WO1993010793A1

WO1993010793A1 - Antivirus agent

Info

Publication number: WO1993010793A1
Application number: PCT/RU1992/000170
Authority: WO
Inventors: Jury Petrovich Vainberg; Elli Nikolaevna Kaplina
Original assignee: Jury Petrovich Vainberg; Elli Nikolaevna Kaplina
Priority date: 1991-12-03
Filing date: 1992-09-09
Publication date: 1993-06-10

Abstract

An antivirus agent consists of an active substance which is a complex of a sodium salt of native DNA with a polyvalent metal selected from metals Ni, Fe, Co, Mn, Mg or Zn taken at a mole ratio of 10-1000:0.5-3.0, respectively, and a pharmaceutical diluent. The claimed agent has an antivirus activity against the herpes virus, the orthomyxo virus - influenza virus, the paramyxovirus - plague virus of carnivores and retroviruses, including the human immunodeficiency virus.

Description

FIELD OF THE INVENTION The present invention is in the medical field, and more precisely, in a new medical device. 5 PREVIOUS LEVEL OF TECHNOLOGY

Various chemo-therapeutic agents are widely known, causing death of the virus or inhibiting their development, which are used for the treatment and treatment of virus infections. The effect of these devices is directed to different stages of the US production of viral particles in human cells. or lively. Separate industrial products are different as far as the mechanism of action is concerned, as is the use of active substances in the case of 15 It is known, for example, that there is an ingestion of an addictive substance (kerocide), which has the potential to include a serious disturbance in the presence of an active substance and, due to this, there is an active disease Наdnaκο ,. this product is highly effective and is used only for 0 for local use in patients with ocular disease; It also has a number of adverse effects (discharge of the conjunctiva, leakage of eyelids).

The computer has been known to interfere with the synthesis and replication of virus particles. Οdnaκο, eτοτ πρeπaρaτ HA 5 χοdiτ ^{'πρimenenie} in οsnοvnοm for προφilaκτiκi gρiππa.

Widely known drugs are derivatives of adamantamines (amantadine, remantadine), which inhibit the earlier stages of the reproduction of the virus of the group. These drugs are most active in relation to the type of virus and find less than 0 For the treatment and treatment of the disease only.

Also known are the drugs that have a beneficial effect on the anti-virus, (anti-virus, anti-virus). 5 The most famous known drug, which is highly active in the presence of drugs, including the human immunodeficiency virus (is 3

DΟΤIΜIDIΗ). . (yϊsϊικa ^η B- _Β,, Ρϊzs.1 ^'_, _ Λ,, Zgχesο:..} - ;., Yιo ο>ϊsχ1; s o £ aζχάο-uπχάχηe (ϋΖΙ) χη 1; ηe 1; gea1 ; those η1; 0 ^ - 2 - ρаϊгϊеιгз ν ^~ 1 ~ ЬЬ ΑΙΒ8 аηά ΑΙΙ) 8-ге1а1; еά сοιаρϊеχ, J.ΕηξΙ. СϊΙеά. 1987, 317: 192-197; ϋ-ви уа уа Η Η, Βгοάегε, 31 ^; ha1 ^; e £ ^χ eε £ οг а -Ηιегρу ± ^η ΑΙБ8, ΙΤз-игиге, 1987, 325: 773-778). This medication is used to support the treatment of patients with impaired human immunodeficiency (HIV) and a long-term illness. However, this product is not efficient enough and efficient. Β sisτeme "χη ν ^ 1 ^{^^^,} azidοτimidin in maκsi- malnο πeρenοsimοy dοze not ποlnοsτyu οsτanavlivaeτ ρaz- viτie viρusοv ΒICH, πρi eτοm ποgibaeτ οκοlο 30 $ nορmal- nyχ κleτοκ. Inτeρtseρebρalnοe treatment of mice with 10 mg / κg eτοgο πρeπaρaτa vyzyvaeτ death of 50 lively.

Owing to a commercially available product, it possesses a commercially viable activity in respect of non-hostile and non-hostile countries.

Ρasκρyτie izοbρeτeniya Β οsnοvu izοbρeτeniya ποyaοzhena task sοzdaniya nοvο- gο προτivοviρusnοgο sρedsτva, οbladayuschegο vysοκοy προτi- vοviρusnοy aκτivnοsτyu κaκ DΗΚ κ-sοdeρzhaschim, τaκ and ΡΗΚ- sοdeρzhaschim viρusam not τοκsichnοgο not imeyuschegο vρednyχ ποbοchnyχ deysτvy.

Task ρeshena τem, chτο zayavlyaeτsya προτivοviρusnοe sρedsτvο, sοdeρzhaschee deysτvuyuschee veschesτvο and φaρmatsevτi- chesκy ρazbaviτel, κοτοροe sοglasnο izοbρeτeniyu in κa- chesτve deysτvuyuschegο veschesτva sοdeρzhiτ κοmπleκs naτρievοy sοli naτivnοi dezοκsiρibοnuκleinοvοϊ κislοτy with ποlivalenτ- nym meτallοm, vybρannym of gρuππy meτallοv w, Ρe, Sο ,, §§ or Ζη, taken in the 10–1000 millennium: 0.5–3, respectively.

In order ποvysheniya προτivοviρusnοy aκτivnοsτi targeted sοοbρaznο, chτοby zayavlyaemοe sρedsτvο in κachesτve deysτ- vuyuschegο veschesτva sοdeρzhalο κοmπleκs naτρievοy sοli HA τivnοy DΗΚ with mοleκulyaρnοi massοy 300-500 κD with zhelezοm or niκelem or κοbalτοm or tsishοm or maρgantsem, vzyaτymi in mοlnοm Ratio 10-1000: 0.5-3.0 respectively. - -

Zayavlyaemοe sρedsτvο mοzheτ byτ isποlzοvanο in ρazlich- nyχ φaρmatsevτichesκi πρigοdnyχ leκaρsτvennyχ φορma ^χ (in vi- de ineκtsy, ρasτvοροv, suπποziτορiev and dρugi). For the purposes of the application, we declare a device in the form of injections, it is only constituent that contains active material in quantities

1.5 wt., And in the form of disinfectants for commercial use in the amount of 3.0 wt. As a pharmaceutical agent, the claimed medication may contain any kind of suitable diluent. The claimed product is preferably in the form of injections or disposable preparations for use in a diluent-supplied water.

Zayavlyaemοe προτivοviρusnοe sρedsτvο οbladaeτ vysο- κοy προτivοviρusnοy aκτivnοsτyu πο οτnοsheniyu κ-DΒΚ sοdeρzhaschim viρusam (viρusu geρπesa) and κ ΡΒΚ-sοdeρzhaschim viρusam (ορτοmiκsοviρusam - vοzbudiτelyam gρiππa, πaρamiκsοviρusu - vοzbudiτelyu πlοτοyadnyχ plague, ρeτροviρusu - vοzbudiτelyu AIDS).

Zayavlyaemοe sρedsτvο, vvedennοe in ορganizm, προniκaeτ πuτem endοtsiτοza in tsiτοπlazmu aκτivnοdelyascheysya κleτκi, naπρimeρ, limφοtsiτa tying therein κaκ οbρaτnuyu τρansκ- ρiπτazu (πuτem οbρazοvaniya sshivοκ cheρez vχοdyaschie in κοmπ- leκs meτally) τaκ and belκi ΒICH, προdutsiροvannye genοmοm zaρazhennοy κleτκi (due to interaction with the deoxyribonuclein part of the complex). The used traditional components in the event of a deactivation of the internal components are subject to permanent disruption of any non-compliant products.

Β As a result of these processes, the development of human immunodeficiency virus in the cell is inhibited in the South.

Best vaρianτ οsuschesτvleniya izοbρeτeniya Zayavlyaezhοe προτivοviρusnοe sρedsτvο sοsτοiτ of In- sτvuyuschegο veschesτva - κοmπleκsa naτρievοy sοli naτivnοy DΗΚ with ποlivalenτnym meτallοm, vybρannym of meτallοv κι, ^ e Sο, Μ ^η, β or ^ζη, vzyaτymi in mοlnοm sοοτnοshenii 10-1000: 0 5-3.0 and pharmaceutical diluent. It is representative that the natrium salt of the naive had a molecular weight of 300-500 kDa. THIS EXCEEDS PROSPECTS - 4 - the nomenclature of the complex in the body of a human organism or living and contributes to the improvement of its digestibility. Compounds of sodium nativity with the indicated fuel metals taken in the ratio of 10-1000: 0.5-3.0, have the advantage of an effective activity. Relationships of the above and the below-mentioned divisions of the divisions adversely affect the activity of the claimed assets. The claimed system is dominated by active activity in the case of the virus of the virus, the virus - the virus, the virus. The plague of carnivores and the plague - the virus of human immunodeficiency.

Tests of the productive activity of the claimed preparation were carried out. Declared a component in the form of a separate product of the complex of the native D-Ms and iron (a small ratio of 250: 1,7) was impaired in the suspension of the vehicle. Virus of the group Ι-τ τ и и а; the plague virus is unpleasant (parasamic virus).

Further, the cages were incubated for 3-4 days in an atmosphere of 5 СΟ ^ and 98 $ of humidity at a temperature of 37 ° С. For each type of virus, at least 2 parallel operations were introduced.

The pressure was divided: a) a complete pressure of the virus group was observed due to the concentration of the claimed 400-500 mkg / ml; b) The total pressure of the Herpes spp. virus is observed at a concentration of 500-600 mcg / ml; c) A complete suppression of the plague virus is completely observed, with a concentration of 800-900 mcg / ml. Pροτivοviρusnaya aκτivnοsτ zayavlyaemοgο sρedsτva πο οτnοsheniyu κ viρusu immunοdeφitsiτa chelοveκa was studied in ρazlichnyχ vaρianτaχ zayavlyaemοgο sρedsτva, πρedsτavlyayu- schiχ sοbοy vοdny ρasτvορ naτρievοy sοli naτivnοy DΗΚ with meτallami ^η or vι or Sο, or ^, or ^Ιη, or K {in mοlnyχ sοοτnοsheniyaχ DΗΚ -v? z / He, ρ equals 10: 3; 350: 0.5 1000: 1, with an active substance concentration of 5 mg / ml. The tests were carried out in comparison with the known - - with a preparation with azidothymidine (aqueous solution with a concentration of 5 mg / ml of the active substance).

We used live human cells C-58 and ΜΤ4. The cells were cultured at a concentration of 0.3-0.5x10 ^e cells in I ml of salmon nnα 1640 with 10 $ fetal calves, 300 g / ml of glutamine, and 100 g / ml of gene were diluted. The life of the cage was shared by a 0.4-share of the market. As a source of the virus, they used the ΒICH-Ι / ИΒ8 strain, highlighted in the Institute of Virology named after D.I.Ivanovsk, and group of highly infectious viruses. The division of the antigenic activity of the preparations was carried out in 96-well plastic panels (the “Dinatec” company, Switzerland) according to the known immunoassay. The study of human immunodeficiency virus products was carried out by taking into account virus-induced syncytia in cell cultures.

For the analysis of the effective activity of the drugs, the suspension of the cassettes were placed in plastic 24-well panels, they treated the patient for various conditions. The infectiousness of infection amounted to 0.01 ΤTsZ ₅₀ / clet. After this culture, the cells were incubated at 37 ° C in atmosphere 5, with С0 ₂ and 98 $ humidity for 5-7 days after taking into account the results.

The study of the cytotoxicity of the drugs was carried out similarly, only a step of infection of the human immunodeficiency was neglected. The efficiency of the devices is based on the following parameters:

I. Οπρedelyali eφφeκτ ποdavleniya πρeπaρaτοm οbρazο- Bani viρusindutsiροvanny ^χ sintsiτiev chuvsτviτelnyχ κ-ΒIYA κleτοchnym κulτuρa.

Φορmiροvanie sinΣ _^ giev - giganτsκiχ κοnglοmeρaτοv κleτοκ - yavlyaeτsl οdnοy of ^ aρaκτeρnyχ οsοbennοsτey ΒIΗ πρi egο ρeπροduκtsii. Οbρazοvanie sintsiτiev svyazanο with vzaimοdeysτviem viρusnοgο belκa & ΡΙ20, ρasποlοzhennο- gο on ποveρ ^χ nοsτi zaρazhennyχ κleτοκ ρetseaτορa Sa and 4, - 6 - uncleaned ^χ cells located on the membrane. The result of the interaction is the merger of the cell membranes and the formation of non-immune! cell-based structures, which include many cell-based nuclei.

2. We assessed the level of antigen in the culture fluid of cell culture, infected with HIV. with the help of immunoassay analysis. This test examines the products of the Virus run for various stages of infection of the cell culture. The results of the experiments are reported in tables 1,2,3,4.

Τablitsa I Issledοvanie anτigenοv viρusa immunοdeφitsiτa chelοveκa meτοdοm immunοφeρmenτnοgο analysis κulτuρalnοy zhidκοsτi κleτοκ, οbρabοτanny researched πρeπaρaτami ^χ and ^χ inφitsiροvanny ΒICH

ππ PREPARATIONS INDICATORS OF OPTICAL ACCESSIBILITY

Oscillation treatment, w τgΑ / ml

200 400 800 1600

1 Declared on the basis of ДΗΚ-υа with Ζη

2 Declared Mediation on the Basis of D-Ya with Μa

3 Declared Mediation on the Basis of Д-Ηа with Ρе

4 Declared mediation on the basis of ДС-иа с з.

5 Declared mediation on the basis of D-Ms with

6 Declared mediation on the basis of ДΗΚ- ^ а with ёё

7 CONTINUOUS VIRUS

8 CONTROL OF UNINFECTED CELLS

9 Azidothymidine 50 g / ml

10 Azidothymidine 500 g / ml

- -

Page 2 Investigation of the cytotoxicity of the claimed media in the S-55 cell culture

^ Preparation Preparation of the preparation, burned / ml ^*

0 200

Thor- Life- Thor- Life- black and white; simple way to clean the cell; in the I ml cell;

1 Declared matter on the basis of Д-Да with ζ ^η 0.78 74.5 Declared matter on the basis of Η-Иа with И 0,84 86, 7 Declared claim on the basis of Де-Дсе 4 се ^ 0 The base of DΗΚ-Ηa with a metric of 0.88 80.5 The claimed composition on the basis of DΗΚ-сa on the basis of 0.90 89.2 The declared claim on the basis of DΗΚ-аa of 0.79 75, 4 Κοntrοl 0,90 -

Table 2

Concentration of the preparation. burn / ml

400 800 1600 3200 life-spares - glue:., $,

8 9 10 II 12 13 14

0.85 81, 7 0.83 76, 9 0.84 87, 1 0.82 32, 8 - 8 -

Table 2

Table 3 Comparison of the study of the known cytotoxicity of azidothymidine in the culture of the S3-ZZ cell

Concentration of the preparation, burning / ml ππ Preparation

0 50

0.78 66.7 0.61 55.2 0.43 33.1 - -

Table 3

7 8 9 10 ϊϊ 12

Table 4 Potential activity and quotient of the claimed medication compared to a known drug (azidothymidine) and potentiol

- 10 -

Table 4

1 7 8

2 0.82 71.0 0.86 79.0 0.85 84.3

3 0.86 78.4 0.84 87.1 0.89 76.3

4 0.87 79.3 0.84 82.1 0.87 81.7

5 0.61 55.2

6 -

7 0.91 86.3

Note: All data are provided for optimal operating efficiency due to received data.

The most efficient activity is noted in the claimed sredstva on the basis of DΗΚ-Κa from the 2nd. A dose of 200–400 g / ml in 5–10 times reduces the amount of viral induced syncytia. At a concentration of 1,600 g / ml, a practically complete disappearance of the virus-induced syncytia is observed. A comparative analysis of this product with a known preparation of azidothymidine (Felka, United States.) Indicates that the consumptions are made of the following;

Issledοvanie uροvnya προduκtsii anτigena Βyϊ meτοdοm immunοφeρmenτnοgο analysis (τablitsa I) vyyavilο ποdavle- of seκρetsii viρusny ^χ bezhοv claimed sρedsτvοm. The level of decrease in the production of antigen for HIV and the use of the claimed drug and azidothymidine was practically identical: the claimed drug 0.147-0.103 and azidothymidine 0.147-0.091. By virtue of the effective effect in the case of ΒJ-Ι, the claimed assets are also available on the basis of DΗΚ-Ms with Ζη or on the basis of DΗΚ-ϊϊa with Сο, ιτϊ, Μη, Ш £. However, the glue protection - - The effect of the virus was lower. As for the claimed preparation, on the basis of Д-Ж, with ζη, an effective effect was observed only in a concentration of 1600 g / ml. With this, the number of viral induced syncnts decreased only 2 times.

The claimed medium on the basis of D-Ms was previously effective in the concentration of 400-1600 g / mp. For doses of 800–1600 g / ml, its potent effect was compatible with the beneficial activity of azidothymidine. The study of cases of neglect indicates the absence of a toxic effect in the case of the use of all variants of the treatment (1,600 cases).

Higher concentrations of the preparations (3200 g / ml) indicate an appreciable significant effect. It is noted that the decrease in the total number of cells (in 1.4-2.5 times), and the decrease in their life costs up to 32.8-43.5 $ (claimed to be for free) ДΗΚ-ιΤа с Сο).

For comparison, note that the use of an effective concentration of the azidothymidine preparation

(500 g / ml) showed the toxic effect on the cells, yielding up to $ 30 each. However, at the same time, there was no convenient effect on the applicants on the basis of Dv-va with, D-va s-s and D-va, and there was an analogous protection.

Ηeοbχοdimο οτmeτiτ, chτο isποlzοvanie in eκsπeρimen- τaχ dvuχ vidοv κleτοchnyχ κulτuρ: ΜΤ-4 and SΕΜ-ss, dvuχ shτammοv ΒICH-Ι, and τaκzhe ρeaκtsii immunοφlyuορestsentsii assay viρusindutsiροvannyχ sintsiτiev not vyyavilο suschesτ- vennyχ ρazlichy in ποluchennyχ dannyχ and ποdτveρdilο univeρ- salnοsι ... found out) The results obtained testify to the fact that the claimed mediocre property is inoperative in the case of ΒICH-Ι.

The most beneficial aspect is noted in the claimed mediation on the basis of the DΗΚ-υа with He. Before the active substance 800–1600 g / ml was detected, the complete disappearance of the virus-induced syncytia was detected. ^Χ Used for distribution of this product, in the absence of azide- - 12 - midina, not soooooo.

The claimed medium is a minor one. When the mice were administered intramuscularly, a maximum dose of 1,500 mg / kg was not observed, there were no significant toxic effects.

The claimed medium has been tested in clinics in Lviv, a large SDS. The drug was prescribed for oral administration in doses of 300 to 1200 mg for administration of 1-2 times per day for 30 days. Observations indicated that there was no other harmful effect in all cases. In this case, the product was reactivated by activating the activity of the immune system, increasing the content of all types of lymphomas in the body. Patients felt better, ^' they had normalized temperature, showed symptoms, they began to lose weight. Table 5 below shows the effect of the claimed drug on the state of the immunological system. ^"

Table 5

Bologna I

Accounts 5000 4600 3600 4600 4300 3900

Total Limits 950 1800 1400 1500

140 550 170 250

420 1210 510 560 τ ₄ ⁸ /, τ ₈ 0.34 0.45 0.33 0.45

women 95 290 290 210 women

Bolnoy 2

Ecommerce 3100 3200 4100 3900 Total Limits 360 480 1700 1000

- -

Table 5

7

260

The inventive medium can be used in any other medicinal formulations, is predominantly used as an industrial solution The medicinal formulations of the claimed medication are prepared by well-known methods. The material in question is the property of the natur- al salt, which is safe for people to eat and has the advantage of being completely non-hazardous.

The claimed medium is used in the form of injectable substances with a content of active substance of 1.5 wt. $ Or of a consumable for direct use of which consumes $ 3. The claimed device does not have any other beneficial effects or indications for use.

INDUSTRIAL APPLICABILITY The claimed inventive medication is used in medicine and veterinary medicine for the treatment of diseases caused by various types of viral infections.

Claims

- 14 - ΦΟΡΜШ. ΟΟΡΕΤΗΗΗ

1. Pροτivοviρusnοe sρedsτvο, sοdeρzhaschee deysτvuyuschee veschesτvο and φaρmatsevτichesκii ρazbaviτel, χaρaκτeρizuyuscheesya τem, chτο in κachesτve deysτvuyuschegο veschesτva οnο sοdeρzhiτ κοmπleκs naτρievοy sοli naτivnοi dezοκsiρibοnuκleinοvοy κislοτy with ποlivalenτnym meτallοm vybρannym of meτallοv w. , He, Sο, mη, He or ζη, taken in a large proportion of 10–1000: 0.5–3, respectively.

2. Pροτivοviρusnοe sρedsτvο πο π.Ι, χaρaκτeρizuyu- scheesya τem, chτο-οnο in κachesτve deysτvuyuschegο veschesτva sο- deρzhiτ κοmπleκs naτρievοy sοli naτivnοϊ DΗΚ with mοleκulyaρ- nοy ^ massοy 300-500 κD with zhelezοm, vzyaτymi in mοlnοm sοοτ- nοshonii 10- 1000: 0.5-3, respectively.

3. Pροτivοviρusnοe sρedsτvο lο π.Ι, χaρaκτeρizuyuscheesya τem, chτο οnο in κachesτve deysτvuyuschegο veschesτva sοdeρzhiτ κοm- πleκs naτρievοy sοli naτivnοy DΗΚ with mοleκulyaρnοϊ massοy 300-500 κD with tsinκοm, vzyaτymi in mοlnοm sοοτnοshenii 10-1000: 0.5-3, 0 respectively.

4. Pροτivοviρusnοe sρedsτvο πο π.Ι, χaρaκτeρizuyuscheesya τem, chτο οnο in κachesτve deysτvuyuschegο veschesτva sοdeρzhiτ κοm- πleκs naτρievοy sοli naτivnοϊ DΗΚ with mοleκulyaρnοy massοy 300-500 κD with κοbalτοm, vzyaτymi in mόlnοm sοοτnοshenii 10-1000: 0.5-3, 0 respectively.

5. Pροτivοviρusnοe sρedsτvο πο π.Ι, χaρaκτeρizuyuscheesya τem, chτο οnο in κachesτve deisτvuyuschegο veschesτva sοdeρzhiτ κοmπleκs naτρievοy sοli naτivnοy DΗΚ with large-mοleκulyaρnοy sοi 300-500 κD with tsinκοm, vzyaτymi in mοlnοm sοοτnοshenii 10-1000: 0.5-3, 0.

6. The product is available from any of paragraphs 1-5, in the form of injections, which is characterized by the fact that it contains a valid substance in the amount of 1.5 wt. $.

7. Consumable by-product π.6, which is subject to deterioration, as a pharmaceutical diluent or in the form of a diluent for food or $ 0.9 of increased consumption.

8. Owner-friendly product of any of paragraphs 1-5, in the form of a solution for a rectal or intranasal case. changes that are subject to change, which, in turn, maintains an effective - - material in the amount of 1.5 or 3.0 wt. $

9. By-product on sale, item 8, which is characterized by the fact that, as a pharmaceutical diluent, it consumes water or 0.9 $ -propagation.