Pharmaceutical preparation for treating benign prostatic hyperplasia

FIELD: medicine, pharmaceutics.

SUBSTANCE: what is presented is using at least one protease for producing a drug preparation for treating and/or preventing benign prostatic hypertrophy/hyperplasia wherein the drug preparation is acceptable for the oral administration with at least one protease being specified in a group consisting of plant and microorganism proteases, and with at least one protease being administered in the amount of 1 to 100 mg/kg of body weight, and a respective pharmaceutical preparation.

EFFECT: what is shown is reducing high vascular endothelial growth factor in patient's blood under the action of a mixture of enzymes that makes it possible to inhibit angiogenic growth factors (VEGF), thereby leading to lower vascularisation, and reducing benign prostatic hypertrophy/hyperplasia, with the enzymatic therapy under the present invention having no effect on the normal VEGF level.

13 cl, 7 dwg

 

The technical field to which the invention relates.

The present invention relates to a medicinal product for the treatment of benign prostate hyperplasia.

The level of technology

Angiogenesis is the process by which changes prior vascularization with the formation of a complex network of new blood vessels to support the growing tissue. First is the local release of ECM proteases by EU-degradation of the basal membrane of blood vessels. Then, the activated endothelial cells (EC) penetrate the stroma, proliferate and form the structure of the tubes, which are combined together. In the end, the basement membrane is synthesized again and pericyte migrate to sites of newly formed capillaries. Angiogenesis is involved in various physiological processes such as embryogenesis, wound healing and contributes to the progression of some pathological conditions (cancer, hyperplasia). In normal endothelial cells in adult mammals are dormant and share only every few years. Under pathological conditions, such as benign hyperplasia of the prostate, breast, rheumatoid arthritis, atherosclerosis, endometriosis, psoriasis, or ocular cancer revascularization, local growth of endothelial cells value is positive increases.

Angiogenesis is regulated by a large number of Pro - and antiangiogenic factors, including the growth of the placenta (P1GF), fibroblast growth factor, angiopoietin and others. The main limiting step in this process, however, is the growth factor of the endothelium of blood vessels (VEGF). Previously described as a factor causing persistent violation of vascular permeability and called vascular permeability factor (Vascular Permeability Factor, VPF), this growth factor later became known factor vascular endothelial growth (VEGF) in the calculation of its mitogenic effect in endothelial cells.

In some models in vitro and in vivo, it was found that VEGF promotes the growth of endothelial cells of blood vessels and form blood vessels. VEGF also serves as a life supporting factor for endothelial cells. In vitro VEGF prevents apoptosis induced by phlebotomy. In vivo VEGF inhibition in neonatal but not in adult mice leads to severe apoptotic changes in the vascular network. Nokautiroval by VEGF-A in mice (or zero [VEGF-A-/-]) or heterozygous [VEGF-A+/-] form) leads to the death of the embryo between 11 and 12 weeks due to developed abnormalities, impaired vascularization in certain organs and a significant reduction in the number containing the kernel of red blood cells in the blood Islands of the yolk SAC, suggesting that VEGF-A controls as vasculogenic, that is and hematomas. In the end, VEGF increases vascular permeability and endothelial fenestration in the endothelial cells of the adrenal glands.

The expression of VEGF may be caused by hypoxia via activation of hypoxia inducible factor-1 (HIF-I). The lack of oxygen, caused, for example, excessive tissue growth, activates the expression of HIF-I. Then complexes with HIF-I accumulate in the cell nucleus, are connected with HIF-I-binding site DNA and initiate respectively unregulated transcription of VEGF mRNA by triggering the angiogenic switch, which can cause sprouting of adjacent blood vessels in hypoxic tissue. Various proinflammatory cytokines can also induce the expression of VEGF, which explains the role of angiogenesis in the first place, in inflammatory diseases, such as psoriasis or rheumatoid arthritis.

With recognition of this key role in the regulation of VEGF and its receptors act as an attractive therapeutic target. Some strategies that affect VEGF-mediated angiogenesis, was used mainly in the treatment of cancer, but also in other diseases characterized by excessive angiogenesis (e.g., macular degeneration or benign prostatic hyperplasia). Investigational medicinal product included antibiotics, toxin conjugates, interfering PE the Chida, soluble receptors and tyrosine kinase inhibitors. Taken together, these results suggest that drugs targeted to VEGF, improve the future treatment of diseases such as cancer, rheumatoid arthritis, psoriasis, ocular neovascularization and other

Benign prostatic hyperplasia (BPH) refers to diseases associated with the increased size of prostate in men of middle and old age. When the prostate reaches a large enough size, it squeezes the urethra and causes a partial, sometimes even complete obstruction of the urethra that prevents the normal flow of urine. In some cases, the gland may grow up to ten times the size of normal. It is considered that BPH is not a precancerous lesion, but may become a significant health issue for older men, mainly due to the negative impact on quality of life, such as difficulty in starting urination, frequent urination, increased risk of infection of the urinary tract, and urinary retention.

BPH is a relatively slowly progressing condition that requires decades to develop symptoms associated with the disease. However, up to 60 years up to 50% of the men has a pathological state. Verhamme et al., Eur Urol. 2002 Oct; 42 (4):323-8 reports that the coefficient Zab the rate has been going BPH at 3/1000 men per year from age 45 to 49 years reaches 38 per 1000 men per year in the age group 75-79 years. In men older than 80 years, the incidence rate remains constant.

Currently, there are three schemes of treatment of BPH.

1. "Waiting on observation lies in the fact that it is recommended if the symptoms are mild and do not affect the quality of life of men. Individuals can choose this treatment if they feel that drug therapy will be more uncomfortable than when the symptoms of BPH. If symptoms and discomfort occurs, changes the activity of everyday life, it is recommended drug therapy.

2. Drugs used in the General symptoms related to the urinary tract associated with BPH, to reduce the size of the prostate or to slow the growth of prostate (Madersbacher et al. Internist (Berl.). 2007 Apr 11). Usually the drugs used to treat the symptoms include alpha-blockers, such as doxazosin (Cardura) side effects: dizziness, drowsiness, slight headache, migraine, constipation, loss of appetite, dry mouth, fatigue, nasal congestion, blurred vision, dry eyes or restless sleep], tamsulosin (Flomax) side effects: reduction in blood pressure resulting in fainting or loss of consciousness, dizziness, migraine, nasal congestion and palpitations], inhibitors of 5-alpha-reductase, such as phynaster the d (Proscar) side effects: decreased number of sperm on aukusti, reduced sexual desire, impotence] or dutasteride (Avodart) [side effects: the coarsening of the mammary glands, decreased libido, problems with aulularia, breast enlargement in men, urticaria, impotence, itching, rash].

3. In case of severe symptoms of BPH, such as recurrent urinary retention, appearance of blood in the urine, recurring infection in the urinary tract or stones in the bladder, the method of choice is surgical removal of the prostate (prostatectomy). Side effects of prostatectomy include additional risk of secondary infections, bladder perforation, deep vein thrombosis and total incontinence.

WO 90/08555 concerns the application of hydrolytic enzymes such as proteases, such as trypsin, chymotrypsin or bromelain, which is used for treating hypertrophy of the prostate. The drug is administered by injection directly into the prostate.

In WO 2006/127431 described a method of treatment of an individual suffering from hypertrophy of the prostate. If this treatment is administered enzymes mammalian type trypsin.

ZA 200002342 discloses the use of plant sterols and their glycosides for the prevention of benign prostatic hypertrophy. To increase the speed of absorption of plant sterols to these pharmaceutical compositions add protease type bromel is on.

US 2006/0024385 discloses compositions which are used to reduce oxidative stress. These compositions may contain plant extracts, which may include, among other substances and also bromelain.

US 2003/0194400 describes compositions containing trombogen, selective for vascular tissue that can be contacted tissue caused by hyperplasia of the prostate.

In WO 94/019005 described pharmaceutical composition, containing one or more proteases. In this document, the proteases are trypsin, chymotrypsin, bromelain, papain and fitsin.

The purpose of the present invention is to provide medicines that are suitable for the treatment or prevention of benign prostate hyperplasia and which eliminate the disadvantages of the commonly used dosage forms and tools.

The present invention relates to the use of at least one proteolytic enzyme protease for the manufacture of medicinal products for the treatment and/or prevention of benign prostatic hypertrophy/hyperplasia of the prostate, when the drug is intended for enteral administration, and at least one protease selected from the group consisting of proteases of plants not related to mammals animals, microorganisms, and at least one protease is administered in an amount of from 1 is about 100 mg/kg of body weight.

It has been unexpectedly discovered that a certain drug contains a combination of at least one protease selected from the group consisting of proteases of plants not related to mammals and microorganisms, provides for the introduction of individual significant reduction factor vascular endothelial growth (VEGF) (at least 40%, preferably 50%, more preferably, 60% or 70%, most preferably at least 80%, and especially 90% compared with the level VEGF specified individual prior to the introduction of the medicinal product in accordance with the present invention and, therefore, suppression of angiogenesis. Inhibition of angiogenesis is especially important for the prevention and treatment of benign hyperplasia/hypertrophy of the prostate in the individual.

This effect is even more unexpected, because the drug containing at least one protease, enter enterline (preferably orally or rectally): the dose of 1-100 mg protease/kg of body weight (equal to 3 to 300 IU/kg body weight) is low. The prior art, in particular WO 90/08555, proposes protease directly into the prostate for destruction of hyperplastic tissue. However, unexpectedly turned out to be an effective non-invasive enteric system introduction Protea the individual in the amount disclosed in the present invention, although, of course, used the dosage is much lower than the dosage used for the destruction of human tissues (WO 90/08555). The combination of more than one protease, particularly at least two (preferably at least three, at least four, at least five, at least six), proteases increases several times therapeutic effect of prevention and/or treatment of benign hyperplasia of the prostate in the individual.

It has been unexpectedly discovered that the combination of at least two proteases provides a more effective treatment of benign hypertrophy/hyperplasia of the prostate than using a single protease.

The prostate is androgen-dependent in the sense that for normal growth, development, differentiation and functioning of required testosterone and a decrease in the content of androgens will reduce the size of the prostate. Antiandrogenna therapy aimed at reducing the proliferation of prostate tissue, has only a limited effect, because it is based on the fact that steroids can influence, rather than cause the growth of the prostate. The data show that the growth of the prostate may be under the direct control of specific growth factors, such as growth factor endo is the men of blood vessels (VEGF) and transforming growth factor-beta (TGF-beta), and can only indirectly be regulated by androgens.

For BPH is characterized by hyperplasia of stromal and epithelial cells of the prostate and increased density of microvessels in the tissue of the prostate (Stefanou et al. In Vivo. 2004 Mar-Apr; 18 (2): 155-60). Androgens stimulate the beginning of hypertrophy of cells in the prostate. Hypertrophy can gradually lead to local hypoxia, which stimulates the expression of hypoxia-induced factors (HIF-1 and HIF-2). HIF stimulates VEGF synthesis in the epithelial cells of the prostate (Walsh et al. Prostatic Dis. 2002; 5 (2): 119-22). Increased expression of VEGF can stimulate angiogenesis, which may result in clinical progression of BPH and the formation of large isolated nodules in periuretralnuu region of the prostate. Interruption of this continuum should reduce hyperplasia.

Therefore, targeting VEGF directly represents treatment regulating pathological overgrowth of prostate tissue and may also decrease the intensity of symptoms associated with benign prostatic hyperplasia. The concentration of VEGF in the serum of patients with BPH increased compared with the control group of healthy people, but significantly lower than in patients with prostate cancer (Trapeznikova et al. Vestn Ross Akad Med Nauk. 2005; (5):14-6).

Angiogenesis elevated in BPH, is more pronounced in malignant tumors of the prostate and aurelium with a concentration of VEGF (Stefanou et at. In Vivo. 2004 Mar-Apr; 18 (2): 155-60). Anti-VEGF therapy reduces angiogenesis in the prostate of castrated rats treated with testosterone (Lissbrant et al. Prostate. 2004 Jan 1; 58 (1): 57-65).

In accordance with the present invention, the terms "benign hypertrophy of the prostate and benign prostatic hyperplasia" refers to one disease and are therefore used in the present invention are interchangeable.

Introduction proteases derived from bacteria, fungi, plants, and non-mammal animals, is particularly suitable, since these proteases do not show significant toxicity when interacting with human cells or animal, especially endothelial cells. Even a combination of proteases, as described here, is not toxic to the animal or human, but it affects angiogenesis.

It should be noted that the protease derived from mammals (i.e. people)are not able to effectively inhibit or prevent angiogenesis. Therefore, a separate introduction of such proteases individual may not be applied for the prevention or treatment of diseases related to neoangiogenesis.

The term "drug"as defined here, includes not only pharmaceutical products but also food additives.

Used here the value of "protease plants" and "protease animal is x" refers to the protease, which are found in nature in plants or animals (non-mammal animals) and which are allocated or received from them. "Protease plants" and "protease animals" are also recombinant proteases, whose encoding DNA (e.g. cDNA) is derived or obtained from a plant or animal (containing in nature specified DNA in its genome), respectively, and cloned into appropriate vectors and expressed in cell culture prokaryotes (e.g., bacterial) or eukaryotic (e.g., insect cell, a cell of a mammal).

"Protease microorganisms, used here the value is a protease, which occur naturally in microorganisms, such as bacteria and fungi (such as yeasts, moulds). These proteases may, however, be isolated from other cells and microorganisms, provided that these cells and organisms contain DNA microbial protease and is capable of producing the specified recombinant protease.

Also preferred is the use of the drug of the present invention for the prevention and/or treatment of individuals suffering from disease due to high levels of VEGF and related angiogenesis, resulting in these diseases are mostly or completely unrelated povyshennoi proliferative activity. At least one protease plants preferably selected from the group consisting of bromelain, papain or fitsin. Protease plants, preferably used in accordance with the present invention, listed above.

These proteases can be obtained by recombinant expression in host or by extraction from plants producing these proteases in a natural way, thereby extract itself can directly be used for the manufacture of a medicinal product in accordance with the present invention. Methods of extraction of proteases are well known in the prior art.

Bromelain: Pineapples since ancient times been used as medicinal plants among the native plants of South and Central America. Bromelain was first introduced in Europe as a secondary drug in 1957 and was primarily used for the treatment of edema and inflammation (review Maurer. Cell Mol Life Sci 58 (9):1234-1245, 2001).

Papain: can be obtained as the crude dry material by collecting the milky juice of the fruit of the papaya tree (papaya). The milky juice is collected after incision of the neck of the fruit, after which it may be dried fruit or packaged in the container. Then this latex can be dried. From this moment he is the driest of Norrbotten the m material. Cleaning stage is necessary to exclude contaminants. Cleaning consists of solubilization and extraction of the enzyme papain.

Fitsin: the milky juice of some species of ficus (Moraceae) is traditionally used as a deworming drugs in Central and South America. It is recognized that antihelminthic activity due to the presence of proteolytic fraction, called fitsin.

Protease plants: bromelain, papain, fitsin, Ananin, balankin, curtain, fustukian, hieronymian, capiain, macrodantin, pingvinen, cusomizing, aktinidin.

Protease bacteria/microorganisms: In accordance with the preferred embodiment of the present invention the protease isolated from bacteria or fungi, which are selected from the group consisting of brinase (from Aspergillus oryzae), nattokinase {Bacillus subtilis Natto), pronase (Streptomyces griseus), sobrosa-S (Aspergillus melleus), serrapeptase (Bacillus Serratia E 15), sphericity (Bacillus sphaericus), subtilisin (Bacillus subtilis) and mut-mutanty (Aspergillus sp.).

Protease microorganisms can be obtained using recombinant technology or can be extracted directly from the microbial culture containing microorganisms that produce these proteases. Nattokinase, for example, derived from natto, a traditional Japanese food made from fermented soybeans, or cultures, the soda is containing organisms of specific subtypes of Bacillus subtilis (Bacillus subtilis var. Natto), which are capable of producing the specified protease.

Serrapeptase, also known as serrapeptidase or serratiopeptidase derived from microorganisms of Serratia E 15 that live in the intestinal wall silk worm, helps digestion and dissolve the crystals. This enzyme can be used as an additional component in the treatment of pain and inflammation and is used in clinical practice in some countries of Asia and Europe. Serrapeptase used as an alternative to non-steroidal anti-inflammatory drugs (NSAIDs)are commonly used to treat arthritis and inflammation.

Protease animals (of memleketim) is preferably extracted from Antarctic krill (crustacea) Euphasia superba, chilensis, batroxobin and lumbrokinase from earthworm, alfimeprase, fibrolite.

Additionally preferred proteases are Matanza, alfimeprase, fibrolite, cusomizing, aktinidin, Ananin, balankin, curtain, gastusin, hieronymian, Karategin, pingvinen and macrodantin. These proteases can be produced recombinante using methods known from the prior art, or can be obtained directly from the relevant animal.

Especially preferred medicines contain bromelain and/or papain as the FR is C plants and if necessary, the nattokinase as protease microorganisms. The preferred ratio between these proteases in the medicinal product in accordance with the present invention can be found in the following table.

BromelainPapainNattokinase
100,00%0,00%0,00%
75,00%25,00%0,00%
16,67%16,67%66,67%
25,00%25,00%50,00%
0,00%75,00%25,00%
75,00%0,00%25,00%
25,00%0,00%75,00%
16,67%66,67%16,67%
50,00%0,00%50,00%
0,00%100,00% 0,00%
0,00%50,00%50,00%
0,00%25,00%75,00%
33,33%33,33%33,33%
25,00%75,00%0,00%
0,00%0,00%100,00%
50,0%25,00%25,00%
25,00%50,00%25,00%
50,00%50,00%0,00%

In addition, the preferred combinations of proteases used in the medicinal product, in accordance with the present invention are:

bromelain and papain (if necessary together with carlesimo, brinzei, fitsin, lumbrokinase, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), bromelain and fitsin (if necessary together with carlesimo, brinzei, papain, lumbrokinase, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), bromeline nattokinase (if necessary together with carlesimo, brinzei, fitsin, lumbrokinase, papain, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), bromelain and Brinza (if necessary together with carlesimo, papain, fitsin, lumbrokinase, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), bromelain and sobrosa (if necessary together with carlesimo, brinzei, fitsin, lumbrokinase, nattokinase, pronasol, papain, serrapeptase, sphericity, subtilisin and/or batroxobin), bromelain and sericata (if necessary in conjunction with carlesimo, brinzei, fitsin, lumbrokinase, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), bromelain and subtilisin (if necessary together with carlesimo, brinzei, fitsin, lumbrokinase, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, papain and/or batroxobin), bromelain and kurilensis (if necessary together with brinzei, fitsin, lumbrokinase, nattokinase, papain, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), bromelain and batroxobin (if necessary together with carlesimo, brinzei, fitsin, lumbrokinase, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or papain, bromelain and lumbrokinase (if necessary the particular together with carlesimo, brinzei, fitsin, papain, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin);

papain and fitsin (if necessary together with carlesimo, brinzei, bromelain, lumbrokinase, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), papain and nattokinase (if necessary together with carlesimo, brinzei, bromelain, lumbrokinase, fitsin, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), papain and Brinza (if necessary together with carlesimo, fitsin, bromelain, lumbrokinase, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and or batroxobin), papain and sobrosa (if necessary together with carlesimo, brinzei, bromelain, lumbrokinase, nattokinase, pronasol, fitsin, serrapeptase, sphericity, subtilisin and/or batroxobin), papain and sericata (if necessary together with carlesimo, brinzei, bromelain, lumbrokinase, nattokinase, pronasol, ciprobay-S, serrapeptase, fitsin, subtilisin and/or batroxobin), papain and subtilisin (if necessary together with carlesimo, brinzei, bromelain, lumbrokinase, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, fitsin and/or batroxobin), papain and kurilensis (when the necessity is together with carlesimo, brinzei, bromelain, lumbrokinase, nattokinase, fitsin, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), papain and batroxobin (if necessary together with carlesimo, brinzei, bromelain, lumbrokinase, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or fitsin), papain and lumbrokinase (if necessary together with carlesimo, brinzei, bromelain, fitsin, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin);

fitsin and nattokinase (if necessary together with carlesimo, brinzei, bromelain, papain, lumbrokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), fitsin and Brinza (if necessary together with carlesimo, nattokinase, bromelain, papain, lumbrokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), fitsin and sobrosa (if necessary together with carlesimo, brinzei, bromelain, papain, lumbrokinase, pronasol, nattokinase, serrapeptase, sphericity, subtilisin and/or batroxobin), fitsin and sericata (if necessary together with carlesimo, brinzei, bromelain, papain, lumbrokinase, pronasol, ciprobay-S, serrapeptase, nattokinase, subtilisin and/or batroxobin), fitsin and subtilisin(if necessary together with carlesimo, brinzei, bromelain, papain, lumbrokinase, pronasol, ciprobay-S, serrapeptase, sphericity, nattokinase and/or batroxobin), fitsin and kurilensis (if necessary together with nattokinase, brinzei, bromelain, papain, lumbrokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), fitsin and batroxobin (if necessary together with carlesimo, brinzei, bromelain, papain, lumbrokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or nattokinase), fitsin and lumbrokinase (when necessary in conjunction with carlesimo, brinzei, bromelain, papain, nattokinase, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin);

nattokinase and Brinza (if necessary together with carlesimo, lumbrokinase, bromelain, papain, fitsin, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), nattokinase and sobrosa (if necessary together with carlesimo, lumbrokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, sphericity, subtilisin and/or batroxobin), nattokinase and sericata (if necessary together with carlesimo, lumbrokinase, bromelain, papain, fitsin, pronasol, ciprobay-S, serrapeptase, brinzei, subtilisin and/or batroxobin), nattokinase and subtilisin (when dealing with carlesimo, lumbrokinase, bromelain, papain, fitsin, pronasol, ciprobay-S, serrapeptase, sphericity, brinty and/or batroxobin), nattokinase and kurilensis (if necessary together with brinzei, lumbrokinase, bromelain, papain, fitsin, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin), nattokinase and batroxobin (if necessary together with carlesimo, lumbrokinase, bromelain, papain, fitsin, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or brinty), nattokinase and lumbrokinase (when necessary in conjunction with carlesimo, brinzei, bromelain, papain, fitsin, pronasol, ciprobay-S, serrapeptase, sphericity, subtilisin and/or batroxobin),

Brinza and sobrosa (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, lumbrokinase, serrapeptase, sphericity, subtilisin and/or batroxobin), Brinza and sericata (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, lumbrokinase, serrapeptase, ciprobay, subtilisin and/or batroxobin), Brinza and subtilisin (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, lumbrokinase, serrapeptase, sphericity, ciprobay and/or batroxobin), Brinza and kurilensis (if necessary the particular together with ciprobay, the nattokinase, bromelain, papain, fitsin, pronasol, lumbrokinase, serrapeptase, sphericity, subtilisin and/or batroxobin), Brinza and batroxobin (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, lumbrokinase, serrapeptase, sphericity, subtilisin and/or ciprobay), Brinza and lumbrokinase (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, ciprobay, serrapeptase, sphericity, subtilisin and/or batroxobin);

sobrosa and sericata (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, lumbrokinase, subtilisin and/or batroxobin), sobrosa and subtilisin (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, lumbrokinase, sphericity and/or batroxobin), sobrosa and kurilensis (if necessary together with sphericity, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, lumbrokinase, subtilisin and/or batroxobin), sobrosa and batroxobin (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, lumbrokinase, subtilisin and/or sphericity), sobrosa and lumbrokinase (when neo is needed together with carlesimo, the nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, sphericity, subtilisin and/or batroxobin);

staricase and subtilisin (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, ciprobay, lumbrokinase and/or batroxobin), staricase and kurilensis (if necessary together with subtilisin, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, sphericity, ciprobay, lumbrokinase and/or batroxobin), staricase and batroxobin (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, sphericity, subtilisin, lumbrokinase and/or ciprobay), staricase and lumbrokinase (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, subtilisin, ciprobay and/or batroxobin);

subtilisin and kurilensis (if necessary together with lumbrokinase, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, sphericity, ciprobay and/or batroxobin), subtilisin and batroxobin (if necessary together with lumbrokinase, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, sphericity, ciprobay and/or carlesimo), subtilisin lumbrokinase (if necessary together with carlesimo, the nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, sphericity, ciprobay and/or batroxobin);

chilensis and batroxobin (if necessary together with substring, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, sphericity, ciprobay and/or lumbrokinase), kurilensis and lumbrokinase (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, sphericity, ciprobay and/or batroxobin),

batroxobin and lumbrokinase (if necessary together with carlesimo, nattokinase, bromelain, papain, fitsin, pronasol, brinzei, serrapeptase, sphericity, ciprobay and/or carlesimo).

These combinations of proteases can optionally be combined with at least one protease which is selected from the group consisting of atanazy, alfimeprase, fibrolite, macumazana, actinidine, Ananina, Balanchine, Carolina, gastusin, hieronymian, karatina, pingvinen and macrodantin.

Especially preferred drug of the present invention contain the bromelain in combination with papain, fitsin, nattokinase, brinzei, ciprobay, sphericity, subtilisin, carlesimo, batroxobin, lumbrokinase.

In the medicine of the present invention, each of the at least d is uh protease is present in a quantity at least 10%, preferably at least 15%, relative to the total content of the proteases contained in the specified drug.

In accordance with another preferred embodiment of the present invention, at least two protease contained in the medicinal product in an amount of from 5 to 10, preferably from 10 to 90 wt%./weight., preferably from 20 to 80 wt%./weight., more preferably from 30 to 70 wt%./weight.

At least one protease is preferably administered to an individual in the amount of 2-50 mg/kg, preferably 5-20 mg/kg of body weight. As mentioned above, the number of input proteases varies from 3 to 300 IU/kg body weight, preferably from 6 to 150 U/kg body weight, more preferably from 15 to 60 U/kg of body weight. Also protease can be administered in an amount of from 3 to 150 U/kg body weight or from 3 to 60 U/kg of body weight. The drug may preferably further comprise at least one pharmaceutically suitable carrier solvents and/or excipients, preferably a binding agent, a filler, a leavening agent, a lubricating agent, a preservative and/or shell.

Depending on pharmaceutical composition of the medicinal product in accordance with the present invention can be applied to other substances such as excipients, shells and other

Drug is the first tool of the present invention is preferably suitable for oral administration.

In accordance with a preferred embodiment of the present invention, the drug is presented in a pharmaceutical form which is selected from the group consisting of drops, spray, tablets, preferably soluble tablets, effervescent tablets, gastroresistant tablets and sublingual tablets, capsules, preferably gastroresistant capsules, powders, granules, liquids for oral administration, drops for oral administration and suppositories.

The drug is suitable for oral administration. This route of administration is non-invasive and, therefore, provides for re-introduction (without causing harm to the patient) of the medicinal product, which has been tested in approximately 50 years (Ransberger. Erfahrungsheilkunde 1:14-17, 1971).

The drug of the present invention may be specially prepared for administration in solid or liquid form, including those suitable for oral administration, for example jelly (aqueous or non-aqueous solutions or suspensions), tablets, boluses, powders, granules, pastes for application to the tongue.

The drug of the present invention may also be provided for other routes of administration:

(1) parenteral administration, for example, by subcutaneous, intramuscular or intravenous injection as, for example, CTE is not think of a solution or suspension;

(2) local application, for example, as a cream, ointment or spray applied to the skin; or

(3) vnutribruchinnogo, for example, as a pessary, cream or foam.

The phrase "pharmaceutically suitable" is used here in the meaning refers to those compounds, materials, compositions and/or dosage forms which are medically suitable for use in contact with the tissues of humans and animals without increased toxicity, irritation, allergic response, or other problems or complications, and their use is commensurate with the appropriate ratio of benefit/risk.

Examples of materials that can be used as pharmaceutically suitable carriers include:

(1) sugars, such as lactose, glucose and sucrose;

(2) starches, such as corn starch and potato starch;

(3) cellulose, and its derivatives, such as carboxymethylcellulose sodium, ethylcellulose and cellulose acetate;

(4) powdered tragant;

(5) malt;

(6) gelatin;

(7) talc;

(8) excipients, such as cocoa butter and candle wax;

(9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil;

(10) glycols, such as propylene glycol;

(11) polyols, such as glycerin, sorbitol, mannitol and polyethylene is glycol;

(12) esters, such as etiloleat and tillaart;

(13) agar;

(14) buffering agents, such as magnesium hydroxide and aluminum hydroxide;

(15) alginic acid;

(16) pyrogen-free water;

(17) isotonic;

(18) ringer's solution;

(19) ethyl alcohol;

(20) phosphate buffer solutions; and

(21) other non-toxic compatible substances used in pharmaceutical compositions.

Moisturizing agents, emulsifying agents and lubricating agents such as sodium lauryl sulfate and magnesium stearate, and colouring agents, agents for lubricating, coating agents, sweeteners, fragrances and flavors, preservatives and antioxidants can also enter into the composition of the medicinal product in accordance with the present invention.

Examples of pharmaceutically suitable antioxidants include:

(1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like;

(2) oil-soluble antioxidants, such as ascorbyl palmitate, bottled oxyanion, bottled hydroxyanisol, lecithin, propylgallate, alpha-tocopherol and the like; and

(3) metalhalide agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid and the like.

The amount of active ingredient which can be combined with a carrier to obtain a dosage form for a single application, will be this number of connections, which provides a therapeutic effect.

The methods of preparation of medicines and formulations in accordance with the present invention include the stage of combining the compounds of the present invention with the carrier and, optionally, one or more additional ingredients. Typically, the compositions are prepared by simultaneous and equal Association of the compounds of the present invention with liquid carriers, or powdered solid carriers, or both, followed, if necessary, shaping the product. The compositions from which bretania, suitable for oral administration may be in the form of capsules, wafers, pills, tablets, lozenges (using a moisturising base, usually sucrose and acacia or tragakant), powders, granules or as a solution or suspension in aqueous or non-aqueous liquid, or as a liquid emulsion oil-in-water or water-in-oil", or syrup, or as lozenges (using an inert base such as gelatin and glycerin, or sucrose and acacia), each dosage form contains a predetermined amount by a combination of this the invention as active ingredients.

The protease of the present invention can also be administered as a bolus, medicinal porridge or pasta. In solid dosage forms of the invention for oral administration (capsules, tablets, pills, coated tablets, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically suitable carriers, such as sodium citrate or dicalcium phosphate, and/or with any of the following:

(1) fillers or diluents, such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid;

(2) binding agents such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or acacia;

(3) humectants, such as glycerol;

<> (4) agents to improve raspadaemosti tablets, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate;

(5) agents that slow grasping for solutions, such as paraffin;

(6) absorption accelerators, such as Quaternary ammonium compounds;

(7) humectants, such as, for example, cetyl alcohol and glycerol monostearate;

(8) absorbents, such as kaolin and bentonite clay;

(9) lubricating agents such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and

(10) coloring agents.

In the case of capsules, tablets and pills, the pharmaceutical compositions may also contain buffering agents. Solid compositions of the same type can also be used as fillers in soft and hard gelatin capsules using such excipients as lactose or milk sugar and high molecular weight polyethylene glycols and the like.

The tablet can be prepared by compressing or molding, optionally with one or more additional ingredients. Compressed tablets can be prepared using a coupling agent (for example, gelatin or hydroxypropylmethylcellulose), lubricating agent, inert the x solvents, the preservative agent for separation (for example, sodium starch glycolate or net carboxymethylcellulose sodium), surface-active or dispersing agent. Molded tablets can be made by molding in a suitable machine, where a mixture of the powdered compound moistened with an inert liquid diluents. Tablets and other solid dosage forms of the pharmaceutical compositions of the present invention, such as tablets, capsules, pills and granules, may optionally be obtained or prepared with coatings and shells, such as intersolubility and other coatings well known in the prior art in the field of pharmacy. They can also be composed so as to provide slow or controlled release of the active ingredient, and their composition is administered, for example, hypromellose in different proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They can be sterilized, for example, by filtration through inhibiting bacteria filter or by the introduction of the sterilizing agents in the form of sterile solid compositions which can be dissolved in sterile water or any other injectable medium immediately before use. These compositions could the t also optionally contain opalescent component and can be such compositions, which release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract or, if necessary, slow way.

Examples of coating compositions which can be used include polymeric substances and waxes. Proteases can also be microencapsulating form optionally together with one or more of the abovementioned excipients. Liquid dosage forms for oral administration of the compounds of the invention include pharmaceutically suitable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the field of engineering, such as water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butyleneglycol, oils (in particular, cottonseed, groundnut, corn, wheat germ, olive, castor and sesame oils), glycerol, glycols and esters of fatty acids and sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include an adjuvant, such as moistening agents, emulsifiers and suspendresume Agay is you, sweeteners, flavors, dyes, fragrances and preservatives.

Suspensions, in addition to the active compounds, may contain suspendresume agents, such as ethoxylated isostearyl alcohols, polyoxyethylenesorbitan and esters sorbitan, microcrystalline cellulose, Metagalaxy aluminum, bentonite, agar-agar and tragakant, and mixtures thereof.

Formulations of the pharmaceutical compositions of the invention for rectal injection can be in the form of suppositories which can be prepared by mixing the protease of the invention with one or more suitable not cause irritation with excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, wax suppositories or a salicylate, and which retains the solid form at room temperature but becomes liquid at body temperature and therefore will melt in the cavity of the rectum and release the active compound. The compositions of the present invention, which is suitable for rectal administration, also include pessaries, tampons, creams, gels, pastes, foams or sprays, while formulations containing such carriers known from the prior art.

Dosage forms for local or transdermal application of the compounds of this invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalers. The protease can be mixed under sterile conditions together with a pharmaceutically suitable carrier, and with any preservatives, buffers, or propellants that may be required.

Ointments, pastes, creams and gels may contain, in addition to the active compound of this invention, excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragakant, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the proteases of this invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbon, such as butane and propane.

Transdermal patches have the added advantage, as they provide a controlled delivery of the protease of the present invention in the body. Such dosage forms can be prepared by dissolving or dispersing proteases in a suitable medium. Amplifiers absorption can also be used to improve the penetration of proteases through the skin. The speed of such penetration can be controlled either by ensuring the ecene controlling the speed of the membrane, or by dispersing the compound in a polymer matrix or gel.

The pharmaceutical compositions of this invention suitable for parenteral administration contain protease of the invention in combination with one or more pharmaceutically suitable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be dissolved in a suitable sterile solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostatic, solutions that represent the composition to be isotonic with the blood of the intended recipient or suspendresume agents or thickeners. Examples of suitable aqueous and nonaqueous carriers which may be used in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like), and suitable mixtures thereof, vegetable oils, such as olive oil and injectable organic esters, such as etiloleat. The desired fluidity can be achieved, for example, through the use of covering materials, such as lecithin, by storing the required particle size in the case of dispersions and by the use of surfactants. These compositions can also contain adjuvant, such as preservatives, humectants, emulsifiers and dispersing agents. Prevention of the action of microorganisms on the claimed compositions can be achieved by inclusion in the composition of various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, and the like. Also desirable is to include isotonic agents such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be due to the inclusion of agents which delay absorption such as aluminum monostearate and gelatin. In some cases, prolongation of drug action is desirable to slow the absorption of drugs from the area of subcutaneous or intramuscular injection.

This can be explained by using a liquid suspension of crystalline or amorphous material having poor water solubility. Alternatively, delayed absorption of parenteral introduced drugs is accomplished by dissolution or suspension of the drug in an oil carrier. Injectable depot forms are made by forming microencapsulating matrices of the desired compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer and the nature of the specific polymer, the rate of release of drug can be controlled. Examples of other biodegradable polymers include poly(orthoevra) and poly(anhydrides).

Injectable depot formulations are also prepared by incorporating the drug in liposomes or microemulsions that are compatible with the tissue of the person. If the protease of the present invention is administered as medicines to people or animals, they can be taken per se or as a pharmaceutical composition containing, for example, of 0.1 to 99.5% (more preferably 0.5 to 90%) protease in combination with a pharmaceutically suitable carrier. The preparations of the present invention can be administered orally, parenterally. They, of course, is in the form of dosage forms appropriate for each route of administration.

For example, they are administered in tablets or capsulerebel form, by injection, Invalidovna, as an eye lotion, ointment, suppository, etc. administered by injection, infusion or inhalation; topical by using a lotion or ointment; and rectal by suppositories. Oral and local injection are preferred. The phrases "parenteral administration" and "administered parenterally" used here is the mean value of the route of administration other than enteral and local administration, usually by injection, and include, without limitation, intramuscular, intraarterial, intrathecal, intracapsular, Intro metallinou, intracardially, intradermal, intraperitoneally, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.

The drug of the present invention can enter humans and animals to treat any suitable route of administration, including oral, nasal, using, for example, a spray, rectally, parenterally, intracisternally or locally, using, for example, powders, ointments or drops, including buccal and subligual.

Regardless of the chosen route of administration of the protease of the present invention, which can be used in a suitable hydrated form, and/or pharmaceutical compositions of the present invention is made in the form of pharmaceutically acceptable dosage forms by traditional methods known to a person skilled in this field.

Specific dose levels of active ingredients in the pharmaceutical compositions of the present invention can be varied to provide the amount of the active ingredient, which is effective to achieve the desired therapeutic response in a particular patient, compositions and routes of administration in the absence of toxicity to the patient. The selected dose will depend on various factors, including the activity of the applied definition is certain protease of the present invention, the route of administration, time of administration, rate of excretion applicable specific connection, duration of the treatment, other drugs, compounds and/or materials used in combination with the applied proteases, age, weight, condition, General health and medical history of the patient, which they were treated, and like factors well known in the prior art in the field of medicine. A physician or veterinarian having a middle level of competence, can readily determine and prescribe the effective amount of the desired pharmaceutical composition. For example, the physician or veterinarian could start assigning dosages of protease of the invention used in pharmaceutical compositions, with a level below what is required to achieve the desired therapeutic effect and gradually increase the dosage to achieve the desired effect.

Although to enter the protease of the present invention are possible, and separately, it is preferable introduction proteases in pharmaceutical formulation (composition).

In accordance with another preferred embodiment of the present invention, the drug further comprises at least one additional active ingredient.

The specified active ingredient can be any, but to contribute to the prevention and treatment of angiogenic diseases together is by proteases in accordance with the present invention. However, it is also possible to add active ingredients exhibiting a different action than the indicated protease.

In accordance with another preferred embodiment of the present invention, the drug contains at least one, preferably at least two (three or four) protease, which is selected from the group consisting of:

proteases of plants: bromelain, papain, fitsin, Ananin, balankin, curtain, faustman, hieronymian, Karategin, macrodantin, pingvinen, cusomizing, aktinidin;

proteases of bacteria: nattokinase, pronase, serrapeptase, staricase and subtilisin;

proteases mushrooms: brinase, sobrosa-S, Matanza;

proteases crustaceans: chilensis;

proteases earthworms: lumbrokinase;

proteases snakes: alfimeprase, fibrolite.

The present invention is additionally illustrated by the following drawings and examples.

Figure 1 shows the activity of LDH in the supernatant of the treated enzyme HUVEC.

Figure 2 shows the analysis based on the MTT with the processed enzyme HUVEC.

Figure 3 (A) shows the inhibition of the formation of tubules induced by VEGF, through a combination of 25% of bromelain, 50% nattokinase and 25% of papain.

Figure 3(B) shows the controlled processing only VEGF. Whereas VEGF-control narrow line formed tubes visible, processing the hydrated enzyme sample shows a broad region is not formed tubes, pointing to the antiangiogenic activity of a mixture of enzymes.

Figure 4 shows the concentration of VEGF in the blood of patients exposed to treatment rotozoom.

Figure 5 shows the MTT-assay with VEGF-stimulated HUVEC and rutoside. The inhibition of proliferation is not observed.

Figure 6 shows the toxic effects of Rutoside on HUVEC. Toxic effects not identified in the inactive HUVEC, whereas VEGF-activated HUVEC showed a small effect.

Figure 7 (left) shows the inhibition of spontaneous formation of tubules in HUVEC. Bromelain, fitsin, nattokinase, papain and serrapeptase, but not chymotrypsin or trypsin, inhibit the formation of tubules.

Figure 7 (right) shows the inhibition of the formation of tubules caused by VEGF in HUVEC. Bromelain, fitsin, nattokinase, papain and serrapeptase, but not chymotrypsin or trypsin, inhibit the formation of tubules almost the same as that of the untreated HUVEC.

EXAMPLES

BPH

Materials:

Bromelain from the stem of pineapple with activity 3,51 U/mg was obtained from Sigma Aldrich, Austria. Nattokinase activity of 10,000 IU/ml was purchased from Japan Bio Science Laboratory Co., Ltd. Papain from Carica papaya activity >3 U/mg was obtained from Sigma Aldrich, Austria

Methods:

The test for the toxicity

Antiproliferative activity of bromelain, nattokinase and papain were evaluated using analysis of activity lackaday the rogenaza deficit (LDH). Endothelial cells of the umbilical vein of a person (HUVEC) from polyethlyene cultures were treated with trypsin, were sown with a density of 2500 cells/well in 96-well microplate pre-coated with human fibronectin. To ensure proper attachment, the cells were incubated for 24 hours in endothelial basal medium 2MV (Cambrex Biochemicals)containing 10% fetal calf serum, 60 μg/ml of growth factor, endothelial cells, hrEGF, hrFGF2, hrIGF, hrVEGF, ascorbic acid and heparin. After attachment, the cells were incubated at 37°C and humidity of 95% in medium 199+10% Fetal calf serum (FCS) without growth factors. After 24 hours the supernatant was replaced with medium 199 containing 10% FCS, VEGF and different concentrations of enzymes. After 48-hour incubation period, the supernatant was collected and performed analysis on the activity of LDH in accordance with the manufacturer's instructions (Promega, Germany): 50 ál aliquot from all wells were transferred to a clean 96-well flat-bottomed (for enzymatic analysis) tablet. Analytical buffer was added to the substrate mixture and gently stirred. 50 μl of the recovered substrate mixture was added to each well. Tablet incubated for 30 minutes at room temperature. Then to each well was added 50 μl of stop solution. Not later than one hour was measured by optical density at DL is the wavelength of 490 nm with a reference wavelength of 620 nm. The results were expressed as % of untreated control.

The results are presented in figure 1. They clearly show that bromelain, nattokinase and papain up to level 25 µg/ml no toxic effects on HUVEC after 2 days of incubation.

Antiproliferative activity

Antiproliferative activity of bromelain, nattokinase and papain and their mixtures were evaluated using analysis based on 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Endothelial cells of the umbilical vein of a person (HUVEC) from polyethlyene cultures were treated with trypsin, were sown with a density of 2500 cells/well in 96-well microplate pre-coated with human fibronectin. To ensure proper attachment, the cells were incubated for 24 hours in endothelial basal medium 2MV (Cambrex Biochemicals)containing 10% fetal calf serum, 60 μg/ml of growth factor, endothelial cells, hrEGF, hrFGF2, hrIGF, hrVEGF, ascorbic acid and heparin. After attachment, the cells were incubated at 37°C and humidity of 95% in medium 199+10% fetal calf serum (FCS) without growth factors. After 24 hours the supernatant was replaced with medium 199 containing 10% FCS, VEGF and different concentrations of bromelain, nattokinase or papain. The cells were additionally incubated for 48 hours at 37°C and humidity of 95%. The TT-analysis was performed using MTT-EZ4U kit (Biomedica, Austria; in accordance with the manufacturer's instructions). The optical density was measured at a wavelength of 490 nm with a reference wavelength of 620 nm. The results were expressed as % of untreated control or % proliferation at 100% of the existing proliferation of VEGF-treated control.

The results of the experiments according to the release from the concentration presented in figure 2. They demonstrate a clear antiproliferative effect of bromelain, nattokinase and papain with bromelain and papain, reaching 75% increase at concentrations of 25 μg/ml together with a test on the determination of the activity of LDH, these data show a clear antiproliferative effect, but not the cytotoxic activity of bromelain, nattokinase or papain on HUVEC.

Antiangiogenic activity

Antiangiogenic activity in HUVEC was evaluated using the analysis of the formation of tubules. Matrigel (Becton Dickinson, Vienna), the inhibitory growth factor, was thawed at 4°C. In wells of 96-well microplates were pietravalle 50 μl per well. The plate was left at 4°C for 24 hours. Before the experiment, the plate was incubated for 30-60 minutes at 37°C for curing gel. HUVEC were incubated for 24 hours with medium 199 containing 2% FCS. The cells were collected by treatment with trypsin and were cultivated in covered with Matrigel 96-well micro is lansey with a density of 10000 cells per well. Drugs were added to the desired concentrations and cells were incubated in the presence or absence of 5 ng VEGF/ml Through the following 6 hours, the wells were photographed. Determined the total size of the tubes, using the software ImageJ (National Institute of Health, Bethesda, USA).

The results:

Example 1: Inhibition of growth of HUVEC using enzymes

The results of the mixtures shown in table 1. Visible clear antiproliferative effect.

Table 1
Inhibition of HUVEC growth using a combination of bromelain, nattokinase and papain in the presence of VEGF
BromelainNattokinasePapain% proliferation
0,00%0,00%100,00%75,45%
75,00%0,00%25,00%89,09%
16,67%16,67%66,67%91,82%
25,00%75,00%0,00% 94,55%
66,67%16,67%16,67%96,82%

Example 2: Formation of HUVEC tubes, incubated with the enzymes bromelain, nattokinase and papain

The results of the analyses of the formation of tubules mixtures are presented in figure 3 and in table 2.

25,00%
Table 2
Inhibition of the formation of tubules in % in HUVEC in the presence of VEGF. The results are shown as % inhibition of the formation of tubes for 0% accepted the formation of tubules in the samples treated only VEGF
BromelainNattokinasePapain% inhibition of the formation of tubules
75,00%25,00%0,00%98,11%
16,67%16,67%66,67%93,52%
25,00%25,00%50,00%87,75%
0,00%75,00%87,42%
75,00%0,00%25,00%86,34%
25,00%0,00%75,00%83,77%
16,67%66,67%16,67%83,65%
50,00%0,00%50,00%82,73%
0,00%50,00%50,00%80,01%
0,00%25,00%75,00%78,26%
33,33%33,33%33,33%74,14%
25,00%75,00%0,00%61,57%
50,00%25,00%25,00%47,78%
0,00%0,00%100,00%3,86%

Table 2 shows the length of the tubes, expressed as % inhibition of the formation of tubules.

The results clearly show the following.

1. The formation of tubules ingibirovalo with a mixture of bromelain, nattokinase and papain.

2. The combination of bromelain, nattokinase and papain has a more pronounced effect than the drugs separately.

Figure 3 (A) shows a combination of 25% of bromelain, 50% nattokinase and 25% of papain. (C) shows the control treated with VEGF. Whereas VEGF-control visible narrow area formed tubules (arrows), the sample is treated with enzymes, demonstrates a wide area without educated tubes, showing the antiangiogenic activity of a mixture of enzymes.

Example 3:

The influence of the drugs, containing bromelain in combination with other enzymes, the endothelial cells of the human prostate (HProsEC) was analyzed as in example 2 with minor modifications. The cells were kept for 24 hours in endothelial basal medium-2 (MV) (Lonza, Verviers), supplemented with ascorbic acid and hydrocortisone. The cells were inoculated to a density of 10,000 per well in covered with Matrigel 96-well microplates and incubated in the presence or absence of 5 ng VEGF/ml with combinations of enzymes (or 100% of bromelain, or 50% of enzyme and 50% of bromelain 12.5 µg/ml total). After 6 hours Proc. of the barrels were photographed and measured, using the software ImageJ (National Institute of Health, Bethesda, USA). The results were as follows:

enzyme + bromelain % inhibition of the formation of tubules

gene enzyme

% inhibition of the formation of tubules

Enzyme + bromelain

Brinase 21,00% 81,00%

Pronase 94,25% to 99.00%

Subtilisin 27,00% 100,00%

Streptokinase 29,00% 80,00%

Protease Type X 96,70% 100,00%

Protease Type XIII 60,60% for 98.00%

Bromelain 68,00%

Table 4: the results clearly show that proteasa combination containing bromelain and additional protease, leads to increased inhibition of the formation of tubules.

Example 4: the Formation of tubules in HUVEC in the presence of proteases from animals

Protease animals

For the study, inhibited the formation of tubes because of the separation of cells, HUVEC were treated with chymotrypsin and trypsin, both proteolytic enzyme widely used in cell separation. The results are shown in table 3. Either chymotrypsin or trypsin showed no significant effect.

Concentration [µg/ml]

Chymotrypsin Trypsin

0 100,00% 100,00%

2,5 112,40% 112,82%

5 of 95.94% 112,23%

10 81,16% 95,22%

20 131,18% 127,87%

Table 3: % of education tubules in HUVEC after treatment VEGF and trypsin or chymotrypsin

A significant reduction of proliferation were detected in the endothelial cells of the umbilical vein after obrabotkaponravilos, the nattokinase and papain individually or in combination in the concentration range from 25 to 100 mcg/ml

Demonstrated relatively low toxicity of bromelain, nattokinase and papain, especially when these levels with the provision antiproliferative effect. Concentrations up to 50 µg/ml did not cause toxic action HUVEC after 2 days incubation at 37°C.

Example 5: Effect of a mixture of enzymes on the concentration of VEGF in the blood

In this example investigated the effect of enzymatic treatment (mixture of nattokinase, bromelain, papain) on the content of VEGF in the blood. Shown (see results below)that enzymatic therapy significantly reduces the concentration of VEGF in human blood.

The test was performed as a randomized, no placebo control, multicenter pilot study on 111 patients with diabetes mellitus type 2, both sexes in two parallel two groups. 54 patients took rotosim™ (nattokinase (of 20,000 FU/g) 25 mg bromelain (2450 GDU/g) 90 mg of papain N. F. (2.400 USP Units/mg). 100 mg, Marlyn Nutraceuticals, USA) for 4 weeks. VEGF concentration in the plasma of the patient was determined before the addition of and immediately after 4 weeks after supplementation.

Patients served themselves as self-control relative to their initial values.

VEGF concentration in the blood was divided into 4 different groups (quartiles; see figure 4): VEGF to the concentrations in the blood of patients before therapy < 50 ng/ml; (i50 = source <50 nd/ml; C50 = end); and 100: VEGF concentration <100 ng/ml before therapy; 200: <200 ng/ml before therapy and I:>200 ng VEGF before therapy.

You can see that the use of the drug in accordance with the present invention, containing protease plants and/or microorganisms, can be used to reduce the level of VEGF in the blood and, therefore, can be used for the treatment of diseases related to neoangiogenesis.

Conclusion

Proteolytic enzymes of plant, bacterial and fungal origin can be used for the treatment of benign hypertrophy of the prostate gland in mammals with a decrease in the progression and regression of hypertrophic prostate tissue and, therefore, provide a means from obstructive symptoms associated with the disease.

Oral enzyme therapy reduces the increased VEGF concentration in diabetic patients, but no effect on normal levels of VEGF.

Proteolytic enzymes reduce the formation of tubules in HUVEC induced VEGF.

Enzymatic therapy inhibits angiogenic growth factors, leading to a decrease in vascularization, and this is based on its effect on the reduction of benign prostatic hypertrophy/hyperplasia of the prostate.

1. Applying at least one Protea the s for the manufacture of a medicinal product for the treatment and/or prevention of hypertrophy/hyperplasia benign prostate where the medicinal product is suitable for oral administration, and at least one protease selected from the group consisting of proteases of plants and microorganisms, and it is administered in an amount of from 1 to 100 mg/kg of body weight.

2. The use according to claim 1, wherein the protease plants are selected from the group consisting of bromelain, papain, fitsin, Ananina, Balanchine, Carolina, Faustina, hieronymian, karatina, macrodantin, pingvinen, macumazana, Actinidia.

3. The use according to claim 1, wherein the protease microorganisms are selected from the group consisting of nattokinase, brinase, seproz, sphericity, subtilisin and atanazy.

4. The use according to claim 1, wherein the protease from a non-mammal animals are selected from the group consisting of celinska, batroxobin, lumbrokinase, alfimeprase and atanazy.

5. The use according to claim 1, wherein at least one protease is contained in a medicinal product in an amount of 5 to 100% weight/weight, preferably from 20 to 80% weight/weight, more preferably from 30 to 70% weight/weight.

6. The use according to claim 1, wherein at least one protease is administered to the individual in an amount from 2 to 50 mg/kg, preferably from 5 to 20 mg/kg of body weight.

7. The use according to claim 1, in which the medicinal product further comprises at least one pharmaceutically acceptable carrier, diluent and/or excipient, preferably the binding agent, a filler, a leavening agent, a lubricating agent, a preservative and/or shell.

8. The use according to claim 1, in which the drug is provided in a pharmaceutical form which is selected from the group consisting of tablets, gastroresistant tablets and capsules, preferably gastroresistant capsules.

9. The use according to claim 1, in which the medicinal product further comprises at least one additional active ingredient.

10. The use according to claim 9, wherein at least one additional active ingredient is a flavonoid and/or antioxidant.

11. Use PP, 10, wherein the additional active ingredient is contained in a medicinal product in an amount of 5 to 35% weight/weight, preferably from 10 to 30% weight/weight, more preferably from 15 to 25% weight/weight.

12. The use according to claim 1, wherein the drug includes at least one protease which is selected from the group consisting of brinase, bromelain, fitsin, lumbrokinase, nattokinase, papain, pronase, seproz-S, serrapeptase, sphericity, subtilisin, celinska and batroxobin, Athanasy, alfimeprase, fibrolite, macumazana, Actinidia, Ananina, Balanchine, Carolina, gastusin, hieronymian, karatina, pingvinen and macrodantin.

13. The pharmaceutical is ski preparation for oral administration, containing at least two protease according to any one of claims 1 to 12 for the treatment of benign hypertrophy/hyperplasia of the prostate.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutics and medicine, and concerns a pharmaceutical composition for treating cancer combined with preventing and/or treating side effects, comprising as active substance at least vincristine, paclitaxel or oxaliplatin and at least oxime cholest-4-en-3-one, or one of a derivatives thereof.

EFFECT: reducing side effects of the anti-cancer therapy.

9 cl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to biochemistry. What is presented is pegylated interferon-α2b (IFN-α2b) having a structure as shown in the patent claim. Pegylated interferon-α2b is prepared by combining IFN-α2b with branched Y-polyethylene glycol (YPEG), wherein YPEG is bound to IFN-α2b by an amide bond formed by ε-amino group of a side of Lys residue in IFN-α2b in the position of 134 in SEQ ID No. 1. A method for preparing and purifying pegylated IFN-α2b involves the stages as follows. In an alkaline medium at pH 9.0, branched Y-polyethylene glycol is reacted with IFN-α2b to prepare pegylated IFN-α2b. The prepared reaction products are recovered by an anion exchange resin, and the given products are eluted by an anion gradient to prepare modified products. The anion exchange resin is Q Sepharose FF, and the anion gradient is a chloride ion gradient. Further, the modified products are eluted by a cation exchange resin with a cation gradient. The cation exchange resin is SP Sepharose FF, and the cation gradient is a sodium ion gradient. Thereafter, each peak is collected separately. The product activity of each peak is determined to choose a peak corresponding to the reaction product having the highest activity. What is also presented is a formulation for treating a disease requiring IFN-α2b to be used, which consists of a pharmaceutically effective amount of said pegylated IFN-α2b and a pharmaceutically acceptable carrier, or an inactive substance. Pegylated IFN-α2b or the above formulation is also used for preparing a medicine for treating various diseases requiring IFN-α2b to be used.

EFFECT: presented pegylated IFN-α2b has a higher specific activity of 2,65±0,185×106 IU/mg and a prolonged serum half-life.

26 cl, 11 dwg, 5 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to tetraazaphenalen-3-one compounds, particularly a compound of formula which inhibit poly(ADP-ribose)polymerase (PARP) and may be used for chemosensitisation in relation to anticancer chemotherapeutic agents.

EFFECT: higher effectiveness of the chemotherapeutic agents, and in treating cancer diseases characterised by the DNA repair defects.

13 cl, 2 tbl, 6 dwg, 3 ex

Cytokine inhibitors // 2485113

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to imidazole compounds of formula wherein the radical values A, X, R1, R2, R3 are presented in clause 1 of the patent claim.

EFFECT: there are disclosed pharmaceutical compositions of sail compounds for cytokine (eg TNFα or IL-1β) reduction.

16 cl, 107 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new compounds of general formula III which possess the properties of JAK pathway inhibitors and JAK-kinase inhibitors. In formula III: X is specified in a group consisting of C1-C10alkyl, amino, halogen, carboxyl, carboxylic acid ester, C2alkynyl, substituted tri-C1-C6alkylsilyl; R represents hydrogen; the cycle A is specified in a group consisting of C6aryl, bicycloheptene, five-and sis-member mono- or 10-member bicyclic heteroaryl including 1 to 3 heteroatoms specified in a group of heteroatoms, including N, O or S, and five- or six-member mono- or 10-member bicyclic heterocycle, including 1 to 2 heteroatoms specified in a group of heteroatoms, including N or O; p means 0, 1, 2 or 3; each of R2 is independently specified in a group consisting of C1-C6alkyl, C1-C4alkyl substituted by 1 to 3 substitutes. The other substitute and radical values are specified in the patent claim.

EFFECT: compounds may be used in preparing a therapeutic agent for T-cell mediated autoimmune disease, for treating or preventing allograft rejection in a recipient, for treating or preventing a type IV hypersensitivity reactions, which includes administering the above agent containing the compound according to cl 1-11, in an amount effective to treat the autoimmune disease or the allograft rejection or the type IV hypersensitivity.

23 cl, 7 dwg, 12 tbl, 43 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to medicine, particularly to a combination for treating cancer comprising at least 1) a fusion protein comprising a portion specifically recognizing EDb-fibronectin representing an antibody, and a portion representing interleukine-2 and 2) a molecule that binds to B-cells, B-cell precursors and/or their cancerous counterpart which is an antibody or antibody or an antibody fragment specifically binding to CD-20. What is also declared is the use of this combination.

EFFECT: group of inventions provides higher clinical effectiveness in treating a tumour.

23 cl, 1 tbl, 7 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to pharmaceutical composition for treatment of oncological diseases based on sodium dichloroacetate, characterised by fact, that it additionally contains theobromine (3,7-dimethylxanthine) or caffeine (1,3,7- threemethylxanthine) or paraxanthine (1,7- dimethylxanthine), with ratio of sodium dichloroacetate to xathine methyl derivative of those mentioned above constitutes 1:1-10:1.

EFFECT: pharmaceutical composition ensures synergetic effect.

9 tbl, 9 ex, 6 dwg

FIELD: medicine.

SUBSTANCE: on the 1st, 5th, 10th and 15th experimental day at 15 to 16 o'clock, cyclophosphan 50 mg/kg is intraperitoneally introduced into white outbread rats suffering grafted sarcoma 45 having reached the size of 3 cm3. That is combined with daily intragastric introduction of a cyclic solution of adenosine-3'-5'-monophosphate in a dose of 0.01 mg three times a day every 2 hours starting from 10 o'clock in the morning.

EFFECT: higher non-specific anti-tumour resistance, intensified action of the cytostatic on the late tumour, and inhibited systemic action of the tumour.

3 tbl

FIELD: biotechnologies.

SUBSTANCE: cell lines of human melanoma, such as Mel-XYl, deposited in DSMZ under No. DSM ACC2830, Mel-XY2, deposited in DSMZ under No. DSM ACC2831, Mel-XY3, deposited in DSMZ under No. DSM ACC2832, and Mel-XX4, deposited in DSMZ under No. DSM ACC2829, are radiated to produce populations with apoptotic and necrotic phenotype. The produced cell lines are used in various combinations for treatment of malignant diseases, namely, for treatment of human melanoma. Also the specified populations of radiated cells are cocultivated with autological dendrite cells for further joint injection to a patient suffering from melanoma.

EFFECT: invention makes it possible to induce a resistant anti-cancer immune response in a mammal suffering from melanoma.

21 cl, 10 dwg, 4 tbl, 5 ex

FIELD: biotechnologies.

SUBSTANCE: medicinal agent for inhibition of MASP-2-dependent complement is an agent containing an antibody or its fragment, bound with a full-size polypeptide MASP-2, but not bound with a MASP-2 N-terminal fragment containing CUBI-EGF-CUBII domains and not bound with a MASP-2 C-terminal fragment, containing of CCPII-SP domains.

EFFECT: invention makes it possible to selectively inhibit MASP-2-dependent activation of complement, at the same time leaving Clq-dependent classic path of complement activation as functionally unaffected.

9 cl, 39 dwg, 7 tbl, 31 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely urology, and may be used for treating suppurative septic complications of prostate adenoma. For this purpose, with underlying administering antibiotics, ozonised saline solution in the concentration of 3 to 6 mg/l in the amount of 200 to 400 ml is additionally intravenously administered; the therapeutic course is 3-5 injections depending on the patient's state. Besides, ozonised saline solution in the concentration of 5 to 8 mg/l is injected into the bladder in the preoperative preparation period, and from the second postoperative day, saline solution is introduced drop-by-drop in the amount of 400.0 ml daily five times with a two-way flushing system.

EFFECT: method enables reducing the risk of urosepsis, eliminating intoxication and accelerating postoperative wound healing of a BPH bed by anti-inflammatory, detoxification, analgesic action, improved blood rheology and microcirculation, increased local immunity with activated regenerative and reparative processes.

2 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula (1), in which Ar is a group of formula (Ar-1) or (Ar-2), in which R1 is a halogen, R2 is hydrogen, R3 is hydrogen, R4 is hydrogen, alkyl or alkenyl, X is a nitrogen atom or CH, R5 and R6 are each hydrogen and h equals 1; 1 equals 1 or 2; m equals 1 or 2; n equals 0, 1 or 2; o equals an integer from 0 to 3, under the condition that n and o are equal to 0 at the same time. Values of group A are as given in claim 1 of the invention. Described also is a pharmaceutical composition having agonistic activity with respect to 7 serotonin (5-HT4-receptors), which contains a compound of formula (1) and an agent which stimulates enterokinesis or improves functioning of the alimentary canal, which contains a compound of formula (1) as an active ingredient.

EFFECT: novel compounds are obtained and described, which have strong affinity towards 4 serotonin receptors, which are useful as an agent which stimulates enterokinesis or an agent which improves functioning of the alimentary canal.

28 cl, 233 ex, 29 tbl

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to urology, and can be used for treatment of chronic abacterial prostatitis in men, which is not connected with dysfunction of urethral sphincter. For this purpose, efficient amount of fispemiphene is introduced to patient who needs it. Introduction of fispemiphene is performed in amount in the range from 0.1 to approximately 100 mg/kg of body weight.

EFFECT: invention ensures efficient treatment of abacterial prostatitis at the early stage of disease where only histological changes of prostate gland are present without presence of urethral sphincter dysfunction, with drug fispemiphene, which demonstrates antagonistic activity to estrogen receptors in particular with respect to prostate tissue.

2 cl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of pharmaceutical industry, in particular to composition, which contains epigallocatechine-3-gallate as active component and target additive. Pharmaceutical composition for peroral introduction, which contains epigallocatechine-3-gallate and block-copolymer of oxyethylene and oxypropylene, in which content of hydrophobic block constitutes not less than 50 wt %, and molecular weight of hydrophilic block constitutes 2250 Dalton and not more, with specified weight ratio of epigallocatechine-3-gallate and block copolymer 1:1-10:1. Application of upper described pharmaceutical composition for treatment of atopic dermatitis, Crohn's disease, adenomyosis, and hyperplastic diseases of prostate.

EFFECT: composition has increased bioavailability in peroral application, improves absorption of active compound epigallocatechine-3-gallate in blood flow.

6 cl, 2 dwg, 4 tbl, 10 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmacy, namely technology of preparation of rectal suppositories. The suppositories with solid licorice extract consist of solid licorice extract 0.75 g on a hydrophilic base, polyethylene glycol 6000 1.25 g and polyethylene glycol polystear 0.25 g.

EFFECT: making the preparations exhibiting anti-inflammatory action, and also a positive effect on the hormonal status and the copulative function.

7 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to novel acid-additive salts of pyrrolopyrimidinone derivative, represented by formula (1) which is selected from gentisate, maleate, citrate, fumarate and semitartrate salts, which possess improved properties in their application, in particular higher stability.

EFFECT: invention also relates to method of obtaining acid-additive salts of pyrrolopyrimidinone derivative, represented by formula (1) and to pharmaceutical composition, containing them, for treatment and prevention of erectile dysfunction, pulmonary arterial hypertension, chronic obstructive lung disease, benign prostate gland hypertrophy and diseases of lower urinary tract.

11 cl, 30 ex, 7 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: declared group of inventions relates to medicine. There are declared methods of treatment and preventive treatment of prostate cancer in males and a method for treatment of prostate cancer in male dogs involving a single agent therapy in the form of introducing a therapeutic amount of N,N'-bis(2,3-butadienyl)-1,4-butane-diamine or its pharmaceutically acceptable salt or solvate. The therapeutic amount is specified within the range from 1-100 mg/kgbody weight and dosed within the range from twice a week to once a day.

EFFECT: group of inventions has high therapeutic efficacy.

26 dwg, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to coated pharmaceutical formulations with a controlled-release active substance and to the use thereof for treating benign prostatic hyperplasia. The formulation contains a therapeutically effective amount of the active substance representing tamsulosin or its pharmaceutically acceptable salt encapsulated in a granulated core, and an insoluble permeable polymer found both in the granulated core and in the coating. Said pharmaceutical formulations may also contain a surfactant and other optional excipients. Besides, the invention refers to a method for preparing the pharmaceutical formulations which involves mixing core ingredients, granulating, pressing and shaping in a sphere, drying and coating.

EFFECT: invention provides the formulations with pH-independent release of the active substance, low inter-individual variability and low biological variability.

14 cl, 7 ex

FIELD: medicine.

SUBSTANCE: The invention relates to pharmacology, in particular, to medication for preventing or treating herpes labialis or herpes genitalis. The homeopathic medication or biologically active additive with anti-viral effect or preventing or treating herpes labialis or herpes genitalis contains: Nisylen, Cepa, Euphrasia, Belladonna and Mericulis Solubilis, furthermore, the components are present with a certain dilution and amount. The combination of the said components is used for production of homeopathic medication or biologically active additive with anti-viral effect or preventing or treating herpes labialis or herpes genitalis.

EFFECT: production of medication which effectively treats herpes labialis or herpes genitalis.

14 cl, 1 ex

FIELD: medicine.

SUBSTANCE: invention refers namely to pharmaceutical industry, and concerns tablets with the slowed down liberation, containing alfuzosin hydrochloride. The tablet includes: the top layer including 0,2 weight parts of alfuzosin HC1, 90 weight. parts of hypromellose, 29. 6 weight parts of polyethylene oxide, 1 weight part of stearyl alcohol, 0.4 weight parts of easy waterless silicon acid and 1 weight part of ethyl cellulose; and the bottom layer including 9.8 weight parts alfuzosin HC1, 36-62 weight parts of microcrystalline cellulose, 24-60 weight parts of polyethylene oxide, 20 weight parts low substituted hydroxypropyl cellulose, 1-6 weight parts of stearyl alcohol and 99 weights parts of hydroxypropyl cellulose.

EFFECT: received tablet is characterized by the liberation of the medicine with the curve of the zero order.

3 cl, 2 tbl, 10 ex, 6 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry and represents a thermoreversible, thermoplastic pharmaceutical composition comprising: (a) a biologically active botulinum toxin and (b) a thermoplastic poloxamer wherein poloxamer stabilises botulinum toxin so that botulinum toxin keeps biological activity with botulinum toxin release from the pharmaceutical compositions in vivo, wherein the pharmaceutical composition can be prepared with no substances contained in blood, such as albumin and therefore without any infectious substances of blood components, such as prion.

EFFECT: invention provides high stability and high percentage of the toxin activity recovery.

18 cl, 6 ex

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