GB2319474A - Anti-viral agents - Google Patents
Anti-viral agents Download PDFInfo
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- GB2319474A GB2319474A GB9804302A GB9804302A GB2319474A GB 2319474 A GB2319474 A GB 2319474A GB 9804302 A GB9804302 A GB 9804302A GB 9804302 A GB9804302 A GB 9804302A GB 2319474 A GB2319474 A GB 2319474A
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- flavin
- derivative
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
- A61K31/525—Isoalloxazines, e.g. riboflavins, vitamin B2
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- General Health & Medical Sciences (AREA)
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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
ANTI-VIRAL AGENTS
The present invention relates to anti-viral agents and their use in the treatment of human and animal patients to alleviate or cure the ill-effects caused by viral infection, especially HIv. A detailed study of compounds according to the invention has been carried out to evaluate their efficacy against infection from several strains of
HIV-1. The compounds have similar activity against HIV in both acutely and chronically infected cells. This is a dual property only ocassionally associated with other compounds which are in current use in the therapy of HIV infection although de nova (acute) infections of cells may be treated by compounds which act early in the replication cycle of HIV to block integration of vDNA into the host chromosome. It is this integration which signifies entry of the infection into the chronic state. Compounds which act post-integration of HIV -are therefore inhibitors of chronically infected cells. Zidovudine (AZT) for example is only active against de nova in,f,ection of H5V-and has no significant activity against chronically infected cells.
Inhibitors of gene expression of HIV (which is a positive strand RNA virus) would therefore be active in HIV chronically infected cells.
HIV is a positive strand RNA virus which affects humans.
The virus attaches to cell membranes by virion adsorption to CD4 surface receptor. The virion then passes through the cell membrane penetratively and enters the cell cytoplasm. Uncoating of the virion then takes place in the cytoplasm whereby the viral envelope and the protein coat of the genome release the viral RNA into the cytoplasm.
Reverse transcription therein produces a double-stranded
DNA transcript from host cell genetic material. This invades the host cell nucleus and integrates with the host cell chromosomal DNA. Transcription follows to produce a vRNA replicate which is translated in the cytoplasm to produce new viral proteins. The latter then assembles with vRNA at the inner cell surface to produce new virus particles which are released from the host cell.
HIV is normally associated with an initial asymptomatic phase. This initial asymptomatic phase may last a number of years before the early signs of HIV disease occur.
A number of ideas causing cell death are proposed.
Apoptosis is one of these. It is a morphologically distinctive form of programmed e~-death. involved in many physiological and pathological processes including cellular processes which seek to maintain appropriate intracellular oxidant-antioxidant balance. Cell death in T-cells is closely associated with this balancing process. Infection with HIV is thought gradually to disturb the balance in favour of cell death. Another critical factor in determining whether cells will grow and divide in a normal fashion is intracellular ATP concentration. Low intracellular levels of ATP are associated with ischemic death. T-lymphocytes are especially vulnerable to depletion of intracellular ATP levels. HIV infection may disturb cellular oxidative phosphorylation which is the cellular process responsible for ATP levels ir. the cell.
Cell death from whatever cause will eventually lead to cell depletion to a level that induces AIDS.
Much of the current work in the field of antiviral research is concerned with targeting specific viral encoded enzymes.
Compounds discovered from this research, in principle, should have low toxicity on cellular processes. The long term use of compounds in clinical trials in HIV infection treatment has not given the degree of benefit initially expected, and new approaches are needed.
Riboflavine is a known compound, which is also variously known as: E101; Lactoflavin;
Riboflavin;
Riboflavinum;
Vitamin B2;
Vitamin G; 7,8-Dimethyl-10-(1'-D-ribityl) isoalloxazine; and 3B10-Dihydro-7,8-dimethyl-1o-(D-ribO-2,3t4 5-tetra- hydroxypentyl) benzopteridine-2,4-dione.
Riboflavine is commercially available as such or as its sodium phosphate or tetrabutyrate salt, typically in the former instance as the dihydrate salt. It is also available in various mixtures with other vitamins, all essentially being for the treatment of, inter alia, vitamin
B deficiency. In such mixtures the dose of riboflavin varies between 0.5 and 10 mg, with a maximum recommended daily dose being 30 mg.
No adverse effects have been reported with the use of riboflavine. However, significant doses of riboflavine result in a bright yellow discoloration of the urine which may interfere with certain laboratory tests.
The riboflavine requirement of humans is often related to the energy intake, but it appears to be more closely related to resting metabolic requirements. A daily dietary intake of about 1.3 to 1.8 mg of riboflavine is recommended that is to say the basic recommended-intake of'?iboflavine is 550 ssg per 4200 kj (1000 kcal) of diet - Report of a
Joint FAO/WHO Expert Group, Tech. Rep. Ser. Wld 111th Org.
No. 362, 1967.
The estimated acceptable daily intake of riboflavine is up to 500 ssg per kg body weight - see Thirteenth Report of
FAO/WHO Expert Committee on Food Additives, Tech. Rep. Ser.
WHO. No. 445, 1971.
Riboflavine, which is a water-soluble vitamin, is essential for the utilisation of energy from food. The active, phosphorylated forms, flavine mono-nucleotide and flavine adenine dinucleotide, are involved as co-enzymes in oxidative/reductive metabolic reactions.
Various other flavins and derivatives thereof are also know, mainly as flavouring agents.
It has now been found surprisingly that the administration of riboflavine, 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, in particular HIV. The structure of the compound indicates involvement in the process of oxidative phosphorylation within cells. It is possible that the compounds of the invention preferentially target the same target as HIV and so resist or prevent the manifestations of infect ion incwlUuding the" "procreative capacity of the virus.
Accordingly, the present invention in one aspect provides the use of a flavin, especially riboflavine, or a derivative thereof for the manufacture of a medicament for the management and treatment of viral infection.
Moreover, insofar as certain flavins and derivatives thereof are not known as pharmaceuticals, even in a general sense as with riboflavine (known as an enzyme co-factor vitamin) , the invention in a second and broader aspect provides such certain flavins or a derivative thereof for use as anti-viral agents.
In the use according to the invention riboflavine or other flavin may be used as such or as a derivative and the flavin derivative may be any derivative which is safe for human or animal use. Preferably, however, in the case of riboflavine the derivative is a riboflavine salt and more preferably the riboflavine salt is riboflavine sodium phosphate or riboflavine tetrabutyrate. Most preferably, the flavin or derivative should be of high purity and contamination with spurious ingredients should be avoided.
In more general terms, the flavin or derivative for use in accordance with the invention may be defined as a compound of the formula (I), namely:
wherein:
OH OH OH 7 111 X is - CH2 -C-C-C- CH2BH I I I . .
H H H (ribottavine) OH OH OH ill - CH2 - C C COOH Iii H H H (scbizoftavinSF1) OH OH OH III -CH2- C-C-C- COH l l l H H H (schizoflavin SF2) OH OH OH H2 0 I I II ONa C H2 C rrCOP OH I O H H HH (riboflavine-5'-phosphate sodium salt dihydrate)
(flavin-adenine dinucleotide) or CH3 (lumiflavin).
In addition, in the above formula (I) the group X may be alkyl, or H or an aromatic or other cyclic hydrocarbon group.
Thus, and furthermore, the use of the invention may be realised with flavins or derivatives such as: (A) lumichrome of the formula:
(B) Roseoflavin of the formula:
(C) B-Hydroxyflavine, alloxazines and other derivatives thereof:
wherein
R is ribityl, alkyl, or H;
X is OH, Br, C1, -SH, OAlk or SAlk.
Some Examples of the above are:
R = alkyl
ribityl or rib-P (8-hydroxy-FMN) R@Rib-P-AMP (8-hydroxy-FAD)
wherein R is as above.
(D) 8a-N(3)-histidylflavins
wherein R denotes the ribityl side chain for the riboflavin derivative.
(E) 8aN(l)-histidylflavins:
wherein R denotes the ribityl side chain for the riboflavin derivative.
(F) 8a-Cysteinylflavin thioethers: (G) 6-S-cysteinylflavin thioethers: (H) Lumiflavins:
wherein R1=R4=H, R2=R3=CH3 for lumiflavin itself.
(I) 5-Deazaflavins:
These may be illustrated by the following formula:
wherein the substituent groups are as defined below:
R1 R2 R3
H CH3 H
H C2H5 H
H n-C3H7 H
H n-C4H9 H
CH3 CH3 H
CH3 C2H5 H
CH3 n-C H H
CH3 n-C4H9 H
H CH3 7,8-(CH3)2
H D-ribityl 7,8-(CH3)2 H C2H5 7 CH3
CH3 C2H5 7-CH3
CH3 D-ribityl 7,8-(CH3)3 and derivatives thereof such as:
(J) 5-Carba-5-deaza and l-carba-l-deaza analogs of
riboflavin, FMN, and FAD.
These may be illustrated by riboflavin analogs (X), 5carba-5-deazariboflavin analogs (XI) and l-carba-ldeazariboflavin analogs (XII), that is:
(K) Flavin 1, N6-Ethyenoadenine dinucleotide
(L) Schizoflavins and derivatives.
CH2OH CHO COOH
HCOH HCOH HCOH
HCOH HCOH HCOH
HCOH HCOH HCOH
CH2 CH2 CH2 7,8-dimethyl- 7,8-dimethyl- 7,8-dimethylisoalloxazine isoalloxazine isolloxazine
Riboflavin SF2 SF1
The above are chemical structures of schizoflavins and show their formation from riboflavin. SF2 and SF1 can be identified as 7,8-dimethyl-10-(2,3,4-trihydroxy-4formylbutyl) isoalloxazine and 7,8-dimethyl-10-(2,3,4trihydroxy-4-carboxybutyl) isolloxazine, respectively.
Other flavins may be illustrated by:
3-carboxymethylriboflavin
3-carboxymethyl FMN
7-amino-10-(1'-D-ribityl)isoalloxazine
8-amino-7,10-dimethylisoalloxazine 8a (5-Mercaptopropionic acid) riboflavin
8a(S-Mercaptopropionic acid) FMN
8a(N-Aminohexyl)FMN
9-Azobenzoyl FMN
10-(w-carboxyalkyl)-7,8-dimethylisoalloxazine
In the use according to the invention the flavin such as riboflavin, or derivative thereof, is preferably employed at a high dose level significantly in excess of the doses currently used or recommended. Thus, typically the riboflavin or other flavin in the clinical trial is used in the present invention at a dosage regime of at least about 1 to about 100 or more (eg 10 or above) mg/kg of body weight per day. In additIon, use according to the invention preferably is one wherein the medicament is in orally administrable form, especially as a capsule (eg two part).
Additionally or alternatively the invention includes a pharmaceutical or veterinary composition for use in the management and treatment of viral infections and in unit dosage form, which composition comprises a unit dose of at least about 35 mg such as 50mg or more (eg 50 to 300 mg, such as 50 to 200 or 50 to iOomg) of a flavin such as riboflavine or derivative thereof as described or defined herein, together with a pharmaceutically or veterinarily acceptable diluent, excipient or carrier.
A composition according to the invention is preferably one wherein the unit dose is from about 35 mg to about 1000 mg.
More preferably, the unit dose is from about 250 to 500 mg.
In addition, a composition according to the invention is preferably in oral or injectable form. Within that context a preferred composition is one as a solution in sterile water.
The invention also includes a pröess- for the manufacture of a medicament for use in the management and treatment of viral infections, which process comprises formulating a flavin such as riboflavine, or a derivative such as the tetrabutyrate salt thereof for anti-viral use.
As will be appreciated, a process according to the above definition may be carried out using one or more of the additional features mentioned herein.
In addition, the invention includes a product containing a flavin such as riboflavine, or a derivative thereof, as an anti-viral agent, together with another compound(s) having anti-viral activity as a combined preparation for simultaneous, separate or sequential use in anti-viral therapy.
Again, a product according to the above definition may be one which includes one or more of the other specific features of the invention defined herein.
The invention further includes a method for the treatment of viral infection, which method comprises orally or parenterally administering an effective amount of a flavin such riboflavine, or a derivation thereof.
Preferably in a method according to the invention, the amount administered is 1 to 100 (eg at least 10) mg/kg of patient body weight. - Furthermore, the method is particulary useful when the virus is human immunodeficiency virus, HIV.
Once again, a method according to the invention may include one or more of the other specific features of the invention defined herein.
Most preferably, the invention is carried out with one or more of riboflavine, riboflavine sodium phosphate, flavinadenine dinucleotide, lumiflavin, lumichrome, or especially riboflavin tetrabutyrate, whose formula is set forth below:
CH2 - 0 butyrate H - C - 0 butyrate H - C - 0 butyrate H - C - 0 butyrate H-C-H H3C < 9 o H3C 9 N9 In Vi tro Assay The following in vitro assays were used to investigate the anti-viral activity against HIV of compounds in accordance with the invention:1 Acute Infection Assays
1.1 Standard Acute Assay
High titre virus stocks of the human immunodeficiency
virus HIV-1 (HTLV-lllB) were grown in H9 cells with
RPMI 1640 supplemented 10% fetal calf serum as growth medium. Cell debris was removed by low speed centrifugation and the supernatant stored at -700C until required. In a typical assay, C8166 Tlymphoblastoid cells were incubated with 10TCID50 HIV1 at 37"C for 90 minutes and then washed three times with phosphate buffer saline (PBS). Aliquots of 2 x 105 cells were resuspended in 1.5ml of growth medium in 6m1 culture tubes, and test compound at log dilutions from 0.2 to 200ssM was added immediately.
The test compound was dissolved in 70% ethanol and the final concentration of alcohol in the culture was < 1%.
Cultures were incubated at 370C for 72 hours in 5% CO2.
200ss1 of supernatant was taken from each culture and assayed by optical density measurement at 450nm for
HIV p24 core antigen (Kinchington et al 1989, Roberts et al 1990) using a commercial ELISA which recognises all the core proteins equally (Coulter Electronics
Ltd, Luton, UK). To determine the IC50 values standard curves were drawn from untreated cultures containing < 1% alcohol. AZT and ddC were used as internal controls. Assays were carried out in duplicate.
1.2 Depleted Medium Assay
In the standard assay system, cell cultures were harvested, split and fed with fresh medium approximately 18 to 24 hours before the start of
assay. Addition of fresh medium stimulates the cells
to enter a log phase of growth. To investigate the
effect of cells reaching confluence in conditions of
depleted media, cell cultures were fed and split at
72, 48 and 24 hours before being used in a standard
acute assay.
1.3 Light Exposure Assay
A freshly dissolved sample of test compound was split
into two aliquots. They were placed either in
daylight or the dark for two hours before being
subjected to standard acute assay.
1.4 Preincubation Assay
Target cells were preincubated with test compound at
log dilutions of 200 to 0.2 M for 18/24 hours before
infection with HIV-1. Each sample concentration was
then treated individual by as in the standard acute
assay.
2 Assays for Chronicallv Infected Cells
2.1 Standard Chronic Assay
H9 cells chronically infected with HIV-lrf (H9rf) were
washed three times with medium to remove extracellular virus and incubated with test compounds (200 to 0.2M) for three days. p24 antigen was then determined by optical density measurement at 450nm as described for the acute infection standard assay. To determine the ICso values standard curves were drawn from untreated cultures containing 1% alcohol. RO 31-8959 (Roche
Proteinase inhibitor) was used as an internal control.
Assays were carried out in duplicate.
2.2 Depleted Medium Assay
In the standard assay, cell cultures were harvested, split and fed with fresh medium approximately 18 to 24 hours before assay. Addition of fresh medium stimulates the cells to enter a log phase of growth.
To investigate the effect of cells reaching confluence in conditions of depleted media, cell cultures were fed and split at 72, 48 and 24 hours before being used in a standard acute assay.
2.3 Light Exposure Assay
A freshly dissolved sample of test compound was split into two aliquots. They were placed either in daylight or the dark for two hours before being subjected to standard chronic assay.
3 Toxicity Assay
To test for compound toxicity, aliquots of 2 x uninfected cells were cultured with test compounds at the same log dilutions for 72 hours (1.1 and 2.1). The cells were then washed with medium and resuspended in 200ss1 of growth medium containing C14 protein hydrolysate. The cells were harvested after 5 or 20 hours and the C14 incorporation measured. Untreated cells were used as controls.
The assays were applied to the compounds identified in
Table 1 below:
Table 1
Code Compound
Fl Riboflavine
5'phosphate
F2 Riboflavine
F3 Flavine adenine
dinucleotide
F4 Lumiflavin
F5 Lumichrome
F6 Riboflavin tetranicotinate
F7 Riboflavin tetrabutyrate
Initial assays were carried out in relation to the various compounds mentioned in Table 2 to achieve preliminary results. The IC50 results in Table 2 are subject to confirmation; they are not consistent with re-run assays conducted to date. Assay results are shown in the graphs forming the following drawings and in Tables 2 to 10 which follow:
Figure 1: Antigen as optical density (OD) for Compounds F2,
F4 (first antigen assay) and F5 at 450 nm versus
concentration (M). The dotted line at OD 0.371
represents IC50 (active).
Figure 2: Antigen optical density (OD) for Compounds Fl and
F3 at 450 nm versus concentration (cm). The
dotted line at OD 0.371 represents IC50 (active).
Figure 3: Toxicity as C14 uptake (dpm) versus concentration
(M) for Compounds F2, F3, F4 (first toxicity
assay) and F5. The dotted line at 6035 dpm
represents CC50 (non-toxic).
Figure 4: Toxicity as C14 uptake (dpm) versus concentration (M) for Compound F1. The dotted line at 6035
dpm represents CC50 (non-toxic).
Figure 5: Antigen optical density (OD) for Compound F4
(second antigen assay) at 450 nm versus
concentration (M). The dotted line at OD 0.371
represents ICso (active).
Figure 6: Toxicity as C uptake (dpm) versus concentration (,fM) for Compound F4 (second toxicity assay)
The dotted line at 6035 dpm represents CC50 (non
toxic).
Figure 7: Antigen optical density (OD) for Compounds F6 and
F7 at 450 nm versus concentration ( M). The
dotted line at OD 0.371 represents IC50 (active).
Figure 8: Toxicity as C14 uptake (dpm) versus concentration
(M) for Compounds F6 and F7. The dotted line at
6035 dpm represents CC50 (non-toxic).
Figure 9: Antigen control (ddC)
As shown by the Tables, the test compounds were evaluated for activity against cells both acutely and chronically infected with HIV. Antiviral (iso) and toxicity (CC50) data is shown below. In another series of experiments, compounds were tested in cell cultures in which fresh media was added at 72, 48 and 24 hours prior to infection. This experiment was carried out to investigate the effects of the compounds on cells in actively dividing and quiescent states. This data indicates that cells may be more sensitive to the test compounds when quiescent. The effect of light on stability, preincubation of target cells, and the activity against an African HIV-1 isolate were also investigated. Exposure to light for two hours had no effect on the activity of the compound. Preincubation with the target cells enhanced its activity and it showed significant activity against the Africa HIV-l isolate.
Table 2 (Figures 1 to 4) - Acute Infection Standard Assay (l.lJ Compound Nol IC CC 0 SI
Assay No (Figures 1 and 2) (Figures 3 and 4)
F1/1 1 to 20 > 200
Fl/2 < 0.4 > 400 > 1000
Fl/3 0.1 (Figure 2) > 800 (Figure 4) > 8000
F2 3 (Figure 1) > 200 (Figure 3) > 60
F3 0.8 (Figure 2) > 200 (Figure 3) > 200
F4 1 (Figure 1) 150 (Figure 3) 150
F5 3 (Figure 1) > 200 (Figure 3) > 60
Table 3 (Figures 7 and 8) - Acute Infection Standard Assay (1.1) Compound No/
Assay No IC50 CC50 SI
F7/1 27 (Figure 7) 130 (Figure 8) 5
F7/2 57 > 200 > 4
F7/3 10 70 7
F7/4 25 140 6
Table 4 - Chronic Infection Standard Assay (2.1)
ComPound No/
Assay No IC50 CC50 SI
F7/l 0.2 7 35
F7/2 > 20 > 20
F7/3 10 > 200 > 20
F7/4 4 75 19
F7/5 26 > 200 > 7
Table 5 - Acute Infection Depleted Medium Assay (1.2)
Compound 72 hours 48 hours 24 hours
No
IC50 CC50 IC50 CC50 IC50 CC50
F7 10 160 21 100 110 160
Table 6 - Chronic Infection Depleted Medium Assay (2.2)
Compound 72 hours 48 hours 24 hours
No
IC50 CC50 IC50 CC50 IC50 CC50
F7 40 75 90 ' 250 60 101
Table 7 - Acute Infection Light Radiation Exposure Assay
(1.3)
Compound No Daylight Darkness IC50 CC50 IC CC50
F7 60 > 200 60 > 200
Table 8 - Acute Infection Preincubation Assay (1.4)
Preincubation of target cells with test compound for 24 hours before infection
ComPound No IC50 CCso SI
F7 5 120 24
Table 9 (Figures 5 to 8) - Acute Infection Standard Assay
(1.1)
Compound No IC CC50 SI
F4 13 (Figure 5) 150 (Figure 6) 12
F6 30 - 60 (Figure 7) > 200 (Figure 8) min 3 -6
Table 10 - Acute Infection Standard Assay (1.1)
Assay applied to C8166 Cells (T-lymphoblastoid cells transformed and immortalized by HTLV) with an African HIV
Isolate (HIV-l CBL4)
Compound No IC50 also SI
F7 4 150 37.5
Table 11 (Figures 10 to 12) - Acute Infection Standard
Assay (1.1)
Compound No IC50 CC50 SI
F7 32 200 6.3 ddC (control) 0.2
The variation in the end points observed with Compound F7 may be due to the properties of the target lymphoblastoid cells. Even in synchronized cultures there may be subtle changes in the metabolic state of sub-populations of cells.
This is reflected in the shift in the end points observed in the paired antiviral and toxicity values from assay to assay (Table 3). The results tabulated in Tables 5 and 6 indicate that cell culture in active or quiescent states may have different sensitivities to the test compound.
Patient Treatment
Thirty-five patients were placed on therapy. Thirty had follow up medical visits. i) General Condition of the Patients
Twenty patients out of thirty who came for follow-up visits reported an improvement in their general condition. The majority of these reported improvement insofar as malaise, appetite and weight gain was concerned. Two patients also reported improvement in skin rash wiLl rzegression of skin lesions while one reported no new skin lesions developed while on therapy. One patient also reported improvement in impotence (which had been present for three months prior to onset of therapy), while two other patients reported cessation of long standing coryza. ii) Sick Visits
Few patients attended clinic for unscheduled sick visits:1. One patient had recurrent abscesses as well as septic arthritis which persisted even on therapy.
2. Two patients had recurrent lower respiratory tract infect ions with one developing frank broncho-pneumonia during second week of therapy. Repeated smears for AAFBS have continued to be negative.
3. Two patients had pyrexia with no localizing signs and repeated blood smear for malarial parasites were negative and no significant growth on blood culture. One of these patients responded well to amoxycillin and is now afebrile.
4. One patient had gastroenteritis during the third week of therapy.
5. Oral and vulvo-vaginal candidiasis were reported by two patients, with the vulvo-vaginal candidiasis being recurrent as soon as a course of Nystatin pessaries and tablets was completed.
6. Two patients also reported recurrent attacks of herpes simplex genitalis. iii) Toxicity
Most of the cases of toxicity reported occurred during the first two weeks of therapy and have been transient.
Two patients experienced pruritus which averaged four days during first week of therapy and cleared spontaneously without any supportive medication.
Four patients reported moderate diarrhoea during the first two weeks of therapy. This has averaged four days. This has been a difficult symptom to attribute as between it being a side effect or a natural manifestation of the HIV infection. However, the consistency of its appearance in the first week of therapy, and its transient nature makes it reasonable to suppose it is a side effect.
One patient reported drowsiness and another reported darkening of her urine. MSU was normal.
Two patients reported abdominal discomfort. iv) Laboratory Results
Three patients had transient rises in liver enzymes during the second to third week of therapy, with no clinical signs of liver disease. However, the enzyme levels always returned to normal.
The above clinic trial reports are the preliminary results of a clinical trial which has currently been in progress for several weeks using Compound F7 administered orally in capsule form (the capsules are as described in Example 4 below) dosage was:
Dose level 1: lmg/kg body weight per day orally in two
divided dosages
Dose level 2: 2mg/kg body weight per day orally in two
divided dosages
Dose level 3: lOmg/kg body weight per day orally in two
divided dosages
Dose level 4: 15mg/kg body weight per day orally in two
divided dosages
Dose level 5: 2Omg/kg body weight per day orally in two
divided dosages
Dose level 6: 30mg/kg body weight per day orally in two to
three divided dosages
Dose level 7: 40mg/kg body weight~per-day orally in two to
three divided dosages
Dose level 8: 50mg/kg body weight per day orally in two to
three divided dosages
Dose level 9: 100mg/kg body weight per day orally in two
to three divided dosages
The following specific Examples illustrate compositions formulated in accordance with the invention:
Example 1
A formulation can be prepared from the following:
riboflavine-5-phosphate 10 mg
sterile water 2 ml to provide a unit dosage of 10 mg of riboflavine for administration once per day in the treatment of viral infect Ion.
Example 2
A formulation can be prepared from the following:
riboflavine-5-phosphate 30 mg
sterile water 2 ml to provide a unit dosage of 30 mg of riboflavine for administration once per day in the treatment of viral
Infection.
Example 3
Similar formulations to those of Examples 1 and 2 can be prepared at doses of:
10 mg per ml,
25 mg per ml, and
50 mg per ml respectively, in either a unit amount of 2 ml or 5 ml of sterile water and based on an active ingredient which is:
Riboflavine 5'phosphate
Riboflavine
Flavine adenine dinucleotide
Lumiflavin
Lumichrome or a mixture thereof.
Example 4
The following capsules were formulated:
Sizes: 25mg 5Omg , 100mg
200mg 400mug Type: 2-part hard gelatin
Composition: Compound F7 in admixture with
microcrystalline cellulose Ph. Eur 166.4/156.7/118.6/108.7/50mg to give capsule weights of 191.4/206.7/218.6/ 308.7/450mg.
Claims (33)
- CLAIMS 1. Use of a flavin, flavin derivative or a mixture comprising two or more thereof for the manufacture of a medicament for the treatment by prophylaxis or therapy of disease caused by viral Infection.
- 2. Use as claimed in Claim 1 wherein the flavin derivative is riboflavin or a riboflavin derivative.
- 3. Use as claimed in Claim 2 wherein the riboflavin derivative is a riboflavin salt.
- 4. Use as claimed in Claim 3 wherein the riboflavin salt is riboflavin sodium phosphate or riboflavin tetrabutyrate.
- 5. Use as claimed in Claim 1 wherein the flavin or flavin derivative is a compound of the general formula:wherein R is hydrogen or alkyl; R1 and R4 are, each independently, hydrogen, alkyl, hydroxy, halo, alkoxy, alkylthio, thio or an optionally substituted aromatic or non-aromatic nitrogen heterocycle, and X is: (i) hydrogen, ribityl, alkyl, hydrogen or an aromatic or nonaromatic carbocycle (ii) a group of the general formula: -CH2- (CHOH) Y in which n is an integer of 3 or 4 and Y is -CH2OH1-COOH or -COH or a group of the formula:wherein R is hydrogen or alkyl; and wherein W1 and W2 are, each independently, alkyl, hydroxy, halo, alkoxy, alkylthio, thio or an optionally substituted aromatic or non-aromatic nitrogen heterocycle.
- 6. Use as claimed in Claim 1 wherein the flavin or flavin derivative is a compound of the general formula:wherein X is (i) hydrogen, ribityl, alkyl, hydrogen or an aromatic or nonaromatic carbocycle (ii) a group of the general formula: -CH2-(CHOH)n-Y in which n is an integer of 3 or 4 and Y is -CH2OH1-COOH or -COH or a group of the formula:wherein R is hydrogen or alkyl; and wherein W1 and W2 are, each independently, alkyl, hydroxy, halo, alkoxy, alkylthio, thio or an optionally substituted aromatic or non-aromatic nitrogen heterocycle.
- 7. Use as claimed in Claim 1 wherein the flavin or flavin derivative is a compound of the general formula:-wherein R1 is hydrogen or an alkyl group, R2 is an alkyl group or a ribityl group, and R3 represents hydrogen or mono- or di-substitution of the outer carbocyclic ring with an alkyl group.
- 8. Use as claimed in Claim 1, wherein the flavin or flavin derivative is lumichrome; roseflavin; a hydroxyflavin; an alloxazine or derivative thereof; an 8a-N(3)-histidylflavin; an 8a N(1)-histidyl flavin; an 8a-cysteinyl thioether; an 6a-S-cysteinyl thioether; a lumiflavin; a 5-deazaflavin; a 5-carba-5-deaza or 1carba-l-deaza analog of riboflavin, FMN or FAD; flavin-1,N6ethenoadenine dinucleotide; 9-methylflavin; 9-phenylflavin; 9benzylflavin; 9-cyclohexylflavin; 6,9-dimethylflavin; 6,7,9trimethylflavin; 9-oxyethylflavin; 9-dioxypropylflavin; 6,8,9trimethylflavin; lacroflavin; flavin-9-carboxylic acid; 6,7dimethylflavin-9-carboxylic acid; or a schizaflavin.
- 9. Use as claimed in any preceding claim at a dosage regime of at least about 10 mg/kg of body weight per day.
- 10. Use as claimed in any preceding çlalm-.therein the medicament is in injectable form.
- 11. A flavin or flavin derivative for use in the manufacture of a medicament useful in the treatment by prophylaxis or therapy of disease caused by viral infection.
- 12. A flavin or flavin derivative as claimed in Claim 11 and as defined in any one of Claims 2 to 8.
- 13. A pharmaceutical composition for the treatment by prophylaxis or therapy of disease caused by viral infect ion, the composition being characterized in that it comprises a flavin or flavin derivative.
- 14. A composition as claimed in Claim 13 wherein the flavin or flavin derivative is as defined in any one of Claims 2 to 8.
- 15. A composition as claimed in Claim 12 or Claim 13 which composition comprises a unit dose of at least about 35 mg of a flavin or flavin derivative together with a pharmaceutically or veterinarily acceptable diluent, excipient or carrier.
- 16. A composition as claimed in Claim 15 wherein the unit dose is from about 35 mg to about 1000 mg.
- 17. A composition as claimed in Claim,1W-wherein the unit dose is from about 250 to 500 mg.
- 18. A composition as claimed in any one of Claims 15 to 17 which is in injectable form.
- 19. A composition as claimed in Claim 18 in the form of a solution in sterile water.
- 20. A receptacle for pharmaceutical containment immediately preadminstration, said receptacle being manipulable in a drug adminstration procedure by medical practitioners and containing a flavin or flavin derivative for discharge from the receptacle to a patient or to an administration device and said receptacle carrying a representation of instruct ions for use of the flavin or flavin derivative as a medicament for the treatment by prophylaxis or therapy of disease caused by viral infection.
- 21. The combination of : (a) a flavin or flavin derivative formulated for pharmaceutical use, and (b) instructions for use of said formulated flavin or flavin derivative for the manufacture of a medicament for the treatment by therapy or prophylaxis of disease cause,d..hy-v1ral inf!Ction or for use thereof for said treatment.
- 22. The combination of Claim 21 wherein the treatment is referred to in the instructions and is the treatment of HIV-infection.
- 23. The combination of Claim 22 wherein the HIV-infection is chronic infection.
- 24. A process for the manufacture of a medicament for use in the management and treatment of viral infection, which process comprises formulating a flavin or flavin derivative for anti-viral use.
- 25. Flavin, or a flavin derivative as an anti-viral agent, together with another compound(s) having anti-vIral activity, as a combined preparation for simultaneous, separate or sequential use in anti-viral therapy.
- 26. A method for the treatment by prophylaxis or therapy of disease caused by viral infection which method comprises administering therapeutically to a patient suffering from such disease an effective amount of a flavin or a flavin derivative or administering prophylactically to a patient at risk of viral infection an effective amount thereof.
- 27. A method as claimed in Claim 26 wherein the amount administered is at least about 1 to about 10 or more mg/kg of patient body weight.
- 28. A flavin or a flavin derivative thereof not known for any pharmaceutical utility for use as an anti-viral agent.
- 29. A flavin or flavin derivative for use in the treatment by prophylaxis or therapy of a disease caused by viral infection.
- 30. A flavin or flavin derivative as claimed in Claim 9 and as defined in any one of Claims 1 to 8.
- 31. An anti-viral agent for use in the treatment of HIV-infection in a mammalian subject at least at a chronic infection stage, the agent having a cellular target and optionally also a viral target and being a flavin or flavin derivative acting intracellularly on cell metabolism in mammalian cells infected chronically or acutely with HIV to block or compensate for the effects of the viral infection on the cell in the asymptomatic and post-asymptomatic phases of the infection b the virus.
- 32. An anti-viral agent as claimed in Claim 31 and which is a riboflavin derivative.
- 33. A method of in vitro diagnostic assay which method comprises sampling the cells of a mammalian patient infected with HIV after treating the patient by a treatment regisme-in which a flavine or flavine derivative is administered to the patient, and performing an assay upon the cell sample externally of and separate from the patients body to determine the activity and/or progress of the viral infection.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB9804302A GB2319474A (en) | 1993-10-19 | 1994-10-19 | Anti-viral agents |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB939321558A GB9321558D0 (en) | 1993-10-19 | 1993-10-19 | Anti-viral agents |
GB9421099A GB2283913A (en) | 1993-10-19 | 1994-10-19 | Anti-viral agents comprising flavins |
GB9804302A GB2319474A (en) | 1993-10-19 | 1994-10-19 | Anti-viral agents |
Publications (2)
Publication Number | Publication Date |
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GB9804302D0 GB9804302D0 (en) | 1998-04-22 |
GB2319474A true GB2319474A (en) | 1998-05-27 |
Family
ID=26303706
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
GB9804302A Withdrawn GB2319474A (en) | 1993-10-19 | 1994-10-19 | Anti-viral agents |
Country Status (1)
Country | Link |
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GB (1) | GB2319474A (en) |
Citations (9)
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US4219545A (en) * | 1979-03-23 | 1980-08-26 | Collins Calvin E | Treatment of infectious keratoconjunctivitis in animals |
US4264601A (en) * | 1979-06-12 | 1981-04-28 | The Board Of Regents Of The University Of Oklahoma | Antihypertensive agents and their use in treatment of hypertension |
US4500524A (en) * | 1982-09-15 | 1985-02-19 | Trustees Of Boston University | Tranquilizing and reducing or preventing seizures |
WO1991002529A2 (en) * | 1989-08-14 | 1991-03-07 | John Bennett Kizer | Product and method for killing abnormal vertebrate cells |
EP0417485A2 (en) * | 1989-09-12 | 1991-03-20 | Dorst Maschinen und Anlagenbau Otto Dorst und Dipl.-Ing Walter Schlegel GmbH & Co. | Press for making dimensionally stable pressed articles from particulate material |
WO1992017173A2 (en) * | 1991-04-02 | 1992-10-15 | Jean Berque | Use of riboflavin for treating hiv-related diseases, herpes, retinitis pigmentosa and malaria |
WO1993005784A1 (en) * | 1991-09-13 | 1993-04-01 | Eisai Co., Ltd. | Immunopotentiating and infection protective agent and production thereof |
US5217716A (en) * | 1990-07-18 | 1993-06-08 | The Beth Israel Hospital Association | Method for treating viral infections using oxidized lipoproteins |
WO1993010784A1 (en) * | 1991-11-25 | 1993-06-10 | University Of Michigan | Therapeutic composition and method for preventing reperfusion injury |
-
1994
- 1994-10-19 GB GB9804302A patent/GB2319474A/en not_active Withdrawn
Patent Citations (9)
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US4219545A (en) * | 1979-03-23 | 1980-08-26 | Collins Calvin E | Treatment of infectious keratoconjunctivitis in animals |
US4264601A (en) * | 1979-06-12 | 1981-04-28 | The Board Of Regents Of The University Of Oklahoma | Antihypertensive agents and their use in treatment of hypertension |
US4500524A (en) * | 1982-09-15 | 1985-02-19 | Trustees Of Boston University | Tranquilizing and reducing or preventing seizures |
WO1991002529A2 (en) * | 1989-08-14 | 1991-03-07 | John Bennett Kizer | Product and method for killing abnormal vertebrate cells |
EP0417485A2 (en) * | 1989-09-12 | 1991-03-20 | Dorst Maschinen und Anlagenbau Otto Dorst und Dipl.-Ing Walter Schlegel GmbH & Co. | Press for making dimensionally stable pressed articles from particulate material |
US5217716A (en) * | 1990-07-18 | 1993-06-08 | The Beth Israel Hospital Association | Method for treating viral infections using oxidized lipoproteins |
WO1992017173A2 (en) * | 1991-04-02 | 1992-10-15 | Jean Berque | Use of riboflavin for treating hiv-related diseases, herpes, retinitis pigmentosa and malaria |
WO1993005784A1 (en) * | 1991-09-13 | 1993-04-01 | Eisai Co., Ltd. | Immunopotentiating and infection protective agent and production thereof |
WO1993010784A1 (en) * | 1991-11-25 | 1993-06-10 | University Of Michigan | Therapeutic composition and method for preventing reperfusion injury |
Non-Patent Citations (2)
Title |
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Journal of Acquired Immune Deficiency Syndromes 6(8), pages 949-958 (1993) * |
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Publication number | Publication date |
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GB9804302D0 (en) | 1998-04-22 |
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