CZ113796A3 - Flavin and derivatives thereof as antiviral active preparations - Google Patents

Abstract

Various flavin derivatives are disclosed for administration to mammalian subjects as an anti-viral agent. Riboflavin and riboflavin derivatives are given as particular examples which may be preferred.

Description

TRAPLQyAXHAKS ♦ XUSÁ ¥ Adypkitic and serif k * ac * li 170 00 Ρπώβ 7, nUMWÉ M

Flavin and its derivatives as antiviral agents

The present invention relates to anti-viral agents and their use in administration to patients in both human and veterinary practice which alleviates or treats disease symptoms caused by viral infection, particularly HIV. A detailed study of the compounds of the invention was performed to evaluate their efficacy in infections caused by various strains of HIV-1. The compounds have similar activity against HIV in both acutely and chronically infected cells.

This is a double property occurring only occasionally with other compounds commonly used in the treatment of HIV infection, although acute cell infections should be treated with compounds that act early in the HIV replication cycle and block the integration of vDNA into host chromosomes. It is this integration that indicates the progress of infection to the chronic stage. Thus, compounds that cause the later integration of HIV are inhibitors of chronically infected cells. For example, zidovudine (AZT) is only active against acute HIV infection and has no significant efficacy in chronically infected cells. Thus, inhibitors of HIV gene expression (an RNA positive strand virus) may be effective in cells chronically infected with HIV.

BACKGROUND OF THE INVENTION

HIV is a positive strand RNA virus that attacks humans. The virus attaches to cell membranes by adsorption of the virion to the CD4 surface receptor. The virion then crosses the cell membrane by infiltration and enters the cytoplasm of the cell. The removal of the virion then takes place in the cytoplasm, whereby the viral envelope and the genome protein envelope release viral RNA into the cytoplasm. Reverse transcription creates double-stranded DNA transcript from the host's genetic material. It invades the nucleus of the host cell and associates with the chromosomal DNA of the host cells. This is followed by transcription producing a vRNA replicate that is transferred to the cytoplasm and produces new viral proteins. These are collected with v3NA at the inner surface of the cell to form new virus particles that release 2 host cells.

HIV is usually accompanied by an initial asymptomatic phase. This initial asymptomatic phase may take several years until early symptoms of HIV disease appear.

More ideas have been proposed about what is causing cell death. Apoptosis is one of them. Morphologically, it is a different form for grammed cell death occurring in many physiological and pathological processes, including cellular processes that seek to maintain a suitable intracellular oxidative-reduction equilibrium. Cell death in T-cells is closely related to this equilibrium process. It is believed that in HIV infection, this equilibrium gradually progresses in favor of cell death. Another critical factor is whether intracellular ATP concentration is determined by whether cells will grow and divide normally. Low intracellular AT levels? are accompanied by ischemic death. Especially vulnerable to lowering intracellular AT levels? are T-lymphocytes. HIV infection violates cellular oxidative phosphorylation, a cellular process that is responsible for AT levels? in the cell. Cell death from any cause may eventually lead to cell depletion to a level that induces AIDS.

Much of the recent work in the field of anti-virus research aims to search for specific encoded viral enzymes. In principle, the compounds discovered by this research should be of low toxicity in cellular processes. Long-term use of the compounds in clinical trials for HIV treatment has not provided an improvement to the degree initially expected and therefore new approaches are needed.

Riboflavin is a well-known compound known by various names such as:

101, lactoflavin, riboflavin, riooflavin, vitamin B?, Vitamin G,

7, 6-dimethyl-10- (1'-D-ribityl) isoalloxazine and 3,10-dihydro-7,8-dimethyl-1C- (D-ribo-2,3,4,5-tetrahydroxypentyl) benzopteridine-2 , 4-dione.

Riboflavin is commercially available as such or as a salt, namely sodium phosphate or tetrabutyrate; initially in particular as a dihydrate. It is also available in various mixtures with other vitamins that. basically all of them are intended for the treatment of other vitamin S deficiency. In these compositions, the doses of riooflavin are in the range of 0.5 to 10 mg, with the highest recommended daily dose being 30 ° C.

No adverse doses of riboflavin were reported with ribcflavin, but caused clear urine, which interfered with laboratory tests.

effects. The color of riboflavin in humans is often related to energy intake, but appears to be more closely related to other metabolic requirements. A daily intake of riboflavin in the diet of about 1.3 to is recommended

1.8 mg, which is, in other words, the basic recommended intake of riboflavin is 550 µg per 4200 kj / 1000 koal / diet / see Report of a Joint FAO / 7EO Sxpert Group, Tech. Rep. Ser. Wldllll Org. No. 362, 1967].

The stated acceptable daily intake of riboflavin is up to 500 µg per kg body weight / see Thirteenth Report of FAO / JhO 3xpert Committee of Food Additives, Tech. Rep. Ser. Ϊ, Ηϋ. No. 445 (1971).

Riboflavin, which is a water-soluble vitamin, is indispensable for utilizing energy from food. The active phosphorylated forms, flavin mononucleotide and flavin adenine dinucleotide, as coenzymes, are involved in redox metabolic reactions.

Various other flavins and their derivatives are also known, mainly as flavorings.

Substance_of the invention

It has now surprisingly been found that administration of riboflavin as well as other flavins and derivatives thereof, at doses far higher than previously used or recommended, can be highly effective in the management and treatment of viral infections, particularly HIV. The structure of the compound indicates participation in the oxidative phosphorylation process within cells. It is possible that the compounds of the invention preferentially target the same target as HIV and thus resist or prevent the manifestation of infection, including the reproductive capacity of the virus.

Accordingly, in one aspect, the present invention relates to the use of flavin, particularly riboflavin, or derivatives thereof, for the manufacture of a medicament for the management and treatment of viral infections.

In addition, when certain flavins and derivatives thereof are not known as pharmaceuticals, generally in the context of riboflavin (known as vitamin-cofactor enzymes), the invention further relates to certain such flavins or derivatives thereof and their use as antiviral agents.

When used according to the invention, riboflavin or other flavin can be administered as such or as a derivative, and the flavin derivative should be a derivative that is safe for human and veterinary application. In the case of riboflavin, however, the preferred derivative is a riboflavin salt, and more preferably the riboflavin salt is riboflavin sodium phosphate or riboflavin tetrabutyrate. It is particularly preferred that the flavin or derivative thereof is highly

- 5 highly clean and prevent contamination by trace elements.

In more general terminology, a flavin or derivative thereof for use in the invention may be defined as a compound of formula I, in particular

in which X stands for:

OH OH OH lil

- CH, -CCC-CH _? 0H ^L III

Η Η H (riboflavin)

OH OH OH I I I

-CH ₂ - C- C- C -COOH

Η Η H / schizoflavin SP · ^ /

OH OH OH I I I

CH ₇ - CCC —COH ^L II ι

Η Η H / schizoflavin SP

OH OH OH H ₇ O lili li - c - c - c - c - O - PII

Η Η H

ONa • OH (riboflavin-5'-sodium phosphate dihydrate)

OH QH ribose - Q - P II o

/ Ib /

HO QH (flavin-adenine dinucleotide) or methyl group (lumiflavin).

In addition, in formula (I) above, X may be an alkyl or hydrogen atom or an aromatic or other cyclic hydrocarbon group.

Thus, further, the use according to the invention can be realized with the aid of flavin or derivatives thereof, such as [A] lumichrome of formula II:

- Ί / 11 /

(Β) roseoflavin of formula III

Η ι

(HO-C-H) -

(G) B-hydroxyflavin, alloxazines and other derivatives of formula 17

/ iv / _row in which

R represents a ribityl group, an alkyl group or a hydrogen atom.

X is hydroxyl. a bromine atom, a chlorine atom, a sulfhydryl group, an alkoxy group or an alkylthio group.

Some examples of the above are:

H = alkyl, ribityl or rib-P (8-hydroxy) -PN M /

R = Rio-P-AM? (8-Hydroxy-PAD)

wherein H is as defined above.

(D) .alpha.N [3] -histidylflavins of formula (V)

H ₃ N-CH-COO in which R denotes the ribityl side chain of the ri boflavin derivatives.

(3) .alpha.-N-.alpha.-histidyl avalanches of the formula (VI)

₃ H H - CH-COO

(vi), wherein R is a ribityl side chain of a ribo flavin derivative.

(R) Thioethers of Salfa-cysteinylflavin.

(G) 6-S-Cysteinylflavin thioethers.

(H) Lumiflavins of formula VII

(VII) in which R 1 = R 1 = hydrogen, R ₂ = R 1 = methyl for lumiflavin alone.

/ 1/5-Deaazaflavins.

These are illustrated by this general formula (VIII)

wherein the substituent groups are defined as follows:

^and i and ₂ -h hydrogen atom methyl group hydrogen atom hydrogen atom ethyl group hydrogen atom hydrogen atom n-propyl group hydrogen atom hydrogen atom n-butyl hydrogen atom methyl group methyl group hydrogen atom methyl group ethyl group hydrogen atom methyl group n-propyl group hydrogen atom methyl group n-butyl hydrogen atom hydrogen atom methyl group 7.9 - hydrogen atom D-ribityl group 7.9- (CH3) p hydrogen atom ethyl group 7-GH ₃ methyl group ethyl group _7-CH3 methyl group D-ribityl group 7.8 - / ^ 3/3

as well as derivatives thereof, for example

O

CHOCOCH3

CHOCOCH3

CH2OCOCH3 [.alpha.] 5-carbadeasa-1-carbba-1-desaza-riboflavin analogs, 7MN and 7 AD.

This is illustrated by the riboflavin analogs of formula X, the analogs of 5-carba-5-desaza-riboflavin of formula XI and the analogs of 1-carba-1-desaza-riboflavin of formula XII, i.

/ XII /

(2) Avalanche, N-ethylenoadenine dinucleotide

H 0

H: What?

l II 1 I / y ^c Y ^c \ A

H ₃ C

CH / XIJT / / L / Schizoflavins and their derivatives

CH-OH 1 CHO AND COOH 1 HCOH AND 1 HCOH AND 1 HCOH 1 1 HCOH 1 1 HCOH J 1 HCOH AND 1 HCOH 1 HCOH AND 1 HCOH ch ₂ 1 ch ₂ 1 ch ₂ 7,8-dimethyl- 7,8-dimethyl- 7,8-dimethyl- isoalloxazine isoalloxazine isoalloxazine riboflavin with? ₂ QT? ^bi l

The chemical structures of schizoflavins are shown here and show their formation from riboflavin. Network ^and SP ^ · ^ l ^of described as 7,8-dimethyl-10- / 2,3,4-trihydroxy-4- formylbutyl / isoalloxazine or fall 7, 8-dimethyl-1-10- / 2.3 4 - trihydroxy-4-carboxybutyl / isoalloxazine.

Other flavins that may be mentioned are:

-a oy xy me thy Ir ib of avalanche,

-carb o xy me thy 1-Fivin,

7-amino-10- (1'-D-ribityl) isoalloxazine,

S-amino-7,10-dimethylisoalloxaine,

8alpha / S-mercaptopropions1 / riboflavin,

8alpha / 3-mercaptopropionyl / FMN,

8alpha / N -aminohexyl / PMN, γ-azobenzoyl-PMN,

10- (α-carboxyalkyl) -7,8-dimethylisoalloxazine.

When used according to the invention, a flavin, such as riboflavin or a derivative thereof, is preferably administered in high doses which significantly exceed those normally administered or recommended. Thus, for example, riboflavin or other flavin in a clinical trial is administered according to the present invention to a dosing regimen ranging from at least about 1 to about 100 or more (for example, 10 or more / m 2 / kg body weight). Moreover, the administration according to the invention is carried out in such a way that the medicament is formulated in a perorally administrable form, in particular in the form of a capsule, e.g.

Additionally or alternatively, the invention encompasses a pharmaceutical or veterinary composition for administration in the treatment and treatment of viral infections in the form of dosage units, the composition comprising a dosage unit containing at least about 35 mg, for example about 50 nv; or more (for example, 50 to 300 m 2, i.e. about 50 to 200 or 50 to 100 m 2) of a flavin such as riboflavin or a derivative thereof as described or defined herein in combination with pharmaceutically or veterinarily acceptable diluents. , excipients or carriers.

The composition of the invention is preferably formulated so that the dosage unit contains from about 35 mg-nsi to 10 mg mg. Even more preferably, the unit dose contains from about 250 to 500 mg.

In addition, the composition of the invention is conveniently formulated into an oral or injectable form. Accordingly, a preferred composition is a composition in the form of a solution in sterile water.

The invention also relates to a process for the manufacture of a medicament for use in the management and treatment of viral infections, comprising treating a flavin, for example a derivative such as its tetrabutyrate salt, for riboflavin, or for use against viral

It is to be realized using the method of definition 1z above; one or more of the aforementioned measures.

The invention further relates to an article of manufacture comprising a flavin, for example riboflavin or a derivative thereof, as an antiviral agent in combination with another agent or agents having antiviral activity, such as a combined preparation for simultaneous, separate or sequential administration in antiviral therapy.

The product as defined above may include one or more of the other specific features of the invention already mentioned.

The invention further relates to a method of treating a viral infection comprising administering orally or parenterally an effective amount of a flavin, for example riboflavin, or a derivative thereof.

In the method of the invention, the amount administered is from 1 to 100, for example at least 10, mg / kg body weight of the patient.

Furthermore, the method is particularly useful when the virus is human immunodeficiency virus (HIV).

Once again, the method of the invention may include one or more of the other specific features of the invention as defined above.

Most preferably, the invention is practiced with one or more riboflavins, riboflavin sodium phosphate, flavin = adenine dinucleotide lumiflavin, lumichrome, or especially riboflavin tetrabutyrate, the formula of which is given below:

CH - O butyrate I _

H - C - O butyrate

H - C - O butyrate

H - C - O butyrate

AND

H-C-H

O

In vitro tests

The following in vitro assays were used to investigate the antiviral activity of the compounds of the invention for those HIV:

Acute infection test

Standard acute test

High titer HIV-1 (HTLV-1113) human immunodeficiency virus strains were cultured in H9 cells with

RPMI 1640 as growth medium supplemented with 10 fetal calf serum. Cell debris was removed by low speed centrifugation and the supernatant was stored at -70 ° C as needed. In a typical assay, C 8166 T-lymphoblastoid cells were incubated with 10TCID, _Q- HIV-1 at 37 ° C for 90 min, and then washed three times with phosphate buffered saline (PES). Aliquots of 2 x 10 ⁷ cells were resuspended in 1.5 ml growth medium in 6 ml culture tubes, and immediately the test compound was given in a logarithmic dilution of 0.2 to 200 µϊί. The test compound was dissolved in 70% ethanol and the final concentration of ethanol in the culture was less than 1 x. The cultures were incubated at 37 ° C for 72 h τ 5 µl of carbon dioxide. 2G0 μ 2 of the supernatant was collected from each culture and tested by measuring the optical density at 450 nm for the HIV p24 / Kinchington se sp.

1989, Hoberts et al. 1990 / using the commercial 3LI · SA assay, which differentiates all core proteins equally (Coulter Hlectronics Ltd., Luton, UK). Standard curves from control cultures containing less than 1 ethanol were constructed to determine IC 50 values. AZT and ddC were used as internal controls. The tests were performed in duplicate.

1.2. Depleted medium test

In a standard assay system, cell cultures were harvested, crushed, and fed with fresh medium, approximately 18 to 24 hours prior to the start of the assay. Addition of fresh medium stimulated the cells to initiate the logarithmic growth phase. To investigate the effect of concomitant cell enrichment under depleted media conditions, cell cultures were fed and crushed for ^7, 43 and 24 h before being used in a standard acute assay.

1.3. Light exposure test

A freshly dissolved sample of the test compound was divided into two aliquots. These were stored both in daylight and in the dark before being subjected to a standard acute test. Exposure time 2 h in both cases.

Preincubation test

Target cells were first incubated with the test compound at a logarithmic dilution of 200 to 0.2 µM for 18/24 h prior to HIV-1 infection. A sample of each concentration was then individually processed as in the standard acute test.

Test with chronically infected cells

Standard chronic test

Chronically infected HIV-1rf (H9rf) cells were washed 3 times with medium to remove extracellular virus and incubated with test compounds (200 to 0.2 juid) for 3 days. The p24 antigen was then determined by measuring the optical density at 450 nm as described in the standard acute infection test. After determination of IC-θ values, standard curves were constructed from control cultures containing 1 ethanol. RO 31-5959 (Roche proteinase inhibitor) was used as internal control. The tests were performed in duplicate.

2.2. Depleted medium test

In a standard assay, cell cultures were harvested, crushed, and fed with fresh medium approximately 18-24 h before. testem. Addition of fresh medium stimulated the cells to initiate the logarithmic growth phase. To investigate the effect of overlapping cell enrichment under depleted media conditions, cell cultures were fed and crushed for 72, 48, and 24 h before being used in a standard acute assay.

2.5 Fresh Rosette Light Exocation Assay A released sample of the test compound was divided into two aliquots. These were stored both in daylight and in the dark before being subjected to a standard acute test. Boba exposure for 2 h in both cases.

Toxicity test

In the toxicity test of the compounds, aliquots of 2 x were each

ΙΟ ⁷ unindicated cells cultured together with test compounds at the same log dilution for 72 h (1.1 and 2.1). The pellets were then washed with medium and resuspended in 2CG µL of growth medium containing the C 4 -protein hydrolyzate. were isolated after 5 or 20 h and mixed and the C? Cells that were not cultured with the test compounds were used as controls.

The tests were applied to compounds that are listed in those bulce 1 below. Table 1 Code Compound BI riboflavin-5'-phosphate P2 riboflavin B3 flavin-adenine-dinucleotide ? 4 lumiflavin 5 * 5 lumi chrom P6 riboflavin tetranicotinate ? 7 riboflavin tetrabutyrate Introductory the tests were carried out with various compounds variable v Table 2 to obtain them Preliminary results. Obtain- kané h IO-q values are subject to k proof; no harm with

tests that have been carried out so far. Test results are shown ;; 1 to 12 and numerically expressed in Tables 2 to 10 following the drawings.

Giant. 1 and 2 show the anti-viral effect (IC-V) test of compounds P1 to P5 of the invention, using an antigen (Ag). The x-axis shows the concentration values of compounds in μλΐ, the y-axis shows the optical density (00) of the antigen.

Giant. 1: solid lines indicate optical density / 00 / antigen at 450 nm to the concentration of compound P2, P4 and P5 in drinking / first antigen assay / dotted line at 0.371 is 00 denotes IC- _N / active.

Giant. 2: relationship of optical density (OD) of antigen at 450 nm to concentration of compounds P1 and P3. The dashed line at 00 0.371 indicates 1C-q (active).

Giant. 3 and 4 show the toxicity test of compounds P1 to P5 (CO-) as C ⁴ incorporation. The x-axis shows the concentration values of compounds in pM, and the y-axis the incorporation value (dpm).

Giant. 3: solid curves indicate the relationship, incorporation of C ⁴ (dpm) to the concentration of compounds P2, # 3, P4 and P6 in the first toxicity test. The dashed line at 6035 dpm indicates CC (non-toxic).

Giant. 4: Analogous data for compound P1 (non-toxic).

Fig. 5: relationship of optical density (OD) of antigen at 450 nm to concentration of compound P4 (second antigen test). The dashed line at 00 0.371 indicates IC-θ (active).

Giant. 6: analogous data to FIG. 3 for compound P4 (second toxicity test). The dashed line at 6035 dpm indicates CC-Q (non-toxic).

Giant. 7: centering denotes the IC relationship, the optical density (OD) of the antigen at 450 nm and the compounds F6 and 77. The dashed line for the values; / OD '0.371 - ₀ (active).

Giant. 8; analogous data to that of FIG. 6 for compounds F6 and F7. The dashed line at 6055 dera indicates CC-q (non-toxic).

Giant. 9: antigen control (ddC).

Giant. Figures 10 to 12 show a standard acute infection assay for compound F7 and control (ddC), similar to Figures 1 and 3.

As shown in the following tables, the activity of both HIV / HIV infected cells was evaluated acutely and chronically for the test compounds. Values of antiviral activity (± C-q) and toxicity (CC-q) are given below. In another experimental series, compounds were tested in cell cultures to which fresh medium was added 72, 45 and 24 h prior to infection. This experiment was carried out to determine the effects of the compounds on the cell at the division and resting stages. The data suggest that cells are more sensitive to test compounds at rest. The effect of light, pre-incubation of target cells and activity against African HIV-1 isolate was also investigated. Exposure to light for 2 h did not affect the efficacy of the compounds. Pre-incubation with target cells increased their efficacy, indicating a significant efficacy against HIV-1.22 African isolate

Table 2 /giant. 1 to 4 / Standard acute test infection / 1 Compound No. * n ± υ Pv ^CC 50 SI Test no. /giant. 1 a 2 / /giant. 3 and 4 / 71/1 1 to 2G > 200 - 71/2 <0.4 > 400 1000 71/3 0.1 / fig. 2 / > 800 / oor. 4 8000 72 3 / oor. 1 / > 200 / fig. 3 / 60 73 0.8 / fig. 2 / > 200 / fig. 3 / 200 74 1 / oor. IN 150 / fig. 3 / 150 ? 5 3 / oor. 1 / > 200 / oor. 3 / . 60 Table 3 / oor. 7 and 8 / Standard acute test infection / 1 Compound No. ΙΟ -.- Λ cc ... after SI Test no. 77/1 27 / fig. 7 / 130 / fig. 8 / 5 77/2 57 > 200 > 4 77/3 10 70 7 77/4 25 140 6 Table 4 Standard te ! St chronic infections /2.1/ Compound No. ^IC 50 ^CC 50 SI Test Nr. 77/1 0.2 7 35 77/2 > 20 > 20 - 77/3 10 > 200 > 20 77/4 4 75 19 Dec 77/5 26 > 200 > 7

IC- _n CC- _n p0 pO

Table 5 Acute infection test with depleted medium /1.2/

Compound No.

h h

h ^IC 50 cc

160 ^IC pO

CC -, -

PU

100 ALIGN!

110

160

Table 6 Acute infection test with depleted medium

Compound No.

h h

h ^IC 50 ^CC 50

IC ^CC 50

250 ^IC 50 ^CC 50

101

Table 7 Acute infection test after light exposure /1.3/

Compound S.

Oenní svetlo

Dark

IC

CC- pu _r ^IC50

CC

200

200

Table 3 Preincubation test of acute infection /1.4/

Centrifugation of target cells with test compound for 24 h from infection

Compound No.

^IC 50 ^CC 50

SI

120

Table y / FIG. 5 to 8 / Standard test of acute infection /1.1/

Combine CC-.λ SI nina δ. ⁵⁰ / fig. 5

- 60 /obr.7/

/giant. 6/12 / fig. 8 / min. 3-6

Table 10 Standard test of acute infection /1.1/

Test applied to C8166 cells (lymphoblastoid T cells, transformed and immortalized with HTLV / HIV African H1V-1 CBL4 isolate)

Merge-IC-q nina ³

SI

150 37,5

Table 11 / FIG. 10 to 12 / Standard test of acute infection /1.1/

ICc _Q nina no.

32 ddC / control / 0.2

CC ~ SiO

200 6.3

The endpoint variations observed with compound 77 may be due to the characteristics of the target lymphoblastoid cells.

Even in synchronized cultures there may be subtle changes in the metabolic state of cell subpopulations. This is reflected in the endpoint shift observed for parallel antiviral activity and toxicity values from one test to another (see Table 3).

The results summarized in Tables 5 and 6 indicate that active or quiescent cell cultures are differently sensitive to test compounds.

Treatment of patients

35 patients were treated. 2 of them 30 underwent medical examinations i / General condition of patients (arrived

Twenty of the thirty patients who had received a medical examination reported an improvement in their general condition. Mostly this improvement was related to restlessness, appetite and weight gain. Two patients also reported an improvement in skin rash with retreated skin lesions, while one patient reported that no new lesions developed during therapy. -One patient also reported a decrease in impotence (which occurred for three months prior to initiation of therapy), while two other patients reported a recurrence of long-running rhinitis.

ii / Patient visits

Several patients have:

arrived at the clinic for an emergency check1. One patient had recurrent abscesses as well as septic arthritis that resisted therapy.

2. Two patients had recurrent lower respiratory tract infections, with one developed open bronchopneumonia, during the second week of therapy. Repeated smears on AAF3S were consistently negative.

3. Two patients forged smear had pyrexia with non-localized symptoms, unlike malaria parasites was negative and was not

- 26 observed significant growth in blood culture. One of these patients responded well to amoxycillin and is now temperature-free.

4. One patient had gastroenteritis during the third week of therapy.

5. Two patients oral and vulvo-vaginal candidiasis, the vulvo-vaginal form reappearing when nystatin treatment with pessaries and tablets was discontinued.

o. Two patients also reported recurrent attacks of herpes simplex genitalis.

iii / Toxicity

The manifestations of toxicity mostly occurred within the first two weeks of therapy and were transient.

Two patients suffered from pruritus for an average of four days in the first week of treatment and healed spontaneously without any supportive medication.

Four patients reported mild diarrhea during the first two weeks of therapy. It lasted an average of four days. The symptoms of the symptoms were how to tell whether it was a side effect or a natural manifestation of HIV infection. However, the duration of their occurrence in the first week of therapy and their transient nature justified the assumption that this was a side effect.

One patient complained of drowsiness and another complained of darkening of urine. MSU was normal.

Two patients complained of abdominal problems.

iv / Laboratory results

Three patients had transient increases in liver enzymes from the second to third weeks of therapy, with no clinical signs of liver disease. Over time, enzyme levels have always returned to normal.

The above clinical trial reports are a preliminary result of a clinical trial that was continued continuously over several weeks using compound 7 administered orally as a capsule (s) as the one described in Example 4 below / at this dosage:

Dosage 1: 1 mg / kg body weight daily orally ve two divided benefits, Dosage 2: 2 mg / kg body weight daily orally ve two divided benefits, Dosage 3: 10 mg / kg body weight daily orally ve two divided benefits, Dosage 4: 15 mg / kg body weight daily orally ve two divided benefits, Dosage 5: 20 mg / kg body weight daily orally ve two divided benefits, Dosage 6: 30 mg / kg body weight daily orally ve two up to three divided doses, Dosage 7: 40 mg / kg body weight daily orally ve two up to three divided doses, Dosage 8: 50 mg / kg body weight daily orally ve two up to three divided doses, Dosage 9: ICC mg / kg body weight per day orally in

two to three divided doses.

The following examples illustrate the compositions prepared according to the invention.

Example 1

The preparation is made from the following components:

riboflavin-5-phosphate 10 mg sterile water 2 ml

A unit dose of 10 mg riboflavin is obtained, for administration once daily for the treatment of viral infection.

Example 2

The preparation is made from the following components:

riboflavin-ρ-phosphate 30 mg sterile water 2 ml

A unit dose of 30 mg riboflavin for once daily administration for the treatment of viral infection is obtained.

Example 3

Formulations similar to those of Examples 1 and 2 may be prepared in mg / ml, mg / ml, and 50 mg / ml doses in a single dose of 2 ml or 5 ml of sterile water based on one active ingredient which is: riboflavin-5-phosphate, riboflavin, flavin-adenine dinucleotide, lumiflavin, lumichrome, optionally based on a mixture thereof.

Example 4

Capsules of the following composition were prepared:

Size: 25mg, mg,

100 mg,

200 mg,

400 mg.

Type: two-piece, hard gelatin.

Composition: Compound P7 mixed with microcrystalline cellulose Ph. Sur. 166. 4 / 156.7 / 118.6 / 108.7 / 50 mg provided capsules weighing 191. 4 / 206.7 / 218. 6 / 308.7 / 450 mg.

^ ^íloválováloválováloválová.nost...

The present invention proposes solid and liquid dosage forms which contain flavins and / or derivatives thereof, particularly riboflavin, including salts, at extremely high doses and which are intended to treat viral infections, particularly HIV-induced infections, both in human and human and veterinary practice.

ING. JAN KUBAT patent attorney

Claims (9)

PATENTS n i a o κ zy CURREjyr TEXT Use of a flavin compound or a mixture comprising two or more of these compounds for the manufacture of a medicament for prophylactic or therapeutic use in a disease caused by HIV infection, wherein the flavin compound is riboflavin-5-phosphate, flavin-adenine dinucleotide, lumiflavin, lumichrome, tetranicotinate riboflavin or riboflavin tetrabutyrate. Use of a flavin, flavin derivative or a mixture comprising two or more of these compounds for the manufacture of a medicament for prophylactic or therapeutic application in a disease caused by a viral infection. Use according to claim 2, characterized in that the riboflavin derivative is a riboflavin salt. Use according to any one of claims 1 to 3, characterized in that the riboflavin salt is riboflavin phosphate sodium or riboflavin tetrabutyrate. Use according to claim 2, characterized in that the flavin or flavin derivative is 5.1 Pi CURRENT TEXT vs which R represents a hydrogen atom or an alkyl group, R ₁ to P <4, independently of one another, represent a hydrogen atom, an alkyl group, a halogen atom, a hydroxyl group, an alkoxy group, an alkylthio group, a thio group or an optionally substituted aromatic or non-aromatic nitrogen heterocycle and X represents: (i) a hydrogen atom, a ribityl residue, an alkyl group, a hydrogen atom or an aromatic or non-aromatic carbocycle; -CH 2 - (CHOH) -Y 2 n wherein n is an integer of 3 or 4 and Y is -C 1 R 4 OH-COOE 1 or -COH or a group of formula Ib CURRENT TEXTILES in which R represents a hydrogen atom or an alkyl group and and, independently of one another, represents an alkyl group, a hydroxyl group, a halogen atom, an alkoxy group, an alkylthio group, a thio group or an optionally substituted aromatic or non-aromatic nitrogen heterocycle, and X is (i) a hydrogen atom, a ribityl residue, an alkyl group, a hydrogen atom or an aromatic or non-aromatic carbocycle, (ii) a group of the general formula -CH - (CHOH) L Π in which n is an integer of 3 or -COOH or -COH or a group of formula - Y, 4 and Y denote the group -CH 2 OH 1b OH OH 5.3. A compound of formula VIII (VIII), (CURRENT TEXT 1) in which it represents a hydrogen atom or an alkyl group; R 1 represents an alkyl group or a ribityl group and R 1 represents a hydrogen atom or a single or double substitution by an alkyl group on the outer carbocyclic ring. Use according to claim 2, characterized in that the flavin or flavin derivative is lumichrome, roseflavin, hydroxyflavin, alloxazine or a derivative thereof, 8alpha-N (3) histidylflavin, 8alpha-N (1) -histidylflavin, 8alpha-cysteinylthioether, 6alpha -S-cysteinylthioether, lumiflavin, 5-desazaflavin, 5-Carba-5-desaza- or 1-Carba-1-desaza-riboflavin analogue, g FMN or FAD, flavin-1, N -ethenoadenine dinucleotide, 9-methylflavin, 9-phenylflavin, 9-benzylflavin, 9-cyclohexylflavin, 6,9-dimethylflavin, 6,7,9-trimethylflavin, 9-oxyethylflavin, 9-dioxypropylflavin, 6,3,9-trimethylflavin, lactoflavin, flavin-9-carboxylic acid, 6,7-dimethylflavin-9-carboxylic acid or schizflavin. A pharmaceutical composition for prophylactic or therapeutic application in a disease caused by a viral infection, which composition is characterized by a content of flavin or flavin derivative. The composition of claim 7, wherein the flavin or flavin derivative is as defined in any one of claims 3 to 6. The composition of claim 7 or 8, wherein the composition comprises a unit dose of at least about 35 mg of flavin or flavin derivative in combination with a pharmaceutically or veterinarily acceptable diluent, excipient, or carrier.