Progesterone compound and use thereof

A progesterone compound represented by the following formula (1): ##STR1## wherein R.sup.1 represents a C1-C23 hydrocarbon group!, and a neovascularization inhibitor containing the same as the active ingredient. The compound (1) has a potent neovascularization inhibitory effect and is hence useful in the treatment of malignant tumors, diabetic retinitis, rheumatism, etc.

This application is a 371 of PCT/JP95/00642 filed Apr. 3, 1995. 
TECHNICAL FIELD 
The present invention relates to asteroid compound, and more particularly 
to a novel progesterone compound which has excellent neovascularization 
inhibitory action, and thus is useful as a remedy for malignant tumors, 
diabetic retinitis, rheumatism, etc. The invention also relates to 
neovascularization inhibitors containing the compound as the active 
ingredient. 
BACKGROUND ART 
Malignant tumors, diabetic retinitis, rheumatism, etc. are known as 
diseases in which neovascularization participates critically in the cause 
of disease or aggravation of pathology. Among such diseases, malignant 
tumors have widely been studied with respect to their relation to 
neovascularization, and it has come to be elucidated that 
neovascularization is crucial in the metastasis and prognosis of malignant 
tumors. 
Recently, some substances exhibiting neovascularization activity have been 
reported. As such substances, there are known, for example, sulfated 
polysaccharides, platelet factor-4 (PF-4), pentosan polysulfates, TNP-470 
(an analog of a mold product) tissue metalloproteinase inhibitors (TIMP), 
and minocycline. However, neovascularization inhibitory actions of these 
substances are not necessarily satisfactory, and there remains a need for 
development of neovascularization inhibitors with more excellent effects. 
Estrogen is closely related to the genesis of endometrial carcinoma and 
mammary carcinoma, and therefore, anti-estrogen agents and high doses of 
progestogen have been used for their treatment. Specifically, 
medroxyprogesterone acetate, a progestogen, has been used as a remedy for 
mammary carcinoma and endometrial carcinoma. It has recently been reported 
that medroxyprogesterone acetate has neovascularization inhibitory action. 
However, when the present inventors examined the neovascularization 
inhibitory action of medroxyprogesterone acetate, they found its action to 
be weak, and thus it is questionable was to whether medroxyprogesterone 
acetate manifests its effect at an ordinary clinical dose. 
Accordingly, an object of the present invention is to provide a novel 
progesterone derivative having excellent neovascularization inhibitory 
action. 
DISCLOSURE OF THE INVENTION 
The present inventors synthesized a huge number of progesterone derivatives 
and screened them not only for their progestogen action but also for their 
neovascularization inhibitory action. As a result, they found that the 
9.alpha.-fluoro-6.alpha.-methylprogesterone derivative represented by the 
following formula (1) exhibits neovascularization inhibitory activity not 
less than 20 times as potent as that of medroxyprogesterone acetate. The 
present invention was accomplished based on this finding. 
Accordingly, the present invention provides a progesterone compound 
represented by the following formula (1): 
##STR2## 
wherein R.sup.1 represents a C1-C23 hydrocarbon group!. 
The present invention also provides a neovascularization inhibitory agent 
containing the progesterone compound of formula (1) as an active 
component. 
The present invention also provides a pharmaceutical composition containing 
the progesterone compound of formula (1) and a carrier for 
pharmaceuticals. 
The present invention also provides use of the progesterone compound of 
formula (1) as a pharmaceutical. 
The present invention also provides a method for the treatment of malignant 
tumors, diabetic retinopathy, and rheumatism, characterized by 
administering an effective amount of the progesterone compound of formula 
(1). 
BEST MODE FOR CARRYING OUT THE INVENTION 
In formula (1), which represents the progesterone compound of the present 
invention, examples of the hydrocarbon group R.sup.1 include C1-C23 linear 
or branched or cyclic alkyl or alkenyl groups, with C1-C17 alkyl groups 
being more preferred. Specific examples of particularly preferred R.sup.1 
groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, n-pentyl, 
n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-dodecyl, n-tridecyl, 
n-tetradecyl, and n-pentadecyl. Of these, methyl and n-pentadecyl are 
particularly preferred. 
The progesterone compound of formula (1) of the present invention may be 
prepared in accordance with either of the following reaction schemes A or 
B. 
##STR3## 
wherein R.sup.1 has the same meaning as defined above!. 
Briefly, a pregnenediol (2) is acetylated to obtain a 11.beta.-acetoxy 
derivative (3). Ethylene glycol is reacted therewith to protect the 
ketone, and is subjected to oxidation and then methylation so as to 
introduce a methyl group at the 6- position. When the resultant 6-methyl 
derivative (6) is hydrolyzed, dehydrated, and then fluorinated, a 
9.alpha.-fluorinated derivative (9) is obtained. Ketalization of 
9.alpha.-fluorinated derivative (9) and hydrolysis of ketal group gave a 
9.alpha.-fluoro-6.alpha.-methyl derivative (11), which is subsequently 
acylated to obtain a compound (1) of the present invention. 
The acetylation reaction of pregnanediol (2) is carried out by reacting an 
acetylating agent typified by acetic anhydride with pregnanediol (2) in 
the presence of a base such as 4-dimethylaminopyridine. Also, ketalization 
of the resultant 11.beta.-acetoxy derivative (3) is carried out by 
reacting ethylene glycol with the derivative (3) in the presence of a 
condensing agent such as p-toluene sulfonic acid. Oxidation of compound 
(4), which is performed using a peracid typified by m-chloroperbenzoic 
acid, produces a 5,6-epoxy derivative (5). Methylation of the 5,6-epoxy 
derivative (5) is preferably performed using a Grignard reagent such as 
magnesium methyl bromide. Hydrolysis of the resultant 6-methylated product 
(6) is preferably performed using a weak base such as potassium hydrogen 
sulfate, sodium hydrogen carbonate, and sodium carbonate. The resultant 
compound (7) is dehydrated with sodium hydroxide, potassium hydroxide, 
etc. Compound (8) is fluorinated by reacting hydrogen fluoride-pyridine 
with compound (8). 6-Methyl-9.alpha.-fluoro derivative (9) may be ketaled 
in a manner similar to that mentioned above using ethylene glycol, and the 
resultant ketal (10) may be hydrolized using a weak acid as described 
above. Acylation of the resultant 9.alpha.-fluoro-6.alpha.-methyl 
derivative (11) is preferably performed by reacting the derivative (11) 
with a reactive derivative of carboxylic acid (R.sup.1 COOH) such as acid 
anhydride, acid halide, etc. in the presence of a condensing agent such as 
p-toluenesulfonic acid. 
##STR4## 
wherein R.sup.1 has the same meaning as defined above!. 
Briefly, the hydroxy group at the 17- position of 
6.alpha.-methylpredonisolone (12) is acylated to obtain a 
17.alpha.-acylated product (13), which is then dehydroxylated to obtain a 
methylated product (14). When the double bond at the 1-position of the 
product (14) is reduced, an enone derivative (15) is obtained. Subsequent 
introduction of a fluorine atom into the 9.alpha.- position of the enone 
derivative affords the compound (1) of the present invention. 
Acylation of the 6.alpha.-methylpredonisolone (12) is performed through a 
two step reaction; reacting the compound (12) with trimethyl ortho- 
acetate or a similar material, and then hydrolyzing the ortho- ester. The 
hydroxyl group of the resultant 17.alpha.-acylated product (13) is removed 
by first reacting the product (13) with methanesulfonyl chloride and then 
refluxing with heat in the presence of methyl iodide. The obtained 
methylated product (14) is reduced via hydrogenation in the presence of a 
metal catalyst such as chlorotris(triphenylphosphine)rhodium (I), etc. 
Fluorination of the enone derivative is performed by the reaction with 
hydrogen fluoride in the presence of pyridine, etc. 
The thus-obtained compound (1) of the present invention exhibits excellent 
neovascularization inhibiting action with hormonal activities being 
eliminated. Therefore, the compound (1) can be used in the treatment of 
malignant tumors, diabetic retinitis, rheumatism, etc. as a 
neovascularization inhibitor. 
When the compound (1) of the present invention is used for the 
above-mentioned purposes, it is preferably used after being prepared into 
a pharmaceutical composition together with an ordinarily used 
pharmaceutical carrier. Such pharmaceutical compositions may be oral 
preparations such as tablets, granules, and capsules; injections; per 
rectum preparations; and transdermal preparations. When these compositions 
are prepared, carriers for pharmaceuticals may be employed, including 
vehicles, binders, disintegrants, solubilizers, and flavoring agents. 
When the compound (1) of the present invention is used as a 
neovascularization inhibitor, the dose is preferably from 0.1 to 600 mg 
per day for an adult/kg/day which is divided in 1 to 5 times, though it 
may differ according to the administration route, patient's condition, 
age, body weight, etc.

EXAMPLES 
The present invention will next be described in detail by way of examples, 
which should not be construed as limiting the invention. 
Example 1 
(1) 11.beta.-Acetoxy-17.alpha.-hydroxy-4-pregnene-3,20-dione (3) 
A mixture of 4-pregnene-11.beta.,17.alpha.-diol-3,20-dione (2) (2 g), 
acetic anhydride (10 ml), pyridine (20 ml) and 4-dimethylaminopyridine (20 
mg) was stirred for 6 hours at room temperature. The reaction mixture was 
diluted with water (50 ml) and extracted with chloroform. The chloroform 
layer was sequentially washed with 10% HCl, a 5% aqueous NaHCO.sub.3 
solution, and brine, dried over Na.sub.2 SO.sub.4, and concentrated. The 
residue was subjected to column chromatography (silica gel, 150 g) using 
70% chloroform/hexane as eluant, thereby obtaining an acetylated 
derivative (3) (2.24 g, 100%). Recrystallization from ethyl acetate/hexane 
afforded white crystals having a melting point of 
159.5.degree.-160.degree. C. 
.sup.1 H-NMR (CDCl.sub.3): .delta. 0.85(3H, s), 2.01 (3H, s), 2.21(3H, s), 
3.04(2H, m), 5.34-5.56(1H, m), 5.65(1H, s). 
MS:m/z388 (M.sup.+). 
Elementary analysis for C.sub.23 H.sub.32 O.sub.5 : Calculated: C 71.10; H 
8.30 Found: C 71.08; H 8.50 
(2) 6-Methyl-4-pregnene-11.beta.,17.alpha.-diol-3,20-dione (8) 
11-Acetate (3) (2 g), ethylene glycol (1.1 g), and p-toluenesulfonic 
acid.1H20 (40 mg) were refluxed in benzene (100 ml) for 7 hours while 
preventing humidity (by the use of a reflux condenser equipped with a 
moisture separator). After the temperature of the reaction mixture was 
returned to room temperature, the mixture was washed with brine, dried 
over Na.sub.2 SO.sub.4, and concentrated. (The residue is used directly in 
the next step.) 70% m-chloroperbenzoic acid (2 g) was added to the residue 
(4) (2 g) in chloroform (150 ml), and the mixture was stirred for 12 hours 
at room temperature. The reaction was sequentially washed with an aqueous 
Na.sub.2 CO.sub.3 solution and brine, and then concentrated. The resultant 
residue (5) is thoroughly dried, and then dissolved in anhydrous 
tetrahydrofuran (30 ml). While vigorously stirring the mixture, magnesium 
methyl bromide (30 ml, 1.02M tetrahydrofuran solution) was added. The 
reaction mixture was refluxed for 18 hours, the temperature returned to 
room temperature, and the contents poured in iced water containing 
ammonium chloride. The mixture was extracted with chloroform, and the 
chloroform layer was washed with saturated brine, dried over Na.sub.2 
SO.sub.4, and concentrated. The residue (6) was refluxed at 70.degree. C. 
for 2 hours in a mixture of acetone (80 ml) and an aqueous 5% KHSO.sub.4 
solution (40 ml). The solvent was evaporated, and the residue was 
extracted with chloroform. The chloroform layer was washed with saturated 
brine, dried over Na.sub.2 SO.sub.4, and concentrated. Subsequently, the 
residue (7) was stirred for 2 hours in a mixture of 0.05N NaOH (100 ml) 
and methanol (100 ml) at room temperature. Acetic acid (1 ml) was added to 
the reaction mixture, and the mixture was concentrated to ca. a half 
volume. Extraction was performed using chloroform. The chloroform layer 
was washed with brine, dried over Na.sub.2 SO.sub.4, and concentrated. The 
residue was subjected to column chromatography (silica gel, 70 g) using 
30% ethyl acetate/hexane as eluant, thereby obtaining the title compound 
(8) (23%, from compound (3)). Recrystallization from ethyl acetate 
afforded white crystals having a melting point of 221.degree.-225.degree. 
C. 
.alpha.! +64.6.degree. (MeOH). 
.sup.1 H-NMR (CDCl.sub.3): .delta. 1.03(3H, s), 1.05(3H, s), 1.06(3H, d, 
J=7.33Hz), 1.27(3H, d, J=7.57Hz), 1.42(3H, s), 1.50(3H, s), 2.29(3H, s), 
2.30(3H, s), 2.68-2.71(2H, m), 3.48(1H, q, J=7.01Hz), 4.47(1H, d, 
J=2.43Hz), 5.73(1H, s). 
MS:m/z360(M.sup.+). 
Elementary analysis for C.sub.22 H.sub.32 O.sub.4 : Calculated: C 73.30; H 
8.95 Found: C 73.40; H 8.99 
(3) 9.alpha.-Fluoro-17.alpha.-hydroxy-6-methyl-4-pregnene-3,20-dione (9) 
Hydrogen fluoride-pyridine 4 ml, HF:pyridine=7:3 (w/w)! were placed in a 
Teflon container. While maintaining the exterior temperature at 
-15.degree. C. under nitrogen, 
11.beta.-hydroxy-6-methyl-4-pregnane-3,20-dione (9) (200 mg) was added. 
The mixture was stirred for 60 hours at the same temperature, and 
subsequently, the mixture was sequentially washed with 3% HCl, 5% 
NaHCO.sub.3, and brine, dried over Na.sub.2 SO.sub.4, and concentrated. 
The residue was subjected to column chromatography (silica gel, 30 g) 
using 30% ethyl acetate/hexane as eluant, thereby obtaining a fluorinated 
compound (9) (47 mg, 23.3%). Recrystallization from ethyl acetate afforded 
white crystals having a melting point of 232.degree.-235.degree. C. 
.sup.1 H-NMR (CDCl.sub.3): .delta. 0.75(3H, s), 0.78(3H, s), 1.11(3H, d, 
J=6.35Hz), 1.20(3H, d, J=4.96Hz), 1.30(3H, s), 1.37(3H, s), 2.27(3H, s), 
2.28(3H, s), 2.69-2.75(2H, m), 2.99-3.07(1H, m), 3.82(1H, d, J=1.84Hz), 
5.85(1H, d, J=3.09Hz), 5.90(1H, d, J=2.00 Hz). 
MS:m/z225(M.sup.+). 
Elementary analysis for C.sub.22 H.sub.31 O.sub.3 F: Calculated: C 72.89; H 
8.62 Found: C 72.95; H 8.75 
(4) 9.alpha.-Fluoro-17.alpha.-hydroxy-6.alpha.-methyl-4-pregnene-3,20-dione 
(11) 
6-Methylpregnene (9) (40 mg), ethylene glycol (0.014 ml), and 
p-toluenesulfonic acid (2 mg) were added to benzene (20 ml), and refluxed 
with heat for 3 hours using a reflux condenser equipped with a moisture 
separator. The solvent was distilled off, and the residue was combined 
with an aqueous 5% KHSO.sub.4 solution (5 ml) and acetone (5 ml). The 
mixture was refluxed for 2 hours at 70.degree. C. The reaction mixture was 
concentrated and extracted with chloroform. The chloroform layer was 
washed with saturated brine, dried over Na.sub.2 SO.sub.4, and 
concentrated. The residue was subjected to column chromatography (silica 
gel, 20 g) using 30% ethyl acetate/hexane as eluant, thereby obtaining an 
.alpha.-methylated derivative (11) (23 mg, 57.5% from compound (9)). 
Recrystallization from ethyl acetate afforded white crystals having a 
melting point of 237.degree.-239.degree. C. 
.alpha.! +23.76.degree. (MeOH). 
.sup.1 H-NMR (CDCl.sub.3): .delta. 0.86(3H, s), 1.10(3H, d, J=6.45Hz), 
1.30(3H, s), 2.69-2.79(2H, m), 2.99-3.03(1H, d, 3.84(1H, d, J=1.80Hz), 
5.89(1H, d, J=1.80Hz). 
MS:m/z362(M.sup.+). 
Elementary analysis for C.sub.22 H.sub.31 O.sub.3 F: Calculated: C 72.89; H 
8.62 Found: C 72.90; H 8.78 
(5) 17.alpha.-Acetoxy-9.alpha.-fluoro-6.alpha.-methylprogesterone (1a) 
9.alpha.-Fluoromedroxyprogesterone (11) (20 mg) was stirred for 5 hours at 
-10.degree. C. in the presence of p-toluenesulfonic acid, acetic anhydride 
(0.4 ml), and methylene chloride (1 ml) under nitrogen. Water (10 ml) was 
added to the reaction mixture, and extraction was performed using 
chloroform. The chloroform layer was washed with an aqueous 5% NaHCO.sub.3 
solution and saturated brine, dried over Na.sub.2 SO.sub.4, and 
concentrated. The residue was subjected to column chromatography (silica 
gel, 20 g) using 30% ethyl acetate/hexane as eluant, thereby obtaining 
17.alpha.-acetoxy-9.alpha.-fluoro-6.alpha.-methylprogesterone (1a) (8 mg, 
35.8%). Recrystallization from ethyl acetate/hexane afforded white 
crystals. 
.alpha.!.sub.D.sup.20 +36.0.degree. (c=0.210, CHCl.sub.3). 
.sup.1 H-NMR (500 MHz, CDCl.sub.3) .delta.: 0.68(3H, s), 1.12(3H, d, 
J=6.5Hz), 1.31(3H, s), 2.05(3H, s), 2.10(3H, s), 2.96(1H, ddd, J=2.5, 
11.3, 14.3Hz), 5.90(1H, d, J=1.5Hz). 
.sup.13 C-NMR (126 MHz, CDCl.sub.3) .delta.: 13.3, 17.9, 21.1, 21.3(d, 
J=5.4Hz), 23.6, 25.0(d, J=24Hz), 26.2, 26.8, 29.1(d, J=5.5Hz), 30.3, 33.1, 
33.6, 34.4(d, J=3.0Hz), 37.6(d, J=21.0Hz), 43.3(d, J=21.8Hz), 44.6, 46.1, 
96.2, 99.4(d, J=181Hz), 123.9, 169.8, 170.6, 198.7, 203.6. 
MS:m/z 404(M.sup.+). 
High MS m/z (M.sup.+) Calcd C.sub.24 H.sub.33 O.sub.4 F:404.2361, 
Found:404.2380. 
IR (KBr) .nu..sub.max 1742, 1709, 1674 cm.sup.-1. 
m.p. 208.degree.-210.degree. C. 
Example 2 
(1) 
17.alpha.-Acetoxy-11.beta.,21-dihydroxy-6.alpha.-methyl-1,4-pregnadiene-3, 
20-dione (13) 
To a solution (60 ml) of 6.alpha.-methylpredonisolone 
(6.alpha.-methyl-11.beta.,17.alpha.,21-trihydroxy-1,4-pregnadiene-3,20-dio 
ne) (12) (2.13 g ) in dichloromethane were added pyridinium 
p-toluenesulfonate (28.0 mg) and trimethyl orthoacetate (1.01 ml). The 
mixture was stirred at room temperature (20.degree.-25.degree. C.) for 5 
hours. When the reaction was completed, the reaction mixture was 
concentrated, and the residue was dissolved in acetone. (30 ml). While 
cooling the mixture on ice, an aqueous solution (30 ml) of 
p-toluenesulfonic acid.H.sub.2 O (1.19 g) was added thereto. The mixture 
was stirred for 30 minutes on ice. Upon completion of reaction, a 
saturated aqueous NaHCO.sub.3 solution was added and extraction was 
performed using ethyl acetate. After steps of washing with brine, drying 
over Na.sub.2 SO.sub.4, evaporation of the solvent, and silica gel column 
chromatography (silica gel, 40 g) using 50% ethyl acetate/hexane as 
eluant, 17.alpha.-acetoxy-11.beta.,21-dihydroxy-6.alpha.-methyl-1,4-pregna 
diene-3,20-dione (13) (2.11 g, 89%) was obtained. 
.alpha.!.sub.D.sup.20 +18.1.degree. (c=0.490, CHCl.sub.3). 
.sup.1 H-NMR (500 MHz, CDCl.sub.3) .delta.: 0.97(3H, s), 1.13(3H, d, 
J=6.0Hz), 1.46(3H, s), 2.02(3H, s), 2.74-2.82(1H, m), 4.24(1H, dd, J=4.0, 
18.0Hz), 4.33(1H, dd, J=3.5, 18.0 Hz), 6.04(1H, s), 6.28(1H, d, J=10.0 
Hz), 7.25(1H, d, J=10.0Hz). 
MS:m/z 416(M.sup.+). 
m.p. 159.degree.-161.degree. C. 
(2) 
17.alpha.-Acetoxy-11.beta.-hydroxy-6.alpha.-methyl-1,4-prednadiene-3,20-di 
one (14) 
To a solution (10 ml) of 
17.alpha.-acetoxy-11.beta.,21-dihydroxy-6.alpha.-methyl-1,4-pregnadiene-3, 
20-dione (13) (557 mg) in pyridine was added methanesulfonyl chloride (0.15 
ml). The mixture was stirred at between 0.degree. C. and 5.degree. C. 
(exterior temperature) for 3 hours. After completion of reaction, iced 
water and ethyl acetate were added to the reaction mixture. The resultant 
mixture was sequentially washed with 10% HCl, a saturated aqueous 
NaHCO.sub.3 solution, and brine, and then dried over Na.sub.2 SO.sub.4. 
After the solvent was evaporated, the residue was dissolved in pyridine 
(10 ml). Sodium iodide (447 mg) was added to the solution, and the mixture 
was refluxed for 50 minutes. Upon completion of the reaction, a saturated 
aqueous Na.sub.2 SO.sub.3 solution was added to the reaction mixture and 
extraction was performed using ethyl acetate. After steps of washing with 
10% HCl, a saturated aqueous NaHCO.sub.3 solution, and saturated brine and 
drying over Na.sub.2 SO.sub.4, the solvent was evaporated, and the residue 
was subjected to silica gel column chromatography (silica gel=16 g) using 
50% ethyl acetate/hexane as eluant, thereby obtaining 
17.alpha.-acetoxy-11.beta.-hydroxy-6.alpha.-methyl-1,4-pregnadiene-3,20-di 
one (14) (316 mg, 59%). 
.alpha.!.sub.D.sup.20 +13.2.degree. (c=0.430, CHCl.sub.3). 
.sup.1 H-NMR (500 MHz, CDCl.sub.3) .delta.: 0.95(3H, s), 1.13(3H, d, 
J=6.5Hz), 1.46(3H, s), 2.06(3H, s), 2.05(3H, s), 2.90-2.98(1H, m), 
6.05(1H, t, J=2.0Hz), 6.29(1H, dd, J=2.0, 10.0Hz), 7.25(1H, d, J=10.0Hz). 
MS:m/z 400(M.sup.+). 
m.p. 138.degree.-140.degree. C. 
(3) 
17.alpha.-Acetoxy-11.beta.-hydroxy-6.alpha.-methyl-4-pregnene-3,20-dione ( 
15) 
To a solution (30 ml) of 
17.alpha.-acetoxy-11.beta.-hydroxy-6.alpha.-methyl-1,4-pregnadiene-3,20-di 
one (14) (316 mg) in dichloromethane-ethanol (1:1), 
chlorotris(triphenyl-phosphine)rhodium (I) (8.51 mg) was added. The 
mixture was stirred for 27 hours at room temperature 
(20.degree.-25.degree. C.) in a stream of hydrogen gas (1.5 kg/cm2). After 
completion of reaction, the reaction mixture was concentrated, and 
subjected to silica gel column chromatography (silica gel=10 g) using 50% 
ethyl acetate/hexane as eluant, thereby obtaining 
17.alpha.-acetoxy-11.beta.-hydroxy-6.alpha.-methyl-4-pregnene-3,20-dione 
(15) (285 mg, 90%). 
.alpha.!.sub.D.sup.20 +55.4.degree. (c=0.773, CHCl.sub.3). 
.sup.1 H-NMR (500 MHz, CDCl.sub.3) .delta.: 0.93(3H, s), 1.06(3H, d, 
J=6.5Hz), 1.43(3H, s), 2.06(3H, s), 2.07(3H, s), 2.92-2.98(1H, m), 
5.73(1H, d, J=1.5Hz). 
MS:m/z 362(M.sup.+). 
m.p. 193.degree.-195.degree. C. 
(4) 17.alpha.-Acetoxy-9.alpha.-fluoro-6.alpha.-methylprogesterone (1a) 
A solution of 
17.alpha.-acetoxy-11.beta.-hydroxy-6.alpha.-methyl-4-pregnene-3,20-dione 
(15) (145 mg) was dissolved in hydrogen fluoride pyridine (4.0 ml) at an 
exterior temperature of -15.degree. C. The solution was stirred for 65 
hours at the same exterior temperature of -15.degree. C. When the reaction 
was completed, iced water was added to the reaction mixture, which was 
then extracted with ethyl acetate (50 ml, 30 ml). The organic layer was 
sequentially washed with 10% HCl, saturated NaHCO.sub.3, and brine, and 
then dried over Na.sub.2 SO.sub.4. After the solvent was evaporated, the 
residue was subjected to silica gel column chromatography (silica gel=6 g) 
using 40% ethyl acetate/hexane as eluant, thereby obtaining 
17.alpha.-acetoxy-9.alpha.-fluoro-6.alpha.-methylprogesterone (1a) (37.9 
mg, 26%). 
Test Example 1 
Neovascularization Inhibitory Activity 
The neovascularization inhibitory activity was tested by a CAM 
(chorioallantoic membrane) method using the chorioallantoic membrane of a 
fertilized chicken egg. Briefly, two holes were made, using a drill, in 
the chorion of a fertilized chicken egg incubated in an incubator at 
37.degree. C. for 4.5 days. From a side hole, about 3 ml of egg white was 
removed by aspiration using an injector. The chorioallantoic membrane was 
separated from the chorion membrane in the lower section of the air cell, 
and subsequently, the chorion and chorion membrane in the upper section of 
the air cell were removed, to thereby expose the embryo and the 
chorioallantoic membrane. A chorioallantoic membrane having a diameter of 
3-5 mm was used in the test. A silicone ring having an inner diameter of 3 
mm was placed in the chorioallantoic membrane, cocentrically with the 
membrane. In the opening of the silicone ring, was placed one of EV 
(ethylene vinyl acetate copolymer) pellets containing various doses of the 
compound of the present invention. In a control group was placed an EV 
pellet containing no specimen. In a positive control group was placed 
medroxyprogesterone acetate (MPA), which is known to have 
neovascularization inhibitory activity. 
The chicken egg was incubated for a further 2 days at 37.degree. C., with 
the upper section of the chicken egg being covered with a metal cap coated 
with Teflon. In order to facilitate observation of the blood vessel 
network on the chorioallantoic membrane, a suitable amount of a fatty milk 
was injected to the inside of the chorioallantoic membrane. 
The neovascularization inhibitory activity was assessed by measuring the 
avascular zone in the chorioallantoic membrane. The neovascularization 
inhibitory activity was assessed as positive when the avascular zone 
exceeded 3 mm in diameter. The frequency of thus-determined positive 
results was counted, and the results are shown in Table 1. 
TABLE 1 
__________________________________________________________________________ 
Number of 
Number of chorioallantoic 
Frequency of emergence 
Dose 
chorioallantoic 
membranes showing fields 
of fields of no blood 
Compound 
(.mu.g/egg) 
membranes used 
of no blood vessels 
vessels (%) 
P value* 
__________________________________________________________________________ 
Compound 
0 26 0 0 -- 
(1a) 0.01 
12 2 17 &gt;0.05 
0.1 12 4 33 &lt;0.05 
1 12 6 50 &lt;0.001 
10 12 9 75 &lt;0.001 
100 12 12 100 &lt;0.001 
MPA 100 12 6 50 &lt;0.001 
300 12 9 75 &lt;0.001 
__________________________________________________________________________ 
*Statistical processing of the data regarding neovascularization 
inhibiting action was performed using a Fischer's accurate randomized 
test. P values satisfying P &lt; 0.05 are considered to represent 
statistically significant results. 
Test Example 2 
Activity of urokinase-type Plasminogen Activator (uPA) Secreted by Vascular 
Endothelial Cells into a Culture Medium 
The effect exerted by the compound of the present invention on uPA secreted 
by vascular endothelial cells into a culture medium was measured by the 
following method. Briefly, vascular endothelial cells (2.times.10.sup.5 
cells/well) were distributed into a 24-multiwell dish (product of Falcon), 
each well containing 1 ml of a basal culture medium (Dulbecco's modified 
Eagle's medium (DMEM)+25 mM HEPES+4.5 mg/ml glucose+0.584 mg/ml 
glutamine+100 units/ml penicillin+100 .mu.g/ml streptomycin) supplemented 
with 10% fetal bovine serum. The dish was cultured in a humidified 
incubator (5% CO.sub.2 --95% air) at 37.degree. C. for 16 hours. The cells 
were then washed using a serum-free basal medium twice, and cultured in 
serum-free basal media (1 ml each) containing the compound of the present 
invention at a variety of concentrations. After incubation for 24 hours, 
the serum-free culture medium was recovered and subjected to 
centrifugation. The activity of uPA contained in the supernatant was 
measured using plasminogen and a synthetic substrate S-2251. The activity 
is indicated by the urokinase unit (U) per ml of culture supernatant. In 
view of the fact that medroxyprogesterone acetate, which exhibits 
neovascularization inhibitory action, suppresses uPA secretion from 
vascular endothelial cells, medroxyprogesterone acetate was used as a 
positive control. The results are shown in Table 2. 
TABLE 2 
______________________________________ 
uPA Activity (U/1 ml Culture 
Compound Concentration 
supernatant)* 
______________________________________ 
Compound 0 0.88 .+-. 0.16 
(1a) 10.sup.-9 0.71 .+-. 0.16 
10.sup.-8 0.61 .+-. 0.12 
10.sup.-7 0.36 .+-. 0.10 
10.sup.-6 0.22 .+-. 0.04 
MPA 10.sup.-6 0.22 .+-. 0.05 
______________________________________ 
*Mean .+-. S.D. 
Example 3 
Tablets each weighing 100 mg were prepared according to the following 
formulation. 
______________________________________ 
Compound Example 1 10 (mg) 
Lactose 70 
Cornstarch 16 
Hydroxypropylcellulose 3 
Magnesium stearate 1 
______________________________________ 
Example 4 
Tablets each weighing 100 mg were prepared according to the following 
formulation. 
______________________________________ 
Compound of Example 1 1 (mg) 
Lactose 75 
Cornstarch 20 
Hydroxypropylcellulose 3 
Magnesium stearate 1 
______________________________________ 
Example 5 
Tablets each weighing 100 mg were prepared according to the following 
formulation. 
______________________________________ 
Compound Example 1 10 (mg) 
.beta.-Cyclodextrin 30 
Lactose 46 
Cornstarch 10 
Hydroxypropylcellulose 3 
Magnesium stearate 1 
______________________________________ 
Example 6 
Tablets each weighing 100 mg were prepared according to the following 
formulation. 
______________________________________ 
Compound of Example 1 1 (mg) 
.beta.-Cyclodextrin 3 
Lactose 64 
Cornstarch 28 
Hydroxypropylcellulose 3 
Magnesium stearate 1 
______________________________________ 
As described above, it was confirmed that the compound of the present 
invention exhibits neovascularization inhibitory action and suppresses 
secretion by vascular endothelial cells of plasminogen activator. 
Moreover, it was found that the neovascularization inhibitory action of 
the compound of the present invention is not less than 20 times as potent 
as that of conventionally known neovascularization inhibitory substance, 
medroxyprogesterone acetate. 
INDUSTRIAL APPLICABILITY 
The compound (1) of the present invention possesses a potent 
neovascularization inhibitory effect, and thus is useful as a remedy for 
malignant tumors, diabetic retinitis, and rheumatism based on the 
neovascularization inhibition.