WO1992018130A1

WO1992018130A1 - Use of guanine derivative or a prodrug thereof in the treatment of hiv-1 infection

Info

Publication number: WO1992018130A1
Application number: PCT/GB1992/000647
Authority: WO
Inventors: Richard Anthony Vere Hodge; Martin David John Kenig
Original assignee: Smithkline Beecham Plc
Priority date: 1991-04-13
Filing date: 1992-04-10
Publication date: 1992-10-29
Also published as: ZA922702B; IE921156A1; JPH06506673A; AU1541292A; MX9201665A; EP0580657A1; CA2108175A1; GB9107896D0; PT100367A

Abstract

A method of treatment of HIV-1 infections in mammals, which mammals are infected with herpesviruses, which method comprises the administration to the mammal in need of such treatment, an effective amount of a compound of formula (A) or a pro-drug, or a pharmaceutically acceptable salt, phosphate ester and/or acyl derivative of either of the foregoing.

Description

USE OF GUANINE DERIVATIVE OR A PRODRUG THEREOF IN THE TREATMENT OF HIV-1 INFECTION

This invention relates to a method of treatment of HIV-1 infection in humans and animals , and to the use of compounds in the preparation of a medicament for use in the treatment of such infection .

EP-A-242482 (Beecham Group p . I . e . ) discloses BRL 44385 , the compound of formula (A) :

HO(CH₂)₃0

(A)

and salts thereof, as antiviral agents.

Pro-drugs of the compound of formula (A) disclosed EP-A-242482 are of formula (B) :

HO(CH₂)₃0

(B)

and salts and derivatives thereof as defined under formula (A); wherein X is C-_j__g alkoxy, NH2 or hydrogen. Alternative pro-drugs are in the form of acetal derivatives of the side chain, described in EP-A-413544 (Beecham Group p.I.e.), especially 2-amino-9- [3- (isopropoxymethoxy)propoxy)purine, which is BRL 55792. The compound of formula (A) and derivatives thereof have been described as useful in the treatment of infections caused by herpesviruses, such as herpes simplex type 1, herpes simplex type 2, varicella-zoster and Epstein-Barr viruses.

It has now been discovered that this compound has potential activity against the human immunodeficiency virus (HIV-1) , in patients also infected with herpesviruses, and are therefore of potential use in the treatment of HIV infections in such patients.

This discovery is related to the ability of the triphosphate derivative of BRL 44385 to inhibit the RNA-directed DNA polymerase (reverse transcriptase) activity of human immunodeficiency virus type 1 (HIV-1) . The reverse transcriptase of HIV-1 is a virus-encoded enzyme essential for the conversion of genomic RNA into proviral ds-DNA, and is therefore an excellent molecular target for antiviral chemotherapy.

The ability of HIV to enter cells previously infected with herpesviruses is known (for example, B-lymphocytes infected with EBV¹) . The presence of both herpes and human immunodeficiency viruses in the same cell has particular consequences.

1. BRL 44385 would be phosphorylated by herpes virus-encoded thymidine kinase leading to a high level of BRL 44385 triphosphate. The triphosphate formed is not only an inhibitor of herpes DNA polymerase, but this work indicates that it also inhibits HIV reverse transcriptase.

2. HIV replication may be enhanced by herpesvirus transactivating factors. A product of HSV, ICP-4

(infected-cell protein) can increase the initiation of HIV transcription. 3. Double infection of herpesviruses and HIV may result in phenotypic mixing and the production of 'pseudotype' HIV p particles bearing herpesvirus envelope glycoprotems . The packaging of HIV genomic RNA with HSV capsid proteins is also believed to occur. Either situation may lead to the infection by HIV of CD4-negative, herpesvirus-permissible cells, previously not susceptible to entry of this virus. This may also result in doubly-infected cells.

Accordingly, the present invention provides a method of treatment of HIV-1 infections in mammals, including humans, which mammals are infected with herpesviruses, which method comprises the administration to the mammal in need of such treatment, an effective amount of a compound of formula (A) :

(A)

or a pro-drug, or a pharmaceutically acceptable salt, of either of the foregoing.

Examples of pro-drugs, pharmaceutically acceptable salts and derivatives are as described in the aforementioned European Patent references, the subject matter of which are incorporated herein by reference.

Particular pro-drugs of interest are as described in EP-A-413544. The compound of formula (A), pro-drugs, salts and derivatives may be prepared as described in the aforementioned European Patent references.

The compound may be administered by the oral route to humans and may be compounded in the form of syrup, tablets or capsule. When in the form of a tablet, any pharmaceutical carrier suitable for formulating such solid compositions may be used, for example magnesium stearate, starch, lactose, glucose, rice, flour and chalk. The compound may also be in the form of an ingestible capsule, for example of gelatin, to contain the compound, or in the form of a syrup, a solution or a suspension. Suitable liquid pharmaceutical carriers include ethyl alcohol, glycerine, saline and water to which flavouring or colouring agents may be added to form syrups.

For parenteral administration, fluid unit dose forms are prepared containing the compound and a sterile vehicle. The compound depending on the vehicle and the concentration, can be either suspended or dissolved. Parenteral solutions are normally prepared by dissolving the compound in a vehicle and filter sterilising before filling into a suitable vial or ampoule and sealing. Advantageously, adjuvants such as a local anaesthetic, preservatives and buffering agents are also dissolved in the vehicle. To enhance the stability, the composition can be frozen after filling into the vial and the water removed under vacuum.

Parenteral suspensions are prepared in substantially the same manner except that the compound is suspended in the vehicle instead of being dissolved and sterilised by exposure to ethylene oxide before suspending in the sterile vehicle. Advantageously, a surfactant or wetting agent is included in the composition to facilitate uniform distribution of the compound of the invention.

Preferred parenteral formulations include aqueous formulations using sterile water or normal saline.

As is common practice, the compositions will usually be accompanied by written or printed directions for use in the medical treatment concerned.

An amount effective to treat the virus infection depends on the nature and severity of the infection and the weight of the mammal.

A suitable dosage unit might contain from 50mg to lg of active ingredient, for example 100 to 500mg. Such doses may be administered 1 to 4 times a day or more usually 2 or 3 times a day. The effective dose of compound will, in general, be in the range of from 0.2 to 40mg per kilogram of body weight per day or, more usually, 10 to 20 mg/kg per day.

The present invention also provides the use of a compound of formula (A) or a pro-drug, or a pharmaceutically acceptable salt of either of the foregoing, in the preparation of a medicament for use in the treatment of HIV-1 infections in mammals, including humans, which mammals are infected with herpesviruses.

Such treatment may be carried out in the manner as hereinbefore described.

The present invention further provides a pharmaceutical composition for use in the treatment of HIV-1 infections in mammals, including humans, infected with herpesviruses, which comprises an effective amount of a compound of formula (A) or a pro-drug, or a pharmaceutically acceptable salt of either of the foregoing, and a pharmaceutically acceptable carrier. Such compositions may be prepared in the manner as hereinafter described.

The compound of formula (A) and its prodrugs show a synergistic antiviral effect in conjunction with interferons; and treatment using combination products comprising these two components for sequential or concomitant administration, by the same or different routes, are therefore within the ambit of the present invention.

It will be appreciated that the treatment of herpesvirus infected patients may include prophylaxis of herpesvirus episode attacks (suppressive treatment) . In patients with HSV infection only, suppressive treatment would probably only be given to those patients with frequent recurrences. In contrast, the aforementioned finding in relation to HIV- 1, indicates the need for continuous suppressive treatment with BRL 44385 or a pro-drug to all HIV-1 infected patients with herpesvirus recurrences, particularly HSV-1, HSV-2 and VZV recurrences, even though these recurrences may be infrequent.

The compound of formula (A) and its prodrugs may also be administered in combination with other antiviral agents, especially anti-HIV agents such as AZT.

The following biochemical data illustrate the invention. BIOCHEMICAL DATA

Materials and Methods

Chemicals [³H]dGTP was purchased from Amersham

International, Aylesbury, U.K. Template primers Poly (rC) . P(dG)₁₂_₁₈ and Poly (dC) . p(dG)₁ _₁₈ ^were obtained from Pharmacia Ltd., Milton Keynes, U.K. BRL 44385 triphosphate (BRL 44385-TP) was prepared in the laboratories of SmithKline Beecham Pharmaceuticals, Great Burgh, United Kingdom.

Reverse transcriptase Purified, E_;_ coli expressed HIV-1 reverse transcriptase (RT) was supplied by the Protein Biochemistry Department of SmithKline Beecham

Pharmaceuticals, Upper Merion, U.S.A. The enzyme was stored and diluted in a buffer containing 50mM Tris-HCl (pH 8.0), lOmM Hepes, llOmM NaCl, 5.7mM DTT, 0.3mM EDTA, 0.06% Triton X-100, 50% glycerol.

Assay for reverse transcriptase activity The reaction mixture for the HIV-1 RT assay contained in a volume of 120μl: 33mM Tris HCl (pH 8.0), lOOmM KCl, 3.3mM MgCl₂, 3.3mM dithiothreitol, 0.2mM glutathione, 0.33mM EGTA (ethylene glycol-bis- (β-aminoethyl ether) N,N-tetra acetic acid), 0.033% Triton X-100, 4.OμM [³H]dGTP (4.16 Ci/mmol, 16.7μCi/ml) 0-50μM BRL 44385-TP, 0.3 A₂₆₀ units/ml Poly (rC) . p(dG)₁₂__l8 or Poly (dC) . P(dG)₁₂_₁₈ and 167ng/ml RT (equivalent to 1.43nM for an equimolar mixture of p66 and p51 polypeptides) . The reaction mixtures without RT were prepared in the microwells of a 96-well plate and preincubated at 37°C before the reactions were started by the addition of 20μl of the enzyme solution. The plates were then incubated for 95-100 minutes at 37°C. The reactions were terminated by the addition of 40μl of EDTA solution (0.2M, pH6.5) . The individual reaction mixtures were transferred to a DEAE filter mat (1205-405, LKB Wallac, Finland), prewetted with 0.3M NaCl/0.03M Na citrate, using a cell harvester (1295-001, LKB Wallac) . The filter mat was washed three times in the NaCl/Na citrate buffer and then once in 95% ethanol. Scintillation fluid (Beta Plate Scint, LKB Wallac) was added to the dried filter, and the reaction mixtures assayed for incorporation of radioactive dGMP in an LKB 1205 Beta Plate Liquid Scintillation Counter.

Results

From the plot of % inhibition against concentration of inhibitor approximate IC^_Q values were obtained for BRL 44385-TP as follows.

Experiment 1

Poly (rC) . ρ(dG)₁₂_₁₈ template - primer used:-

IC₅₀ BRL 44385-TP = 3.7μM.

Experiment 2

Poly (rC) . P(dG)₁₂-i8 ^temPlate - primer used:-

IC₅₀ BRL 44385-TP = 1.9μM.

Poly (dC) . p(dG)₁₂_₁₈ template - primer used:-

IC₅₀ BRL 44385-TP = 1.2μM. Conclusion

These results indicate that the concentration of BRL 44385 triphosphate required to give 50% inhibition of HIV-1 5 reverse transcriptase using a RNA template is in the range 1.9 - 3.7μM. When DNA was used as the template for the reverse transcriptase, the IC^Q was 1.2μM. These levels of BRL 44385-TP should be obtained in the herpes-infected cell.

10

References

151. Complement Receptor 2 Mediates Enhancement of Human Immunodeficiency virus infection in Epstein-Barr virus-carrying B cells. Tremblay et a_l., J. Exp. Med. 171, 1791 (1990).

202. Phenotypic mixing between human immunodeficiency virus and vesicular stomatitis virus or herpes simplex virus. Zhu et al., J. of AIDS, 3 , 215 (1990).

Claims

1. A method of treatment of HIV-1 infections in mammals, which mammals are infected with herpesviruses, which method comprises the administration to the mammal in need of such treatment, an effective amount of a compound of formula (A) :

HO(CH₂)₃0

(A)

or a pro-drug, or a pharmaceutically acceptable salt, phosphate ester and/or acyl derivative of either of the foregoing.

Use of a compound of formula (A) :

HO(CH₂)₃0

(A)

or a pro-drug, or a pharmaceutically acceptable salt, phosphate ester and/or acyl derivative of either of the foregoing; in the manufacture of a medicament for use in the treatment of HIV-1 infections in mammals, which mammals are infected with herpesviruses.

3. A pharmaceutical composition for use in the treatment of HIV-1 infections in mammals, which mammals are infected with herpesviruses, comprising a compound of formula (A) :

HO(CH₂)₃0

(A)

or a pro-drug, or a pharmaceutically acceptable salt, phosphate ester and/or acyl derivative of either of the foregoing; and a pharmaceutically acceptable carrier.

4. A method, use or composition according to claim 1, 2 or 3 wherein the compound is BRL 44385, of formula (A) .

5. A method, use or composition according to claim 1, 2 or 3 wherein the compound is a pro-drug of the compound of formula (A) , as described in EP-A-413544 (Beecham Group p.I.e. ) .

6. A method, use or composition according to claim 5 wherein the pro-drug of the compound of formula (A) is

2-amino-9- (3- (isopropoxymethoxy)propoxy)purine, known as BRL 55792. 7. A method, use or composition according to claim 5 or 6 wherein the medicament is adapted for oral administration.

8. A method, use or composition according to claim 1, 2 or 3 wherein the compound administered is in a 50mg to lg unit dose.

9. BRL 44385, or a pro-drug therefor, for use in continuous suppressive treatment of HIV patients with HSV or VZV recurrences.