Synthesized .beta.-adrenergic blockers derivatives of guaiacol

Compounds of formula I ##STR1## in which OR.sup.1 is --OR.sub.3 NR.sub.4, in which R3 is a secondary alcohol group with 1.0 to 6 carbon atoms; a cyclic oxygen containing group with two carbon atoms; a pyridylmethyl or a piperidyl-ethyl group, R.sub.2 is alkyl, an ester group, an aldehyde group, a carboxylic acid group or a ketone group with 3 to 6 carbon --O--CONHR.sub.9 atoms, the group O--R.sub.7 NCOR.sub.8 or the group --O--CONHR.sub.9 in which R.sub.4 is an alkyl group with 1 to 8 carbon atoms, R.sub.7 is an alkylene group with 1 to 6 carbon atoms, each of R.sub.8 and R9 is an alkyl group with 1 to 12 carbon atoms and their pharmaceutically acceptable acid addition salts, are selective .beta.-blockers, antagonists of platelet aggregation, and .beta.-receptor binding. The processes of preparation, compositions, method of treatment are also described.

FIELD OF THE INVENTION 
This application relates to .beta.-adrenergic blockers containing the 
guaiacol ring. 
BACKGROUND OF THE INVENTION 
Chinese crude drugs or spices eg. Zingiber officinale, Eugenia 
caryophyllata, Allium sativum, have been used in medicine and in flavoring 
foods. Crude ginger is used as an antiemetic and expectorant, an 
antitussive and accelerator of the digestive organs. Semidried old crude 
ginger is also used for stomachache, chest pain, low back pain, cough, 
common cold and as a cure for a form of edema being called "stagnate of 
water". Zingerone is the major component which accounts for the spicy 
character of ginger; gingerol and shogaol are other pungent components in 
ginger. Gingerol has cardio-tonic action, suppresses the contraction of 
isolated portal veins in mice, and modulates the eicosanoid-induced 
contraction of mouse and rat blood vessels. Shogaol exhibits pressor 
response. Both gingerol and shogaol are mutagenic, whereas zinger and 
zingerone have been found to exhibit antimutagenic activity. Shogaol has 
inhibitory activity on the carrageenin-induced paw edema and platelet 
aggregation. 
It is known that drug-induced physiological responses are mediated through 
the binding of drugs to their specific receptors in various tissues and 
.beta.-receptor blockade has been clinically used in cardio-vascular 
diseases. The pharmacological effects of .beta.-blockers are evaluated 
based on (1) cardioselectivity, (2) .alpha.-adrenergic blocking action, 
(3) intrinsic sympathomimetic activity, (4) local anesthetic-activity, and 
(5) pharmacokinetic parameters related to the metabolism and distribution. 
Atenolol, Metoprolol, Acebutolol . . . etc. are clinically used as 
.beta.-blockers and possess cardioselectivity. Some non-selective 
.beta.-blockers are suitable for treating bronchial spasm and 
insulin-dependent diabetes. The selectivity of .beta.-blockers is 
determined by comparing the dosage of each agent needed to produce 
inhibitory effects on .beta..sub.1 and .beta..sub.2 receptors in tissues. 
In addition to in vivo tests, cardiac tissue, adipose tissue, the smooth 
muscle of trachea, and uterus from animals or human lymphocyte are also 
used to examine the selectivity of .beta.-blockers. It was reported first 
by Weksler, B. B. (1977), then by Greer, I. A. (1985) that .beta.-blockers 
affect the platelet function. 
Further, Srivastava, K. C. reported that lipid soluble .beta.-blockers with 
membrane stabilizing activity inhibited the platelet coagulation and 
thromboxane synthesis. About the action mechanism of .beta.-blockers on 
the platelet function, it has been suggested that .beta.-blockers inhibit 
the activity of platelet phospholipase A.sub.2 (Greer, I. A. et al, 
Thromb. Haemost., vol. 54, 480-84) or interfere with intracellular 
Ca.sup.+2 mobilization (Weksler, B. B. et al, Blood, vol. 49, 185-96). 
.beta.-blockers, such as Propanolol also inhibit platelet release and the 
adherence of platelets on collagen, but propanolol does not inhibit the 
shape change of platelets. The structure-activity relationship between 
many .beta.-blockers and phenoxypropanolamine has been shown by Kierstead, 
R. W. et al (J. Med. Chem. vol. 26, 1561-69, 1983). They have also shown 
that replacement of the hydrogen atoms with isopropyl group in 
phenylethylamine produce better affinity with .beta.-blockers. 
SUMMARY OF THE INVENTION 
An object of the present invention is to provide new guaiacoxy 
propanolamine derivatives of formula of, I, their related pharmaceutically 
acceptable salts and compositions comprising the same which are selective 
.beta.-blockers, antagonists of platelet aggregation, and .beta.-receptor 
binding 
##STR2## 
in which R.sub.1 is alkyl with 1 to 6 carbon atoms, hydrogen, the group 
R.sub.3 NR.sub.4, or the group R.sub.5 R.sub.6 ; in which R.sub.3 is a 
secondary alcohol group with 1 to 6 carbon atoms, R.sub.2 is an alkene, an 
ester, an aldehyde group, a carboxylic acid group or a ketone group with 3 
to 6 carbon atoms, the group R.sub.7 N--COR.sub.8, or the group 
--CONHR.sub.9 in which R.sub.7 is an alkyl group with 1 to 6 carbon atoms, 
R.sub.8, R.sub.9 are alkyl group with 1 to 12 carbon atoms R.sub.4 is an 
alkyl group with 1 to 8 carbon atoms, R.sub.5 is an alkyl group with 1 to 
4 carbon atoms, R.sub.6 is a cyclic oxygen containing group with 2 to 4 
carbon atoms, provided that when R.sub.1 is hydrogen, R.sub.2 is a ketone 
group with 3-6 carbon atoms. 
Another object of the invention is to provide .beta.-adrenergic blockers 
compositions and a method of treatment of patients in need of treatment. 
Another object of the invention is to provide processes for the preparation 
of the novel compounds and to pharmaceutical compositions comprising the 
same. 
This invention has shown that even a minor chemical modification of the 
substituents on the aromatic ring of guaiacol-based compounds may result 
in a marked reduction of pain-producing potency and lead to a complete 
loss of antinociceptive activities, better) .beta.-blockers activity, and 
less toxicity than that of natural pungent substances. 
The reaction schemes are illustrated hereinbelow. The synthetic reactions 
are preferably carried out in the alkaline solution of starting materials 
like vanillin, and eugenol (formula V) illustrated only with respect to 
vanillin, and further reacted with compounds (formula II IV) to produce 
compound of formula I. According to the reaction scheme hereinbelow 
compound 2 (dehydrozingerone) was synthesized from vanillin of formula V, 
then the compound 2 was hydrogenated and converted to compound 
1(zingerone). Treating compound 2 with the compound of formula II gave 
compound 3; amination of compound 3 with the compounds of formula IV in 
alcoholic solution produced the compound 4 or 6. The compound 5 was 
synthesized from compound 1 or hydrogenated from compound 4. R.sub.5 is 
alkyl group with 1 to 4 carbon atoms, R.sub.6 is a cyclic oxygen 
containing group with 2 to 4 carbon atoms and preferably the total number 
of carbon atoms of R.sub.5 and R.sub.6 is lower than six. 
##STR3## 
The R.sub.3 is a secondary alcohol group with 1 to 6 carbon atoms. R.sub.4 
is an alkyl group with 1 to 8 carbon atoms and preferably the total number 
of carbon atoms of R.sub.3 and R.sub.4 is lower than six. As a suitable 
base, one may include an inorganic base such as an alkali metal hydride, 
for instance sodium hydride, or an alkali metal hydroxide, eg. sodium 
hydroxide, potassium hydroxide, or an alkali metal carbonate, eg. sodium 
carbonate, or an alkaline earth metal hydroxide, for instance magnesium 
hydroxide, or the like. 
In formula I, the O R.sub.1 group may be a methoxy group in the meta 
position, and R.sub.2 on the para position with respect to the O R.sub.1 
group. The structure of all compounds were supported by data derived from 
melting points, infrared(IR) and nuclear magnetic resonance (NMR) spectra, 
mass spectra. 
The guaiacoxy- propanolamine derivatives according to the present invention 
and also their salts display useful pharmacological properties. When 
R.sub.1 is hydrogen, the salts or the compounds are the sodium salts, 
potassium salts, calcium salts, or magnesium salts. The formula I 
compounds and their pharmaceutically acceptable salts are selective 
.beta.-adrenergic blockers, antagonists of platelet aggregation, and 
.beta.-receptor binding. 
The compounds according to this invention are useful as a medicine for the 
prevention of thrombosis. The following tests are given for the purpose of 
illustrating the pharmacological activity. 
TEST 1 
Test Method 
The wiping test was performed as described by Szolcsanyiand, Jancso-Gabor. 
Briefly, the solution or suspension of the test compound and its analogs 
was made up in 10% ethanol, 5% Tween 80 and then diluted with saline to 
the required concentrations. Each solution or suspension of 10.sup.-4 M, 
10.sup.-3 M, 10.sup.-2 M was dropped into the right eye (vehicle being 
administered to the left eye as negative control) of male Wistar rats 
weighing 180-250 g and the total number of protective movements 
(scratching, wiping of the eye with the foreleg) was counted for 30 
minutes. Each test was carried out with a total of 6 rats, and a 
dose-response curve was obtained from the mean value of each group. MPPs 
(the concentrations having a moderate pain-producing potency) were 
calculated from the dose-response curve and those concentrations inducing 
equal reactions of 32 scratchings were recorded. 
Test Compounds 
4-(4'-hydroxy-3'-methoxyphenyl)-3-butan-2-one (zingerone,1) 
4-(4'-hydroxy-3'-methoxyphenyl)-3-buten-2-one (Dehydrozingerone, DZ,2) 
4-4'-(2,3-Epoxypropoxy)-3'-methoxyphenyl!e-3-buten-2-one (DZE,3) 
4-4'-(2-hydroxy-3-isopropylaminopropoxy)-3'-methoxyphenyl!-3-buten-2-one 
(DZN, 4) 
4-4'-(2-hydroxy-3-tertbutylaminopropoxy)-3'-methoxyphenyl!-3-buten-2-one(D 
ZBN,6) 
Test Result 
Based on the obtained MPP values, RPP (relative pain producing potency) 
values were determined with respect to the pain-producing potency of 1, 
which was taken as 1000. As shown in FIG. 1, the pain producing potency of 
compounds 4 and 6 of DZN and DZBN is lower than that of DZ and zingerone. 
TEST 2 
Test Method 
Following the method described by Koster antinociceptive tests were carried 
out in male mice after intraperitoneal administration of test solution. 
Briefly, four groups of eight male mice (ddk strain) weighing 18-22 g were 
brought to the laboratory on the day prior to study, and housed overnight 
with free access to food and water. Solutions of compound 1 and its 
analogs as well as indomethacin were made up in 10% ethanol, 10% Tween 80 
and 80% saline, and then diluted with saline to the required 
concentrations. The test solution was administered by intraperitoneal 
injection of a single dose 0.2 ml (vehicle administered as control). 
Twenty minutes after injection, 0.2 ml of 0.7% acetic acid was injected 
intraperitoneally to induce writhing. Following the injection, the mice 
were placed in separate clear glass cages and the number of writhes was 
counted for 18 consecutive 5 minute periods beginning 5 minutes after the 
acetic acid injection, a writhe being defined as a sequence of arching of 
the back followed by pelvic rotation and hind limb extension. 
The compounds tested were: 
4-(4'-hydroxy-3'-methoxyphenyl)-3-butan-2-one (zingerone,1) 
4-(4'-hydroxy-3'-methoxyphenyl)-3-buten-2-one(Dehydrozingerone DZ,2) 
4-4'-(2,3-Epoxypropoxy)-3'-methoxyphenyl)!-3-buten-2-one (DZE,3) 
4-4'-(2-hydroxy-3-isopropylaminopropoxy)-3'-methoxyphenyl!-3-buten-2-one 
(DZN, 4) 
The ED50 values reported in Table 1 show significant variations in 
antinociceptive effect, zingerone is 3 times more potent than DZ. ED50 is 
the effective dose for 50% of the animals. 
TABLE 1 
__________________________________________________________________________ 
Comparison of antinociceptive effects of 
zingerorie analogues on acetic acid-induced 
writhing syndrome. 
Dose Writhes No. 
Protection 
ED.sub.50 (95% C.I.) 
Potency 
Compound 
(mg/kg) 
N (Ave. .+-. S.E.) 
(%) (mg/kg) ratio 
__________________________________________________________________________ 
Saline 24 80.14 .+-. 5.13 
0 
Zingerorie 
0.25 
8 47.13 .+-. 4.49 
10.82 
0.25 (1.06-0.06) 
1.50 
1.25 
8 20.75 .+-. 3.40 
71.10 
2.50 
8 4.00 .+-. 2.20 
95.01 
DZ 0.25 
8 58.13 .+-. 5.71 
27.46 
0.38 (0.82-0.17) 
1.00 
1.25 
8 29.60 .+-. 3.75 
63.06 
2.50 
8 2.33 .+-. 0.83 
97.09 
DZE 0.25 
8 46.67 .+-. 2.09 
41.76 
0.23 (0.88-0.06) 
1.70 
1.25 
8 27.50 .+-. 3.32 
65.69 
2.50 
8 13.86 .+-. 1.82 
132.71 
DZN 0.25 
8 29.80 .+-. 5.73 
62.82 
0.11 (0.60-0.02) 
3.50 
1.25 
8 11.17 .+-. 2.36 
86.06 
2.50 
8 3.20 .+-. 0.23 
96.01 
__________________________________________________________________________ 
a: Writhes were counted for 30 min after acetic acid injection (i.p.) 
b: Protection (%) = 100 - (experimental/control .times. 100) 
c: ED.sub.50 's and 95% confidence intervals were calculated by the 
Litchfield and Wilcoxon method 
FIG. 2 shows the total numbers of writhes of the compounds tested. 
TEST 3 
Test Method 
Anesthesed rats were administered intravenously DZN (compound 4) in the 
amounts of 0.1, 0.5, 1.0 mg/kg, Propranolol and isoproterenolol were 
administered as control. The effect of DZN on heart rate and blood 
pressure were monitored. 
Test Compound 
4-4'-(2-hydroxy-3-isopropylaminopropoxy)-3'-methoxyphenyl!-3buten-2-one 
(DZN,4) 
Test Results 
(1) As shown in FIGS. 3-6 DZN induced dose-dependent bradycardia and this 
effect lasted over 1 hr. DZN and propranolol slightly reduced the blood 
pressure for 5-10 minutes after injection. Statistical significance was 
not found. 
(2) As FIGS. 6(A) and 6(B) show the 0.5 mg/kg isoproterenolol increased the 
heart rate and decreased blood pressure of rats. Propranolol antagonized 
the effects of isoproterenolol. These results are in agreement with the 
findings of Baird, J. R. C. et al (J. Pharm. Pharmac., vol. 24, 880-85, 
1972). DZN compound 4 not only reduced resting heart rate but also lowered 
the blood pressure induced by isoproterenolol. 
(3) The ID.sub.50 value of propranolol induced heart rate change was 0.14 
mg/kg, and that of DZN was 0.22 mg/kg (Table 2) calculated by the method 
of Litchfield, J. L. et al (J. Pharmacol. Exp. Ther., vol. 96, 99-113). 
The ID.sub.50 value is the response of a .beta. drug which produces a 
response in 50% of a drug. 
TABLE 2 
______________________________________ 
ID.sub.50 Heart Rate response of various .beta.-adrenergic 
blocking agents 
______________________________________ 
.beta.- Adrenergic ID.sub.50 : heart rate 
blocking agent response (95% C.I.) 
(mg/kg) 
Propranolol 0.14 (0.96-0.02) 
DZH (compound 4) 0.22 (1.29-0.04) 
______________________________________ 
TEST 4 
Test Method 
(1) The test was conducted according to the method described by Malta, E. 
(BR.J.Pharmac., vol. 85, 179-87, 1985) 0.50 .mu.M of phenoxybenzamine was 
first applied to the 10 ml organ bath of isolated guinea pig right atrium 
to block the response of .beta.1 adrenergic receptor. A series of doses of 
isoproterenolol from 10.sup.-10 -3.times.10.sup.-8 M were then introduced 
into the bath. A bath as described in Gen. Pharmacology 25 p 652 is used 
to put in the solution of the drug and the organ is hung within the bath 
and the contraction or relaxation response is shown on a transducer 
(2) The isolated guinea pig right atrium was pretreated with DZN, and then 
given isoproterenolol described as above. 
Test Compound 
4-4'-(2-hydroxy-3-isopropylaminopropoxy)-3'-methoxyphenyl!-3-buten-2-one 
(DZN, compound 4) 
Test Results 
(1) The effects of isoproterenolol on contraction force and beating rate of 
right atrium are dose-dependent. The maximum effect was obtained at 
3.times.10.sup.-8 M isoproterenolol (FIG. 7). 
(2) To obtain the maximum effect of beating rate, a concentration of 
isoproterenolol is required higher than that of control group (FIGS. 7 and 
8). After the treatment with DZN, the sigmoid curve 
isoproterenolol-dependent concentration effect was partially shifted to 
the right as shown in FIG. 9. A very close phenomenon was seen when DZN 
was replaced with propranolol. The estimated pA.sub.2 values of DZN and 
propranolol were 8.16 and 8.36, respectively. Regression analysis revealed 
a strong correlation between the doses of antagonists and isoproterenolol 
(FIG. 10). 
As described in "Manual of pharmacological calculation with computer 
programs, New York: Springer Verlag: 1987. by Tallarida, R. J.; Murray, R. 
B.", the pA.sub.2 is a measure of the affinity of a competitive antagonist 
for its receptor. The determination of the pA.sub.2 is made from 
experiments in which a fixed concentration of the antagonist is used along 
with graded concentrations of an agonist acting on the same receptor. The 
presence of the antagonist shifts the agonist dose-response curve to the 
right. 
TEST 5 
Test Method 
(1) The method developed by Piercy, V. et al (J. Pharmac. Methods., vol. 
20, 125-33, 1988) was employed for this experiment. Female rats were 
intraperitonealy injected with stilbestrol (1 mg/kg) to elevate the 
sensitivity of .beta..sub.2 adrenergic receptor in uterus horns. The rats 
were subsequently administered reserpine to deplete the catecholamine in 
uterus. The isolated uterus horns were treated with phenoxybenzamine to 
block the neuronal uptake and the interaction between neuron cells and the 
adrenergic receptor. 
Test Compound 
4-4'-(2-hydroxy-3-isopropylaminopropoxy)-3'-methoxyphenyl!-3-buten-2-one 
(DZN,4) 
Test results (1) When a krebs' solution was used as the rinsing solution, a 
fast contraction in the uterus smooth muscle was observed after 
depolarization, then the muscle relaxed and reached a steady state. 
Isoproterenolol ranging from 10.sup.-8 -10.sup.-6 M was added. A 
dose-related decrease in relaxation force was observed in the smooth 
muscle of isolated uterus horns (FIG. 11). 
(2) Pretreatment of the isolated uterus horns with 10.sup.-8 -10.sup.-6 M 
DZN caused the concentration of isoproterenolol needed to reach the 
maximum relaxation effect to be increased as shown in FIGS. 11 and 12. 
Pretreatment with DZN led the concentration effect curve of 
isoproterenolol to partially shift to the right (FIG. 13); the pA.sub.2 
value of DZN was 7.53. The estimated slope of the regression analysis was 
0.99.+-.0.11 (FIG. 14). 
(3) Compared to isoproterenolol and DZN, propranolol produced a regression 
slope of 0.99.+-.0.12 and a pA.sub.2 values of 8.33 in its 
concentration-effect curve. As summarized in Table 3, the 
cardioselectivity of DZN was 4.26 according to the pA.sub.2 values 
obtained from the log concentration-effect curve. 
TABLE 3 
______________________________________ 
.beta.-Adrenoceptor blocking potency and cardioselectivity of 
propranolol and DZN on guinea-pig and rat in vitro 
preparations. pA2 values and slopes of regression calculated 
from Schild plots are shown. Cardioselectivity ratio is 
obtained from the antilogarithm of the difference between 
the mean pA2 values obtained from different tissues. 
.beta.1 .beta.2 Cardiose- 
.beta.-Adrenoceptor 
guinea-pig right atrium 
rat uterus 
lectivity 
blocking agent 
(slope) (slope) ratio 
______________________________________ 
Propranolol 
8.36 .+-. 0.02 
8.33 .+-. 0.19 
1.07 
(0.97 .+-. 0.10) 
(0.99 .+-. 0.12) 
DZN 8.16 .+-. 0.03 
7.53 .+-. 0.10 
4.26 
(0.96 .+-. 0.05) 
(0.99 .+-. 0.11) 
______________________________________ 
TEST 6 
Test Method 
Method of receptor binding assay. After punching the heads of guinea pigs, 
the blood was drained and the heart ventricle immediately removed. The 
ventricles were placed in 10 vols. of ice-cold buffer (250 mM sucrose, 1 
mM Magnesium Chloride, 50 mM Tris-HCl, pH 7.5) and all subsequent 
procedures were carried out at 4.degree. C. The ventricles were 
homogenized using a homogenizer 3-4 times. The period of each 
homogenization took 15 seconds. The initial homogenate was filtered, and 
the filtrate was centrifuged at 700 g for 12 minutes. The supernatant was 
further centrifuged at 10,000 g and the final pellet was resuspended in a 
small volume of 75 mM Tris buffer containing 25 mM Magnesium Chloride. The 
membrane protein content was determined by a protein assay dye. The 
receptor binding assay was initiated by incubating 100 .mu.l of, a 
membrane protein solution with 50 .mu.l of .sup.3 
H!-dihydroalprenolol(.sup.3 H-DHA) and the competing drugs. Additional 
buffer was added to increase the reaction volume up to 250 .mu.l. The 
binding reaction was activated at 25.degree. C. water by constantly 
shaking for 60 minutes. The binding reaction was stopped by diluting it 
with 1 ml of ice-cold assay buffer. The solution was then filtered on a 
Whatman GF/C glass fiber filter paper under vacuum suction. The filters 
were washed 3 times with ice-cold assay buffer and dried at 60.+-.C. in an 
oven for 2 hrs. Then 4 ml of scintillation fluid was added and the 
radioactivity was counted on .beta.-counter. 
Test Compounds 
4-4'-(2-hydroxy-3-isopropylaminopropoxy)-3'-methoxyphenyl!-3-buten-2-one 
(DZN,4) 
4-4'-(2-hydroxy-3-tert-butylaminopropoxy)-3'-methoxyphenyl!-3-buten-2-one 
(DZBN,6) 
4-4'-(2-hydroxy-3-isopropylaminopropoxy)-3'-methoxyphonyl!-2-butanone 
(ZPN,9) 
Test Results 
As shown in FIG. 15 and Table 4, the IC.sub.50 values of DZBN6 and DZN4 are 
close to the IC.sub.50 values of the well known .beta.-adrenergic agonist 
Atenolol. The term IC.sub.50 is Inhibitory Concentration at 50%, which 
means the effective concentration of antagonist (or blocker) such as DZBN 
to inhibit (or decrease) the maximal efficacy of agonist such as 
isoproterenolol to the half of this maximal efficacy. 
TABLE 4 
______________________________________ 
EC.sub.50 (M) value osf various .beta.-adrenergic 
antagonists 
.beta.-antagonists EC.sub.50 (M) 
______________________________________ 
(-)Propranolol 1.1 .times. 10.sup.-8 
(.+-.)DZBN 3.6 .times. 10.sup.-6 
(.+-.)ZPN 1,1 .times. 10.sup.-5 
(.+-.)DZN 1,3 .times. 10.sup.-6 
(.+-.)Atenolol 3.5 .times. 10.sup.-5 
______________________________________ 
TEST 7 
Test 
Effect of addition of isoproterenolol in different concentration to the 
isolated rat right atrium pretreated with reserpine. 
Test Compounds 
4-(4'-hydroxy-3'-methoxyphenyl)-3-buten-2-one(Dehydrozingerone DZ,2) 
4-4'-(2-hydroxy-3-isopropylaminopropoxy)-3'-methoxyphenyl!-3-buten-2-one 
(DZN,4) 
Test Results 
(1) FIG. 16 shows that isoproterenolol increases the heart rate and 
contraction force. 
(2) The concentration effect curve indicated that the maximum effect was 
achieved at 3.times.10.sup.-6 M isoproterenolol (FIG. 17). 
(3) The effect of DZN was different from that of isoproterenolol; DZN 
reduced the heart rate at concentration about 10.sup.-6 M. On the other 
hand, DZ compound 2 did not affect the heart rate within 10.sup.-9 
-10.sup.-5 M. 
TEST 8 
Test 
The platelet aggregation test was conducted by the method reported by Yeh, 
H. I. et al (Thromb.Res., vol. 45,39-49, 1987), or Srivastava, K. C. et al 
(Leuk.Med., vol. 29, 79-84, 1987) as reference. 
Test Compounds 
4-(4'-hydroxy-3'-methoxyphenyl)-3-butan-2-one (zingerone,1) 
4-(4'-hydroxy-3'-methoxyphenyl)-3-buten-2-one(Dehydrozingerone DZ,2) 
4-4'-(2,3-Epoxypropoxy)-3'-methoxyphenyl!-3-buten-2-one (DZE,3) 
4-(4'-(2-hydroxy-3-isopropylaminopropoxy)-3'-methoxyphenyl!-3-buten-2-one 
(DZN, 4) 
Test Results 
A monophasic aggregation curve was observed when 10 .mu.g/ml of collagen 
was added to platelet rich plasma (PRP). Pretreatment of the platelet rich 
plasma with propranolol, Zingerone, DZ, DZE, and DZN caused a 
dose-dependent inhibition of platelet aggregation induced by collagen. As 
seen in FIG. 18, the platelet aggregation was completely inhibited by 250 
.mu.M propranolol, 100 .mu.M Zingerone, 250 .mu.M DZ-2, 100 .mu.M DZE-3, 
250 .mu.M DZN-4. 
TEST 9 
Test Method Statistical Methods 
The test was conducted according to the method described by Litchfield, J. 
L. et al (J. Pharmacol.Exp.Ther., vol. 96, 99-113). 
The compounds tested were: 
4-(4'-hydroxy-3'-methoxyphenyl)-3-buten-2-one(Dehydrozingerone DZ,2) 
(4-4'-(2-hydroxy-3-isopropylaminopropoxy)-3'-methoxyphenyl!-3buten-2-one 
(DZN,4) 
Test Results 
The LD.sub.50 's values are shown in Table 5. 
TABLE 5 
______________________________________ 
Acute toxicity of propranolol, DZH and DZ in 
mice. 
LD.sub.50 (95% C.L.).sup.a 
Compound Route (mg/kg) 
______________________________________ 
Propranolol p.o. 446.92 (303.08-659.04) 
i.p. 288.79 (200.13-416.72) 
DZN p.o. &gt;1000 
i.p. &gt;1000 
DZ p.o. &gt;1000 
i.p. &gt;1000 
______________________________________ 
a: LD.sub.50 's and 95% confidence limits were calculated by Litchfield 
and Wilcoxon method. 
The compounds of formula I and their pharmaceutically acceptable acid 
addition salts can be used as medicaments, eg. in the form of 
pharmaceutical preparations for enteral, parenteral, topical or oral 
application. The manufacture of the pharmaceutical compositions may be 
carried out in a manner which is familiar to any person skilled in the art 
by using the described compounds of formula I and their pharmaceutically 
acceptable acid addition salts, optionally in combination with other 
therapeutically valuable subcutaneous substances such as corn starch, 
starch, lactose, sodium carboxymethylcellulose, ethanol. The 
.beta.-adrenergic blocker compositions to be used orally contain about 60 
mgs. per dose of a compound according to the present invention. 
Vaninolol of formula hereinbelow 
##STR4## 
has been shown to lower the intraocular pressure (IOP) and is useful in 
the treatment of glaucoma in the form of an eye solution. 
This 
4-4'-(2-hydroxy-3-tert-butylaminopropoxy)-3'-methoxyphenyl!-3-buten-2-one 
(DZBN) compound 6 is watersoluble and exhibits lower pungency so that it 
is preferred for parenteral and eye solution. The preferred individual 
dosage is 0.25 %, 0.5% eye solution and may be administered up to twice 
daily.