WO2003053439A1 - Use of benzofuroxan derivatives as antiplatelet agents - Google Patents
Use of benzofuroxan derivatives as antiplatelet agents Download PDFInfo
- Publication number
- WO2003053439A1 WO2003053439A1 PCT/IB2002/005353 IB0205353W WO03053439A1 WO 2003053439 A1 WO2003053439 A1 WO 2003053439A1 IB 0205353 W IB0205353 W IB 0205353W WO 03053439 A1 WO03053439 A1 WO 03053439A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- benzofuroxan
- carbonyl
- lower alkyl
- hydrogen
- pharmaceutically acceptable
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Ceased
Links
- QKGBRANQIWBMED-UHFFFAOYSA-N COCCN(CCC1)C1=O Chemical compound COCCN(CCC1)C1=O QKGBRANQIWBMED-UHFFFAOYSA-N 0.000 description 1
- JAEQOSKUYPMJAT-UHFFFAOYSA-N COCCN1CCOCC1 Chemical compound COCCN1CCOCC1 JAEQOSKUYPMJAT-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/4245—Oxadiazoles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
- A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
Definitions
- the present invention relates to the use of benzofuroxan compounds of general formula (I) as defined herein, their derivatives, their analogues, their tautomeric forms, their stereoisomers, their polymorphs, their hydrates, their pharmaceutically acceptable salts, their pharmaceutically acceptable solvates and pharmaceutically acceptable compositions containing them, as antiplatelet agents.
- the compounds are found to be potent inhibitors of platelet aggregation in the treatment of cardiovascular diseases.
- Antiplatelet agents' are those compounds, which either prevent adhesion, activation or aggregation of platelets.
- Antiplatelet drug therapy can be used in treatment or prevention of myocardial infarction, unstable angina, venous thromboembolism, coronary artery bypass graft (CABG), percutaneous transluminal angioplasty (PTCA), cerebrovascular disease like stroke, atrial fibrillation, prosthetic heart valve, valvular heart disease, acute peripheral occlusive disease, primary and secondary prevention of arterial thromboembolism, clot prevention in extra corporeal devices as the aetiology in all the above disease state is platelet activation and subsequent thrombus formation.
- CABG coronary artery bypass graft
- PTCA percutaneous transluminal angioplasty
- cerebrovascular disease like stroke atrial fibrillation
- prosthetic heart valve valvular heart disease
- acute peripheral occlusive disease primary and secondary prevention of arterial thromboembolism
- Pharmacologic therapy with antiplatelet agents such as aspirin provide clinical benefit in acute ischemic syndromes.
- Prostaglandin Ei has been reported as a powerful endothelium derived inhibitor of platelet aggregation.
- PGEi Prostaglandin Ei
- PDE PGEi phosphodiesterase III inhibitors
- inotropic agents, amrinone and milrinone increase cAMP by inhibition of PDE III and inhibit platelet aggregation in vitro.
- the other anti-platelet aggregatory drug developed included cyclo-oxygenase and thromboxane synthase inhibitors, prostaglandin derivatives, thromboxane A 2 receptor antagonists, and GPIIb/IIIa antagonists, which apart from PDE inhibitors have been widely used in clinical situation (Schafer, 1996, Anti platelet therapy, Am. J. Med, 101, 199-209).
- the benzofuroxan compounds of the instant invention which can exhibit dual activity viz. antianginal as well as antiplatelet activity make them most suitable for cardiovascular applications. .
- the invention provides a group of benzofuroxan compounds, their derivatives, analogues, tautomers, stereoisomers, polymorphs, their pharmaceutically acceptable salts or solvates as antiplatelet agents.
- the invention also provides a method of treatment or prevention of disease conditions caused by formation of platelets in human and non-human mammals by administration of said benzofuroxan compounds, their derivatives, analogues, tautomers, stereoisomers, polymorphs, their pharmaceutically acceptable salts or solvates.
- the invention further provides a pharmaceutical composition
- a pharmaceutical composition comprising the said benzofuroxan compounds, their derivatives, analogues, tautomers, stereoisomers, polymorphs, their pharmaceutically acceptable salts or solvates in admixture with a pharmaceutically acceptable diluent, carrier or excepient effective as an antiplatelet agent.
- the invention provides for use of said benzofuroxan compounds, their derivatives, analogues, tautomers, stereoisomers, polymorphs, their pharmaceutically acceptable salts or solvates in the manufacture of a medicament useful as an antiplatelet agent.
- the present invention relates to the new use of benzofuroxan compounds of formula (I),
- R is -C(O)-R ⁇ , halogen, acetoxy, -X-R 2 or -C(O) NR 3 R 4 , or C(O)Cl R ⁇ is -O- (CH 2 )
- n 1 to 6
- X is oxygen, sulfur, -C (O)-, or -C (O) O-
- R 2 is hydrogen, straight chain or branched lower alkyl (C ⁇ -C 8 ), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen wherein substitution on nitrogen is with hydrogen or lower alkyl;
- R 3 and are independently hydrogen, straight chain or branched lower alkyl (Ci-
- R 3 and R 4 are linked together with or without a heteroatom selected from the group consisting of oxygen and nitrogen wherein substitution on nitrogen is with hydrogen or lower alkyl;
- Y is -NHC (O)- or oxygen
- R 5 is lower alkyl (C ⁇ -C 8 ), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen wherein substitution on nitrogen is with hydrogen or lower alkyl;
- R 6 is hydrogen, nitro, lower alkyl or -C (O)-R ;
- R 7 is hydrogen, lower alkyl or aryl
- R 8 is hydrogen ; or "Ri" is selected from the group consisting of
- the substituent R 8 in the said compounds of formula is a halogen selected from the group F, CI, Br and I.
- the compounds of the present invention can be prepared according to the methods disclosed in US6,248,895 (Sankaranarayanan, 2001) and US 6,232,331 (Sankaranarayanan, 2001).
- PRP platelet rich plasma
- PPP platelet poor plasma
- PRP The anticoagulated blood sample was centrifuged at 250g for 10 minutes and the supernatant (PRP) removed carefully into a plastic tube, and retained at 18 - 26°C for the duration of the test.
- Preparation of PPP The sediment (cells) from the above contrifugation is again re-centrifuged at 1500g for 10 minutes and the supernatant (PPP) is collected and kept at 18 -26°C for the duration of the test.
- the compounds 31 and 10 were tested for their antiplatelet activity using E 840 aggregometer.
- the inhibition of platelet aggregation by the test compounds was studied according to the modified method of Nishikawa M, et al.1981 using three different platelet aggregation reagents viz. (a) Adenosine -5' - diphosphate (ADP) : 10 ⁇ M, (b) Collagen: 0.2 mg/ml and (c) Epinephrine: 10 ⁇ M. (All agonists are obtained from Sigma-Aldrich - 'platelet aggregation reagents'). Assay Procedure:
- test drug solution final cone, of 46 ⁇ M or 57 ⁇ M OR saline (for control) was added to the PRP cuvettes and incubated for 1 min. 5.
- the aggregant was brought to room temperature (18 - 26°C) and swirled to mix.
- the aggregation response was measured for 5 min or till the max response is reached. 8.
- the extent of aggregation was expressed by the maximum change of light transmission expressed as a percentage, taking the difference between light transmission for PRP and PPP as 100.
- test compound 31 showed good inhibitory activity of 92%, 88% and 84% respectively at concentration (46 ⁇ M) with the three aggregants ADP, epinephrine and collagen.
- the test compound 10 showed about 25% inhibition with epinephrine as aggregant, while at higher concentration it showed marked increase in inhibitory activity.
- Platelet aggregation induced by an aggregating agent has also measured using a whole blood aggregometer which monitors the change in the electrical-impedance. It is the first simple method by which aggregation of human platelets can be measured in their natural environment (Zwierzina and Kunz, 1985).
- the tests were performed using venous blood sample of normal healthy male volunteers, (non smokers, non alcoholics) who were not on any medication known to affect platelet aggregation for at least 2 weeks prior to the study.
- the anticoagulant used was 3.8% tri sodium citrate, in the ratio of 9:1.
- the samples were kept at room temperature until tested and the experiments were completed within 2-3 hrs of blood- collection (Zwierzina and Kunz, 1985; Mutsuhito kikura et al., 2000).
- the test compounds and the reference compound Nicorandil were tested for their antiplatelet activity using chronolog whole blood aggregometer.
- the inhibition of platelet aggregation by the test compounds was studied using the aggregate collagen (chrono-log corp, chrono-par 385 collagen reagent).
- benzofuroxan compounds useful for the invention may be prepared by the process as disclosed in US 6,248,895 and US 6,232,331.
- This compound can be prepared by the same method as described in Example 3 infra for Step 2 of Compound No. 37 using 5(6)-azidocarbonyl benzofuroxan (0.50 gm, 0.0024 mole) and N-(2-hydroxyethyl-2-pyrrolidinone)(0.32 gm, 0.0024 mole). Yield: 0.60 gm. m. p.. : 164-165°C
- Step 1 Preparation of 5(6)-azidocarbonyl benzofuroxan
- Step 2 Preparation of 5(6)- ⁇ 2-(3-pyridylcarbonylamino)-ethyl ⁇ oxy carbonyl amino benzofuroxan
- a solution of azido comp. (0.41 g, 0.002 mole, as prepared in step 1) in toluene (20 ml) was refluxed for 2 hrs.
- Added N-(2-hydroxyethyl)-nicotinamide (0.33 gm, 0.002 mole) to the above solution and refluxed for another 2 hr., cooled to the room temp, precipitated product filtered and dried.
- Step 2 Preparation of 4-azido-3-bromo-5-nitro benzoic acid.
- Step 4 Preparation of 5(6)-[4-bromo]-isopropyloxycarbonyl benzofijroxan
- Compound of Formula (I) is dissolved in propylene glycol and warmed to about 40 °C to obtain a clear solution.
- the volume of the above solution is made up with 0.9% sodium chloride solution.
- a tablet can be prepared with the compounds of the invention and following ingredients together in the proportions by weight specified below:
- Compound of Formula (I) is mixed with Starch and Microcrystalline Cellulose and granulated with Starch mucilage (prepared by gelatinizing starch in water). The wet mass is dried at 50°C and the dried granules are sized through ASTM #20 sieve. The sized granules are blended with Sodium Starch Glycolate and Magnesium Stearate and the blend was compressed into tablets using 7 mm punches to contain 25 mg of
- Compound of Formula (I) Other tablets may be compressed to contain 50, 75,100,
- the pharmaceutical composition can be in the range of 0.5 % to 90 % by weight of the total composition.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to the use of benzofuroxan compounds, their derivatives, their analogues, their tautomeric forms, their stereoisomers, their polymorphs, their hydrates, their pharmaceutically acceptable salts, their pharmaceutically acceptable solvates and pharmaceutically acceptable compositions containing them, as anti-platelet agents for prevention or treatment of disease conditions resulting from platelet aggregation in a human or non-human mammal and also method and composition for such use.
Description
USE OF BENZOFUROXAN DERIVATIVES AS ANTIPLATELET AGENTS
This application claims benefit of U.S. Provisional Application No.06/341,283 filed December 20, 2001, the disclosure of which is incorporated herein by reference in its entirety.
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to the use of benzofuroxan compounds of general formula (I) as defined herein, their derivatives, their analogues, their tautomeric forms, their stereoisomers, their polymorphs, their hydrates, their pharmaceutically acceptable salts, their pharmaceutically acceptable solvates and pharmaceutically acceptable compositions containing them, as antiplatelet agents. The compounds are found to be potent inhibitors of platelet aggregation in the treatment of cardiovascular diseases.
2. Description of the Related Art
The past two decades have witnessed a widespread understanding of the important role that platelets play in the clinical manifestations of cardiovascular diseases, such as unstable angina which cause sudden death. Rather than a single factor being responsible, the interplay of platelet aggregation, thrombosis and coronary artery vasomotion on the substrate of atherosclerotic disease often leads to cardiac failure. The central role played by platelets has led to successful use of 'anti- platelet therapy' in such patients.
'Antiplatelet agents' are those compounds, which either prevent adhesion, activation or aggregation of platelets. Antiplatelet drug therapy can be used in treatment or prevention of myocardial infarction, unstable angina, venous
thromboembolism, coronary artery bypass graft (CABG), percutaneous transluminal angioplasty (PTCA), cerebrovascular disease like stroke, atrial fibrillation, prosthetic heart valve, valvular heart disease, acute peripheral occlusive disease, primary and secondary prevention of arterial thromboembolism, clot prevention in extra corporeal devices as the aetiology in all the above disease state is platelet activation and subsequent thrombus formation. The complications associated with the formation of platelets in these conditions results not only from their central role in initiation of a thrombus, but also from their secretion of potent vasoactive substances on aggregation, such as Thromboxane A2. Hence, a need exists for new and improved treatment options for individuals, suffering from vascular diseases.
Pharmacologic therapy with antiplatelet agents such as aspirin provide clinical benefit in acute ischemic syndromes.
The less widely recognised organic nitrates have been documented to manifest antiplatelet effects in, in vivo and in vitro studies (Jaraki et al 1994).
Prostaglandin Ei (PGEi) has been reported as a powerful endothelium derived inhibitor of platelet aggregation. In vitro tests have shown that PGEi inhibit platelet aggregation by increasing the level of cyclic adenosine monophosphate (cAMP). In contrast to PGEi phosphodiesterase (PDE) III inhibitors (inotropic agents, amrinone and milrinone) increase cAMP by inhibition of PDE III and inhibit platelet aggregation in vitro.
The other anti-platelet aggregatory drug developed included cyclo-oxygenase and thromboxane synthase inhibitors, prostaglandin derivatives, thromboxane A2 receptor antagonists, and GPIIb/IIIa antagonists, which apart from PDE inhibitors have been widely used in clinical situation (Schafer, 1996, Anti platelet therapy, Am. J. Med, 101, 199-209).
However, none of them discloses the potential of benzofuroxn or its derivatives as antiplatelet agent. The inventor of the present invention who earlier found that a group of benzofuroxan derivatives exhibit antianginal activity and are
tolerant resistant unlike organic nitrates, has now found that the same compounds are also potential antiplatelet agent.
PB Ghosh et al. (Journal of Medicinal Chemistry, 1968) disclosed the method of synthesis of various benzo-2,l,3-oxadiazoles (benzofurazans) and their N-oxides (benzofuroxans) and their potential as antileukemic and immuno-suppressive drugs in vitro.
PB Ghosh et al. (Journal of Medicinal Chemistry, 1972) tested 4-nitro benzofurazans and 4-nitrobenzofuroxans bearing electron withdrawing substituents in the 5 and 6 position (relative to NO2) as potential antileukemic and immuno suppressive drugs in vitro.
PB Ghosh et al (Journal of Medicinal Chemistry, 1974) tested benzofuroxan and its derivatives for their vasodilation activities and found furazanobenzofuroxan, furazanobenzothiadiazole and their N-oxides as potent vasodilators.
The benzofuroxan compounds of the instant invention have been earlier disclosed in US 6,248,895 (Sankaranarayanan,2001) and US 6,232,331 (Sankaranarayanan, 2001) for their antianginal activity.
The benzofuroxan compounds of the instant invention which can exhibit dual activity viz. antianginal as well as antiplatelet activity make them most suitable for cardiovascular applications. .
SUMMARY OF THE INVENTION
The invention provides a group of benzofuroxan compounds, their derivatives, analogues, tautomers, stereoisomers, polymorphs, their pharmaceutically acceptable salts or solvates as antiplatelet agents.
The invention also provides a method of treatment or prevention of disease conditions caused by formation of platelets in human and non-human mammals by administration of said benzofuroxan compounds, their derivatives, analogues,
tautomers, stereoisomers, polymorphs, their pharmaceutically acceptable salts or solvates.
The invention further provides a pharmaceutical composition comprising the said benzofuroxan compounds, their derivatives, analogues, tautomers, stereoisomers, polymorphs, their pharmaceutically acceptable salts or solvates in admixture with a pharmaceutically acceptable diluent, carrier or excepient effective as an antiplatelet agent.
Additionally, the invention provides for use of said benzofuroxan compounds, their derivatives, analogues, tautomers, stereoisomers, polymorphs, their pharmaceutically acceptable salts or solvates in the manufacture of a medicament useful as an antiplatelet agent.
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to the new use of benzofuroxan compounds of formula (I),
their derivatives, their analogues, their tautomeric forms, their stereoisomers, their polymorphs, their hydrates, their pharmaceutically acceptable salts, their pharmaceutically acceptable solvates and pharmaceutically acceptable compositions containing them as antiplatelet agents, wherein, R is -C(O)-Rι, halogen, acetoxy, -X-R2 or -C(O) NR3R4, or C(O)Cl Rι is -O- (CH2)„-Y-R5, n = 1 to 6 X is oxygen, sulfur, -C (O)-, or -C (O) O-;
R2 is hydrogen, straight chain or branched lower alkyl (Cι-C8), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen wherein substitution on nitrogen is with hydrogen or lower alkyl;
R3 and are independently hydrogen, straight chain or branched lower alkyl (Ci-
C8) or R3 and R4 are linked together with or without a heteroatom selected from the group consisting of oxygen and nitrogen wherein substitution on nitrogen is with hydrogen or lower alkyl;
Y is -NHC (O)- or oxygen;
R5 is lower alkyl (Cι-C8), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen wherein substitution on nitrogen is with hydrogen or lower alkyl;
R6 is hydrogen, nitro, lower alkyl or -C (O)-R ;
R7 is hydrogen, lower alkyl or aryl;
R8 is hydrogen ; or "Ri" is selected from the group consisting of
In another embodiment, the substituent R8 in the said compounds of formula is a halogen selected from the group F, CI, Br and I.
The following compounds suggested are by way of example alone of the representative compounds of the general formula (I) as defined above and in no way restrict the invention:
5 (6)-(2 -nicotinamide ethyloxycarbonyl) benzofuroxan hydrochloride (compound number 1),
5(6)-(2-isonicotinamide ethyloxy carbonyl) benzofuroxan hydrochloride (compound number 2), 5(6)-(2-pyrolidinone ethyloxy carbonyl) benzofuroxan (compound number 3), 5(6)-(2-hydroxy propyloxy carbonyl) benzofuroxan (compound number 4), 5(6)-(2-morpholino ethyloxy carbonyl) benzofuroxan (compound number 5), 5(6)-(2-methyloxy ethyloxy carbonyl) benzofuroxan (compound number 6), 5(6)-(2,3-dihydroxy propyloxy carbonyl) benzofuroxan (compound number 7), 5(6)-(2-ethoxy ethyloxy carbonyl) benzofuroxan (compound number 8),
5(6)-(3-pyridine methoxy carbonyl) benzofuroxan hydrochloride (compound number
9),
5(6)-(±)-2,2-dimethyl-l , 3-dioxalane-4-methyloxycarbonyl) benzofuroxan (compound number 10), 5(6)-(isosorbide mononitrateoxycarbonyl) benzofuroxan (compound number 11), 5(6)-chloro benzofuroxan (compound number 12),
5(6)-n-propylmercapto benzofuroxan (compound number 13),
5(6)-carboxy benzofuroxan (compound number 14),
5(6)-methylmercapto benzofuroxan (compound number 15),
5(6)-acetoxy benzofuroxan (compound number 16),
5(6)-chlorocarbonyl benzofuroxan (compound number 17),
5(6)-formyl benzofuroxan (compound number 18),
5 (6)-methoxy carbonyl benzofuroxan (compound number 19),
5(6)-hydroxy benzofuroxan hydrochloride (compound number 20),
5(6)-amino carbonyl benzofuroxan (compound number 21),
5(6)-ethoxy carbonyl benzofuroxan (compound number 22),
5(6)-n-propoxycarbonyl benzofuroxan (compound number 23),
5(6)-isopropoxycarbonyl benzofuroxan (compound number 24),
5(6)-tertiary butoxy carbonyl benzofuroxan (compound number 25),
5(6)-dimethylamino carbonyl benzofuroxan (compound number 26),
5(6)-morpholino carbonyl benzofuroxan (compound number 27),
5(6)-((4-methyl) piperazine-1-yl) carbonyl benzofuroxan (compound number 28),
5(6)-isopropyl amino carbonyl benzofuroxan (compound number 29),
5(6)-tertiary butyl amino carbonyl benzofuroxan (compound number 30),
5(6)-(2-nicotinamide ethyloxycarbonyl) benzofuroxan mesylate (compound number
31),
5(6)-(3-pyridine methoxy carbonyl) benzofuroxan tartarate(compound number 32),
5(6)-(S)(2-hydroxy)propyloxycarbonyl-benzofuroxan (compound number 33)
5(6)[3-(3-pyridyl)propyloxycarbonyl benzofuroxan] (compound number 34),
5(6)-{2-(pyrrolidin-2-one-l-yl)}ethyloxycarbonylamino-benzofuroxan (compound number 35),
5(6)-(l-pyrrolidinyl)carbonyl benzofuroxan (compound number 36),
5 (6)- {2-(2-pyridylcarbonylamino)ethyl} oxycarbonyl benzofuroxan (compound number 37),
5(6)-{2-(3-pyridyl carbonylamino)ethyl}oxycarbonyl amino benzofuroxan (compound number 38),
5(6)-nitro- benzofuroxan -(compound number 39),
benzofuroxan - (compound number 40), and
5(6) (4-bromo)-isopropyloxycarbonyl benzofuroxan (compound number 41).
Table 1
: Compound 33 is (S) isomer
(I) (m)
Inhibition of platelet aggregation (in vitro studies)
For study of inhibition of platelet aggregation by the compounds of the invention following methods were used :
Method - 1
Venous blood samples from healthy male human volunteers who were not on any medication for atleast two weeks were used for preparation of platelet rich plasma (PRP) and platelet poor plasma (PPP). 4.5 ml of venous blood were collected from human volunteers into tubes containing 0.5 ml of 3.8% sodium citrate, care was taken to avoid stasis and contamination with tissue fluids and PRP and PPP were prepared immediately and used within 4 hrs.
Preparation of PRP: The anticoagulated blood sample was centrifuged at 250g for 10 minutes and the supernatant (PRP) removed carefully into a plastic tube, and retained at 18 - 26°C for the duration of the test.
Preparation of PPP: The sediment (cells) from the above contrifugation is again re-centrifuged at 1500g for 10 minutes and the supernatant (PPP) is collected and kept at 18 -26°C for the duration of the test.
The compounds 31 and 10 were tested for their antiplatelet activity using E 840 aggregometer. The inhibition of platelet aggregation by the test compounds was studied according to the modified method of Nishikawa M, et al.1981 using three different platelet aggregation reagents viz. (a) Adenosine -5' - diphosphate (ADP) : 10 μM, (b) Collagen: 0.2 mg/ml and (c) Epinephrine: 10 μM. (All agonists are obtained from Sigma-Aldrich - 'platelet aggregation reagents').
Assay Procedure:
1. 450 μl of PRP was added to an aggregation cuvette containing stirring bar and incubated at 37°C for 3 minutes. 2. 500 μl of PPP was added to an aggregation cuvette without stirring bar.
3. PRP and PPP cuvettes were placed in corresponding instrument sampling wells and the baseline was set.
4. 50 μl of the test drug solution (final cone, of 46 μM or 57 μM) OR saline (for control) was added to the PRP cuvettes and incubated for 1 min. 5. The aggregant was brought to room temperature (18 - 26°C) and swirled to mix.
6. 50 μl of epinephrine/ collagen or 25 μl of ADP (aggregants) was added to the PRP cuvette.
7. The aggregation response was measured for 5 min or till the max response is reached. 8. The extent of aggregation was expressed by the maximum change of light transmission expressed as a percentage, taking the difference between light transmission for PRP and PPP as 100.
Results:
The test compound 31 showed good inhibitory activity of 92%, 88% and 84% respectively at concentration (46 μM) with the three aggregants ADP, epinephrine and collagen. The test compound 10 showed about 25% inhibition with epinephrine as aggregant, while at higher concentration it showed marked increase in inhibitory activity.
A summary of the % inhibition of platelet aggregation is shown in the Table - 2.
Table - 2
*Values are mean of 3-4 observations.
The results show that compounds of the invention are potent inhibitors of platelet aggregation induced by a variety of aggregatory agents and hence would be useful for anti- platelet therapy and in treatment of cardiovascular diseases. Method - 2
Methodology:
Platelet aggregation induced by an aggregating agent has also measured using a whole blood aggregometer which monitors the change in the electrical-impedance. It is the first simple method by which aggregation of human platelets can be measured in their natural environment (Zwierzina and Kunz, 1985).
Principle:
In 1962, Born described the aggregation of platelets (ref. Sudo Toshiki et.al. British Journal of Pharmacology, 1396 - 1404, 2001) by ADP and modified a colorimeter to monitor this aggregation in Platelet Rich Plasma (PRP). A very small AC voltage in the mV range is passed between two electrodes, which are mainly made-up of two precious metal wires. During initial contact with the blood, the electrodes become coated with a monolayer of platelets resulting in a stable impedance value. This stable baseline of impedance is assigned a value of zero (ohms) of resistance. When an aggregating agent is added, platelets aggregate on the monolayer and increasing the impedance. This increase in impedance is measured
and quantified in ohms. The increase in impedance is directly proportional to the mass of the platelet aggregate.
Procedure:
The tests were performed using venous blood sample of normal healthy male volunteers, (non smokers, non alcoholics) who were not on any medication known to affect platelet aggregation for at least 2 weeks prior to the study. The anticoagulant used was 3.8% tri sodium citrate, in the ratio of 9:1. The samples were kept at room temperature until tested and the experiments were completed within 2-3 hrs of blood- collection (Zwierzina and Kunz, 1985; Mutsuhito kikura et al., 2000). The test compounds and the reference compound Nicorandil were tested for their antiplatelet activity using chronolog whole blood aggregometer.
The inhibition of platelet aggregation by the test compounds was studied using the aggregate collagen (chrono-log corp, chrono-par 385 collagen reagent).
900μl of citrated blood was taken in a fresh siliconized sampling cuvette along with siliconized stir bar and was preheated for approximately 10- 15 -min at 37°C in the incubation well of the aggregometer. Then lOOμl of prewarmed (37°C) vehicle/reference/test drug was added and allowed to incubate for 10 min. After ensuring that the base line was stable the aggregate, lOμl of collagen (final cone, of
2μg/ml) was added and response recorded for 8-min. After that the impedance was computed in ohms.
Initially maximum soluble concentration of the test compounds / nicorandil in dimethyl sulfoxide (DMSO) was prepared and screened for the antiplatelet activity by using the above mentioned procedure. The anti-platelet activity of these compounds were repeated from blood samples collected from various volunteers. The percent inhibition was calculated using the formula below and the mean percent inhibition was calculated.
(Collagen induced aggregation (Collagen induced aggregation in in presence of vehicle) presence of reference/test comp)
% Inhibition = x 100
Collagen induced aggregation in presence of vehicle
A summary of the % inhibition of platelet aggregation is shown in the Table - 3 below. Parallel tests with same set of compounds were also conducted at same concentration under Method 1 and the results also shown in this table.
Table - 3
Conclusion of the test results:
Inhibitory effects of drugs on platelet aggregation has been thus tested either in PRP (platelet rich plasma) or whole blood. Further, the compounds exhibit sufficient inhibitory activity as determined by both the methods.
The results show that the compounds of the invention are potent inhibitors of platelet aggregation induced by collagen. A comparison with known inhibitors like Nicorandil showed that some of these molecules are much more potent even at much lower concentration and hence would be useful for anti-platelet therapy. Thus, these compounds may be administered as a therapeutic agent, to an individual, to promote platelet inhibition, and to treat or prevent cardiovascular disorders using the advantage of the possible dual action of these compounds as platelet inhibitor and anti-anginal.
In the following section preferred embodiments are described by way of examples to illustrate the process of carrying out this invention. However, this is not intended in any way to limit the scope of the present invention.
As already indicated the benzofuroxan compounds useful for the invention may be prepared by the process as disclosed in US 6,248,895 and US 6,232,331.
The method of preparation of some specific compounds of the invention are given below :-
EXAMPLE -1
Preparation of 5(6)-[(S)(2-hydroxy) propyloxycarbonyl-benzofuroxan] (compound number 33)
To a stirred solution of benzofuroxan carboxylic acid (1.2 g, 0.0066 mole) in dichloromethane (30 ml) was added 5-(+)-propane-l,2-diol (0.50g, 0.0066 mole), dimethylaminopyridine (0.13 g) and dicyclohexylcarbodiimide (DCC, 2.0g, 0.0097 mole) sequentially in ice-cooled condition. Reaction mixture stirred at room temperature for 3 hrs., filtered, evaporated in vacuo to give a sticky crude material, which was purified by column chromatography (ethylacetate: hexanes 1 :5) to give pure product as white solid. Yield: 0.62 gm. m. p.: 82-83°C
1HNMR(CDC13, 400MHz): δ 7.64-8.23 (m, 3H), 5.28-5.34 (m, 1H), 4.21-4.26
(m,2H), 1.33-1.34 (d, 3H, J=6HZ);
Mass: 237 (M+- 1), 179, 163 and 75;
IR (KBr, cm"1): 3345, 3270, 1721, 1613 and 1590;
EXAMPLE - 2
Preparation of 5(6)-{2-(pyrrolidin-2-one-l-yl)}ethyIoxycarbonyIamino- benzofuroxan (compound number 35)
This compound can be prepared by the same method as described in Example 3 infra for Step 2 of Compound No. 37 using 5(6)-azidocarbonyl benzofuroxan (0.50 gm, 0.0024 mole) and N-(2-hydroxyethyl-2-pyrrolidinone)(0.32 gm, 0.0024 mole). Yield: 0.60 gm. m. p.. : 164-165°C
JHNMR (CDC13): δ 8.25 (GS, 1H), 7.16-7.86 (m, 3H), 4.34 - 4.36 (t, J=5.2HZ, 2H), 3.64-3.66 (t, J=5.2 Hz, 2H), 3.52-3.55 (t, J= 7HZ, 2H), 2.41-2.45 (t, J=8.0HZ, 2H), 2.04-2.14 (m, 2H);
Mass: 307 (M++l) and 329 (M++ Na); IR (KBr, cm"1): 3226, 3024, 1751, 1675 and 1573;
EXAMPLE -3
Preparation of [5(6)-{2-(2-pyridylcarbonylamino)ethyl}oxycarbonyl benzofuroxan] (compound number 37)
Step 1 : Preparation of 5(6)-azidocarbonyl benzofuroxan
To a solution of 5(6)-benzofuroxan acid chloride (2.7gm, 0.13 mole) in acetone (30 ml) was added sodium azide (1.5 gm, 0.023 mole) in water (8ml) and stirred at room temperature for 1 hr. Acetone evaporated in vacuo, ice was added to it, separated solid filtered, dried under suction to get 5(6)-azidocarbonyl benzofuroxan. Yield: 2.4 gm.
Step 2: Preparation of 5(6)-{2-(3-pyridylcarbonylamino)-ethyl}oxy carbonyl amino benzofuroxan
A solution of azido comp. (0.41 g, 0.002 mole, as prepared in step 1) in toluene (20 ml) was refluxed for 2 hrs. Added N-(2-hydroxyethyl)-nicotinamide (0.33 gm, 0.002 mole) to the above solution and refluxed for another 2 hr., cooled to the room temp, precipitated product filtered and dried.
Yield: 0.58 gm. m. p.: 118-119° C
1HNMR (DMSO d6): δ 9.18-9.23 (t, J=6HZ, IH), 8.64-8.65 (d, J=4.8 Hz, IH), 7.97- 8.06 (m, 2H), 7.64-7.95 (m, 2H), 7.58-7.62 (m, IH); 4.43-4.45 (t, J=5.2 Hz, 2H), 3.70-3.75 (m, 2H); Mass: 351 (M+ + Na); IR (KBr, cm"1): 3374, 1720, 1664, 1617, 1594 and 1529
EXAMPLE - 4
Preparation of 5(6) [(4-bromo)-isopropyloxy carbonyl benzofuroxan] (compound number 41),
Step 1 : Preparation of 4-amino-3-bromo-5-nitro benzoic acid
To a stirred solution of 4-amino-3-nitro benzoic acid (15 g, 0.082 mole) in acetic acid (150 ml) was added 0.50 gm. of aluminum chloride. After 30 min. 15 gm of liquid bromine (0.093 mole) in acetic acid (100 ml) was added drop wise under cooling to the above solution and stirred at room temperature for 1 hr. Reaction mixture was poured into water; solid was separated out by filtration and dried to get 4-amino-3-bromo-5-nitro benzoic acid. Yield: 18 gm
Step 2: Preparation of 4-azido-3-bromo-5-nitro benzoic acid.
To the acetic acid (100 ml) solution of 4-amino-3-bromo-5-nitro benzoic acid (12 gm, 0.046 mole) as prepared in Step 1, was added cone. H2SO4 (100 ml) and cooled to 0°C. A slurry of sodium nitrite (9.5 gm) in cone. H SO (50 ml) was added slowly while keeping the temperature below 10 °C and the reaction mixture was
allowed to stir at 0 °C for 1 hr. Reaction mixture was poured on ice, the solid was filtered and to the clear filtrate, a solution of sodium azide (9.5 gm) in water was added and stirred at 0-5°C for 1 hr. The precipitated solid was filtered and dried to get 4-azido-3-bromo-5-nitro benzoic acid. Yield: 8 gm
Step 3: Preparation of 4-bromo-benzofuroxan 5(6) carboxylic acid
Solution of 4-azido-3-bromo-5-nitro benzoic acid (5.2 gm, 0.018 mole) (prepared in Step - 2) in toluene (150 ml) was refluxed for 4 hr. The reaction mixture was cooled to room temperature to get the yellow colored precipitate. The solid obtained was filtered and dried under suction to get 4-bromo-benzofuroxan 5(6) carboxylic acid. Yield: 4.2 gm
Step 4: Preparation of 5(6)-[4-bromo]-isopropyloxycarbonyl benzofijroxan
To a stirred solution of bromo benzofuroxan acid (1 gm, 0.0038 mole) in dichloromethane (30 ml) was added isopropyl alcohol (0.3g, 0.005 mole), dimethylaminopyridme (0.13 g) and dicyclohexylcarbodumide (DCC, 0.94 gm, 0.0045 mole), sequentially in ice-cooled condition.
Reaction mixture stirred at room temperature for 3 hrs., filtered, evaporated in vacuo to give a sticky crude material, which was purified by column chromatography (ethylacetate: hexanes 1 :5) to give pure product as white solid.
This product was prepared by, using and in presence of dicyclohexylcarbodumide (DCC) Yield: 0.75 gm. m. p..: 112-113 °C JHNMR (CDC13): δ 8.19 (S, IH), 8.14 (S, IH), 5.26-5.32 (m, IH), 1.41-1.42 (d,
J=6.4 Hz, 6H);
Mass: 301 (M++ 1);
IR (KBr, Cm"1): 3062, 1707, 1603 and 1588;
EXAMPLE - 5
As given below, a parenteral formulation of the following composition can be prepared with the compounds of the invention :
Compound of Formula (I) is dissolved in propylene glycol and warmed to about 40 °C to obtain a clear solution. The volume of the above solution is made up with 0.9% sodium chloride solution.
EXAMPLE - 6
A tablet can be prepared with the compounds of the invention and following ingredients together in the proportions by weight specified below:
Compound of Formula (I) is mixed with Starch and Microcrystalline Cellulose and granulated with Starch mucilage (prepared by gelatinizing starch in water). The wet mass is dried at 50°C and the dried granules are sized through ASTM #20 sieve. The
sized granules are blended with Sodium Starch Glycolate and Magnesium Stearate and the blend was compressed into tablets using 7 mm punches to contain 25 mg of
Compound of Formula (I). Other tablets may be compressed to contain 50, 75,100,
150 and 200 mg of Compound of Formula (I).
The pharmaceutical composition can be in the range of 0.5 % to 90 % by weight of the total composition.
While the present invention has been described in terms of its specific embodiments, certain modifications and equivalents will be apparent to these skilled in the art and intended to be included within the scope of the present invention.
Claims
1. Use of a benzofuroxan compound represented by the formula (I)
its derivatives, analogues, tautomeric forms, stereoisomers, polymorphs, hydrates, pharmaceutically acceptable salts, pharmaceutically acceptable solvates or pharmaceutically acceptable compositions containing them, for treatment of diseases resulting from platelet aggregation wherein:
R is -C (O)-Rι, halogen, acetoxy, -X-R2 or -C (O) NR3R4, or C (O) CI 
n = 1 to 6, X is oxygen, sulfur, -C(O)- or -C(O)O-
R2 is hydrogen, straight chain or branched lower alkyl (Ci - C8), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen wherein substitution on nitrogen is with hydrogen or lower alkyl; R3and R4 are independently hydrogen, straight chain or branched lower alkyl (Ci -
C8) or R and R4 are linked together with or without a heteroatom selected from the group consisting of oxygen and nitrogen wherein substitution on nitrogen is with hydrogen or lower alkyl;
Y is -NHC (O)- or oxygen; R5 is lower alkyl (Cι-C8), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen, wherein substitution is with lower alkyl;
R6 is hydrogen, nitro, lower alkyl or -C (O)-R7; R is hydrogen, lower alkyl or aryl; R8 is hydrogen or "Ri" is selected from the group consisting of
2. Use of a benzofuroxan compound represented by the formula (I)
its derivatives, analogues, tautomeric forms, stereoisomers, polymorphs, hydrates, pharmaceutically acceptable salts, pharmaceutically acceptable solvates or pharmaceutically acceptable compositions containing them, for treatment of diseases resulting from platelet aggregation wherein: R,Rι, n, X, R2, R3, R4, Y, R5, R and R are as defined in claim 1 and R8 is a halogen selected from the group F, CI, Br and I.
3. The use as claimed in claim 1, wherein the said compound is selected from the group consisting of :
(a) 5 (6)-(2 -nicotinamide ethyloxycarbonyl) benzofuroxan mesylate,
(b) 5(6)-(2-isonicotinamide ethyloxy carbonyl) benzofuroxan hydrochloride,
(c) 5(6)-(2-pyrolidinone ethyloxy carbonyl) benzofuroxan,
(d) 5(6)-(2-hydroxy propyloxy carbonyl) benzofuroxa , (e) 5(6)-(2-morpholino ethyloxy carbonyl) benzofuroxan,
(f) 5(6)-(2-methyloxy ethyloxy carbonyl) benzofuroxan ,
(g) 5(6)-(2,3-dihydroxy propyloxy carbonyl) benzofuroxan,
(h) 5(6)-(2-ethoxy ethyloxy carbonyl) benzofuroxan,
(i) 5(6)-(3-pyridine methoxy carbonyl) benzofuroxan hydrochloride, (j) 5(6)-(isosorbide mononitrateoxycarbonyl) benzofuroxan,
(k) 5(6)-chloro benzofuroxan ,
(1) 5(6)-carboxy benzofuroxan ,
(m) 5(6)-rnethylmercapto benzofuroxan,
(n) 5(6)-acetoxy benzofuroxan, (o) 5(6)-chlorocarbonyl benzofuroxan,
(p) 5(6)-formyl benzofuroxan,
(q) 5 (6)-methoxy carbonyl benzofuroxan,
(r) 5(6)-hydroxy benzofuroxan hydrochloride,
(s) 5(6)-amino carbonyl benzofuroxan, (t) 5(6)-ethoxy carbonyl benzofuroxan , (u) 5(6)-n-propoxycarbonyl benzofuroxan ,
(v) 5(6)-isopropoxycarbonyl benzofuroxan,
(w) 5(6)-tertiary butoxy carbonyl benzofuroxan,
(x) 5(6)-dimethylamino carbonyl benzofuroxan,
(y) 5(6)-morpholino carbonyl benzofuroxan,
(z) 5(6)-((4-methyl) piperazine-1-yl) carbonyl benzofuroxan ,
(aa) 5(6)-tertiary butyl amino carbonyl benzofuroxan,
(ab) 5 (6)-(2 -nicotinamide ethyloxycarbonyl) benzofuroxan hydrochloride and (ac) 5(6)-(3-pyridine methoxy carbonyl) benzofuroxan tartarate.
4. The use as claimed in claim 1, wherein said compound is selected from the group consisting of :
(ad) 5(6)-(S)(2-hydroxy)propyloxycarbonyl-benzofuroxan,
(ae) 5(6)[3-(3-pyridyl)propyloxycarbonyl benzofuroxan], (af) 5 (6)- {2-(pyrrolidin-2-one- 1 -yl) } ethyloxycarbonylamino-benzofuroxan,
(ag) 5(6)-(l-pyrrolidinyl)carbonyl benzofuroxan,
(ah) 5(6)-{2-(2-pyridylcarbonylamino)ethyl}oxycarbonyl benzofuroxan ,
(ai) 5(6)-{2-(3-pyridyl carbonylamino)ethyl}oxy carbonyl amino benzofuroxan,
(aj) 5(6)-nitro- benzofuroxan, and
(ak) benzofuroxan.
5. The use as claimed in claim 2 wherein said compound is 5(6) (4-bromo)- isopropyloxycarbonyl benzofuroxan.
6. A Pharmaceutical composition useful as antiplatelet agent for treatment of diseases resulting from platelet aggregation comprising a benzofuroxan compound represented by the formula (I)
its derivatives, analogues, tautomeric forms, stereoisomers, polymorphs, hydrates, pharmaceutically acceptable salts or pharmaceutically acceptable solvates, in admixture with pharmaceutically acceptable diluent, carrier or excepient wherein: R is -C (O)-Rι, halogen, acetoxy, -X-R2 or -C (O) NR3R , or C (O) CI 
n = 1 to 6, X is oxygen, sulfur, -C(O or -C(O)O-
R2 is hydrogen, straight chain or branched lower alkyl ( - C8), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen wherein substitution on nitrogen is with hydrogen or lower alkyl; R3and R4 are independently hydrogen, straight chain or branched lower alkyl (Ci -
C8) or R3 and R4 are linked together with or without a heteroatom selected from the group consisting of oxygen and nitrogen wherein substitution on nitrogen is with hydrogen or lower alkyl;
Y is -NHC (O)- or oxygen; R5 is lower alkyl (Cι-C8), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen, wherein substitution is with lower alkyl;
R6 is hydrogen, nitro, lower alkyl or -C (O)-R7;
R is hydrogen, lower alkyl or aryl; R8 is hydrogen or "Ri" is selected from the group consisting of
7. A Pharmaceutical composition useful as antiplatelet agent for treatment of diseases resulting from platelet aggregation comprising a benzofuroxan compound represented by the formula (I)
its derivatives, analogues, tautomeric forms, stereoisomers, polymorphs, hydrates, pharmaceutically acceptable salts or pharmaceutically acceptable solvates, in admixture with pharmaceutically acceptable diluent, carrier or excepient wherein R,Rι, n, X, R2, R3, R-t, Y, R5, Re and R7 are as defined in claim 6 and R8 is a halogen selected from the group F, CI, Br and I.
8. The Pharmaceutical composition as claimed in claim 6 wherein the said compound is selected from the group consisting of :-
(a) 5 (6)-(2 -nicotinamide ethyloxycarbonyl) benzofuroxan mesylate, (b) 5(6)-(2-isonicotinamide ethyloxy carbonyl) benzofuroxan hydrochloride,
(c)5(6)-(2-pyrolidinone ethyloxy carbonyl) benzofuroxan,
(d)5(6)-(2-hydroxy propyloxy carbonyl) benzofuroxan ,
(e)5(6)-(2-morpholino ethyloxy carbonyl) benzofuroxan,
(f) 5(6)-(2-methyloxy ethyloxy carbonyl) benzofuroxan , (g)5(6)-(2,3-dihydroxy propyloxy carbonyl) benzofuroxan,
(h)5(6)-(2-ethoxy ethyloxy carbonyl) benzofuroxan,
(i) 5(6)-(3-pyridine methoxy carbonyl) benzofuroxan hydrochloride,
(j) 5(6)-(isosorbide mononitrateoxycarbonyl) benzofuroxan,
(k)5(6)-chloro benzofuroxan , (1) 5(6)-carboxy benzofuroxan ,
(m) 5(6)-methylmercapto benzofuroxan,
(n)5(6)-acetoxy benzofuroxan,
(o)5(6)-chlorocarbonyl benzofuroxan,
(p)5(6)-formyl benzofuroxan, (q) 5 (6)-methoxy carbonyl benzofuroxan,
(r) 5(6)-hydroxy benzofuroxan hydrochloride,
(s)5(6)-amino carbonyl benzofuroxan, (t) 5(6)-ethoxy carbonyl benzofuroxan ,
(u)5(6)-n-propoxycarbonyl benzofuroxan ,
(v) 5 (6)-isopropoxycarbonyl benzofuroxan, (w) 5 (6)-tertiary butoxy carbonyl benzofuroxan,
(x)5(6)-dimethylamino carbonyl benzofuroxan,
(y)5(6)-morpholino carbonyl benzofuroxan,
(z) 5(6)-((4-methyl) piperazine-1-yl) carbonyl benzofuroxan ,
(aa) 5(6)-tertiary butyl amino carbonyl benzofuroxan,
(ab) 5 (6)-(2 -nicotinamide ethyloxycarbonyl) benzofuroxan hydrochloride and
(ac) 5(6)-(3-pyridine methoxy carbonyl) benzofuroxan tartarate.
9. The pharmaceutical composition as claimed in claim 6 wherein the said compound is selected from the group consisting of :
(ad) 5(6)-(S)(2-hydroxy)propyloxycarbonyl-benzofuroxan,
(ae) 5(6)[3-(3-pyridyl)propyloxycarbonyl benzofuroxan],
(af) 5 (6)- {2-(pyrrolidin-2-one- 1 -yl) } ethyloxycarbonylamino-benzofuroxan,
(ag) 5(6)-(l-pyrrolidinyl)carbonyl benzofuroxan, (ah) 5(6)-{2-(2-pyridylcarbonylamino)ethyl}oxycarbonyl benzofuroxan ,
(ai) 5(6)-{2-(3-pyridyl carbonylamino)ethyl}oxycarbonyl amino benzofuroxan,
(aj) 5(6)-nitro- benzofuroxan and
(ak) benzofuroxan.
10. The pharmaceutical composition as claimed in claim 7 wherein the said compound is 5(6) (4-bromo)-isopropyloxycarbonyl benzofuroxan.
11. A pharmaceutical composition as claimed in claim 7,8,9 or 10 in the form of a parenteral formulation.
12. A pharmaceutical composition as claimed in claim 11, wherein said pharmaceutically acceptable carriers is selected from propylene glycol and other glycols, alcohols, oils and sterile aqueous diluents such as isotonic saline solutions, buffer solutions and water for injection.
13 A pharmaceutical composition as claimed in claim 6,7,8,9 or 10 in the form of an oral formulation.
14. A pharmaceutical composition as in claim 13, wherein said pharmaceutically acceptable carrier is selected from lactose, dicalcium phosphate, starches, cellulose derivatives, polyvinyl pyrrolidone, cellulose derivatives, starch derivatives, talc, fumed silica and magnesium stearate.
15. A method of preventing or treating diseases resulting from the platelet aggregation in a human or non-human mammal which comprises administering an effective therapeutic dose of a benzofuroxan compound represented by the formula
(I)
its derivatives, analogues, tautomeric forms, stereoisomers, polymorphs, hydrates, pharmaceutically acceptable salts, pharmaceutically acceptable solvates and pharmaceutically acceptable compositions containing them, to a subject in need thereof wherein:
R is -C (O)-Rι, halogen, acetoxy, -X-R2 or -C (O) NR3R4, or C (O) CI
R1 is -O- (CH2)„-Y-R5, n = 1 to 6, X is oxygen, sulfur, -C(O)- or -C(O)O-
R2 is hydrogen, straight chain or branched lower alkyl (Ci - C8), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen wherein substitution on nitrogen is with hydrogen or lower alkyl; R3and R4 are independently hydrogen, straight chain or branched lower alkyl ( -
C8) or R3 and R4 are linked together with or without a heteroatom selected from the group consisting of oxygen and nitrogen wherein substitution on nitrogen is with hydrogen or lower alkyl;
Y is -NHC (O)- or oxygen; R5 is lower alkyl (Cι-C8), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen, wherein substitution is with lower alkyl;
R6 is hydrogen, nitro, lower alkyl or -C (O)-R ;
R is hydrogen, lower alkyl or aryl; R8 is hydrogen or "Ri" is selected from the group consisting of
16. A method of preventing or treating diseases resulting from the platelet aggregation in a human or non-human mammal which comprises administering of an effective therapeutic dose of a benzofuroxan compound represented by the formula (I)
its derivatives, analogues, tautomeric forms, stereoisomers, polymorphs, hydrates, pharmaceutically acceptable salts or pharmaceutically acceptable solvates and pharmaceutically acceptable compositions containing them, to a subject in need thereof wherein:
R,Rι, n, X, R2, R3, R4, Y, R5, R6 and R7 are as defined in claim 1 and R8 is a halogen selected from the group F, CI, Br and I.
17. The method for preventing or treating diseases resulting from the platelet aggregation in a human or non-human mammal as claimed in claim 15, wherein the said compound is selected from the group consisting of :
(a)5(6)-(2-nicotinamide ethyloxycarbonyl) benzofuroxan mesylate, (b) 5(6)-(2-isonicotinamide ethyloxy carbonyl) benzofuroxan hydrochloride,
(c)5(6)-(2-pyrolidinone ethyloxy carbonyl) benzofuroxan,
(d)5(6)-(2-hydroxy propyloxy carbonyl) benzofuroxan ,
(e)5(6)-(2-morpholino ethyloxy carbonyl) benzofuroxan, (f) 5(6)-(2-methyloxy ethyloxy carbonyl) benzofuroxan ,
(g)5(6)-(2,3-dihydroxy propyloxy carbonyl) benzofuroxan,
(h)5(6)-(2-ethoxy ethyloxy carbonyl) benzofuroxan,
(i) 5(6)-(3-pyridine methoxy carbonyl) benzofuroxan hydrochloride,
(j) 5(6)-(isosorbide mononitrateoxycarbonyl) benzofuroxan, (k)5(6)-chloro benzofuroxan ,
(1) 5(6)-carboxy benzofuroxan ,
(m) 5(6)-methylmercapto benzofuroxan,
(n)5(6)-acetoxy benzofuroxan,
(o)5(6)-chlorocarbonyl benzofuroxan, (p)5(6)-formyl benzofuroxan,
(q)5(6)-methoxycarbonyl benzofuroxan,
(r) 5(6)-hydroxy benzofuroxan hydrochloride,
(s) 5(6)-amino carbonyl benzofuroxan,
(t) 5(6)-ethoxy carbonyl benzofuroxan , (u) 5 (6)-n-propoxy carbonyl benzofuroxan ,
(v)5(6)-isopropoxycarbonyl benzofuroxan,
(w) 5(6)-tertiary butoxy carbonyl benzofuroxan,
(x)5(6)-dimethylamino carbonyl benzofuroxan,
(y)5(6)-morpholino carbonyl benzofuroxan, (z) 5(6)-((4-methyl) piperazine-1-yl) carbonyl benzofuroxan ,
(aa) 5(6)-tertiary butyl amino carbonyl benzofuroxan,
(ab) 5(6)-(2 -nicotinamide ethyloxycarbonyl) benzofuroxan hydrochloride and (ac) 5(6)-(3-pyridine methoxy carbonyl) benzofuroxan tartarate.
18. The method for preventing or treating diseases resulting from the platelet aggregation in a human or non-human mammal as claimed in claim 15, wherein the said compound is selected from the group consisting of :
(ad) 5(6)-(S)(2-hydroxy)propyloxycarbonyl-benzofuroxan,
(ae) 5(6)[3-(3-pyridyl)propyloxycarbonyl benzofuroxan],
(af) 5(6)-{2-(pyrrolidin-2-one- 1 -yl)} ethyloxycarbonylamiiio-benzofuroxan,
(ag) 5(6)-(l-pyrrolidinyl)carbonyl benzofuroxan, (ah) 5(6)- {2-(2-pyridylcarbonylamino)ethyl}oxycarbonyl benzofuroxan ,
(ai) 5(6)-{2-(3-pyridyl carbonylamino)ethyl}oxycarbonyl amino benzofuroxan,
(aj) 5(6)-nitro- benzofuroxan and
(ak) benzofuroxan .
19. The method for preventing or treating diseases resulting from the platelet aggregation in a human or non-human mammal as claimed in claim 16, wherein the said compound is 5(6)(4-bromo)-isopropyloxycarbonyl benzofuroxan.
20. Use of a benzofuroxan compound represented by the formula (I)
its derivatives, analogues, tautomeric forms, stereoisomers, polymorphs, hydrates, pharmaceutically acceptable salts or pharmaceutically acceptable solvates in the manufacture of a medicament useful as antiplatelet agent in disease conditions resulting from the platelet aggregation wherein:
R is -C (O)-Rι, halogen, acetoxy, -X-R2 or -C (O) NR3R4, or C (O) CI
Rι is -O- (CH2) „-Y-R5, n = 1 to 6,
X is oxygen, sulfur, -C(O)- or -C(O)O- R2 is hydrogen, straight chain or branched lower alkyl (Ci - C8), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen wherein substitution on nitrogen is with hydrogen or lower alkyl;
R3and R4 are independently hydrogen, straight chain or branched lower alkyl ( - C8) or R3 and R4 are linked together with or without a heteroatom selected from the group consisting of oxygen and nitrogen wherein substitution on nitrogen is with hydrogen or lower alkyl;
Y is -NHC (O)- or oxygen;
R5 is lower alkyl (Cι-C8), aromatic, heteroaromatic, and substituted or unsubstituted saturated heterocyclic ring with one or two heteroatoms such as nitrogen or oxygen, wherein substitution is with lower alkyl;
Re is hydrogen, nitro, lower alkyl or -C (O)-R ;
R is hydrogen, lower alkyl or aryl;
R8 is hydrogen or "Ri" is selected from the group consisting of
21. Use of a benzofuroxan compound represented by the formula (I)
its derivatives, analogues, tautomeric forms, stereoisomers, polymorphs, hydrates, pharmaceutically acceptable salts or pharmaceutically acceptable solvates in the manufacture of a medicament useful as antiplatelet agent in disease conditions resulting from the platelet aggregation wherein:
R,Rι, n, X, R2, R3, R-j, Y, R5, Re and R7 are as defined in claim 1 and R8 is a halogen selected from the group F, CI, Br and I.
-
22. The use of a benzofuroxan compound as claimed in claim 20, wherein the said compound is selected from the group consisting of :
(a) 5(6)-(2 -nicotinamide ethyloxycarbonyl) benzofuroxan mesylate,
(b) 5(6)-(2-isonicotinamide ethyloxy carbonyl) benzofuroxan hydrochloride,
(c) 5(6)-(2-pyrolidinone ethyloxy carbonyl) benzofuroxan,
( 5(6)-(2-hydroxy propyloxy carbonyl) benzofuroxan ,
(e) 5(6)-(2-morpholino ethyloxy carbonyl) benzofuroxan,
(f) 5(6)-(2-methyloxy ethyloxy carbonyl) benzofuroxan ,
(g) 5(6)-(2,3-dihydroxy propyloxy carbonyl) benzofuroxan,
(h) 5(6)-(2-ethoxy ethyloxy carbonyl) benzofuroxan,
(i) 5(6)-(3-pyridine methoxy carbonyl) benzofuroxan hydrochloride,
0) 5(6)-(isosorbide mononitrateoxycarbonyl) benzofuroxan,
(k) 5(6)-chloro benzofuroxan ,
(1) 5(6)-carboxy benzofuroxan ,
(m) 5(6)-methylmercapto benzofuroxan,
(n) 5(6)-acetoxy benzofuroxan,
(o) 5(6)-chlorocarbonyl benzofuroxan,
(P) 5(6)-formyl benzofuroxan,
(q) 5(6)-methoxycarbonyl benzofuroxan,
5(6)-hydroxy benzofuroxan hydrochloride,
(s) 5(6)-amino carbonyl benzofuroxan,
(t) 5(6)-ethoxy carbonyl benzofuroxan , (u) 5(6)-n-propoxycarbonyl benzofuroxan ,
(v) 5(6)-isopropoxycarbonyl benzofuroxan,
(w) 5(6)-tertiary butoxy carbonyl benzofuroxan,
(x) 5(6)-dimethylamino carbonyl benzofuroxan, (y) 5(6)-morpholino carbonyl benzofuroxan,
(z) 5(6)-((4-methyl) piperazine-1-yl) carbonyl benzofuroxan ,
(aa) 5(6)-tertiary butyl amino carbonyl benzofuroxan,
(ab) 5(6)-(2-nicotinamide ethyloxycarbonyl) benzofuroxan hydrochloride and (ac) 5(6)-(3-pyridine methoxy carbonyl) benzofuroxan tartarate.
23. The use of a benzofuroxan compound as claimed in claim 20, wherein the said compound is selected from the group consisting of :
(ad) 5(6)-(S)(2-hydroxy)propyloxycarbonyl-benzofuroxan,
(ae) 5(6)[3-(3-pyridyl)propyloxycarbonyl benzofuroxan],
(af) 5(6)-{2-(pyrrolidm-2-one-l-yl)}ethyloxycarbonylamino-benzofuroxan,
(ag) 5(6)-( l-pyrrolidinyl)carbonyl benzofuroxan,
(ah) 5(6)-{2-(2-pyridylcarbonylamino)ethyl}oxycarbonyl benzofuroxan , (ai) 5(6)-{2-(3-pyridyl carbonylamino)ethyl}oxy carbonyl amino benzofuroxan, (aj) 5(6)-nitro- benzofuroxan and (ak) benzofuroxan .
24. The use of a benzofuroxan compound as claimed in claim 21, wherein the said compound is 5(6)(4-bromo)-isopropyloxycarbonyl benzofuroxan.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2002366756A AU2002366756A1 (en) | 2001-12-20 | 2002-12-13 | Use of benzofuroxan derivatives as antiplatelet agents |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US34128301P | 2001-12-20 | 2001-12-20 | |
| US60/341,283 | 2001-12-20 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2003053439A1 true WO2003053439A1 (en) | 2003-07-03 |
Family
ID=23336943
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/IB2002/005353 Ceased WO2003053439A1 (en) | 2001-12-20 | 2002-12-13 | Use of benzofuroxan derivatives as antiplatelet agents |
Country Status (2)
| Country | Link |
|---|---|
| AU (1) | AU2002366756A1 (en) |
| WO (1) | WO2003053439A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2515413C2 (en) * | 2012-07-05 | 2014-05-10 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Московский государственный университет имени М.В. Ломоносова" (МГУ) | 1,2,5-oxadiazole derivatives, having anti-hiv activity, pharmaceutical composition, method of inhibiting hiv-1 integrase |
| CN114920722A (en) * | 2021-06-02 | 2022-08-19 | 何黎琴 | A kind of 7-hydroxy-3-acetyl coumarin oxime compound, its preparation method and medical use |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0431944A2 (en) * | 1989-12-08 | 1991-06-12 | Merck & Co. Inc. | Benzofurazan derivatives having antiarrhythmic properties |
| EP0574726A1 (en) * | 1992-06-10 | 1993-12-22 | Hoechst Aktiengesellschaft | Annelated 1,2,5-oxadiozole-2 oxides and their use as pharmacological agents |
| US5424326A (en) * | 1992-06-20 | 1995-06-13 | Cassella Aktiengesellschaft | Phenyl-1,2,5-oxadiazolecarboxamide-2-oxides, their preparation and their use |
| WO1998035950A1 (en) * | 1997-02-13 | 1998-08-20 | Cortex Pharmaceuticals, Inc. | Benzofurazan compounds which enhance ampa receptor activity |
| US6232331B1 (en) * | 1998-05-22 | 2001-05-15 | Torrent Pharmaceutical Ltd. | Benzofuroxan derivatives, their therapeutic uses and pharmaceutical compositions |
-
2002
- 2002-12-13 WO PCT/IB2002/005353 patent/WO2003053439A1/en not_active Ceased
- 2002-12-13 AU AU2002366756A patent/AU2002366756A1/en not_active Abandoned
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0431944A2 (en) * | 1989-12-08 | 1991-06-12 | Merck & Co. Inc. | Benzofurazan derivatives having antiarrhythmic properties |
| EP0574726A1 (en) * | 1992-06-10 | 1993-12-22 | Hoechst Aktiengesellschaft | Annelated 1,2,5-oxadiozole-2 oxides and their use as pharmacological agents |
| US5424326A (en) * | 1992-06-20 | 1995-06-13 | Cassella Aktiengesellschaft | Phenyl-1,2,5-oxadiazolecarboxamide-2-oxides, their preparation and their use |
| WO1998035950A1 (en) * | 1997-02-13 | 1998-08-20 | Cortex Pharmaceuticals, Inc. | Benzofurazan compounds which enhance ampa receptor activity |
| US6232331B1 (en) * | 1998-05-22 | 2001-05-15 | Torrent Pharmaceutical Ltd. | Benzofuroxan derivatives, their therapeutic uses and pharmaceutical compositions |
Non-Patent Citations (3)
| Title |
|---|
| GHOSH P B ET AL: "Potential antileukemic and immunosuppressive drugs. Preparation and in vitro pharmacological activity of some benzo-2,1,3-oxadiazoles (benzofurazans) and their N-oxides (benzofuroxans)", JOURNAL OF MEDICINAL CHEMISTRY, vol. 11, no. 2, 1968, pages 305 - 11, XP002113125 * |
| GHOSH, P. B. ET AL: "Potential antileukemic and immunosuppressive drugs. 3. Effects of homocyclic ring substitution on the in vitro drug activity of 4-nitrobenzo-2,1,3-oxadiazoles (4-nitrobenzofurazans) and their N-oxides (4-nitrobenzofuroxans)", JOURNAL OF MEDICINAL CHEMISTRY, vol. 15, no. 3, 1972, pages 255 - 260, XP002232282 * |
| ZHANG W ET AL: "Synthesis and hypoxia-selective cytotoxicity of benzofuraxans and quinoline di-N-oxides", CHEMICAL ABSTRACTS + INDEXES, AMERICAN CHEMICAL SOCIETY. COLUMBUS, US, vol. 1, no. 125, 1 July 1996 (1996-07-01), pages 1182, XP002113146, ISSN: 0009-2258 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2515413C2 (en) * | 2012-07-05 | 2014-05-10 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Московский государственный университет имени М.В. Ломоносова" (МГУ) | 1,2,5-oxadiazole derivatives, having anti-hiv activity, pharmaceutical composition, method of inhibiting hiv-1 integrase |
| CN114920722A (en) * | 2021-06-02 | 2022-08-19 | 何黎琴 | A kind of 7-hydroxy-3-acetyl coumarin oxime compound, its preparation method and medical use |
| CN114920722B (en) * | 2021-06-02 | 2024-03-08 | 何黎琴 | 7-hydroxy-3-acetyl coumarin oxime compound, preparation method and medical application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| AU2002366756A1 (en) | 2003-07-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR0159517B1 (en) | Heterocyclic Derivatives, Method of Preparation and Therapeutic Use | |
| HU208124B (en) | Process for producing pyridazinone derivatives and pharmaceutical compositions comprising such derivatives as active ingredient | |
| PL214701B1 (en) | Pyrrolidinedione substituted piperidine-phthalazones as pde4 inhibitors | |
| EP0281459A1 (en) | Dextrorotatory enantiomer of alpha-(4,5,6,7-tetrahydrothieno[3,2-c]pyrid-5-yl)(2-chlorophenyl)methyl acetate, process for its preparation and pharmaceutical compositions containing it | |
| WO1996014843A2 (en) | Pharmaceutical pyrazole compositions useful as inhibitors of protein kinases | |
| JPH0641466B2 (en) | Pyrazolotriazine derivative and xanthine oxidase inhibitor containing the same | |
| KR20130089264A (en) | Heterocyclic derivates, preparation processes and medical uses thereof | |
| UA46821C2 (en) | SUBSTITUTED N - [(AMINOMINOMETHYL OR AMINOMETHYL) PHENYL] PROPYLAMIDES, PHARMACEUTICAL COMPOSITION, TREATMENT METHODS AND INTERMEDIATES | |
| EP0187977B1 (en) | Tetrahydroquinoline derivatives, process for preparing the same and anti-peptic ulcer compositions containg the same | |
| HU198021B (en) | Process for producing new antiallergic and antitrombotic heterocyclic compounds and pharmaceutical compositions containing them | |
| EP1442026B9 (en) | Triazole derivatives as cyclooxygenase (cox) inhibitors | |
| HU214627B (en) | Process for the preparation of 2,4- and 2,5-substituted pyridine-N-oxides and pharmaceutical compositions containing them. | |
| EP0240158B1 (en) | Benzimidazole derivatives | |
| SK6012002A3 (en) | Imidazole derivative as phosphodiesterase vii inhibitor, use thereof and pharmaceutical composition containing the same | |
| US4619938A (en) | Fatty acid derivatives of aminoalkyl nicotinic acid esters and platelet aggregation inhibitors | |
| NZ260505A (en) | Cyclic indole derivatives and medicaments thereof | |
| US9630915B2 (en) | Thromboxane receptor antagonists | |
| HU209645B (en) | Process for producing heteroaryl-sulphonamido-carbazole derivatives | |
| JPS61172861A (en) | Isoquinolinethromboxane synthetase inhibitor | |
| EP0159652B1 (en) | 4-phenylphthalazine derivatives, their preparation and pharmaceutical compositions containing them | |
| US5405848A (en) | Substituted thiazolylaminotetrahydropyridopyrimidines derivatives useful as platelet aggregation inhibitors | |
| JP2005527518A (en) | Novel chalcone derivatives and their use | |
| WO2003053439A1 (en) | Use of benzofuroxan derivatives as antiplatelet agents | |
| JPS5944370A (en) | Fused ring 4h-1,4-benzothiazine, manufacture and use as medicine | |
| PL144341B1 (en) | Method of obtaining new heterocyclic compounds |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| AK | Designated states |
Kind code of ref document: A1 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ OM PH PL PT RO RU SC SD SE SG SK SL TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW |
|
| AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR IE IT LU MC NL PT SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG |
|
| DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
| WWE | Wipo information: entry into national phase |
Ref document number: 914/KOLNP/2004 Country of ref document: IN |
|
| 122 | Ep: pct application non-entry in european phase | ||
| NENP | Non-entry into the national phase |
Ref country code: JP |
|
| WWW | Wipo information: withdrawn in national office |
Ref document number: JP |