GB2247620A - The use of macrolide compounds for cytomegalovirus infection - Google Patents

Abstract

The use of macrolide compounds of the following formula in the prevention and/or treatment of cytomegalovirus infection:- <IMAGE> wherein [R<1> and R<2>], [R<3> and R<4>], [R<5> and R<6>] are hydrogen or form a direct bond or R<2> is alkyl; R<7> is H, OH, protected hydroxyl or O-alkyl, or R<1> and R<7> is=O; R<8> and R<9> are H or OH; R<10> is H, alkyl, or alkenyl; X is O, (H, OH), (H, H) or -CH2O-; Y is O, (H, OH), (H, H), N-NR<11>R<12> or N-OR<13>; R<11> and R<12> are H, alkyl, aryl or tosyl; R<13>, R<14>, R<15>, R<16>, R<17>, R<18>, R<19>, R<22> and R<23> are H or alkyl; R<20> and R<21> are O, or OH, or O-alkyl; or R<20> and R<21> form an epoxide ring; n is 1, 2, or 3; or Y, R<10> and R<23>, together with the carbon atoms to which they are attached, form a 5- or 6- membered N-, S- or O-containing heterocyclic ring, which is optionally saturated or substituted;

Description

NEW USE OP KACROLIDE COMPOUNDS FOR CYTOMEGALOVIRUS INFECTION This invention relates to a new use of macrolide compounds for cytomegalovirus infection.

Accordingly, this invention provide. a new us. of the ma cro lide compounds for preventing or treating cytomegaloirus infection.

Further, this invention provides a prophylactic or therapeutic agent for cytomegalovirus infection, which comprises the macrolide compounds.

Still further, this invention provides a method for preventing or treating cytomegalovirus infection, which comprises administering sad macrolide compounds to memmals.

Some of the macrolide compounds used in this invention are known and disclosed, for example in Eiaropean Patent Publication No. 0184162 and International Patent Application WO 89/05304.

Those known macrolide compounds include the fermentation products, PR-900506, FR-900520, PR-S00523 and FR-900525 isolated from microorganisms belonging to genus Stretomvces and their related compounds prepared from these fermentation products. Further, new macrolide compounda can be prepared from the above known macrolide compounds in a conventional manner.

These macrolide compounds were indicated L alia for use in the treatment of rejection to transplantation, autoimmune diseases and infectious diseases.

The inventors of this invention have surprisingly found that the inacrolide compounds mentioned hereinbelow are useful for preventing or treating cytomegulovirus infection, particularly human cytomegalovirus (sCKV) infection.

The macrolide compounds used in this invention can be represented by the following general formula (r).

wherein each vicinal pair of substituents [R1 and R2, (3 and R43, [R5 and R6] independently a) represent two vicinal hydrogen atoms, or b) form a second bond between the vicinal carbon atoms to which they are attached; in addition to its significance above, R2 aay represent an alkyl group; R7 represents H, Ox, protected hydroxy or O-alkyl, or in conjunction with R1 it may represent -0; R8 and R9 independently represent H or OH; R10 represents R, alkyl, alkyl substituted by one or more hydroxyl groups, alkenyl, alkyl substituted by one or more hydroxyl groups, or alkyl substituted by =O;; X represents 0, (H,OH), (R,R) or -CH2O-; Y represents 0, (H,OH)1 (H,H), N-NR11R12 or N-OR13; R11 and R12 independently represent H, alkyl, aryl or tosyl; R13, R14, R15, R16, R17, R18, R19, R22 and R23 independently represent H or alkyl; R20 and R21 independently represent 0, or they may independently represent (R20a,H) and (R21a,H) respectively; R20a and R21a independently represent OH, O-alkyl or OCH2OCH2CH2OCH3 or R21a is protected hydroxy;; in addition, R20a and R21a may together represent an oxygen atom in an epoxide ring; n is 1, 2 or 3t in addition to their significances above, y, R10 and R23, together with the carbon atoms to which they are attached, may represent a 5- or 6- membered N- S- or 0containing heterocyclic ring, which may be saturated or unsaturated, and which may be substituted by one or more groups selected from alkyl, hydroxy, alkyl subitituted by one or more hydroxyl groups, Alkyl, benzyl and -CH2Se(C6H5); and pharmaceutically acceptable derivatives thereof.

The term lower used in the specification is intended to mean 1 to 6 carbon atoms, unless otherwise indicated.

Suitable "alkyl" means straight or branched saturated aliphatic hydrocarbon residue and may include lower alkyl such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, neopentyl, hexyl, and the like.

Suitable "alkenyl" means straight or branched unsaturated aliphatic hydrocarbon residue having one double bond and may include lower alkenyl such as vinyl, propenyl, butenyl, methylpropenyl, pentenyl, hexenyl, and the like.

Suitable arylw may include phenyl, tolyl, zylyl, cumenyl, mesityl, naphthyl, and the like.

Suitable hydroxy-protective group in the "protected hydroxy" may include 1-(lower alkylthio)(lower)alkyl, trisubstituted silyl and acyl as exemplified in European Patent publication No. 0184162.

Suitable "5- or 6-membered N-, S- or O-contining haterocyclic ring" may include pyrrolyl, tetrahydrofuryl, and the like.

Preferred embodiments of the Symbols R1 to R10, R14 to R23, X, Y and n are as follows.

R1 and R2 are each hydrogen or combined to form a second bond; R3 and R4 are combined to form a second bond; R5 and R6 are combined to form a second bond; R7 is hydrogen, hydroxy, Lower alkyl such as methoxy or protected hydroxy; R8 is hydrogen; R9 is hydroxy; R10 is methyl, ethyl, propyl or ally; R14, R15, R16, R17, R18 and R19 are each methyls R20 is oxo or (R20a,), wherein R20a is hydroxy or methoxy; R2l is [R21a,H), wherein R21a is hydroxy or protected hydroxy; R23 is hydrogen; X is oxo, (H,OH) or (H,H); Y is oxo; and n is 1 or 2.

Particularly, the most interesting compound is PR-90OS06 of the following formula.

FR-900506 ( FK06 With respect to the macrolide compounds (I) of this invention, it is to be understood that there may be one or more conformers or stereoisomeric pairs such as optical and grometrical isomers due to asymmetric carbon atoms and double bonds, and such isomers are also included within the scope of the present invention.

Salt. of the macrolide compounds of the present invention include all pharmaceutically acceptable salts without limitation.

As example for showing phArmacological activity, the pharmacological test data of the macrolide compounds are illustrated in the following.

Test Effect of the macrolide compounds (I) on the growth of HCXV.

Intracellular HCMV growth (yield) was assessed by one step growth in cells treated with FK506.

Human embryonic lung (HEL) cells in plastic flasks (25 cm2) were infected with 2 PFU/cell of HCMV AD169 for 1 hour. HCMV-infected cells were then washed three times with maintenance medium, i.e. Eagle's minimal essential medium supplemented with 3% fetal bovine serum, and incubated in this medium containing the indicated concentration of FK506 for three days. Then the cells were washed three times, changed to 5 ml of fresh maintenace medium, frozen and thawed three times, and centrifuged at 3,000 rpm for 10 minutes.Serially diluted samples of the supernatanta were inoculated onto HEL cells in plastic dishes (60 mm) and the cells were overlaid with nutrient agarose. After fixation by formaldehyde (58 aqueous solution and staining by methylene blue (0.03%) aqueous solution, the number of plaques was counted and intracellular HCMV yield in the presence of FX506 was determined.

The result is shown in Table 1, in which the data are expressed as man SD of three experiments.

Table 1 t Effect of FX506 on the growth of HCNV Concentration Growth of HCMV of FK506 (%)* ( M) 1 58.5 6.8 10 37.6 t 9.3 yield of HCMV in the presence of FK 506 * ... x 100 yield of SCMV in th. absence of FK506 The macrolide compounds of the present invention may be administered as pure compounds or mixtures of compounds or preferably, in a pharmaceutical vehicle or carrier.

The pharmaceutical compositions of this invention can be used in the form of a pharmaceutical preparation, for example, in solid, semisolid or liquid form, which contains the macrolide compounds of the present invention, as an active ingredient, in admixture with an organic or inorganic carrier or excipient suitable for external, enthral, intravenous, intramuscular, or parenteral applications. The active ingredient may be compounded, for example, with the usual non-toxic, pharmaceutically acceptable, carriers for tablets, pellets, capsules, suppositories, solutions (saline, for example) emulsion, suspensions (olive oil, for example), and any other form suitable for use.The carriers which can be used are water, glucose, lactose, gum acacia, gelatin, mannitol, starch paste1 magnesium trisilicate, talc, corn starch, keratin, colloidal silica, potato starch, urea and other carriers suitable for use in manufacturing preparations, in solid, sQmisolid, or liquid form, and in addition Auxiliary stabil$zing, thickening and coloring agents and perfumes may be used. The active object compound is included in the pharmaceutical composition in an effective amount sufficient to produce tho desired effect upon the process or condition of the disease.

Mammals which may be treated using the method of the present invention include livestock mammals such as cows, horses, etc., domestic animals such as dogs, cat, rats, etc. and humans.

For applying this composition to a human, it is preferable to apply it by oral, parenteral, zte:nal, enteral, intravenous, or intramuscular admistration.

While the dosage of therapeutically effective amount of the macrolide compounds varies from and also depends upon the age and condition of each individual patient to be treated, a daily dose of about 0,01-1000 mg, preferably 0.1-500 mg and more preferably 0.5-100 mg. of the active ingredient is generally givon for treating diseases, and an average single dose of about 0.2-0.5 mg, 1 mg, 3 mg, 10 mg, 50 mg, 100 mg, 250 mg and 500 mg is generally administered.

Daily doses for chronic administration in humans will be in the range of about 0.3 mg/kg/day.

The following examples are given for the purpose of illustrating the present invention.

Example 1 FK 506 ig Hydroxypropyl methylcellulose 29lO (TC-SR) 1 g Lactose 2g Croscarmellose sodium (Ac-D4-Sol) I g The FK 506 (1 g) was dissolved in ethanol (10 ml), and thereto was added hydroxypropyl methylcellulose 2910 (TC-SR) (1 g) to prepare a suspension. To this suspension was added dichloromethane (5 ml) to prepare a homogeneous solution.

Lactose (2 g) and croscarmellose sodium (Trade Xarkt Ac-Di-Sol, maker: Asahi Chemical Industry) were homogeneously suspended to this solution, and then the organic solvent was removed by evaporation. The residual product was dried under reduced pressure for 10 hours by vacuum dryer, milled for 2 minutes by coffee mill and then passed through a sieve (32 mesh) to give the solid dispersion composition of FK 506 (5 g). This composstion was capsulated by a conventional manner to provide capsules containing 1 mg or S mg of FK 506 per each capsule.

Claims (7)

What we claim is:

1. A use of macrolide compounds of the formulas

wherein each vicinal pair of substituents [R1 and R2], [R3 and R4), (R5 and R6] independently a) represent two vicinal hydrogen atoms, or b) form a second bond between the vicinal carbon atoms to which they are attached; in addition to its significance above, R2 may represent an alkyl group; R7 represents H, OH, protected hydroxy or O-alkyl, or in conjunction with R1 it may represent -0; R8 and R9 independently represent H or OH; R10 represents H, alkyl, alkyl substituted by one or more hydroxyl groups, alkenyl, alkenyl substituted by one or more hydroxyl groups, or alkyl substituted by =O;; x represents 0, (H,OH), (H,H) or -CH2O-; Y represents 0, (H,OH), (H,H), N-NR11Rl2 or N-oR13; Rll and R12 independently represent X, alkyl, aryl or tosyl; R13, R14, R15, R16 R17, R18, R19, R22 and R23 independently represent B or alkyl; R20 and R21 independently represent 0, or they may independently represent (R20a,H) and (R21a,H) respectively; R20a and R21a independently represent OH, O-alkyl or OCH2OCH2CH2OCH3 or R2la is protected hydroxy;; in addition, R20a and R21a may together represent an oxygen atom in an epoxide ring n is l, 2 or 3; in addition to their significances above, Y, R10 and R23, together with the carbon atoms to which they are attached, may represent a a 5- or 6 membered N-, S- or 0- containing hoterocyclic ring, which may be saturated or unsaturated, and which may be substituted by one or more groups selected from alkyl, hydroxy, alkyl substituted by one or more hydroxyl groups, Alkyl, benzyl and -CH2Se(C6H3); or a pharmaceutically acceptable salt thereof, for preventing or treating cytomega lovirus infection.

2. A use of the macrolide compounds (I) defined in Claim 1 as a prophylactic or therapeutic agent for cytomegalo virus induction.

3 A prophylactic or therapeutic agent for cytomegalovirus infection, which comprises the macrolide compounds (I) defined in Claim 1.

4. A method for preventing or treating cytomegalovirus infection, which comprises administering the macrolide compounds (I) defined in claim 1 to mammals.

5. A use of the macrolide compounds (Z) defined in claim 1 for manufacturing a medicament for preventing or treating cytomegalovirus infection.

6. A pharmaceutical composition for cytomegalovirus infection, which comprises the macrolide compounds (t) defined in Claim 1 in admixture with a carrier or excipient.

7. A process for preparing the pharmaceutical cmposition of Claim 6, which is characterized by admixing the macrolide compounds (I) with a carrier or excipient.