CA2301852C

CA2301852C - Angiogenesis promoters and angiogenesis potentiators

Info

Publication number: CA2301852C
Application number: CA002301852A
Authority: CA
Inventors: Yasuhiro Egi; Hideaki Kido; Kazutaka Hayashi; Yoshiji Kubo; Norifumi Nakamura
Original assignee: Nissan Chemical Corp
Current assignee: Nissan Chemical Corp
Priority date: 1997-08-28
Filing date: 1998-08-26
Publication date: 2007-07-10
Anticipated expiration: 2018-08-26
Also published as: EP1025847B1; CN1142779C; US6284758B1; TW490303B; ES2247716T3; DK1025847T3; CN1277553A; AU8886298A; CA2301852A1; ATE307584T1; WO1999011268A1; JP4352613B2; KR20010023331A; DE69832089T2; DE69832089D1; EP1025847A1; KR100559192B1; EP1025847A4

Abstract

An angiogenesis promoter and angiogenesis potentiator containing a pyridazinone compound of the formula (I) (see formula I) wherein each symbol is as defined in the specification, or a pharmacologically acceptable salt thereof, as an active ingredient.
The pyridazinone compound (I) and a pharmacologically acceptable salt thereof in the present invention promote angiogenesis and potentiate the angiogenic effect of a drug having such effect, and are useful as an angiogenesis promoter and angiogenesis potentiator.

Description

SPECIFICATION

ANGIOGENESIS PROMOTERS AND ANGIOGENESIS POTENTIATORS
Technical Field The present invention relates to angiogenesis promoters and angiogenesis potentiators, which contain, as an active ingredient, a specific pyridazinone compound or a pharmacologically acceptable salt thereof.
Background Art The below-noted specific pyridazinone compound in the present invention is known to have superior platelet aggregation inhibitory effect, cardiotonic effect, vasodilating effect, anti-SRS-A (Slow Reacting Substances of Anaphylaxis) effect, thromboxane A2 synthase inhibitory effect and the like (JP-B-7-107055, JP-A-7-285869), and is a drug expected to be an antiplatelet agent and the like.
However, there has not been any report on the effect of the pyridazinone compound on angiogenesis.
Disclosure of the Invention The present inventors have conducted various studies of the effect of the pyridazinone compound on the angiogenesis and found that the pyridazinone compound promotes angiogenesis and potentiates an angiogenic effect of a drug having such effect, which resulted in the completion of the present invention.
Accordingly, the present invention provides the following.
An angiogenesis promoter containing a pyridazinone compound of the formula (I) R,, X
N

i R2 N (I) HC-A
Y

wherein Rl, R 2 and R3 are each independently a hydrogen atom or a lower alkyl, X is a halogen atom, cyano or a hydrogen atom, Y is a halogen atom, trifluoromethyl or a hydrogen atom, and A is C1 - CB alkylene optionally substituted by hydroxyl group, or a pharmacologically acceptable salt thereof (hereinafter to be referred to as pyridazinone compounds) as an active ingredient.
A method of promoting angiogenesis, which comprises administering a pyridazinone compound.
Use of a pyridazinone compound for the production of an angiogenesis promoter.
A pharmaceutical composition for promoting angiogenesis, which comprises a pyridazinone compound and a pharmaceutically acceptable carrier.
A commercial package comprising the above-mentioned pharmaceutical composition, and a written matter associated therewith, the written matter stating that the pharmaceutical composition can or should be used for promoting angiogenesis.
A potentiator of a drug having an angiogenic effect, which contains a pyridazinone compound as an active ingredient.
A method of potentiating an angiogenic effect of a drug having such effect, which comprises administering a pyridazinone compound.
Use of a pyridazinone compound for the production of a potentiator of a drug having an angiogenic effect.
A pharmaceutical composition for potentiating an angiogenic effect of a drug having such effect, which contains a pyridazinone compound and a pharmaceutically acceptable carrier.
A commercial package comprising the above-mentioned pharmaceutical composition, and a written matter associated therewith, the written matter stating that the pharmaceutical composition can or should be used for potentiating an angiogenic effect of a drug having such effect.
The pyridazinone compound in the present invention is preferably a compound of the formula (I), wherein R' and R 2 are each hydrogen atom, R3 is hydrogen atom or alkyl having 1 to 4 carbon atoms, X is halogen atom, Y is halogen atom or hydrogen atom and A is C1 -C5 alkylene optionally substituted by hydroxyl group.
Particularly preferable pyridazinone compound of the formula (I) (hereinafter to be referred to as pyridazinone compound (I)) is, for example, 4-bromo-6-[3-(4-chlorophenyl)-propoxy]-5-(3-pyridylmethylamino)-3-(2H)-pyridazinone.
Angiogenesis means that endothelial cells bud from an existing blood vessel and form a new blood vessel. The process of the formation is complicated and is an important phenomenon observed in various aspects in the living body, such as angiogenesis for development and growth, pathologic angiogenesis (e.g. growth of tumor, diabetic retinopathy) and the like.
Administration of an angiogenesis promoter to the patients with cancer or diabetic retinopathy is prohibited because it promotes pathologic angiogenesis.
However, an angiogenesis promoter is useful in that it complements and potentiates the efficacy of a pharmaceutical agent that directly acts on the mainly diseased artery (artery mainly causing the disease), because it forms a collateral circulatory path irrespective of the mainly diseased artery.
The symbols used in this specification are explained in the following.
The lower alkyl at Rl, R2 and R3 has 1 to 6 carbon atoms and may be linear or branched. Examples thereof include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, pentyl, hexyl and the like.
Rl and R2 are each preferably a hydrogen atom and R3 is preferably a hydrogen atom or alkyl having 1 to 4 carbon atoms.
The alkyl having 1 to 4 carbon atoms at R3 is exemplified by methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl and the like.
The halogen atom at X and Y means fluorine atom, chlorine atom, bromine atom or iodine atom.
Preferable X is a halogen atom and preferable Y is a halogen atom and a hydrogen atom.
The C1 - C. alkylene optionally substituted by hydroxyl group at Amay be linear or branched and is exemplified by methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, 2,2-dimethylethylene, 2, 2 -diethyl ethylene, 2,2-di-n-propylethylene, hydroxymethylene, 1-hydroxyethylene, 2-hydroxyethylene, 3-hydroxypropylene and the like.
Preferable A is C1 - C5 alkylene optionally substituted by hydroxyl group.
In the formula (I), methylene group and pyridine ring may be bonded at any position, but preferably bonded at the 3-position relative to the nitrogen atom of the pyridine ring.
Y may be substituted at any position on the benzene ring, but preferably at the 4-position.
Particularly, the pyridazinone compound of the formula (I) wherein R' and RZ are hydrogen atoms, R3 is hydrogen atom or alkyl having 1 to 4 carbon atoms, X is halogen atom, Y is halogen atom or hydrogen atom and A is C1 - C5 alkylene optionally substituted by hydroxyl group is preferable.
More preferable pyridazinone compounds (I) include 4-bromo-6-(3-phenylpropoxy)-5-(3-pyridylmethylamino)-3(2H)-pyridazinone, 4-chloro-6-(3-phenylpropoxy)-5-(3-pyridylmethylamino)-3(2H)-pyridazinone, 4-chloro-6-[3-(4-chlorophenyl)propoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone, 4-bromo-6-[3-(4-chlorophenyl)propoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone, 4-bromo-6-(2,2-dimethyl-3-phenylpropoxy)-5-(3-pyridylmethylamino)-3(2H)-pyridazinone, 4-chloro-6-(2,2-dimethyl-3-phenylpropoxy)-5-(3-pyridylmethylamino)-3(2H)-pyridazinone, 4-bromo-6-[3-(4-chlorophenyl)-2,2-dimethylpropoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone, 4-chloro-6-[3-(4-chlorophenyl)-2,2-dimethylpropoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone, 4-bromo-6-[3-(4-chlorophenyl)-3-hydroxypropoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone, 4-chloro-6-[3-(4-chlorophenyl)-3-hydroxypropoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone, 4-bromo-6-[3-(4-chlorophenyl)-2-hydroxypropoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone and 4-chloro-6-[3-(4-chlorophenyl)-2-hydroxypropoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone.
The pyridazinone compound (I) in the present invention encompasses stereoisomers and optical isomers.
The pyridazinone compound (I) can be produced by a method disclosed in, for example, JP-B-7-107055, US-A-5314883, EP-A-482208, JP-A-7-252237, US-A-5750523 and EP-A-742211.
The pharmacologically acceptable salts of pyridazinone compound (I) include salts with inorganic acid (e.g., hydrochloride, hydrobromide, phosphate, sulfate and the like), salts with organic acid (e.g., acetate, succinate, maleate, fumarate, malate, tartrate and the like), and the like.
The pyridazinone compound (I) can be converted to the above-mentioned salts by known methods.

The pyri.dazinone c.ompound ( I)<-ind ptlannacologically acceptable salts thereof can be used as an angiogenesis promoter by themselves.
They may be also used as a pot:ent.iator of the ang:iogenic effect of a drug having such effect by concurrently using the drug.
The drug having an anq i..ogenic r:~f fect may be a qr_owth factor such as a basic fibroblast grooath factor;b-FGF, an endothelial cell growth factor;ECGF, an epidermal growth factor;EGF, a tr.ansforming growth factor-(t;TGF-Jr, a platea.et-derived endothelial cell growth factor;PDGF, a vascular endothelial growth factor;VEGF, a vascular perrneability factor;VPF zand the like, hepari.n, adenosine and the like.
The pyridazinone compound (I) and pharmacologically acceptable salts thereof are superior as a potent.i_at.or of the angiogenic effect of a growth factor, particularly b-FGF.
The effects of the preserit invention can be confirmed by any known method which is free of limitation.
The pyridazinone compound (I) and pharmacologically acceptable salts thereof, which are the active :ingredi.ents in the present invention, are extremely law toxic arid show an angioc:lenesis-promoting effect and potentiation of ttie angioqeni.c effect of a drug in mammals such as human, dog, cow, horse, rabbit, mouse, rat and the like.
The pyridazinone compound ( I) and pharmacologically acceptable salts thereof can be admirristered parenterally in the form of injection (subcutaneous, intravenous, intramuscular, intraperitoneal injections), ointment, suppository, aerosol agent and the like, or orally i_n the form of tablet, capsul e, granule, pill,, syrup, liquid, emulsion, suspension and the 1ike.
The pyridazinone compound (1) and salts thereof can be formulated into a preparation for administration, according to conventional methods of drug production.
The tablet, c:apsu.l..e, granule arid pil.l for oral administration can be prepared using excipient (e.g., sucrose, lactose, glucose, starch, mannitol and the .L.:ike ), binder ( e. g., syrup, acacia, gelatin, sorbitol, tragacanth, methylcellulose, polyvinylpyrrolidone and the like), disintegrant (e.g., starch, c:arboxymethylcellulose or calcium salt thereof, microcrystalline cellulose, polyethylene glycol and the like), lubricant (e.g., talc, magnesium stearate, calcium stearate, silica and the like), gli..dant (e.g., sodium lauryl. sulfate, glycerol and the like), and the like.

The injection, aerosol agent, syrup, liquid, emulsion and suspension can be prepared using a solvent for the active ingredient (e.g., water, ethyl alcohol, isopropyl alcohol, propylene glycol, 1,3-butylene glycol, polyethylene glycol and the like), a surfactant (e.g., sorbitan fatty acid ester, polyoxyethylenesorbitan fatty acid ester, polyoxyethylene fatty acid ester, polyoxyethylene ether of hydrogenated caster oil, lecithin and the like), a suspending agent (e.g., cellulose derivatives such as carboxymethylcellulose sodium salt, methylcellulose and the like, natural gums such as tragacanth, acacia and the like), a preservative (e.g., p-hydroxybenzoate, benzalkonium chloride, sorbate and the like), and the like. A
suppository can be prepared using, for example, polyethylene glycol, lanolin, coconut oil and the like.
The dose of the pyridazinone compound (I) and a salt thereof can be determined as appropriate according to the age, body weight, disease state and the like of the patient. It is generally 0.001 mg - 5 g/day, preferably 0.005 - 1000 mg/day, for an adult ( human ), which is administered in one to several doses a day.
When the pyridazinone compound (I) or a pharmacologically acceptable salt thereof is used as a potentiator of the angiogenic effect of a drug having such effect, the pyridazinone compound in the present invention and the drug having an angiogenic effect are administered in such a manner that they are both present in the body during the same period of time. The use and dose of the drug having an angiogenic effect are free of limitation as long as they fall within the known ranges. They may be prepared into a single pharmaceutical preparation or separately into individual preparations. When they are separate preparations, the administration route and dose may be the same or different.
Brief Description of the Drawings Fig. 1 is a photograph showing the neovascularized blood vessel at day 7 of the test in a rat sponge model group administered with a vehicle.
Fig. 2 is a photograph showing the neovascularized blood vessels at day 7 of the test in a rat sponge model group administered with compound A.
Fig. 3 shows the amount of hemoglobin in sponge in a rat sponge model, indicating the angiogenesis-promoting effect of compound A.

Fig. 4 is a photograph showing the neovascularized blood vessels at day 4 of the test in a rat sponge model group 1 administered with a BSA-physiological saline solution.
Fig. 5 is a photograph showing the neovascularized blood vessels at day 4 of the test in a rat sponge model group 2 administered solely with a basic fibroblast growth factor (b-FGF).
Fig. 6 is a photograph showing the neovascularized blood vessels at day 4 of the test in a rat sponge model group 3 co-administered with b-FGF and compound A.
Fig. 7 shows the amount of hemoglobin in sponge in a rat sponge model, indicating angiogenic potency of b-FGF by compound A.
Fig. 8 shows the relationship between the dose and angiogenic efficacy by compound A in a rat sponge model.
Examples The present invention is explained in detail in the following Examples and Experimental Examples. The invention is not limited by these Examples in any way.
Experimental Example 1: promotion of angiogenesis by compound A
Method:
Rats were anesthetized by intraperitoneal administration of sodium pentobarbital (50 mg/kg), and the dorsal median line was incised for about 1 cm and an air pocket was made subcutaneously at about 2.5 cm toward the tail side with a Kocher clamp. A hemostatic gelatin sponge ( Spongel , 10 mmXlO mmX7 mm; manufactured by Yamanouchi Pharmaceutical Co., Ltd.) impregnated with physiological saline was embedded therein. The opening was sutured and antisepticized to give a test animal model.
As a reagent, compound A(4-bromo-6-[3-(4-chlorophenyl)-propoxy]-5-(3-pyridylmethylamino)-3-(2H)-pyridazinone hydrochloride, 100 mg) produced by a conventional method was suspended in 0.5% methylcellulose solution (100 ml) in a mortar and used.
As a control, a vehicle (0. 5% methylcellulose solution) was used.
The vehicle was obtained by dissolving methylcellulose (5 g, manufactured by Kishida Chemical Industries, Ltd.) in distilled water (1000 ml).
The reagent (compound A) and the vehicle were orally administered repeatedly from the day of sponge embedding at a dose of 10 ml/kg once a day for 4 days or 7 days. The administration was performed 30 minutes before anesthetizing the animal on the very day of embedding, and the sponge was removed on the next day of the final administration.
After 4 days or 7 days from the embedding, an excess amount of pentobarbital was intravenously injected to the animals for euthanasia.
The back was opened, and the tissue surrounding the embedded sponge was removed and the surface of the sponge was photographed. The sponge was taken out and placed in a 0. 1M aqueous ammonia (2 ml ), which was stood for 4 hr to extract hemoglobin in the sponge. The extract (100 Eil) was taken, and hemoglobin was quantitated using an assay kit (hemoglobin B-TESTWAKO; manufactured by Wako Pure Chemical Industries, Ltd.) and used as the index of angiogenesis. The amount of hemoglobin in the sponge was calculated by the following formula.

Amount of hemoglobin in sponge (mg/sponge) _ hemoglobin (mg) in extract (100 l) X 20 (total extract 2 ml) The obtained data were expressed in mean standard error. For the evaluation of the angiogenic effect, an unpaired t-test was performed using the animals administered with the drug for 4 days or 7 days and respective vehicle groups as control to examine significant difference. The significance was ascribed at less than 5% risk rate.
Results:
1. Observation of photograph Both the reagent (compound A) administration group and vehicle administration group showed an increase of neovascularized blood vessels on the sponge surface at day 4 and day 7 of the test in proportion to the number of days lapsed from the initiation of the test. At day 7 of the test, the reagent (compound A) administration group (Fig.
2) showed more neovascularized blood vessels than in the vehicle administration group (Fig. 1).
2. Amount of hemoglobin in sponge The results are shown in Table 1 and Fig. 3. Both the reagent (compound A) administration group and vehicle administration group showed an increase in the amount of hemoglobin in sponge at day 4 and day 7 of the test, in proportion to the number of days lapsed from the initiation of the test. At day 7 of the test, the reagent (compound A) administration group showed a significant increase as compared to the vehicle administration group.

Table 1 Drug Hemoglobin in sponge (mg/sponge) 4 days after embedding 7 days after embedding Vehicle 6.216 0.903 ( 9) 9.585 0.774 (10) Reagent (compound A) 6.255 0.807 (10) 16.351 1.836 ** (9) ( ): number of animals **:p<0.01 relative to vehicle administration group, unpaired t-test Experimental Example 2 : potentiation of angiogenic effect of b-FGF
by compound A
Method:
The test animal models were prepared in the same manner as in Experimental Example 1.
As a reagent, compound A(4-bromo-6-[3-(4-chlorophenyl)-propoxy]-5-(3-pyridylmethylamino)-3-(2H)-pyridazinone hydrochloride, 50 mg) produced by a conventional method was suspended in 0.5% methylcellulose solution (100 ml) in a mortar and used.
The vehicle (0.5% methylcellulose solution) was obtained by dissolving methylcellulose (5 g, manufactured by Kishida Chemical Industries, Ltd.) in distilled water (1000 ml).
A 0.1% bovine serum albumin (BSA) -physiological saline solution was prepared by dissolving BSA (0.1 g, Sigma) in physiological saline (100 ml).
A b-FGF solution was prepared by dissolving recombinant b-FGF
(20 g,manufactured by BTI) in 0.1% BSA-physiological saline solution (2 ml).
A hemostatic gelatin sponge was the same as that used in Experimental Example 1.
The test group was divided into 3 groups. A sponge wetted with 100 Eil of 0.1% BSA-physiological saline solution was embedded in group 1. A sponge (1 Eig b-FGF/sponge) wetted with b-FGF solution (100 111) was embedded in group 2 and group 3.
The vehicle was orally administered to group 1 and group 2, and the reagent (compound A) was orally administered to group 3 repeatedly frorn the day of sponge ernbedding at the dose of 10 nil/kq twice a day for 4 clays.
In every test group, the adnii_nist.rat_i.on began f.rom the evening (single administratiori) c:if: the very day of enibeddirig. The time of administration of the drt.7g was around 9 a.m. for the fi-r.st administratiori and around. 7 p.m. for the second administ:ration. In consideration of the effect of the druq, the drug was not adniinistered on the veL-y day of sponge .removal (7 times of admiriistration in total ).
After 4 days frorn the embedding, an excess amount of pentobarbital was intravenous]_y injected to the animals for euthanasia.
The back was opened, and the tissue surrounding the embedded sponge was removed and the surfacE:-> o f_ the sponge was photographed. The sponge was takerr out and hemoglobin =in the sporiqe was calculated in the same manner as in Exper:i.mental Example l.
The obtalned data were expressed in niean= _:standard error. For the evaluation of the angiogeni.c effect, a multiple comparison test by Tukey rnethod was performed to examino s:iqnif icant di_f_ference.. The signifacance was ascribed at. less than S , risk rate.
Result-1. Observation of photog.raph The results are shown in Fi.q. 4, Fi_q . 5 and Fiq . 6. At day 4 of the test, group 2 (b-FGF single admi_ni_stration group) visually showed more neovasculari.zed blood vesse:l.s than i.n group 1(0.10 BSA-physi_ological. sal.inc--! solution qroup), and group 3 (b-FGF and reagent ( compound A, 5 mg /kg ) co-admirii-stration group) showed still more neovascularized blc:,,od vessels ttiari i_n qr_oup 2.
2. Amount of hemog].obin in sponge The results are shown in Table 2 and Fig. 7. The group 3 (b-FGF
and reagent (compound A, 'i mg/ kg ) co-administration group ) showed a significant increase iri hemoglobin content as compared t:o group 1 (0. 1%
BSA-physioloyical sali_ne solution qroup) and group 2 (b-FGF single admini..stration group).

- r, Ta~)le 2 Test Hemoglobin in group Drug sponge (mg/sponge) at 4 Oral. administration In-sponge days after _ administration embeddin Group 1 Vehicle BSA-I.hysi_ologic:al 5. 503+0. 562 (10) s a.l ine Group 2 Vehicle b-FGF 7.285 0.715 (10) 14.995 2.307 Group 3 Reagent (compound A) b-FGF ++ ** 10 ( ) : number of an:imals, ++:p<0.01 relative to group 1., **: p<0 . 01. relati.ve to group ~' (Tukey niethod ) Experimental Example 3 : relationship between dose and angiogenesis proinotion by compound A
MethQdi The test. anirnal models were prepared in the same manner as in Experimental Example 1.
As a reagent, coml:~ound A(4-brorno-6-[3-(4-chlorophenyl)-propoxy]-5-(3-pyri(-iylmethylamino)-:3-(2H)--pyridazinone hydrochloride, 35 mg) produced by a convent.ional method was suspended in 0. 5% methylcellulose solution ("7 0 ml) in a mortar and used as a 5 mg/kg administration reagent.
This suspension was diluted two-fold and 5-fold with a vehicle (0.5% methylcellulose solution) to give 2.5 mg/kg administration reagent and 1mg/kg administration reagent (respective dose 10 m1./kg).
The vehicle (0.5% methyl.cellulose :_,olut:ion) was used as a control.
The vehicle was obtained by dissolving inethylcell.ulose (5 g, manufactured by Kishida C.hemical.Industries, Ltd.).in distilled water (1000 ml).
Each reagent (compound A) and the vehicle were orally administered repeatedly (7 tirnes in total) fr_om the day of sponge embedding at a dose of 10 ml/kg twice a day for 4 days. The sponge was removed on the next day of the final administration, and hemoglobin in the sponge was calcu::Lat.ed in the same manner as in Experimental Example 1.
Each group contained 8-9 animals and the obtained data were expressed iri mean standard error. For the evaluation of the :i 1 angiogenic effect, a Dunnett test was performed using the vehicle administration group as a control to examine significant difference.
The significance was ascribed at less than 5% risk rate.
Results-The results are shown in Fig. 8. As compared to the vehicle administration group, 2.5 mg/kg administration group and 5 mg/kg administration group showed a significant increase in hemoglobin amount.
In the 5 mg/kg administration group, the effect tended to weaken somewhat, though a significant difference was found. This is postulated to be attributable to a decrease in blood flow due to hypotensive action, since the dose (5 mg/kg) of compound A causes hypotensive action in rat, based on vasodilating action.
This has clarified that compound A shows superior angiogenesis promotion effect at a dose substantially free from hypotensive action.
Experimental Example 4 : toxicity The acute toxicity (LD50) of compound A was not less than 2 g/kg by the oral administration to rat and dog, and the compound was found to be extremely low toxic.
From the above test results, it is evident that the pyridazinone compound (I) and a salt thereof show superior angiogenesis-promoting effect and potentiation of the angiogenic effect of a drug having such effect, and are low toxic.
Example 1 (tablet) The following ingredients were mixed by a conventional method and prepared into sugar-coated tablets containing 50 mg of the active ingredient per tablet.
Compound A 10 g lactose 20 g starch 5 g magnesium stearate 0.1 g calcium carboxymethylcellulose 7 g total 42.1 g Example 2 (capsule) The following ingredients were mixed by a conventional method and filled in gelatin capsules to give capsules containing 50 mg of the active ingredient per capsule.

Coiilpound A 10 g lactose 20 g inicrocrystall_irie cellulose 10 g magnesiuni stearate 1 g total 41 g Example 3 (ointment) 'i'he following ingredients were niixed by a conventional inethod to give a 1 wt o ointmerit.
Compound A I g olive oil 20 g white petrolatum 79 g total 100 g Example 4 (aerosol suspension) Ttie following ingredients (A) were mixed arid the obtairied inixed so.lutiori was charged iri a container equipped with a valve. A
propellant (B) was pressed therein from a valve nozzle at 20"C to about 2.46 - 2.81 mg/cmz gauge pressure to give an aerosol suspension.
(A)Compound A 0.25 wt%
isopropyl iiiyristate 0.10 wt%
ethanol 26.40 wt%
(B)1,2-dichlorotetrafluoroethane and 1-chloropentafluoroethane (60-40 wt%) 73.25 wt%
Industrial Applicability The pyridazinone compound (I) and pharrnacologically acceptable salt thereof in the present invention promote angiogenesis and potentiate the angiogenic effect of a drug having such effect, and :30 are useful as ari angiogeriesis prolnoter and potentiator. 'I'tierefore, they are effective for the promotion of healing of various diseases wherein angiogenesi_s plays an iniportant role, suclr as promotion of wound healing, proinotiori of adhesiori after skin implantation, promotiori of healing after re-suture on quadruple amputation, trichogenous promotion and the like.

Claims

CLAIMS:

1. A pharmaceutical composition for promoting angiogenesis, which comprises:

a pyridazinone compound of the formula (I):
wherein R1, R2 and R3 are each independently a hydrogen atom or a lower alkyl group; X is a halogen atom, a cyano group or a hydrogen atom; Y is a halogen atom, a trifluoromethyl group or a hydrogen atom; and A is a C1-C$
alkylene group optionally substituted by a hydroxyl group, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

2. The pharmaceutical composition of claim 1, wherein R1 and R 2 are each a hydrogen atom; R3 is a hydrogen atom or C1-C4 alkyl group; X is a halogen atom; Y is a halogen atom or a hydrogen atom; and A is a C1-C5 alkylene group optionally substituted by a hydroxyl group.

3. The pharmaceutical composition of claim 1, wherein the pyridazinone compound of the formula (I) is 4-bromo-6-[3-(4-chlorophenyl)-propoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone.

4. A pharmaceutical composition for potentiating an angiogenic effect of a drug having such effect, which comprises:

a pyridazinone compound of the formula (I):
wherein R1, R2 and R3 are each independently a hydrogen atom or a lower alkyl group; X is a halogen atom, a cyano group or a hydrogen atom; Y is a halogen atom, a trifluoromethyl group or a hydrogen atom; and A is a C1-C8 alkylene group optionally substituted by a hydroxyl group, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

5. The pharmaceutical composition of claim 4, wherein R1 and R2 are each a hydrogen atom; R3 is a hydrogen atom or a C1-C4 alkyl group; X is a halogen atom; Y is a halogen atom or a hydrogen atom; and A is a C1-C5 alkylene group optionally substituted by a hydroxyl group.

6. The pharmaceutical composition of claim 4, wherein the pyridazinone compound of the formula (I) is 4-bromo-6-[3-(4-chlorophenyl)-propoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone.

7. The pharmaceutical composition of any one of claims 4 to 6, wherein the drug having the angiogenic effect is a growth factor.

8. The pharmaceutical composition of claim 7, wherein the growth factor is a basic fibroblast growth factor.

9. Use of a pyridazinone compound of the formula (I):
wherein R1, R2 and R3 are each independently a hydrogen atom or a lower alkyl group; X is a halogen atom, a cyano group or a hydrogen atom; Y is a halogen atom, a trifluoromethyl group or a hydrogen atom; and A is a C1-C8 alkylene group optionally substituted by a hydroxyl group, or a pharmaceutically acceptable salt thereof, for promoting angiogenesis.

10. The use of claim 9, wherein R1 and R2 are each a hydrogen atom; R3 is a hydrogen atom or a C1-C4 alkyl group;
X is a halogen atom; Y is a halogen atom or a hydrogen atom;
and A is a C1-C5 alkylene group optionally substituted by a hydroxyl group.

11. The use of claim 9, wherein the pyridazinone compound of the formula (I) is 4-bromo-6-[3-(4-chlorophenyl)-propoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone.

12. Use of a pyridazinone compound of the formula (I):
wherein R1, R2 and R3 are each independently a hydrogen atom or a lower alkyl group; X is a halogen atom, a cyano group or a hydrogen atom; Y is a halogen atom, a trifluoromethyl group or a hydrogen atom; and A is a C1-C8 alkylene group optionally substituted by a hydroxyl group, or a pharmaceutically acceptable salt thereof, for potentiating an angiogenic effect of a drug having such effect.

13. The use of claim 12, wherein R1 and R2 are each a hydrogen atom; R3 is a hydrogen atom or a C1-C4 alkyl group;
X is a halogen atom; Y is a halogen atom or a hydrogen atom;
and A is a C1-C5 alkylene group optionally substituted by a hydroxyl group.

14. The use of claim 12, wherein the pyridazinone compound of the formula (I) is 4-bromo-6-[3-(4-chlorophenyl)-propoxy]-5-(3-pyridylmethylamino)-3(2H)-pyridazinone.

15. The use of any one of claims 12 to 14, wherein the drug having the angiogenic effect is a growth factor.

16. The use of claim 15, wherein the growth factor is a basic fibroblast growth factor.

17. A commercial package which comprises:

the pharmaceutical composition of any one of claims 1 to 3; and a written matter associated therewith, the written matter stating that the pharmaceutical composition is used for promoting angiogenesis.

18. A commercial package which comprises:

the pharmaceutical composition of any one of claims 4 to 8; and a written matter associated therewith, the written matter stating that the pharmaceutical composition is used for potentiating an angiogenic effect of a drug having such effect.

19. The pharmaceutical composition of any one of claims 4 to 8, which further comprises the drug having the angiogenic effect.

20. The pharmaceutical composition of claim 19, wherein the drug is a growth factor.

21. The pharmaceutical composition of claim 20, wherein the growth factor is a basic fibroblast growth factor.

22. The pharmaceutical composition according to any one of claims 1 to 3, which is used for promoting wound healing, for promoting adhesion after skin implantation, for promoting healing after re-suture on quadruple amputation or for trichogenous promotion.

23. The pharmaceutical composition according to any one of claims 4 to 8, which is used for promoting wound healing, for promoting adhesion after skin implantation, for promoting healing after re-suture on quadruple amputation or for trichogenous promotion.

24. The use according to any one of claims 9 to 11, which is for promoting wound healing, for promoting adhesion after skin implantation, for promoting healing after re-suture on quadruple amputation or for trichogenous promotion.

25. The use according to any one of claims 12 to 16, which is for promoting wound healing, for promoting adhesion after skin implantation, for promoting healing after re-suture on quadruple amputation or for trichogenous promotion.