-Maf.
Route of administration
The antagonist can be delivered directly into the eye (for example, using eye drops or ointments for topical use; devices for slow release in a blind pouch or implanted near the sclera or within the eye; periocular, conjunctival, subaerially, cell injection, injection into Steklov the aqueous body or inside the channels) or systemically (for example, oral, intravenous, subcutaneous or intramuscular injections; parenteral, dermal delivery) ways, well known to specialists in this field. Additionally considered getting antagonists according to the invention in the form of an inner liner or implantiruemy devices. Cell injection can be performed through the cornea into the anterior chamber to allow the agent to reach the trabecular network. Injection inside the channel can be injected into the venous channels of the collector, drainage channel Slamma, or in the channel Slamma.
The patient: Patient, treatment of primary open-angle glaucoma or steroid glaucoma, described here, may be a person or animal is at risk of developing primary open-angle glaucoma or steroid glaucoma or have symptoms of primary open-angle glaucoma or steroid glaucoma.
Compositions and dosage: Antagonists according to the invention can be administered as solutions, suspensions or emulsions (dispersions) in a suitable ophthalmic carrier. The following are examples of possible compositions embodied in this invention.
|The amount in wt.%|
|The inhibitor of the transcription factor c-Maf|
|The benzalkonium chloride||0,01%|
|NaOH/HCl||qs pH 7,4|
|Purified water||qs 100 ml|
|The amount in wt.%|
|Antagonist transcription of c-Maf||0,00005-0,5; 0,0003-0,3;|
|Buffered phosphate saline||1,0|
|The benzalkonium chloride||0,01|
|Purified water||qs to 100%|
|The amount in wt.%|
|Antagonist transcription of c-Maf||0,001|
|Monobasic sodium phosphate||0,05|
|Dibasic sodium phosphate (anhydrous)||0,15|
|The benzalkonium chloride||0,01|
|HCl and/or NaOH||pH 7.3 to 7.4|
|Purified water||qs to 100%|
|The amount in wt.%|
|Antagonist transcription of c-Maf||0,0005|
|Buffered phosphate saline||1,0|
|Purified water||qs to 100%|
In the following embodiment, the ophthalmic composition is prepared for receiving the intraocular concentration of approximately 0.1-100 nm, or in an additional embodiment, the wasp is estline, 1-10 nm antagonist. Compositions for topical application are delivered to the surface of the eye one to four times per day at the discretion of the Clinician. the pH of the composition should be 4-9 or 4.5 to 7.4. Compositions for system reception can contain approximately 10-1000 mg of the antagonist.
The term "effective amount" refers to the amount of the antagonist of c-Maf, which is able to interfere with the expression or activity of a short form c-Maf. This violation leads to the lowering of intraocular pressure and relieving symptoms of glaucoma in a patient with symptoms of primary open-angle glaucoma or steroid glaucoma. Such breach delays or prevents the occurrence of symptoms in a patient at risk of developing glaucoma. An effective amount of the composition may depend on such factors as age, race and gender of the subject or, for example, on the severity of glaucoma. In one of the embodiments, the antagonist is delivered locally in the eye and reaches the trabecular network of the retina or optic nerve head in a therapeutic dose, thereby weakening, the pathological process of glaucoma.
Although the exact scheme is the introduction remains at the discretion of the attending physician, the resulting solution or solutions are preferably administered by instillation of one drop of each solution (each solution in each eye one to four times per day or as defined Leche is brilliant doctor.
Acceptable media: Ophthalmically acceptable carrier called carriers that cause at least slight irritation or not call it at all, provide suitable preservation if needed, and deliver one or more antagonists of c-Maf according to the invention in a homogenous dosage. For ocular delivery of an inhibitor of transcription of c-Maf can be combined with ophthalmically acceptable preservatives, co-solvents, surfactants, amplifiers viscosity, penetration enhancers, buffers, sodium chloride or water for the formation of aqueous, sterile ophthalmic suspension or solution. The composition of the ophthalmic solutions can be prepared by dissolving the inhibitor in a physiologically acceptable isotonic aqueous buffer. In addition, the ophthalmic solution may contain ophthalmically acceptable surfactant to facilitate dissolution inhibitor. To improve the retention of the compounds in the compositions according to the invention can be added to create the viscosity agents, such as hydroxymethylcellulose, hydroxyethylcellulose, methylcellulose, polyvinylpyrrolidone, or the like
To prepare sterile ophthalmic composition in the form of ointments antagonist of c-Maf combined with a preservative in an appropriate medium such as the miner is inoe oil, liquid lanolin, or white petrolatum. Sterile ophthalmic composition can be prepared by suspendirovanie antagonist of c-Maf in the hydrophilic base prepared, for example, the combination of the
CARBOPOL®-940 (BF Goodrich, Charlotte, NC) or the like, in accordance with methods known in this field for other ophthalmic compositions. VISCOAT ® (Alcon Laboratories, Inc. Fort Worth, TX) can be used, for example, for intraocular injection. Other compositions of this invention may contain a reinforcing penetration agents, such as Cremaphor and tween® 80 (monolaurate of polyoxyethylenesorbitan, Sigma Aldrich, St. Louis, MO), in the case where the antagonists of c-Maf are less penetrating eyes.
Isolation of RNA from tissues and cells of the trabecular network of the person
Cells of the trabecular network of the person (TM) received from the donor eye (Central Florida Lions Eye and Tissue Bank, Tampa, Fl) and were cultured as described previously (Steely, et al. (1992), Invest Ophtalmol Vis Sci 33(7): 2242-50; Wilson, et al. (1993), Curr Eye Res 12(9): 783-93; Clark, et al. (1994), Invest Ophtalmol Vis Sci 35(1): 281-94; Dickerson, et al. (1998), Exp. Eye Res 66(6): 731-8; Wang, et al. (2001), Mol Vis 7: 89-94). The TM cells were obtained from pools of four, each from a normal cell lines or cell lines of glaucoma. Total RNA was isolated from cells TM each pool using the reagent TRIZOL® according to the manufacturer's instructions (Invitrogen, Carlsbad, CA).
Analysis using Affyetrix GeneChip
Reverse transcription, the synthesis of the second chain cDNA and Biotin tagging RNA was carried out according to standard Affymetrix protocols. Chips genes have also been® U133A and U133B (Affymetrix, Santa Clara, CA) human genome hybridized, washed and scanned according to standard Affymetrix protocols. Hybridisable arrays GENECHIP® scanned using a GENEARRAY®scanner (Agilent Technologies, Palo Alto, CA). Raw data were collected and analyzed using software Affymetrix Microarray Suite.
Filtering data micromission performed using GENESPRING® software (Silicon Genetics, Redwood City, CA). For each experiment, data were normalized on a single chip by dividing each dimension at the 50th percentile of all measurements of the intensities of the signals for this chip. The ratio of expression for each gene was calculated by dividing the normalized signal for a gene in the treated or pathological sample at the median for this gene in the control sample for each experiment. Genes were selected on the expression level higher than a statistical background, using the Cross-Gene Error Model and set a background equal to the proportion of unique basis for each experiment. For the analysis we considered only genes that were marked as present/marginal on GENECHIP® U133A Affymetrix in all experimental in the circumstances. Gene short form c-Maf is represented only once on the GENECHIP® U133A in the form of a set of probes 209348_s_at. Gene short form c-Maf expressively at least two times more in case of illness or treatment in comparison with control conditions.
cDNA first chain was generated from 1 µg of total RNA using random hexamers and Reverse Transcription reagents TAQMAN ® according to the manufacturer's instructions (Applied Biosystems, Foster City, CA). Then the reaction mixture of 100 μl was diluted 20 times with obtaining efficient cDNA concentration of 0.5 ng/μl.
Measurement of gene expression of short form c-Maf were performed using quantitative RT-PCR real-time (QPCR) using system sequence detection ABI PRISM® 7700 (Applied Biosystems)essentially as described Shepard, et al. (2001) Invest Ophtalmol Vis Sci 42(13): 3173-81. Primers for specific against short forms amplification of c-Maf (GenBank accession #AF055376) were designed using the program PRIMER EXPRESS® (Applied Biosystems). Sequences of forward and reverse primers were TTGGGACTGAATTGCACTAAGATATAA, SEQ ID NO:1 (nucleotides 3773-3799) and GCGTTCTAAACAGTTTTGCAATTTT, SEQ ID NO:2 (nucleotides 3823-3847) and the sequence of probe binding minor groove was CTGCAAGCATATAATACA, SEQ ID NO:3 (nucleotides 3801-3818). 6 F linked to the 5'-end of the probe binding minor groove, and he is the type of flu is refore, attached to the probe TAQMAN®. Another possible fluorophore fluorophore is JOE™ (Applied Biosystems) or a fluorophore VIC™ (Applied Biosystems). "Binding minor groove afluorescent quencher" was linked to the 3'end of the probe and used to absorb fluorescence from 6FAM. Amplification of c-Maf-amplicon 75 gel were normalized relative to the levels of 18S rRNA using 1x pre-obtained set of primer 18S rRNA/probe (20x 18S MASTER MIX®; (Applied Biosystems). QPCR c-Maf consisted of Universal Mixture of 1x TAQMAN® (Applied Biosystems), concentrations of 900 nm and 100 nm primers and probes, respectively, and 2.5 ng of cDNA in a final volume of 50 µl. Conditions of thermal cycles consisted of 50°C, 2 min, 95°C 10 min followed by 40 cycles at 95°C, 15 s, 60°C, 1 min. Quantitative determination of the concentration of cDNA was performed using the method of obtaining the curves of relative standards described in PE Biosystems User Bulletin #2, ABI PRISM® 7700 Sequence Detection System, 2001 (Applied Biosystems) and MS Excel 97 (Microsoft). Reference total RNA human (Stratagene, La Jolla, CA) was used to obtain the curve standards. Data quantitative PCR (QPCR) are presented as mean values ± SEM of c-Maf/18S-normalized relations.
TGFβ2-induced expression of c-Maf in the cells of the trabecular network
An example using quantitative PCR analysis demonstrates that the short form c-Maf gender differential is positive regulated in induced transforming growth factor beta 2 glaucomatous cells.
Gene expression of short form c-Maf were analyzed using Affymetrix U133 GENECHIP®analysis described in example 2, pool glaucomatous cells of the trabecular network, called SGTM2697. These glaucomatous cells were treated for 16 hours with 5 ng/ml transforming growth factor beta 2 (TGFβ2) for the induction of gene expression. Gene expression of short form c-Maf identified as positively regulated. Proof positive regulation of c-Maf was performed using QPCR as described in example 3, using cDNA derived from RNA of the United ± TGFβ32-treated cells SGTM2697 used for Affymetrix NIP®analysis. Short form c-Maf positively regulated with TGFβ2 increased 16-fold in comparison with control, as shown in figure 1. The data of figure 1 are presented as normalized ratio of c-Maf to the levels of ribosomal 18S mRNA (Mean ± SEM (standard error of the mean), n=3)).
Dexamethasone-induced expression of c-Maf in the cells of the trabecular network
This is an example using quantitative PCR analysis demonstrates that the short form c-Maf differential is positively regulated in induced by dexamethasone glaucoma cells.
Gene expression of short form c-Maf were analyzed using Affymetrix U133 GNI®analysis described in example 2, for locomotory cells of the trabecular network, named TMA. These cells were treated for 1 day or 14 days 10-7M dexamethasone (Dex). Gene expression of short form c-Maf identified as positively regulated. Proof positive regulation of c-Maf was performed using QPCR as described in example 3, using cDNA derived from RNA ± Dex-treated cells TMA used for Affymetrix GNI®analysis. Short form c-Maf positively regulated TGFβ2 with increase in 2 times a day 1 and 3.2-fold at day 14, Dex treatment compared to the control, as shown in figure 2. Data are presented as normalized ratio of c-Maf to the levels of 18S ribosomal RNA (Mean ± SEM (standard error of the mean), n=3).
Inhibition by a small molecule basal and TGFβ2-induced gene expression of c-Maf in the cells of the trabecular network
This example demonstrates that the cdk2 inhibitor is an antagonist of the expression of the short form c-Maf.
The action of inhibiting small molecule for the expression of short form c-Maf were analyzed using QPCR analysis as described in example 2, in glaucomatous cells of the trabecular network (6th passage), named SGTN2697. These cells were treated with 5 ng/ml GFβ2 or without TGFβ2 and inhibitor complex cdk2/cyclin a by purvalanol And for 16 hours (Hardcastle, et al. (2002) Annu Rev Pharmacol Toxicol 42: 325-348). Basal levels of c-Maf negative is regulated 2.6 times the processing purvalanol And, as shown in figure 3. TGFβ2-treated c-Maf (positive adjustable 17 times) were completely eliminated by simultaneous treatment with purvalanol And, as shown in figure 3. The data of figure 3 are presented as normalized ratio of c-Maf to the levels of 18S ribosomal RNA (Mean ± SEM (standard error of the mean)), n=6). the y-axis of figure 3 has a lower scale of 0.00-0.03 and the upper scale of 0.08-0,48.
As confirmed by the inhibition of purvalanol And gene expression of short form c-Maf, presented above, this invention provides additional inhibitors of cyclin-dependent kinase 2, as described herein, for use as antagonists of the expression of the short form c-Maf. Such antagonists applicable as a preventive or therapeutic agents for protection from damage or treatment of damage caused by the disease process glaucoma.
Short form of the transcription factor c-Maf in glaucomatous tissue of the optic nerve
Short form of the transcription factor c-Maf is present in elevated levels in glaucomatous tissue of the optic nerve compared to normal tissue of the optic nerve, as shown using the Affymetrix GENECHIP® microarray analysis. The fabric of the optic disc were obtained from pools of eyes or four normal, or five glaucomatous donors. Total RNA separation is of fabric disks of optic nerves using the reagent TRIZOL® according to the manufacturer's instructions (Invitrogen). Expression of c-Maf short form under these conditions additionally indicates a causal or effector role on the part of this factor in the pathogenesis of glaucoma. Provided antagonism in relation to the expression and/or activity of the transcription factor c-Maf short form for the inhibition or attenuation of the pathogenesis of glaucoma and to ensure neurotoxity for retina and optic nerve.
Cited here are links to the extent that they provide examples of procedures or other details supplementary to the examples presented here, given here as a reference.
Specialists in this field will understand that can be made obvious modifications described herein of embodiments without deviating from the idea and scope of this invention. All options described here implement can be made and executed without undue experimentation in light of this description. The full scope of the present invention presented in this description and its equivalent variants of implementation. This description should not be construed as unnecessarily narrowing the full amount of protection that is right this invention.
1. The method of suppressing positive regulation of short forms of the transcription factor c-Maf in the treated with steroids or transform the dominant growth factor β2 (TGF β2) cells of the trabecular network (TM), including the introduction of an antagonist of c-Maf, where specified antagonist possesses inhibitory activity against cyclin-dependent kinase cdk2.
2. The method of suppressing positive regulation of short forms of the transcription factor c-Maf in the treated with steroids or transforming growth factor β2 (TGF β2) cells of the trabecular network (TM), including the introduction of the inhibitor of cyclin-dependent kinase cdk2.
3. The method according to claim 1 or 2, where the positive regulation of short forms of the transcription factor c-Maf causes primary open-angle and steroid glaucoma.
4. The method according to claim 2, where the inhibitor of cyclin-dependent kinase cdk2 is purvalanol A.
SUBSTANCE: invention relates to novel compounds of general formula (I) , in which A is selected from one or several X and/or Y groups; X represents methylene group; Y represents C2-alkinylene group; n represent integer number from 1 to 5; R1 represents group R2, optionally substituted with one or several R3 and/or R4 groups; R2 represents group selected from pyridinyl, pyrimidinyl, pyridazinyl, imidazolyl, oxazolyl, pyrazolyl, isoxazolyl, oxadiazolyl, naphtyl, chinolinyl, isochinolinyl, dihydroisochinolinyl, 2-oxo-3,4-dihydrochinolinyl, indolyl, benzimidazolyl, pyrrolopyridinyl; R3 represents group selected from halogen atoms, groups C1-6-alkyl, C3-7-Cycloalkyl, C1-6-alkoxy, NR5R6 and phenyl; R4 represents group selected from groups: phenyl, naphtyl, pyridinyl; R4 group or groups can be substituted with one or several R3 groups, similar or different from each other; R5 and R6 independently on each other represent C1-6-alkyl group; R7 represents hydrogen atom or C1-6-alkyl group; R8 represents hydrogen atom or group C1-6-alkyl, C3-7-cycloalkyl, C3-7-Cycloalkyl- C1-3-alkylene; in form of base, acid-additive salt, hydrate or solvate. Invention also relates to methods of obtaining formula (I) compound by any of ii. 1-3, to compounds, determined by general formula (IV), (VII), to pharmaceutical composition, as well as to application of formula (I) compounds by any of ii. 1-3.
EFFECT: obtaining novel biologically active compounds possessing activity of enzyme FAAH inhibitors.
10 cl, 5 ex, 1 tbl
SUBSTANCE: invention relates to medicine, particularly to ophthalmology. Invented is a composition for reducing intraocular pressure, treating glaucoma or ocular hypertension, containing from 0.005 to 0.02 wt %/ bimatoprost mass and from 100 mln-1 to 250 mln-1 benzalkonium chloride, where the said composition is a water based liquid, with content suitable for application into eyes. Invented also is a method which is useful in treating glaucoma or related ocular hypertension.
EFFECT: reduction of active component in the composition while retaining effectiveness of the composition and design of a method of its application for reduction of intraocular pressure when treating glaucoma or ocular hypertension.
17 cl, 5 ex, 2 dwg
SUBSTANCE: invention relates to substituted amides of benzoic acid of formula (I), their isomers and salts as inhibitors of KDR and FLT tyrosine kinase, as well as to medication based on them. Compounds can find application as medications for treating diseases caused by persisting angiogenesis. In general formula (I) (I), A represents group =NH, W represents oxygen, Z represents bond or branched or non-branched C1-C12alkyl, R1 represents branched or non-branched C1-C6alkyl or optionally single- or multi-substituted with C1-C6alkyl C3-C10cycloalkyl, or unsubstituted or optionally single- or multisubstituted with halogen, C1-C6alkyl, single- or multisubstituted with halogen C1-C6alkyl phenyl, naphtyl, quinolyl, isochinolyl, indenyl, R2 and R3represent hydrogen, OH-group or group XR11, X represents C2-C6alkyl, C2-C6alkenyl or C2-C6alkinyl, R11 represents unsubstituted or optionally single- or multisubstituted with halogen, C1-C6alkoxygroup or hydroxy group phenyl ot pyridyl, R4, R5, R6 and R7represent hydrogen, R2 and R3 simultaneously do not represent hydrogen, and if R7 represents OH-group, R3 does not represent hydrogen, and if R3 represents OH-group, R2 does not represent hydrogen.
EFFECT: obtaining compounds which can be applied as medications for treating diseases caused by persisting angiogenesis.
3 cl, 160 ex
SUBSTANCE: invention concerns ophthalmology and can be used for improvement of visual functions at primary open angle glaucoma with the normalised intraocular tension. Parabulbar introduction of the 5% Mexidol solution is performed within 10-12 days. In addition nimodipine is administered intravenously in a dose of 10 mg once a day within 10-12 days. Noopept is administered perorally in a dose of 10 mg 2 times a day within 1 month.
EFFECT: method allows strengthening antioxidising activity and circulation in eye tissues, increasing regenerating effect of nervous fibers.
8 tbl, 4 ex
SUBSTANCE: invention concerns medicine, namely to ophthalmology. It is offered to use a composition containing, at least one agonist BMP-4 for treatment of glaucoma. Thus administer the composition with immediate delivery into an eye of the patient, for example using local eye drops; an oculentum; devices of the slowed down release implanted into a cul-de-sac of an eye either about a sclera of an eye or in an eye; by means of an injection. Concentration of BMP-4 in the composition preferably makes from 0.01 to 2%. The method is based on property of BMP-4 to reduce an ophthalmotonus.
EFFECT: creation of a composition for glaucoma treatment.
8 cl, 15 dwg, 2 tbl, 1 ex
FIELD: medicine; ophthalmology.
SUBSTANCE: cytostatic agent is subconjuctive introduced. Agent introduced is added with 0.08% trypan blue solution in amount 0.05 ml. Introduced agent localisation is visualised. In case agent becomes localized in anterior chamber or escapes through conjunctive suture introduction is stopped. In case agent spreads under conjunctiva dose in required amount is introduced.
EFFECT: lower rate of complications caused by cytostatics introduction.
FIELD: medicine; ophthalmology.
SUBSTANCE: carbonic anhydrase inhibitor is instilled. One day after medicamentous therapy started. Azopt therapy is combined with vasotonic massage of anterior ciliary arteries (ACA) that is transconjunctiva ACA effect using multiple alternation of their mechanical compression and decompression by vasotonometer.
EFFECT: higher efficiency of treatment of primary open-angle glaucoma and blood flow improvement of eye great vessel.
1 tbl, 1 ex
SUBSTANCE: Invention relates to ophthalmology, and can be used for treatment of glaucoma optic neuropathy. Dibicor in dose 0.5g two times a day during 1 month, then cortexin and retinalamin in dose 10 mg intramuscular every second day during 20 days are prescribed. Method allows to increase electrophysiological activity of visual analyser in patients with primary open-angular glaucoma due to pathogenically grounded successive application of said medications, on the first stage of treatment intracellular exchange in ganglionar cells of retina improves, then synaptic lesions and transmission of nerve impulse from retina to cortical centre of visual analyser are activated.
EFFECT: increase of electrophysiological activity of visual analyser in patients with primary open-angular glaucoma.
1 tbl, 1 ex
SUBSTANCE: method involves introducing anecortavi acetas at a dose of 3-15 mg in posterior juxtascleral injection or injection into vitreous body or as implant.
EFFECT: enhanced effectiveness of treatment.
18 cl, 5 dwg, 4 tbl
FIELD: organic chemistry, medicine, biochemistry pharmacy.
SUBSTANCE: invention relates to using the known 2-phenyl-substituted imidazotriazinone of the formula (I): (vardenafil) possessing improved properties as compared with other known inhibitors of phosphodiesterase-5 (PDE-5), such as sildenafil and tadalafil. Proposed compound is used for preparing drugs used in treatment of cardiac insufficiency.
EFFECT: valuable medicinal and biochemical properties of compound.
SUBSTANCE: invention relates to hepatic fibrosis inhibitor containing pyrazolopyrimidinon derivative of the following formula (1) as active ingredient, pharmaceutical composition for liver cirrhosis treatment and prevention, portal hypertension inhibitor comprising pyrazolopyrimidinon formula derivative (1), pharmaceutical composition as prevention care for treatment of complications caused by portal hypertension and containing pyrazolopyrimidinon formula (1) derivative as active ingredient.
EFFECT: improved medical compliance in patients suffering from liver diseases.
SUBSTANCE: therapy of the patients suffering from chronic lower limb ischemia involves sampling the autogenic blood cell mass in amount 6-8% of total blood volume by discrete plasmapheresis. Then the autogenic cell mass is incubated with pentoxifylline 10 ml during 20 min at room temperature with ATP 2 ml added. The prepared mass is reinfused to the patient. Thereafter, Vasaprostan is introduced in a dose 40 mkg/day with physiologic saline 200 ml as intravenous infusion during 3 hours. Therapeutic course is 10-15 daily procedures.
EFFECT: higher clinical effectiveness with regard to the pathology due to correction of hemodynamic disorders in an ischemic focus and improvement of rheological properties blood in the absence of absolute indications to surgical procedure or inability thereof.
SUBSTANCE: invention refers to pharmaceutical compound to improve renal function that contains KW-3902 30 mg or its pharmaceutically acceptable salt, ester, amide, and furosemide. Also there are disclosed method to induce diuretic effect, method to improve renal function, method to support renal function and method to recover renal function in a patient.
EFFECT: effective recovery of renal function in patients with congestive heart failure.
13 cl, 2 tbl, 7 ex
SUBSTANCE: medicinal agent contains alpha and/or beta and/or gamma human recombinant interferon, tocopherol acetate or other tocopherol derivatives, ascorbic acid and/or its salts, pantothenic acid or calcium pantothenate, or dexapanthenol, riboflavin or levocarnitine, and orotic acid and/or ornithine or its derivative - citrulline malate, or Glutoxim, antibacterial, antimycotic agents, additives: emulsifiers, stabilisers, preservatives, antioxidants and base. The formulation is suppositories in certain component ratio.
EFFECT: efficiency for severe infectious diseases and mixed infections.
9 cl, 18 ex
SUBSTANCE: invention refers to medicine, namely to immunology and otorhinolaryngology and can be used for treatment of adenoid hypertrophy and chronic adenoiditis. Laboratory examination is combined with the relevant antiviral therapy. And the antiviral therapy involves treatment of all children with viricides as combined with conventional classical local treatment. Herewith considering age and examination data, there are prescribed either Acyclovir taken 0.2 g 4 times a day during 10 days, Viferon 1-150000 UN 2 suppositories daily every twelve hours during 7 days, then two suppositories a day every twelve hours three times a week within 14 days; or Valcyclovir (Veltrex) 0.25 g twice a day every twelve hours during 10 days; or Famcyclovir (Famvir) 0.25 g three times a day every eight hours during 10 days. The local treatment involves nasal toilet with 5% aminocapronic acid three times a day 3-5 ml in each nasal passage within the course 10 days.
EFFECT: method provides higher clinical effectiveness for adenoid hypertrophy in children.
SUBSTANCE: invention relates to medicine, namely to stomatology, and can be used for treatment of herpetic stomatitis. For this purpose lesion focuses of mouth mucosa are cooled by rolling on them roller from permeably-porous titanium nickelide, treated with liquid nitrogen. Rolling is performed without pressing, in reciprocal mode during 2-3 seconds. Then on said focuses Acyclovir ointment is applied.
EFFECT: reduction of terms of disease treatment to 2-3 days due to combined effect of deep cooling of focuses and anti-virus impact of Acyclovir ointment.
2 ex, 3 dwg
SUBSTANCE: present invention refers to substituted 8-heteroarylzantines of general formula where R represents hydrogen, (C1-C5)alkyl or halogen(C1-C8)alkyl; R1 is chosen from (C3-C6)cycloalkyl or (C3-C6)cycloalkyl(C1-C4)alkyl-; R2 is chosen from (C1-C8)alkyl, (C3-C8)alkenyl, (C3-C8)alkinyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl(C1-C8)alkyl- or (C6-C10)aryl(C1-C8)alkyl-; X represents 3-pyridyl substituted in 6th position with Z; Z represents -NR4R5 or (C4-C10)heterocycle where heterocycle is optionally substituted with 1, 2, 3 or 4 substitutes independently chosen from (C1-C8)alkyl; each Z1 independently represents halogen or -NR7R8; R5 is chosen from -C(O)R6, -CO2R6 or -C(O)NHR7; R4 is chosen from hydrogen, (C1-C8)alkyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl(C1-C8)alkyl-, (C3-C10)heterocycle(C1-C8)alkyl-, (C6-C10)aryl, (C6-C10)aryl(C1-C8)alkyl-, (C5-C10)heteroaryl, (C5-C10)heteroaryl(C1-C8)alkyl-, -((CH2)2-4)Y)q-(CH2)2-4-X1, -C(O)R6, -CO2R6 or -C(O)NR7R8; or R4 and R5 together with atoms whereto attached form saturated mono-or bicyclic ring with 5, 6, 7 or 8 ring atoms and optionally containing 1 or 2 heteroatoms chosen of non-peroxide oxy (-0-) and amine -N(R9)- in the ring where the ring is optionally substituted by 1, 2, 3 or 4 substitutes independently chosen from -C(O)Ra and -C(O)NRbRc; X1 represents -OR6; and Y represents oxy (-O-); where alkyl, alkenyl, cycloalkyl, alkinyl, aryl, heterocyclic or hetero aryl groups from R1, R2, R3, R4 and R5 groups are optionally substituted by one or more substitutes independently chosen from (C1-C8)alkyl, -ORa, (C6-C10)aryl, hydroxy(C1-C8)alkyl and RbRcN(C1-C8)alkyl; where R6 represents (C1-C8)alkyl or (C4-C10)heteroaryl; where heteroaryl is optionally substituted by 1, 2, 3 or 4 substitutes independently chosen from halogen, -ORa and halogen(C1-C8)alkyl; where R7, R8 and R9 independently represent (C1-C8)alkyl, RaO(C1-C8)alkyl, (C6-C10)aryl or (C4-C10)heteroaryl; where heteroaryl or aryl are optionally substituted by 1, 2, 3 or 4 substitutes independently chosen from halogen and -ORa; Ra represents hydrogen or (C1-C6)alkyl; each Rb and Rc independently represents hydrogen or (C6-C10)aryl; and where n is equal to 0, 1 or 2; and q is equal to 1; or its pharmaceutically acceptable salt. In addition, the invention concerns pharmaceutical composition based on compound of formula I.
EFFECT: new substituted 8-heteroarylxantines are selective antagonists of A2B adenosine receptors.
38 cl, 1 tbl, 1 ex
SUBSTANCE: invention refers to chemical-pharmaceutical industry, medicine and cosmetology, and concerns an agent for liver pathologies, as well as for improved skin colour and structure, lassitude relief, higher immune protection function, intensified sexual function. The composition contains Laennec, a solution containing ammonium glycyrrhizinate, glycine, L cysteine, 20% dextrose solution ("ничифарген"), vitamin B1, B2, B6, B12, C solutions, physiologic saline. Another version of the composition contains Laennec, 20% dextrose solution, vitamin B1, B2, B6, B12, C solutions, physiologic saline. The compositions are intravenously droplet introduced while providing complex action practically without by-effects.
EFFECT: composition effectiveness is ensured with synergetic action of all the components.
2 cl, 9 ex
SUBSTANCE: invention refers to new compounds of formula I where R1 stands for H, CN, halogen, -COR2, -S(O)xR2, C1-C12alkyl, C2-C12alkenyl, C3-C8dicloalkyl, aryl group, heteroaryl group standing for 5- or 6-merous aromatic mono- or bicyclic heterocyclic group with 1-2 heteroatoms, chosen of N or S, C3-C8cycloalkyl-(C1-C3)alkyl or group aril-(C1-C3)alkyl; alkyl, alkenyl, cycloalkyl, aryl and heteroaryl groups can be optionally substituted with halogen, C1-C6alkyl, group-COR2; R2 stands for -N(R3,R3'), C1-C6alkyl, C3-C8cycloalkyl, aryl, heteroaryl which stands for 5- or 6-merous aromatic mono- or bicyclic heterocyclic group with 1-2 heteroatoms chosen of N, C3-C8cycloalkyl-(C1-C3) alkyl or aril-(C1-C3)alkyl; C1-C6alkyl, C3-C8cycloalkyl, aryl, heteroaryl can be optionally substituted with halogen, C1-C6alkyl; R3 and R3' independently stands for hydrogen or (C1-C3)alkyl; x stands for 0, 1 or 2; and also to their esters, hydrolyzed in physiological environment, and to their pharmaceutically acceptable salts. The invention also concerns a medical product.
EFFECT: production of new biologically active compounds active as COMT inhibitor.
17 cl, 19 ex, 1 tbl
SUBSTANCE: invention relates to field of medicine, in particular to operative gynecology and concerns prevention of complications in hysteromyoma patient in case of hysterectomy. For this purpose during 14 days before and 14 days after operation mediation selmevit is introduced perorally. Dose is 1 tablet a day.
EFFECT: performing supravaginal amputation or uterus extirpation against the background of chronic posthemorrhagic anemia, reduction of intraoperative hemorrhage and prevention of DIC syndrome development.
3 tbl, 2 ex
SUBSTANCE: invention relates to new a compound of formula I or formula II, or to its pharmaceutically acceptable salts, I II, where X is S; R1 is H or C1-C6alkyl; R2 is NR5R6; R3 is aryl, substituted with a halogen; R4 is H; R5 is H; R6 is H; R7 is CH2NR8R9 where R8 is H, C1-C10alkyl, C3-C8cycloalkyl, aryl, aryl(C1-C6alkyl), aryl(C2-C6alkenyl), heterocycle(C1-C6alkyl), heterocycle(C2-C6alkenyl), hydroxyl(C1-C6alkyl), hydroxyl(C2-C6alkyl), C1-C6alkoxycarbonyl, aryl(C1-C6alkoxy)carbonyl, carbamoyl(C1-C6alkyl); where the above mentioned aryl is an aromatic ring and is not substituted or substituted with one to three substituting groups, each of which, independently from the others, is chosen from: methylenedioxy, hydroxy, C1-C6-alkoxy, halogen, C1-C6alkyl, trifluoromethyl, trifluoromethoxy, NO2, NH2, NH(C1-C6alkyl), N(C1-C6alkyl)2, NH-acyl, N(C1-C6alkyl)-acyl, hydroxy(C1-C6alkyl), dihydroxy(C1-C6alkyl), CN, C(=O)O(C1-C6alkyl), phenyl, phenyl(C1-C6alkyl), phenyl(C1-C6alkenyl), phenoxy and phenyl(C1-C6alkoxy), R9 is H, C1-C10alkyl, heterocycle(C1-C6alkyl) or heterocycle(C2-C6alkenyl); where the above mentioned heterocycle represents a 5-member saturated monocyclic ring system, consisting of carbon atoms, as well as heteroatoms, chosen from a group comprising N, O, and S, which can be unsubstituted or have one to three substituting groups, independently chosen from a list which includes NO2, aryl(C1-C6alkyl), arylsulphonyl; or R8 and R9 together with nitrogen, to which they are bonded, form a heterocycle, which represents a 5 - 7-member saturated monocyclic ring system, consisting of carbon atoms, as well as one to three heteroatoms, chosen from a group comprising N, O and S, which can be unsubstituted or have one to three substituting groups, independently chosen from a list which includes C1-C6alkoxy, hydroxy, C1-C6alkyl, C2-C6-alkenyl, C(=O)O(C1-C6alkyl), C(=O)NH2, C(=O)NH(C1-C6alkyl), C(=O)N(C1-C6-alkyl)2, hydroxy(C1-C6alkyl), dihydroxy(C2-C6alkyl), aryl, aryl(C1-C6alkyl), aryl(C2-C6alkenyl), aryl(C1-C6alkoxy) and pyrimidin-2-yl; and m equals 0. The invention also relates to a pharmaceutical composition, as well as to use of formula I or formula II compounds.
EFFECT: obtaining new biologically active compounds, with inhibitory properties towards casein kinase 1ε.
32 cl, 3 tbl