Use of macrolide compounds for eye diseases

Macrolide compounds such as the FR-900506 and its related compounds are provided for the prevention or treatment of eye diseases, particularly, allergic conjunctivitis. Composition containing such compounds is also disclosed.

DESCRIPTION 
NEW USE 
This invention relates to a new use of macrolide compounds for eye 
diseases. More specifically, this invention relates to a new use of 
macrolide compounds for eye diseases, particularly, allergic 
conjunctivitis. 
Accordingly, this invention provides a new use of the macrolide compounds 
for preventing or treating eye diseases as mentioned above. 
Further, this invention provides a prophylactic or therapeutic agent for 
eye diseases as mentioned above, which comprises the macrolide compounds. 
Still further, this invention provides a method for preventing or treating 
eye diseases as mentioned above, which comprises administering said 
macrolide compounds to mammals. 
Some of the macrolide compounds used in this invention are known and 
disclosed, for example, in European Patent Publication No. 0184162 and 
International Patent Application WO 89/05304. 
Those known macrolide compounds include the fermentation products, such as 
FR-900506, FR-900520, FR-900523 and FR-900525, isolated from 
microorganisms belonging to genus Streptomyces, such as Streptomyces 
tsukubaensis No. 9993 (FERM BP-927) or Streptomyces hygroscopicus. subsp. 
yakushimaensis No. 7238 (FERM BP-928), and their related compounds 
prepared from these fermentation products. And new macrolide compounds can 
be prepared from the above known macrolide compounds in a conventional 
manner. 
These macrolide compounds were indicated inter alia for use in the 
treatment of rejection to transplantation, autoimmune diseases and 
infectious diseases caused by pathogenic microorganisms, such as various 
fungi (Aspergills fumigatus, Fusarium oxysperum, Trichophyton asteroides, 
etc)(e.g.J. Antibiotics, XL(9), 1249-1255, 1987). 
The inventors of this invention have surprisingly found that the macrolide 
compounds mentioned hereinbelow are useful for preventing or treating eye 
diseases, particularly, allergic conjunctivitis, and also other allergic 
diseases such as food allergy, allergic rhinitis, etc. 
The macrolide compounds used in this invention can be represented by the 
following general formula (I). 
##STR1## 
wherein each vicinal pair of substituents R.sup.1 and R.sup.2 !, R.sup.3 
and R.sup.4 !, R.sup.5 and R.sup.6 ! 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, R.sup.2 may represent an alkyl 
group; 
R.sup.7 represents H, OH, protected hydroxy or O-alkyl, or in conjunction 
with R.sup.1 it may represent =O; 
R.sup.8 and R.sup.9 independently represent H or OH; 
R.sup.10 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 O,s (H, OH), (H,H) or --CH.sub.2 O--; 
Y represents O, (H, OH) (H,H) N--NR.sup.11 R.sup.12 or N--OR.sup.13 ; 
R.sup.11 and R.sup.12 independently represent H, alkyl, aryl or tosyl; 
R.sup.13, R.sup.14, R.sup.15, R.sup.16, R.sup.17, R.sup.18, R.sup.19, 
R.sup.22 and R.sup.23 independently represent H or alkyl; 
R.sup.20 and R.sup.21 independently represent O, or they may independently 
represent (R.sup.20 a,H) and (R.sup.21 a,H) respectively; R.sup.20 a and 
R.sup.21 a independently represent OH, O--alkyl or OCH.sub.2 OCH.sub.2 
CH.sub.2 OCH.sub.3 or R.sup.21 a is protected hydroxy; 
in addition, R.sup.20 a and R.sup.21 a may together represent an oxygen 
atom in an epoxide ring; 
n is 1, 2 or 3; 
in addition to their significances above Y, R.sup.10 and R.sup.23, together 
with the carbon atoms to which they are attached, may represent a 5- or 6- 
membered N-, S- or O-containing heterocyclic 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, O--alkyl, benzyl and --CH.sub.2 Se(C.sub.6 H.sub.5); and 
pharmaceutically acceptable derivatives thereof. 
The specific examples of the definitions of compound (I) and the preferred 
working modes of the invention are described in detail below. 
The term "lower" as used in this specification means, unless otherwise 
indicated, any number of carbon atoms between 1 and 6, inclusive. 
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 "aryl" may include phenyl, tolyl, xylyl, cumenyl, mesityl, 
naphthyl, and the like. 
Suitable examples of the protective group in the "protected hydroxyl group" 
may include: 
1-(lower alkylthio)(lower)alkyl groups such as lower alkylthiomethyl groups 
(e.g. methylthiomethyl, ethylthiomethyl, propylthiomethyl, 
isopropylthiomethyl, butylthiomethyl, isobutylthiomethyl, hexylthiomethyl, 
etc.), more desirably C.sub.1 --C.sub.4 alkylthiomethyl groups, and most 
desirably methylthiomethyl; 
tri-substituted silyl groups such as tri(lower)alkylsilyl groups (e.g. 
trimethylsilyl, triethylsilyl, tributylsilyl, tert-butyl-dimethylsilyl, 
tri-tert-butylsilyl, etc.); 
lower alkyl-diarylsilyl groups (e.g. methyldiphenylsilyl, 
ethyldiphenylsilyl, propyldiphenylsilyl, tert-butyldiphenylsilyl, etc.), 
more desirably tri(C.sub.1 --C.sub.4)alkylsilyl and C.sub.1 --C.sub.4 
alkyldiphenylsilyl groups and most desirably tert-butyldimethylsilyl and 
tert-butyldiphenylsilyl; and 
acyl groups such as aliphatic acyl groups, aromatic acyl groups and 
aliphatic acyl groups substituted by aromatic groups, which are derived 
from carboxylic acids, sulfonic acids or carbamic acids. 
The aliphatic acyl group may includes lower alkanoyl groups which may 
optionally have one or more suitable substituents such as carboxy (e.g. 
formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, 
pivaloyl, hexanoyl, carboxyacetyl, carboxypropionyl, carboxybutyryl, 
carboxyhexanoyl, etc.), cyclo(lower)alkoxy(lower)alkanoyl groups which may 
optionally have one or more appropriate substituents such as lower alkyl 
(e.g. cyclopropyloxyacetyl, cyclobutyloxypropionyl, cycloheptyloxybutyryl, 
menthyloxyacetyl, menthyloxypropionyl, menthyloxybutyryl, 
menthyloxypentanoyl, menthyloxyhexanoyl, etc.), camphorsulfonyl, lower 
alkylcarbamoyl groups having one or more suitable substituents such as 
carboxy or protected carboxy, for example carboxy(lower)alkylcarbamoyl 
groups( e.g. carboxymethylcarbamoyl, carboxyethylcarbamoyl, 
carboxypropylcarbamoyl, carboxybutylcarbamoyl, carboxypentylcarbamoyl, 
carboxyhexylcarbamoyl, etc.), protected carboxy(lower)alkylcarbamoyl 
groups such as 
tri(lower)alkylsilyl(lower)alkoxycarbonyl(lower)alkylcarbamoyl groups(e.g. 
trimethylsilylmethoxycarbonylethylcarbamoyl, 
trimethylsilylethoxycarbonylpropylcarbamoyl, 
triethylsilylethoxycarbonylpropylcarbamoyl, 
tert-butyldimethylsilylethoxycarbonylpropylcarbamoyl, 
trimethylsilylpropoxycarbonylbutylcarbamoyl, etc.) and so on. 
The aromatic acyl group may include aroyl groups which may optionally have 
one or more suitable substituents such as nitro (e.g. benzoyl, toluoyl, 
xyloyl, naphthoyl, nitrobenzoyl, dinitrobenzoyl, nitronaphthoyl, etc), 
arenesulfonyl groups which may optionally have one or more suitable 
substituent(s) such as halogen (e.g. benzenesulfonyl, toluenesulfonyl, 
xylenesulfonyl, naphthalenesulfonyl, fluorobenzenesulfonyl, 
chlorobenzenesulfonyl, bromobenzenesulfonyl, iodobenzenesulfonyl, etc.), 
and so on. 
The aromatic group-substituted aliphatic acyl group may include 
ar(lower)alkanoyl groups which may optionally have one or more suitable 
substituent(s) such as lower alkoxy and trihalo(lower)alkyl (e.g. 
phenylacetyl, phenylpropionyl, phenylbutyryl, 
2-trifluoromethyl-2-methoxy-2-phenylacetyl, 
2-ethyl-2-trifluoromethyl-2-phenylacetyl, 
2-trifluoromethyl-2-propoxy-2-phenylacetyl, etc.), and so on 
Among the above-mentioned acyl groups, the more desirable acyl groups are 
C.sub.1 --C.sub.4 alkanoyl groups which may optionally be substituted by 
carboxy, cyclo(C.sub.5 --C.sub.6)alkyloxy-(C.sub.1 --C.sub.4)alkanoyl 
groups having two (C.sub.1 --C.sub.4)alkyl groups in the cycloalkyl 
moiety, camphorsulfonyl, carboxy(C.sub.1 --C.sub.4)alkylcarbamoyl groups, 
tri(C.sub.1 --C.sub.4)alkylsilyl(C.sub.1 --C.sub.4)alkoxycarbonyl(C.sub.1 
--C.sub.4)alkylcarbamoyl groups, benzoyl which may have one or two nitro 
groups, halogen-substituted benzenesulfonyl groups, phenyl(C.sub.1 
--C.sub.4)alkanoyl groups having C.sub.1 --C.sub.4 alkoxy and 
trihalo(C.sub.1 --C.sub.4)alkyl groups. Of these groups, the most 
desirable are acetyl, carboxypropionyl, menthyloxyacetyl, camphorsulfonyl, 
benzoyl, nitrobenzoyl, dinitrobenzoyl, iodobenzenesulfonyl and 
2-trifluoromethyl-2-methoxy-2-phenylacetyl. 
Suitable "5- or 6-membered N-, S- or O-containing heterocyclic ring" may 
include pyrrolyl, tetrahydrofuryl, and the like. 
Preferred embodiments of the Symbols R.sup.1 to R.sup.10, R.sup.14 to 
R.sup.23, X, Y and n are as follows. 
R.sup.1 and R.sup.2 are each hydrogen or combined to form a second bond; 
R.sup.3 and R.sup.4 are combined to form a second bond; 
R.sup.5 and R.sup.6 are combined to form a second bond; 
R is hydrogen, hydroxy, 0-lower alkyl such as methoxy or protected hydroxy; 
R.sup.8 is hydrogen; 
R.sup.9 is hydroxy; 
R.sup.10 is methyl, ethyl, propyl or allyl; 
R.sup.14, R.sup.15, R.sup.16, R.sup.17, R.sup.18 and R.sup.19 are each 
methyl; 
R.sup.20 is oxo or R.sup.20 a,H!, wherein R.sup.20 a is hydroxy or 
methoxy; 
R.sup.21 is R.sup.21 a,H!, wherein R.sup.21 a is hydroxy or protected 
hydroxy; 
R.sup.23 is hydrogen; 
X is oxo, (H,OH) or (H,H); 
Y is oxo; and 
n is 1 or 2. 
The pharmaceutically acceptable salt of the compound (I) is a nontoxic 
salt, which may be the corresponding salt with an inorganic or organic 
base such as alkali metal salts (e.g. sodium salt, potassium salt, etc.), 
alkaline earth metal salts (e.g. calcium salt, magnesium salt, etc.), 
ammonium salt and amine salts (e.g. triethylamine salt, 
N-benzyl-N-methylamine salt, etc.) and so on. 
Referring to compound (I), there may exist conformers or one pair or more 
of stereoisomers such as optical and geometrical isomers due to the 
asymmetric carbon or the double bond. Such conformers and isomers also 
fall within the scope of the invention. 
Particularly, the most interesting compound is FR-900506 of the following 
formula. 
##STR2## 
As example for showing pharmaceutical activity, the pharmacological test 
data of the macrolide compounds (I) is illustrated in the following. 
Test: Effect of FK 506 on passive anaphylaxis in rat conjunctiva 
The diluted rat antiserum (IgE) to ovalbumin in a volume of 50 .mu.l was 
injected into both palpebral conjunctivas of male Wistar/ST rats aged 6 
weeks. Two days later, the rats were challenged intravenously with 
physiological saline (3 ml/kg) containing 1% ovalbumin and 0.5% Evans 
blue. The rats were sacrificed 30 min after challenge, and the eye tissues 
(eyelids and eyeballs) were removed. The intensity of anaphylactic 
reaction was assessed by measuring the amount of Evans blue extracted from 
the eye tissues. 
Test drug (eye drop) prepared by Example 2 mentioned below was administered 
topically to the rats 5 and 15 min (Test 1), or 2, 4 and 6 hours (Test 2) 
before challenge. Control groups were similarly given vehicle. The effect 
of drug was expressed as percent inhibition of the optical density at 620 
nm of the control groups. The result was expressed as the mean .+-.S.E. 
and statistical analysis was performed by Dunnett's multiple comparison 
test. 
TABLE 
______________________________________ 
Effect of FK 506 on passive anaphylaxis 
in rat conjunctiva 
Optical Density Inhibition 
Sample Control Test Sample (%) 
______________________________________ 
Test 1 0.300 .+-. 0.035 
0.116 .+-. 0.014** 
61.4 
(n = 12) (n = 10) 
Test 2 0.601 .+-. 0.047 
0.177 .+-. 0.028** 
70.5 
(n = 16) (n = 10) 
______________________________________ 
**: p &lt;0.01 
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(topical), enteral, 
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, eye 
drops, suppositories, solutions (saline, for example), emulsion, 
suspensions (olive oil, for example), ointment and any other form suitable 
for use. The carriers which can be used are water, glucose, lactose, gum 
acacia, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn 
starch, keratin, colloidal silica, potato starch, urea and other carriers 
suitable for use in manufacturing preparations, in solid, semisolid, or 
liquid form, and in addition auxiliary, stabilizing, 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 the 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, cats, rats, etc. and humans. 
For applying this composition to a human, it is preferable to apply it by 
external(topical) administration, particularly in the form of eye drops. 
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 0.0001-1000 mg, 
preferably 0.001-500 mg and more preferably 0.01-100 mg. of the active 
ingredient is generally given for treating diseases, and an average single 
dose of about 0.001-0.01mg, 0.2-0.5 mg, 1 mg, 5 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 0.1-0.3 mg/kg/day.

EXAMPLE 1 
______________________________________ 
FK 506 1 g 
Hydroxypropyl methylcellulose 2910 (TC-5R) 
1 g 
Lactose 2 g 
Croscarmellose sodium (Ac-Di-Sol) 
1 g 
______________________________________ 
The FK 506 (1 g) was dissolved in ethanol (10 ml), and thereto was added 
hydroxypropyl methylcellulose 2910 (TC-5R) (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 Mark: 
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 composition was capsulated by a conventional manner to provide 
capsules containing 1 mg or 5 mg of FK 506 per each capsule. 
EXAMPLE 2 
______________________________________ 
FK 506 (fine powder) 
1 mg 
Polysorbate 80 0.5 
Polyvinyl alcohol 2.8 
Benzalkonium chloride 
0.1 
Sodium chloride 8.6 
pH5.25 Phosphate buffer 
to 1 ml 
______________________________________ 
An aqueous suspending eye drop containing the above-mentioned ingredients 
is prepared according to a conventional manner.