HK1119571B - Use of a benzoyl derivative of 3-aminocarbazole in manufacture of a medicament in the treatment of a disorder associated with the production of prostaglandin e2 (pge2) - Google Patents

Description

Application of benzoyl derivative of 3-aminocarbazole in preparation of medicine for treating diseases related to generation of prostaglandin E2 (PGE 2)

Technical Field

The present invention relates to the use of benzoyl derivatives of 3-aminocarbazole, as shown in Table 1 below, for the preparation of a medicament for the treatment of prostaglandins E₂(PGE₂) To produce associated diseases (e.g. inflammatory processes, pain, fever, tumours, alzheimer's disease and atherosclerosis).

Background

Para prostaglandin E₂(PGE₂) The interest arises from their function as bioregulators together with other prostaglandins produced by the arachidonic acid metabolic pathway, and as mediators of inflammationThe function of (c). Prostanoids are known to be a class of compounds including prostaglandins, thromboxanes and prostacyclins. Prostanoids are lipid modulators that have a role as local hormones for cells adjacent to their site of release. Prostanoids are mainly produced by enzymatic oxidation of arachidonic acid by cyclooxygenase activation. Cyclooxygenase (prostaglandin G/H synthase) catalysis of the sequential production of PGG from arachidonic acid₂And PGH₂. Then PGH₂Conversion to different prostanoids by various specific enzymes. This pathway forms prostaglandin D₂(PGD₂) Prostaglandin E₂(PGE₂) Prostaglandin F_2α(PGF_2α) Prostaglandin I₂(PGI₂) And thromboxane A₂(TXA₂)。

Prostanoids do not accumulate except in the seminal fluid. Following various stimuli (inflammation, immunity, hormonal stimuli, ultraviolet light, tumoral agents and mechanical stimulation), they are synthesized and released into the extracellular space from where they pass into the plasma, urine and other biological fluids.

Prostanoids play an important role in mechanisms that protect organ function and the integrity of the body. This can be confirmed by the fact that: their cytoprotective function in the gastrointestinal tract, their regulation of renal function and microcirculation, their regulation of platelet aggregation and blood coagulation, their association with the differentiation of immune cells and the repair of wounds, and their association with bone metabolism and ovulation.

In particular, it must be emphasized that PGI₂For maintaining vascular tone and preventing thromboembolism and atherosclerosis at the endothelial level, and a metabolite of 15d-PGJ₂PGD of₂Are important, among them 15d-PGJ₂Can activate nuclear PPAR_γ(peroxisome proliferator-activated receptor-. gamma.) receptors exhibit anti-inflammatory effects (Inoue et al, "Feedback control of cycloxygenase-2 expression of deep PPARga)mma″，J.Biol.Chem.2000，275(36)：28028-28032)。

Prostaglandins are therefore bioregulators, but they are also important mediators of inflammation and other diseases.

In particular, PGE₂Is abundant at sites of inflammation and plays a role in the pathology of various acute and chronic inflammations, such as edema, erythema formation, inflammatory pain, joint inflammation and fever. In fact, PGE₂Are powerful pro-inflammatory and analgesic agents. anti-PGE₂The antibody has anti-inflammatory activity and is deficient in PGE₂The recipient animal shows a reduced response to inflammatory stimuli (Portanova et al, "Selective neutralization of prostagladin 2 blocks in fluorescence, hypergesia, and interleukin 6production in vivo", J.Exp.Med.1996, 184 (3): 883; Ueno et al, "Major roles of prostad receptors IP and EP (3) induced enhancement of pain perception", biochem. Pharmacol.2001, (62) (2): 157 and 160), and does not produce a fever response to thermal stimuli (Uhikubi et al, "Paired immune response in microbial degradation expression vector expression, 3", Nature 1998: 284). Because of their inhibitory effects on cyclooxygenase 1 and 2 (FitzGerald and Patrono, "The coxibs, selective inhibitors of cyclooxingene-2", N.Engl. J.Med.2001, 345 (6): 433-₂、PGD₂、PGF_2α、PGI₂And TXA₂) To reduce symptoms associated with inflammation.

In particular, COX-2-selective drugs, now available on the market, can reduce gastrointestinal toxicity compared to conventional non-steroidal anti-inflammatory (NSAI) drugs. However, these COX-2-selective drugs would decrease vascular Prostacyclin (PGI)₂Which is produced mainly by COX-2), alters the normal balance between pro-thrombotic and anti-thrombotic eicosanoids, such thatIt is prone to pre-thrombotic eicosanoids (TXA)₂Which is produced primarily by COX-1) and increases the risk of developing thrombotic cardiovascular events (s.malhotra, MD, DM; shafiq, MD; pandhi, MD, Medscape General Medicine6(1), 2004; mukherjee and e.j.topol, cardiovacular riskand COX-2 inhibitors, artritis res.ther.2003, 5: 8-11-2002).

Patent applications WO 01/07409a1 and WO02/096902a1 relate to a number of carbazole derivatives represented by the general formula in general, comprising A3-aminocarbazole base group substituted in any position, including a nitrogen atom, and one or more aliphatic and/or aromatic organic groups or residues. According to these documents, the carbazole derivatives described above are capable of selectively binding to the human Y5 receptor and modulating its activity. Thus, these carbazole derivatives may be useful in the treatment of hunger and metabolic disorders such as obesity, bulimia nervosa, anorexia nervosa, sleep disorders, morphine dependence and seizures.

Disclosure of Invention

It has now surprisingly been found that certain benzoyl derivatives of 3-aminocarbazole are capable of selectively inhibiting prostaglandin E₂(PGE₂) Is generated.

Compounds useful according to the invention are capable of reducing PGE₂Is thus generated in PGE₂Active in all pathological conditions that act as mediators (e.g. pain, fever and inflammatory response).

Compounds useful according to the invention selectively inhibit PGE₂And (4) synthesizing. Selective inhibition of PGE₂It is advantageous for inhibiting powerful mediators of inflammation, pain and fever, while allowing other prostanoids (e.g., PGF's) to be produced from the arachidonic acid cascade_2α、TXA₂、PGI₂And PGD₂) Is produced without change. Therefore, all mechanisms that protect organ function and body integrity are not affected, whereThe mechanism is typical of other prostanoids.

Similar to conventional non-steroidal anti-inflammatory drugs, the compounds useful according to the invention have anti-inflammatory, antipyretic and analgesic properties and are therefore active in diseases such as inflammation, pain, fever, rheumatoid arthritis and arthropathy. In addition to this, since PGE is known from the literature₂The effects on tumors, Alzheimer's disease and atherosclerosis, the compounds useful according to the invention can also be used in the prevention and treatment of these diseases.

Advantageously, however, the compounds useful according to the invention have little negative effect compared to NSAIs and COX-2 selective drugs that inhibit cyclooxygenase, which are indistinguishable from various prostanoids in inhibiting cyclooxygenase. In particular, the compounds useful according to the invention reduce gastrointestinal, renal and vascular toxicity.

Detailed Description

In a first aspect, the present invention thus relates to the use of a compound selected from the group comprising benzoyl derivatives of 3-aminocarbazole of table 1 below for the preparation of a medicament for the prophylactic or therapeutic treatment of a disease selected from the group comprising inflammatory processes, pain, fever, tumours, alzheimer's disease and atherosclerosis.

In addition, in a second aspect, the present invention relates to a method for the prophylactic or therapeutic treatment of a disease selected from the group comprising inflammatory processes, pain, fever, tumors, alzheimer's disease and atherosclerosis, wherein a therapeutically effective amount of a compound selected from the group comprising benzoyl derivatives of 3-aminocarbazole of table 1 below is administered to an individual.

Typical examples of inflammatory processes that may benefit from the present invention are: edema, erythema, inflammation of joints, rheumatoid arthritis, arthropathy, and the like.

Typical examples of tumors that may benefit from the present invention are carcinomas and adenocarcinomas of the colon-rectum and lung.

Typically, the benzoyl derivative of 3-aminocarbazole according to the invention is administered to a mammal. Preferably, they are administered to humans.

The compounds useful according to the invention are selected from the group comprising benzoyl derivatives of 3-aminocarbazole of general formula (I) shown in table 1 below:

TABLE 1

Compound (I)	R1	R2	R3	R4	R5
						1	Cl	H	H	H	H
2	CH3	H	H	H	H
						3	Br	H	H	H	H
4	I	H	H	H	H
						5	NO2	H	H	H	H
6	Cl	H	H	H	Cl
						7	Cl	H	H	NO2	H
8	Cl	H	H	Cl	H

The compounds useful according to the invention can be prepared according to methods known from the literature, for example as described in patent application WO02/096902A 1.

Preferably, the medicament according to the invention is prepared in a suitable administration form.

Examples of suitable administration forms are tablets, capsules, coated tablets, granules, solutions and syrups for oral administration, creams, ointments and medicated ointments for topical administration, suppositories for rectal administration and sterile solutions for administration by injection, aerosol or ocular means.

Advantageously, these dosage forms are formulated in a manner that ensures controlled release of the compounds of table 1 over time. In fact, depending on the nature of the treatment, the desired release time may be very short, normal or extended.

These administration forms may also contain other conventional ingredients, such as preservatives, stabilizers, surfactants, buffers, salts for regulating the osmotic pressure, emulsifiers, sweeteners, colorants, flavors, and the like.

Furthermore, the dosage form according to the invention may comprise other pharmaceutically active ingredients which can be used for simultaneous administration, if required for a particular treatment.

In the above dosage forms, the amount of the compound according to the invention may vary within wide limits depending on known factors, such as the type of disease to be treated, the severity of the disease, the weight of the patient, the dosage form, the chosen route of administration, the number of daily administrations and the potency of the chosen compound. Nevertheless, the optimal amount can be readily determined by one skilled in the art in a routine manner.

Typical amounts of the compounds in the dosage form for administration according to the invention are those which ensure an administration level of from 0.0001 to 100 mg/kg/day. Even more preferably 0.01 and 10 mg/kg/day.

Dosage forms for administration of the pharmaceutical compositions according to the invention may be prepared by techniques known in pharmaceutical chemistry, including mixing, granulating, tabletting, dissolving, sterilizing, and the like.

Detailed Description

The following experimental section will explain the invention in more detail, without limiting it in any way.

Experimental part

In vitro activity assay

This assay was used to evaluate the inhibition of PGE by each compound tested₂Generation and targeting of PGF_2αThe ability to produce selectivity.

The cell line A549, human lung adenocarcinoma, was used against an proinflammatory cytokine (e.g., IL-1)_β) Is particularly sensitive to and responds to that stimulus (particularly the two Prostaglandins (PGEs)₂And PGF_2α) Production and release) are particularly sensitive (Thoren s., Jakobsson p.j. "coordination up-and down-regulation of glassutathione-dependent prostaglandin E synthase andcyclooxygenase-2in A549 cells.Inhibition by NS-398 andleukotriene C4″，Eur.J.Biochem.2000，267(21)：6428-6434))。

With IL-1_β(1ng/ml) the cells were stimulated simultaneously with 5% CO at 37 ℃₂In an incubator with the test compound in a suitable medium (DMEM-Dulbecco's modified Eagles medium) enriched with 5% bovine fetal serum and L-glutamic acid (finally 4 mM) for 18 hours.

After the culture, PGE produced and released into the supernatant was determined by using EIA kit (manufactured and sold by Cayman Chemicals, Ann Arbor, MI, USA)₂And PGF_2αThe amount of (c).

Indomethacin (Sigma-Aldrich) was used as a reference compound, a non-steroidal anti-inflammatory drug with the same inhibitory effect on both prostanoids tested.

As comparative compounds, compounds A, B and C having the same structure of the above general formula (I) and substituents shown in Table 2 were used. The preparation of these comparative compounds is analogous to the preparation of the compounds according to the invention.

TABLE 2

Compound (I)	R1	R2	R3	R4	R5
						A	OCH3	H	H	H	H
B	H	H	H	H	H
						C	H	H	CH3	H	H

The results shown in Table 3 are expressed for PGE at a concentration of 10 μm₂And PGF_2αPercent inhibition of production of (a).

TABLE 3

Compound (I)	Inhibition at 10 μm%
			PGE2	PGF2α
1	82	16

2	83	44
			3	81	0
4	84	0
			5	27	0
6	89	51
			7	70	0
8	93	37

A	98	95
			B	46	40
C	43	49
			Indometacin	100	100

By way of example only, table 4 shows the pIC of some compounds according to the invention₅₀Value wherein pIC₅₀Is shown as IC₅₀Inverse of logarithm of (1), IC₅₀Showing the PGE in stimulated cells not treated with the compound₂Or PGF_2αThe concentration of the compound that inhibits 50% of the production.

TABLE 4

Compound (I)	pIC50
			PGE2	PGF2α
1	5.7	4.1
			2	5.3	4.3
3	5.5	＜4
			4	5.4	＜4
Indometacin	8.3	8.6

In vivo Activity assay

This test makes it possible to evaluate the activity of the compounds according to the invention in a nociceptive test of inflammatory origin. Compound 1 was used for this purpose in a test model that elicits painful behavior in mice by acetic acid (Stock, J.L. et al, J.Clin.Inv.2001, 107: 325-331). Female CD-1 mice weighing 25-30g were used for this experiment.

These animals were treated orally with compound 1(30mg/kg) suspended in Methylcellulose (MTC). Control animals were treated orally with vehicle (MTC) alone.

One hour after treatment, acetic acid (0.7% v/v physiological solution, 16 μ l/g body weight) was injected intraperitoneally into the animals to induce inflammatory pain and to examine the effect of the treatment on nociceptive responses.

The number of stretches was determined immediately after the administration of acetic acid and within the following 20 minutes, and represents an evaluation parameter for the nociceptive response. The results obtained (mean. + -. SD) are shown in Table 5.

TABLE 5

Compound (I)	Number of stretches in 20 minutes
		MTC	58±5
MTC + Compound 1	40±5

Assay for human primary endothelial cells (HUVEC)

The assay was used to evaluate the compounds in Table 2 for PGI inhibition₂The ability to generate.

Lack of inhibitory activity on these prostaglandins ensures maintenance of PGI₂And provides useful pharmacological information that it has no deleterious effect on the endothelium.

Compound 1 was used in this assay.

The effect of test compounds was evaluated in HUVEC cells under both basal and stimulatory conditions (J.Immunol.1989 June 1; 142 (11): 3993-9).

The results are shown in table 6, which are expressed as percent inhibition compared to control enzyme activity.

Indomethacin was used as a control compound.

TABLE 6

Compound (10 μm)	PGI2Testing of secretion	% inhibition
			1	Foundation (HUVEC)	0
1	Stimulus (HUVEC)	0
			Indometacin	Stimulus (HUVEC)	100

Claims (6)

1. A compound selected from the group consisting of benzoyl derivatives of 3-aminocarbazole of general formula (I) shown in Table 1 below

TABLE 1

Compound (I) R₁ R₂ R₃ R₄ R₅ 1 Cl H H H H 2 CH₃ H H H H 3 Br H H H H 4 I H H H H 5 NO₂ H H H H 6 Cl H H H Cl 7 Cl H H NO₂ H 8 Cl H H Cl H

For the preparation of a medicament for the prophylactic or therapeutic treatment of a disease selected from the group comprising inflammatory processes, fever, tumors, Alzheimer's disease and atherosclerosis with PGE₂The use of the compound in the preparation of medicaments for treating diseases.

2. Use according to claim 1, wherein the inflammatory process is selected from the group comprising inflammatory pain, edema, erythema and arthropathy.

3. Use according to claim 2, wherein the arthropathy is an inflammation of the joint.

4. Use according to claim 3, wherein the inflammation of the joints is rheumatoid arthritis.

5. Use according to claim 1, wherein the tumour is selected from the group comprising carcinomas.

6. Use according to claim 5, wherein the cancer is selected from the group consisting of colon-rectal and lung adenocarcinoma.