Method of treating diseases by vegf antagonists
SUBSTANCE: invention refers to medicine and concerns methods of treating diseases with using VEGF antagonists. Substance of the invention involves application of a VEGF antagonist containing VEGFR1R2-FcΔ C1 (a) SEQ ID NO:4 in preparing a drug for hypertension reduction, in treating the diseases associated with administering the VEGF antagonist where the treatment is conducted by subcutaneous introduction.
EFFECT: advantage of the invention consists in reducing side effects associated with treating the diseases by administering the VEGF antagonist.
8 cl, 3 ex, 1 tbl
The level of technology
The technical field to which the invention relates
The field of invention relates to therapeutic methods for treating diseases people are antagonists of growth factor vascular endothelial (VEGF), which minimizes side effects such as high blood pressure. This method is intended for the treatment of patients in which it is desirable to minimize the rise in blood pressure.
Description of the prior art
Growth factor vascular endothelial (VEGF) is recognized as the main factor stimulating angiogenesis under pathological conditions. Approaches to methods of blocking VEGF include construction of soluble receptors, antisense molecules, RNA aptamers and antibodies. See, for example, PCT WO/0075319 to describe antagonists trap based VEGF receptors.
In patients receiving anti-VEGF humanitariannet monoclonal antibody, bevacizumab, and hypertension were more frequent and more severe (Hurwitz, et al, (2004) N. Engl. J. Med. 350:2335-42).
Brief description of the invention
On the one hand, the invention relates to a method of lowering elevated blood pressure associated with the introduction of the antagonist of the growth factor vascular endothelial (VEGF), including subcutaneous administration of a VEGF antagonist to the human, which it is desirable to minimize the increase krovenapolnenia.
More specifically, the studies described below indicate that systolic and diastolic blood pressure associated with intravenous administration of VEGF antagonists, significantly reduced after subcutaneous administration. The method of the invention is particularly suitable for patients where it is desirable to prevent hypertension.
In the method of the invention can be applied to any antagonist of VEGF, which is associated with high blood pressure when administered to the patient. In one embodiment, the VEGF antagonist is Vysocany dimer fused protein (or "trap"), including fused polypeptide having an immunoglobulin-like (Ig) domain 2 of the VEGF receptor Flt1 and Ig domain 3 of the VEGF receptor Flk1 or Flt4, and multimediali component. More specifically, the VEGF antagonist comprises fused polypeptide selected from the group consisting of Flt1D2.Flk1D3.FcΔC1 (a) (SEQ ID NOs:1-2), VEGFR1R2-FcΔC1 (a) (SEQ ID NOs:3-4), or its functional equivalent. Functionally equivalent molecules include dimeric proteins consisting of two fused polypeptides that are expressed in cells of the host mammal and contain post-translational modifications such as glycosylation, truncation of C-terminal lysine and/or signal peptide, etc.
In the first aspect of the invention "immune" patient is subjected to subcutaneous treatment is conducting an antagonist of the growth factor vascular endothelial (VEGF) in therapeutically effective amounts, and the re-introduction after a therapeutically effective period of time. In accordance with the present invention "immune" to include individuals in need of treatment with VEGF antagonists, but in those cases, when the patient was not allowed to enter a sufficient amount of a VEGF antagonist intravenously, so that the drug was effective, because this introduction led to undesirable rise in blood pressure of the patient. Accordingly, such impervious include those who initially suffer from high blood pressure, which was not sufficiently regulated, so that the increase in blood pressure creates a medical risk to the patient and, in addition, include those patients with normal blood pressure or blood pressure, adjustable within normal levels, which when treated with VEGF antagonist blood pressure rises to levels at which there may be health risks for the patient.
The method according to the invention covers disease and/or condition, or relapses, which are improved, inhibited or reduced by treatment with a VEGF inhibitor. Such conditions include, for example, cancer, diabetes, vascular permeability, edema or inflammation, such as swelling of the brain associated with trauma, stroke, or tumor, edema associated vospalitelnye disorders, such as psoriasis or arthritis, asthma, edema associated with burns, ascites and pleural effusion associated with tumors, inflammation or trauma, chronic airway inflammation, leaky capillaries, sepsis, kidney, associated with increased leakage of protein, vision disorders such as age-related degeneration yellow spots and diabetic retinopathy, abnormal angiogenesis, such as disease polycystic ovaries, endometriosis and endometrial cancer. The VEGF inhibitor can also be used to induce regression or reduce the size of an existing tumor or metastatic cancer, diabetes, decreasing the formation of new blood vessels in tumors, improve time to engraftment of the transplanted cornea, inhibiting rejection of transplanted corneal or lymphangiogenesis and angiogenesis in the cornea.
A patient who undergoes treatment, is patient with one of the above States, who suffers from hypertension at risk of developing high blood pressure or for which the desired prevention or suppression of hypertension, for example the patient at risk for cardiovascular disease, the patient is over 65 years of age or patient, which may not be subjected to treatment with a suitable dose of a VEGF antagonist in any way without the development of hypertension.
During the second aspect, the invention relates to a method of preventing the development of hypertension during treatment with the inhibitor of growth factor vascular endothelial (VEGF) in patients at risk of developing high blood pressure, including the introduction of a VEGF antagonist to the patient by subcutaneous injection.
Other objectives and advantages will become clear from a review in the following detailed description.
Before describing these methods and compositions should be noted that the present invention is not limited to the described specific ways and experimental conditions, since methods and conditions may vary. It should also be noted that the terminology used here is intended only to describe the specific options and is not intended to be limiting, since the scope of claims of the present invention will be limited only by the attached claims
As used in this description and the accompanying claims, the singular number also include the plural, if the context clearly dictates otherwise. For example, "method" means one or more methods, and/or the steps described here type and/or which will become apparent to the person skilled in the art after reading this description, and the like.
If not specified in the che, all technical and scientific terms used in this description have the same meaning as commonly understood by a person skilled in the art to which this invention relates. Although any methods and materials similar or equivalent to those described in the present description, can be used in practice or testing of the present invention, the preferred methods and materials are now described.
In normal mammalian blood pressure is tightly regulated by a complex system of physiological factors. This is important for survival because of high blood pressure (hypertension) can lead to a number of undesirable health events and conditions, such as, for example, stroke, acute coronary syndrome, myocardial infarction and renal failure. Studies show that VEGF temporarily dilates coronary arteriesin vitro(Ku et. al. (1993) Am J Physiol 265:H585-H592) and causes hypertension (Yang et. al. (1996) J Cardiovasc Pharmacol 27:838-844). Known methods of treatment of eclampsia and pre-eclampsia, such as the publication of the patent application U.S. 2003/0220262, WO 98/28006, WO 00/13703, which describes a method of treating hypertension comprising administration to a patient an effective amount of an angiogenic factor such as VEGF or its antagonist. Publication of the patent application US 2003/0144298 shows that the introduction is the high levels of tyrosine kinase inhibitor of VEGF-receptor leads to a sustainable increase in blood pressure with a long reception.
Antagonists of VEGF and VEGF-specific fused polypeptide traps
The method according to the invention can be used with any antagonist of VEGF, which is associated with high blood pressure when administered to the patient. In a preferred embodiment, the VEGF antagonist is a dimeric protein that can bind VEGF with high affinity, consisting of two fused polypeptide receptor-Fc fusions, consisting mainly of the ligand-binding portions of the extracellular domains of the receptor VEGFR1 and VEGFR2 person, merged with Fc part of IgG1 (“VEGF-trap). Specifically, VEGF-trap consists of a domain 2Ig of VEGFR1, which merged with the domain 3Ig of VEGFR2, which in turn merged with the Fc domain of IgG1.
In a preferred embodiment of the invention expressing a plasmid that encodes a VEGF-trap, transfairusa CHO cells, which secrete VEGF-trap in the cultural environment. The resulting VEGF-trap is a dimeric glycoprotein with a molecular mass of 97 kDa and contains ~15% of glycosylation, leading to a total molecular mass of 115 kDa. Polypeptides merge, forming a dimer, excision are modified by glycosylation at one or more residues Asn and/or removing terminal Lys.
Because VEGF trap binds its ligands, using high-affinity binding domains of cocktail recipes. is s, it has a higher affinity against VEGF than monoclonal antibodies. VEGF trap binds VEGF-A (KD= 1.5 pM), PLGF1 (KD= 1.3 nM) and PLGF2 (KD= 50 pM); the binding of other members of the VEGF family is not yet fully characterized.
A group of diseases
Patients are preferably treated using the described method, are people who have it is desirable to prevent or reduce one or more side effects caused by antiangiogenic treatment means, such as hypertension, proteinuria. Especially preferred this method for those suffering from hypertension, which are more than 65 years of age, or persons for whom the reduction or prevention of unwanted side effects allows the use of optimal therapeutic dose antiangiogenic agent, which otherwise could not be used without the risk of an adverse medical outcome for the patient. Patients suffering from kidney cancer, pancreatic cancer, breast cancer in the later stages, colorectal cancer, malignant mesothelioma, multiple myeloma, ovarian cancer or melanoma can be treated with the combined therapy according to the invention. Disease and/or condition or a recurrence, which improved, inhibited or reduced by treatment with combin, is focused therapy, described here include: cancer, diabetes, vascular permeability, edema or inflammation, such as swelling of the brain associated with trauma, stroke, or tumor, edema associated with inflammatory disorders such as psoriasis or arthritis, asthma, edema associated with burns, ascites and pleural effusion associated with swelling, inflammation or trauma, chronic airway inflammation, leaky capillaries, sepsis, kidney, associated with increased leakage of protein, vision disorders such as age-related degeneration yellow spots and diabetic retinopathy, abnormal angiogenesis, such as disease polycystic ovaries, endometriosis and endometrial cancer. The VEGF inhibitor may also be used to stimulate regression or reduction in size of an existing tumor or metastatic cancer; diabetes, reduce the formation of new blood vessels in tumors, improve the time of engraftment of the transplanted cornea, inhibiting rejection of transplanted corneal or lymphangiogenesis cornea and angiogenesis.
In many embodiments of the invention, the VEGF antagonists can be entered in combination with one or more additional components or therapies, including a second molecule antagonist of VEGF and/or protivogipertonicheskoe the th agent. Combination therapy involves the introduction of a single pharmaceutical dosage form that contains a VEGF antagonist and one or more additional agents; as well as the introduction of a VEGF antagonist and one or more additional agents in their own separate pharmaceutical dosage form. For example, the VEGF antagonist and a cytotoxic agent, chemotherapeutic agent, or growth inhibitor can be administered to the patient together in a single dosage form, such as the combined composition, or each agent can be introduced in a separate dosage form. In the case of separate dosage forms, VEGF-specific protein according to the invention and one or more additional agents can be introduced simultaneously or with the passing of time, that is, sequentially.
The term "cytotoxic agent" used herein refers to a substance that inhibits or blocks the function of cells and/or causes destruction of cells. The term also implies the inclusion of radioactive isotopes (I131I125, Y90and Re186), chemotherapeutic agents, and toxins such as enzymatically active toxins of bacterial, fungal, plant or animal origin, or fragments thereof.
"Chemotherapeutic agent" submitted is a chemical compound that suitable for cancer treatment. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (Cytoxan®); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines, such as benzodepa, carboquone, matureup and uredepa; ethylenimines and methylmelamine including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and triethylenemelamine; nitrogen mustards such as chlorambucil, chlornaphazine, chlorpropamide, estramustine, ifosfamide, mechlorethamine, hydrochloride oxide mechlorethamine, melphalan, novobiocin, finestein, prednimustine, trofosfamide, orally mustard; nitrosoanatabine, such as carmustine, chlorozotocin, fotemustine, lomustin, nimustine, ranimustine; antibiotics such as aclacinomycin, actinomycin, autralian, azaserine, bleomycin, actinomycin, calicheamicin, carubicin, karminomitsin, casinopolis, chromomycin, dactinomycin, daunorubicin, demoralizing, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esrimizin, idarubitsin, marcelain, mitomycin, mycofenolate acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, colomycin, radiobeacon, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolite, such as methotrexate and 5-fluorouracil (5-FU); analogs of folic acid, that is their as deeperin, methotrexate, peripherin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, triadapin, tioguanin; pyrimidine analogues such as, ancitabine, azacytidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens, such as calusterone, propionate dromostanolone, epitiostanol, mepitiostane, testolactone; antiotensin, such as aminoglutetimid, mitotane, trilostane; Supplement of folic acid, such as prolinnova acid; Eagleton; aldophosphamide; aminolevulinic acid; amsacrine; astroball; bisantrene; edatrexate; defaming; demecolcine; diazinon; alternity; elliptinium; etoposide; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitrogen; pentostatin; penomet; pirarubicin; podofillina acid; 2-acylhydrazides; procarbazine; PSK®; razoxane; sizofiran; spirogermanium; tinoisamoa acid; triaziquone; 2, 2', 2"-trihlortrietilamin; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; Galitsin; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxanes, such as paclitaxel (Taxol®, Bristol-Myers Squibb Oncology, Princeton, N.J.) and docetaxel (Taxotere®; Aventis, Antony, France); chlorambucil; gemcitabine; 6-tioguanin; mercaptopurine; methotrexate; platinum analogues, such as cisplatin and carboplatin; winbl the Steen; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; Novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; deformational (DMFO); retinoic acid; espiramicina; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the above. This definition also includes antihormonal agents, which are used for the regulation or inhibition of hormone action on tumors such as antiestrogens, such as tamoxifen, raloxifene, 4(5)-imidazoles that inhibit aromatase, 4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone and toremifene (Fareston); and anti-androgens such as flutamide, nilutamide, bikalutamid, leuprolide, goserelin; and pharmaceutically applicable salts, acids or derivatives of any of the above.
The term "agent that inhibits axonal growth", when used in the present description, refers to the compound or compositions that inhibit the growth of cells, particularly cancer cells, orin vitroorin vivo.Examples of agents inhibiting growth include agents that block the development of the cell cycle (phase other than S phase), such agents that cause a delay in G1-phase and delay of the M-phase. Classic blockers M-phases include periwinkle (winks the Steen and vinblastine), Taxol®and topo II inhibitors such as doxorubicin, epirubicin, daunorubicin, etoposide, and bleomycin. The agents that cause a delay in G1 are in the delayed S-phase, such as DNA elitelimousine agents, such as tamoxifen, prednisone, dacarbazine, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil and Aga-C.
The term "protivogipertonicheskoe agent" when used in this description includes calcium channel blockers, inhibitors of angiotensin-converting enzyme (ACE inhibitors), antagonists of angiotensin II receptors (A-II antagonists, diuretics, β blockers-adrenergicheskih receptors, vasodilator and inhibitor of α-adrenergicheskih receptors.
Calcium channel blockers include amlodipine; bepridil; clentiazem; diltiazem; fendilin; gallopamil; mibefradil; prenilamin; remotedir; terodiline; verapamil; aranidipine; barnidipine; benidipine; cilnidipine; efonidipine; elhadidy; felodipine; isradipine; lacidipine; lercanidipine; manidipine; nicardipine; nifedipine; nilvadipine; nimodipine; nisoldipine; nitrendipin; Cinnarizine; flunarizin; lidoflazine; lomerizine; benzilan; athenon and perhexiline.
Inhibitors of angiotensin-converting enzyme (ACE-inhibitors) include alacepril; benazepril; captopril; ceronapril; delapril; enalapril; fosinopril; imidapril; lisinopril; multiperil; pairing the app; quinapril; ramipril; spirapril; temocapril and trandolapril.
Antagonists of angiotensin II receptor include, but are not limited to: candesartan (US 5196444); eprosartan; irbesartan; losartan and valsartan.
β-blockers include, but are not limited to: acebutolol; alprenolol; amosulalol; arotinolol; atenolol; befunolol; betaxolol; bevantolol; bisoprolol; bopindolol; Bussola; buterol; bufuralol; bunitrolol; bupranolol; buterin hydrochloride; butoverall; carazolol; carteolol; carvedilol; celiprolol; atemolol; koronrodilatatia; epanolol; indenolol; labetalol; levobunolol; mepindolol; metipranolol; metoprolol; moprolol; nadolol; ngoxolo; nebivolol; nipradilol; oxprenolol; penbutolol; pindolol; practolol; pronethalol; propranolol; sotalol; sulfinol; talinolol; tetrathalon; tilisolol; timolol; celiprolol and xianya.
α-blockers include, but are not limited to: amosulalol; arotinolol; dapiprazole; doxazosin; fenspirid; indoramin; labetalol; naftopidil; nicergoline; prazosin; tamsulosin; tolazoline; trimazosin and yohimbine.
Vasodilator include cerebral vasodilator, coronary and peripheral vasodilator vasodilator. Cerebral vasodilator include benzilan; Cinnarizine; citicoline; cyclandelate; eclinical; diisopropylaminoethyl; Bornemann; fasudil; phenoxides; flunarizin; ibudilast; ifenprodil; lomerizine; anafranil; nizamettin; nicergoline; nimodipine; papaverine; tieferen; vincamine; Vinpocetine and vikuiti.
Coronary vasodilator include, but are not limited to: matrifen; bendazol; benfenati hemisuccinate; benziodarone; hloratsizin; chromone; cleanfuel; canitrot; cloakroom; dilazep; dipyridamole; droprenilamine; evoxac; erythritol TETRANITRATE; athenon; fendilin; foradil; gangler; genestra bis(β-diethylaminoethyl)new ether; geksobendin; itrain toilet; chillin; lidoflazine; mannitol hexanitrate; megabasin; nitroglycerin; pentaerythritol TETRANITRATE; pencentra; perhexiline; Pineville; prenilamin; propanil nitrate; trapidil; trisomy; Trimetazidine; trolnitrate phosphate; visnadin.
Peripheral vasodilator include, but are not limited to: aluminum nicotinate; Bamyan; benzilan; betahistine; bradykinin; provincein; bufend; buflomedil; butylamine; cetiedil; cyclonical; cinematic; Cinnarizine; cyclandelate; Diisopropylamine dichloroacetate; eledoisin; phenoxides; flunarizin; Gipronickel; ifenprodil; iloprost; Inositol niacinate; isoxsuprine; kallidin; kallickrein; moxisylyte; anafranil; Nikolett; nicergoline; Nicobarese; nylidrin; plentifully; p is Taxifolin; piribedil; prostaglandin E1; suloctidil; tolazoline and xantinol niacinate.
Diuretics include, but are not limited to, diuretic bestialitymovies derivatives, diuretic organo-mercury compounds, diuretic purines, diuretic steroids, diuretic sulfonamidnuyu derivatives, diuretic orally and other diuretics, such as aminosyn; amiloride; arbutin; chloranil; ethacrynic acid; ettlin; hydrocarbon; isosorbide; mannitol; metalcon; muzolimine; perhexiline; ticrynafen; triamterene; and urea.
The routes of administration
The invention provides compositions and methods of treatment using antagonists of VEGF, which avoid, reduce or eliminate high blood pressure, which is associated with the introduction of a VEGF antagonist. Thus, in the method according to the invention, the VEGF antagonist is administered subcutaneously to a patient in need of such treatment.
The composition is suitable to carry out in practice the method according to the invention may be a liquid containing the agent according to the invention in solution, suspension or both. The term "solution/suspension" refers to a liquid composition, where one portion of the active agent is present in solution and the second portion of the active agent exists as particles in suspension in a liquid matrix. Liquid composition which also includes a gel. The liquid composition may be water or in the form of ointment.
Aqueous suspension or solution/suspension suitable for implementation in practice of the methods according to the invention may contain one or more polymers as suspendida agents. Suitable polymers include water-soluble polymers such as cellulosic polymers and water-insoluble polymers, such as cross-crosslinked carboxyl-containing polymers. Aqueous suspension or solution/suspension according to the present invention is preferably viscous or mucoadhesives or that it is even more preferable, and viscous, and mucoadhesives.
Other features of the invention will become apparent from the further description of illustrative embodiments, which are given to illustrate the invention and not intended to be limiting thereof.
The following example is presented in order to provide specialists in this field a complete disclosure and description of how to make and use the methods and compositions according to the invention, and is not intended to limit the scope of what the authors consider it their invention. Efforts were made to ensure the accuracy in terms of numbers (for example, quantity, temperature, etc.) but some experimental errors and is lonene should be taken into account. Unless otherwise specified, the term part is the term part by weight, the molecular weight is an average molecular weight, temperature is in degrees Celsius and the pressure is close to atmospheric.
Example 1. Subcutaneous administration of the VEGF antagonist
The study 0103. The initial group of 3 patients was introduced subcutaneously 25 mg/kg VEGF antagonist-based receptors ("VEGF-trap) (SEQ ID NO:4) a series of six weekly injections. Four weeks later, 3 additional patients were added to the group and received six weekly injections of 50 µg/kg in four weeks, 3 additional patients were added to the group and received six weekly injections of 50 µg/kg This schedule was repeated, drawing in 3 patients in each group with doses of 100 ág/kg 200 ág/kg 400 ág/kg and 800 mg/kg group was incorporated and received 800 μg/kg twice a week.
To join the group, the patients had to meet the following conditions: (1) the patient with recurrent or refractory solid tumor (non-epidermoid lung cancer), or refractory non-Hodgkin lymphoma in relation to at least two standard chemotherapeutic methods and rituximab and at least one metastasis, measurable, or the remainder of the primary tumor; (2) the ez history of tumors or metastases in the Central nervous system; (3) patients who have failed all medical chemotherapeutic regimes because of their underlying disease and who cannot use the methods of standard chemotherapy, immunotherapy, anti-tumor therapy or radiotherapy; (4) with the proper function of organs; (5) with suitable laboratory parameters, including coagulation profile and samples of kidney function.
The blood pressure in the supine position, and standing was controlled in each research visit in the Protocol. In the case of new hypertension or worsening of previous documented hypertension, blood pressure easily regulated by one or two protivogipertonicheskoe agents.
Exploratory analyses of blood pressure "dose-effect".Group dose level were merged into 4 groups doses, in order to facilitate the use of statistical techniques for the study of potential trends. The combined group dose 1 includes patients who received doses of 25 µg/kg 50 µg/kg and 100 mcg/kg; the combined group doses 2 includes patients who received doses of 200 mg/kg and 400 mg/kg; the combined group doses 3 includes patients receiving 800 mg/kg; and the combined group doses 4 includes patients who received doses of 800 mg/kg twice a week. Using the combined group doses, the difference between groups at each time point was assessed with use what Itanium analysis of variance (ANOVA) as a descriptive method to indicate potential trends. Simple linear regression analysis was also used to test linear trends. Separate analyses were conducted for lying position and a standing position, systolic and diastolic blood pressure.
Systolic pressure in the standing position.For changes in systolic pressure in standing position ANOVA at each time point revealed no dozozavisimoe trends in the increase in systolic pressure in standing position; only on day 3 of the 14 evaluated showed a nominal p-value less than 0.05. The average deviation from the baseline obtained in the first 2 weeks of introduction, are given in table 1.
Diastolic blood pressure in standing position.For changes in diastolic blood pressure in standing position, ANOVA on the mean value at each point again revealed no dozozawisimy trends. None of the 14 interim analyses had nominal p-values lower than 0.05, but 4 of the interim analyses had a nominal p-value between 0.05 and 0.1. The average deviation from the baseline obtained in the first 2 weeks of introduction, are given in table 1.
Example 2. Intravenous administration of a VEGF antagonist
The study 0202.Patients with refractory solid tumors or non-Hodgkin lymphoma who did not receive concurrent treatment in which the compared cancer, was subjected to treatment with VEGF trap (SEQ ID NO:4) according to the following procedure. The initial group of 3 patients received a single dose of 0.3 mg/kg VEGF trap administered intravenously. As a single dose has been well transferred, patients received one additional infusion dose of the same magnitude after a 2-week interval. An additional group of 3 patients received 1.0 mg/kg VEGF-trap intravenously according to the same schedule. This pattern was repeated with groups of 3-6 patients treated at 2.0, 3.0 and 4.0 mg/kg VEGF-trap. Blood pressure and tumor weight was measured at the beginning and the end of the week dosing period; patients with stable disease, partial or complete response, you can continue with the introduction of up to 6 additional months of continuous study. The average change on the 15th day from baseline 0-day are shown for each dose group in table 1.
The study 0305.The second study was conducted in accordance with the methodology of the study 0202. Patients selected for the study had relapsed or refractory solid tumors (other than squamous cell carcinoma of the lung), which was not expected positive effect of the standard treatment, or with non-Hodgkin lymphoma that is resistant to at least two standard chemotherapy regimens and rituximab is, and at least one metastasis or the remainder of the primary tumor measured size, and completed the study 0202 without taking the limiting dose toxicity.
Measurement of systolic and diastolic blood pressure were obtained as described in example 1. The mean change from the baseline obtained in the first 2 weeks of introduction, are given in table 1 for each study group.
Basically, subcutaneous administration of VEGF trap led to an increase in blood pressure of 3.9 to 8.2% at the highest dose. Intravenous led to an increase in blood pressure by more than 20% at the highest dose. At comparable doses while subcutaneous injection of 800 mg/kg VEGF-trap diastolic blood pressure increased by 1.5%, while intravenous 1.0 mg/kg VEGF-trap it grew by 3.7-13.3 per cent.
Results subcutaneous (0103) and intravenous
(0202, 0305) research
|0103||25, 50, 100 ág/kg||200-400 mcg/kg||800 ug/kg||1600 mg/kg|
|0202||0.3 mg/kg||1.0 mg/kg||2.0 mg/kg||3.0 mg/kg||4.0 mg/kg|
|0305||0,0||0.3 mg/kg||1.0 mg/kg||3.0 mg/kg|
1. The use of a VEGF antagonist containing VEGFR1R2-FcΔCl (a) SEQ ID NO:4, in the manufacture of drugs to reduce hypertension related to administration of the VEGF antagonist, where the treatment is subcutaneous injection of people suffering from the disease or condition that can be treated with a VEGF antagonist, in which it is desirable to minimize the rise in blood pressure.
2. The use according to claim 1, where using the optional agent together with a VEGF antagonist.
3. The use according to claim 2, where the additional agent is protivogipertonicheskoe tool.
4. The use according to claim 3, where protivogipertonicheskoe tool BBO is seeking simultaneously or sequentially.
5. The use according to any one of the preceding paragraphs, where the disease or condition selected from the group consisting of cancer, diabetes, vascular permeability, edema, ascites and pleural effusion associated with tumors, inflammation or trauma, chronic airway inflammation, leaky capillaries, diseases of sepsis kidney associated with increased leakage of protein, eye disorders and pathological angiogenesis.
6. The use according to claim 1, where the patients over 65 years of age or patients cannot be otherwise subjected to treatment with appropriate dose of a VEGF antagonist without the development of hypertension.
7. The method of treatment, including
(a) identifying the patient as not susceptible in relation to intravenous administration of the VEGF antagonist containing VEGFR1R2-FcΔCl (a) SEQ ID NO:4;
(b) introduction to the patient a therapeutically effective amount of a VEGF antagonist subcutaneous injection;
(c) monitoring the blood pressure of the patient during and after subcutaneous administration of a VEGF antagonist and
(d) repeated administration of the VEGF antagonist subcutaneous injection, it is not necessarily the introduction of an additional agent, where the VEGF antagonist contains VEGFR1R2-FcΔCl (a) (SEQ ID NO:4).
8. The method according to claim 7, in which an additional agent such as defined in claim 3.
SUBSTANCE: aprotinin analogue with a TFFYGGSRGKRNNFKTEEY sequence is obtained, as well as a conjugate based thereon.
EFFECT: invention enables delivery of a compound or medicinal agent through the hematoencephalic barrier in mammals.
6 cl, 9 dwg, 6 tbl, 3 ex
SUBSTANCE: invention concerns nucleic acid molecules including the glycolising fusion designs containing catalytic domain of beta-1,4-N-acetylglucosaminyl transferase III or beta-1,4-galactosyl transferase, and resident polypeptide domain of Golgi complex responsible for localisation in Golgi complex, as well as their applications in host cell glycolisation modification.
EFFECT: invention allows producing polypeptides with the improved therapeutic properties, including antibodies with higher Fc-receptor binding and enhanced effector function.
21 cl, 37 dwg, 2 tbl, 7 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: invention relates to biotechnology and specifically to obtaining versions of glycoprotein IV alpha polypeptide of human thrombocytes (GPIbalpha) and can be used in medicine to treat vascular disorders. Using a recombinant technique, a polypeptide is obtained, which contains substitutes in SEQ ID NO:2 selected from: Y276F K237V C65S; K237V C65S; Y276F C65S; or Y276F Y278F Y279F K237V C65S. The obtained polypeptide is used to inhibit bonding of leucocytes to biological tissue or for treating disorders associated with activation of thrombocytes.
EFFECT: invention enables to obtain GPIbalpha polypeptide which bonds with von Willebrand factor with affinity which is at least 10 times higher than in natural GPIbα polypeptide, and also has low affinity for bonding with alpha-thrombin, lower aggregation and/or high resistance to proteolysis relative the polypeptide with SEQ ID NO:2.
41 cl, 3 dwg, 8 ex
SUBSTANCE: there are produced multipurpose and polyvalent inhibitors prepared of fused proteins containing polypeptide which identifies and blocks a functional site of molecule involved in angiogenesis, and polypeptide containing oligomerisation domain and a functional site modulating or inhibiting angiogenesis and separated by a proteinase sensitive site.
EFFECT: inhibitors are suitable for treatment and prevention of the pathologies caused by angiogenesis, eg a malignant tumour, rheumatoid arthritis or psoriasis.
25 cl, 6 dwg, 1 tbl, 1 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: invention relates to biotechnology, specifically to obtaining genetically engineered vaccines and can be used in medicine. A recombinant gene structure containing a series of human SLC gene, antigen gene and gene Fc-fragment of IgG 1 is obtained.
EFFECT: invention considerably increases efficiency of the immune response of the body to the introduced antigen compared to existing antigen structures.
10 cl, 15 dwg, 1 tbl, 12 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: invention relates to genetic and protein engineering and can be used in biomedical industry. A genetic make up is proposed, which codes a peptide in which two domains bonding the growth hormone (GH) receptor are bonded into a tandem by a "semirigid" or "rigid" linker, which consists of at least 1-4 copies of the A(EAAAK)A amino acid sequence.
EFFECT: as a result of expression of the nucleotide sequence coding the said tandem, GH-linker-GH polypeptides are obtained, which exhibit growth hormone receptor agonist properties, which determine the possibility of their use in medicinal agents for treating diseases related to the need to administer the growth hormone.
10 cl, 31 dwg, 5 ex
SUBSTANCE: recombinant DNA is produced, which codes functionally active hybrid protein (BrdGl7ACA-cbd), consisting of amino-acid sequence of acylase glutaryl-7- aminocephalosporanic acid of strain Brevundimonas diminuta All-Russian collection of industrial microorganisms B-1297 and chitin-binding domain of chitinase Al Bacillus circulans. Recombinant plasmid pSVH0108 is constructed for expression of BrdGl7ACA-cbd in cells E.coli, containing sequence of recombinant DNA that codes hybrid protein under control of promotor and terminator of RNA-polymerase of phage T7. As a result of E.coli strain tranformation with this recombinant plasmid and selection of transformed clones, a new strain E.coli BL21(DE3)/pSVH0108 cbd is produced - producer of hybrid protein BrdGl7ACA-cbd.
EFFECT: high yield of recombinant ferment.
4 dwg, 2 tbl, 7 ex
SUBSTANCE: invention represents a combination containing VEGF Trap and 5-fluorouracil to be applied in treatment of neoplasms.
EFFECT: higher effectiveness of the combination ensured by therapeutic synergism of its components, and reduced toxicity.
4 cl, 1 ex, 1 tbl
SUBSTANCE: plants are transformed with application of nucleic-acid constructs, which contain the first nucleotide sequence that codes γ-sein, or its fragment, which is able to direct and retain protein in endoplasmic reticulum of plant cell, the second sequence of nucleic acid, which codes aminoacid sequence, which is specifically split by ferment or chemical compounds, and the third sequence of nucleic acid, which codes target peptide or protein.
EFFECT: transformation of plant by such constructs makes it possible to produce fused proteins accumulated in endoplasmic reticulum of cells in the form of protein bodies, from which target proteins may be extracted, in particular calcitonin.
47 cl, 19 dwg, 1 ex
SUBSTANCE: there is offered molecule of nucleic acid inducing CEA immune response, containing a nucleotide sequence that codes a fused protein on a basis of carcinoembryonal antigen (CEA) or its functional version fused with a subunit B of thermolabile enterotoxin E coli. There are described versions thereof, as well as the related purified protein. There is disclosed an expression vector containing said molecule of nucleic acid, and a host-cell containing specified vector. There are described adenoviral vaccinal vector for inducing the immune response and a vaccinal plasmid on the basis of the specified molecule.
EFFECT: application of the invention allows to inducing the immune response in a mammal which is stronger, than that induced with natural CEA that can find application in medicine for cancer treatment.
20 cl, 62 dwg, 20 ex
SUBSTANCE: invention relates to novel versions of peptides IL-21, where amino acids are delegated and/or replaced in region, consisting of amino acids No 83-96.
EFFECT: possibility of manufacturing medication for cancer treatment.
8 cl, 2 dwg, 1 ex
SUBSTANCE: thymus-specific protein T101 consisting of 84 amino acids is recovered from human thymus. The recovered full-length peptide T101 includes the signal peptide consisting of 33 amino acids and the peptide sequence T101 consisting of 51 amino acids and exhibiting immunomodulating activity. The full-length peptide T101, and also peptide fragments and derivatives are used as a part of a pharmaceutical composition for treating autoimmune and inflammatory diseases.
EFFECT: invention allows preparing polypeptide capable to stimulate lymphocyte proliferation of human peripheral blood, to inhibit the tumour growth and to modulate the immune system.
10 cl, 15 dwg, 1 tbl, 11 ex
SUBSTANCE: invention is related to nucleic acids and multidomain proteins, which are able to bind vessel endotheliocyte growth factor (VEGF), and may be used in medicine. Recombinant method is used to produce polypeptide, which consists of component (R1R2)X and, unnecessarily, multidomain component (MC), which represents aminoacid sequence with length from 1 to 200 of amino acids, having at least one remainder of cysteine, where X≥1, R1 means antibody-like (Ig) domain 2 of VEGF receptor Llt-1, and R2 means Ig-domain 3 of VEGF receptor Flk-1. Produced fused polypeptide does not contain multidomain component in case, when X=2, and in case when X=1, multidomain component represents aminoacid sequence with length from 1 to 15 amino acids. Produced polypeptide is used in composition of pharmaceutical compound for VEGF-mediated disease or condition.
EFFECT: invention makes it possible to produce highly efficient trap of VEGF, special structure of which is suitable for local introduction into specific organs, tissues or cells.
16 cl, 3 tbl, 7 ex
SUBSTANCE: invention refers to biotechnology and can be used for screening of compounds with properties of agonists or antagonists of leptin receptor. There is disclosed method for detecting leptin receptor definition that provides contact of a candidate compound and cells expressing after contransfection with two expressing vectors, respectively, two fused proteins, one of which consists of a short isoform of leptin receptor (OBRs) and an energy donor protein (preferentially luciferase), and the second - of OBRs and an energy acceptor protein (preferentially GFP or its mutant form); measurement of energy transfer (BRET) between the fused proteins; comparison of its value to the relevant indicator measured in the same system, however without tested compound, and estimated result where higher energy transfer indicates binding of the candidate compound-candidate with leptin receptor. Prospective application of the invention is related to development of the preparations for prevention or treatment of disease wherein leptin or its receptor are involved.
EFFECT: development of effective method for detecting leptin receptor ligands.
4 cl, 15 dwg, 3 ex
FIELD: medicine; pharmacology.
SUBSTANCE: variants of the combined protein which contain the extracellular domain of a human receptor of a hormone of growth and the domain which includes alarm sequence for joining glycosylphosphatidylynozyte (GPI) anchors are offered.
EFFECT: effective medical product for acromegalia and gigantism treatment.
8 cl, 16 dwg
SUBSTANCE: invention relates to field of biotechnology, namely to genetic engineering. New interferon-binding proteins, which modulate activity of different interferon-α subtypes, as well as interferon-β activity, are obtained. Described is cloning of DNA fragment, which codes interferon-α/β, binding protein IFNAB-BPI, and expression of obtained DNA fragment in host-cells both with formation of respective polypeptide and in form of fused proteins. Practical application of obtained proteins as components of pharmaceutical compositions for inhibiting activity of IFN-α or IFN-β is suggested. Invention can be applied in medicine for inhibiting undesirable impact of IFN-α or IFN-β.
EFFECT: obtaining new interferon-binding proteins, which can find application in medicine for inhibiting undesirable impact of IFN-α or IFN-β.
13 cl, 10 dwg, 6 tbl, 17 ex
SUBSTANCE: invention refers to medicine and concerns angiogenesis-preventing immunotherapy. Invention substance includes immunogenic compositions for treatment of disorders associated with angiogenesis intensification, containing oligonucleotides, coding polypeptides VEGFR2, introduced as a part of plasmid or viral vectors, as well as polypeptides VEGFR2, oligonucleotides, coding autologous VEGF with damaged function of receptor activation, polypeptides VEGF and their combinations. Immunogenic compositions can be used for treatment of malignant neoplasms and metastasises, at benign neoplasm and chronic inflammatory and autoimmune diseases. Advantage of invention lies in humoral and cellular immunity induction by means of specified compositions.
EFFECT: development of effective method angiogenesis preventive immunotherapy.
19 cl, 11 ex, 7 tbl
FIELD: technological processess.
SUBSTANCE: oligomeric polypeptide dimmers include ligand-binding leptyne domains. Domains are connected by means of flexible polypeptide linker molecules. Linker molecules might possibly include sites of sensitivity to proteases.
EFFECT: releases biologically active cytokines when injecting to human being or animal.
31 cl, 13 dwg
FIELD: genetic engineering, medicine.
SUBSTANCE: invention proposes chimeric protein (ANTH1) consisting of a sequence corresponding to 60 amino acids of N-terminal region of interleukin-2 and extracellular moiety of N-terminal moiety of human gamma-interferon alpha-chain that are bound each with other by peptide Ala-His-Met-Met. Protein ANTH1 possesses capacity to inhibit biological activity of gamma-interferon and/or interleukin-2 and therefore it can be used in preparing drugs designated for treatment of diseases mediated by activity of interleukin-2, gamma-interferon or both these cytokines, in particular, in treatment of cerebrospinal sclerosis, erythematosus lupus, proliferative arthritis, diabetes mellitus and others. Invention can be used in medical-biological industry.
EFFECT: valuable medicinal and biological properties of protein.
13 cl, 1 tbl, 10 dwg, 11 ex
FIELD: genetic engineering, medicine.
SUBSTANCE: invention relates to interleukin-20 receptors and can be used in treatment of inflammatory diseases. Soluble receptors for interleukin-20 (IL-20) are constructed consisting of two polypeptide subunits IL-20RA and IL-20RB. Subunits as components of IL-20 receptor are bound by disulfide bonds. Also, variants of IL-20 receptor are prepared wherein one subunit is fused with the constant region of immunoglobulin light chain, and another subunit is fused with the constant region of immunoglobulin heavy chain, and these chains of immunoglobulin are bound by a disulfide bond. Invention provides using new IL-20 receptors for down-regulation of IL-20.
EFFECT: valuable medicinal properties if IL-20 receptor.
9 cl, 8 dwg, 13 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to medicine, namely to an agent exhibiting antihypertensive activity which represents 1-alkyl-2-alkylcarbamoylglycerins of general formula I . In formula I R means hydrocarbon radical -(CH2)nCH3 (n=10-18), R1 means methyl or ethyl. Also, the invention refers to a method for preparing the compounds of formula I. The method consists in the fact that parent 1-alkylglycerins of general formula II react with trimethylchlorosilane with using triethylamine in toluene medium at temperature -20°C to 0°C, then the reaction mass is added with appropriate alkylisocyanate and processed with ammonium bifluoride in methanol medium at room temperature.
EFFECT: preparation of the agent exhibiting antihypertensive activity.
2 cl, 7 ex