KR20050074493A - Derivatives of n-phenylanthranlic acid and 2-benzimidazolon as potassium channel and/or cortical neuron activity modulators - Google Patents

Abstract

Compounds, compositions and methods are provided which are useful in the treatment of diseases through the modulation of potassium ion flux through voltage-dependent potassium channels and/or depressing cortical neuron activity. More particularly, the invention provides derivatives of N-phenylanthranilic acid or 2-benzimidazolone, compositions and methods that are useful in the treatment of central or peripheral nervous system disorders and as neuroprotective agents.

Description

Derivatives Of N-Phenylanthranlic Acid And 2-Benzimidazolon As Potassium Channel And / Or Cortical Neuron Activity Modulators} Useful as Potassium Channel and / or Cortical Neuron Action Modulators N-Phenylanthranilic Acid and / or 2-Benzimidazolone

The present invention relates to the field of medicament and to derivatives of N-phenylanthranilic acid and / or 2-benzimidazolone for the pathological treatment associated with pathogens, in particular potassium ion flux via voltage-dependent potassium channels and / or cortical neuronal action. It is about.

Ion channels are cellular proteins that regulate ions, including calcium, potassium, sodium, and chloride, into and out of cells. These channels are present in all animal cells and affect processes such as neurotransmission, muscle contraction, and cell detachment. Among ion channels, potassium channels are the most common and diverse and are found in various animal cells such as nerves, muscles, glands, immunity, regenerative and epithelial tissue. These channels allow potassium to enter and exit the cell under certain conditions. For example, outward flow of potassium ions upon opening of these channels makes the interior of the cell more negative and impedes the polarization voltage applied to the cell. These channels are regulated by calcium sensitivity, voltage-gating, second messenger, specific cell ligand and ATP-sensitivity, for example.

Potassium channels are involved in many physiological processes, including heart rate, arterial dilatation, insulin secretion, regulation of neuronal excitability, and regulation of the transport of kidney electrolytes. Potassium channels consist of alpha subunits belonging to at least eight families based on expected structural and functional similarities (Wei et al., Neuropharmacology 35 (7): 805-829 (1997)). Three of these families (Kv, eag-related, and KQT) share the common motif of the six transmembrane domains and are gated primarily by voltage. The two other families, CNG and SK / IK, also contain this motif, but are gated by cyclic nucleotides and calcium, respectively. The other three families of potassium channel alpha subunits have different patterns of transmembrane domains. The Slo family potassium channel, or BK channel, has seven transmembrane domains (Meera et al., Proc. Natl. Acad. Sci. USA 94 (25): 14066-71 (1997)), both by voltage and calcium Or gated by pH (Schreiber et al., J. Biol. Chem. 273: 3509-16 (1998)). Another family, the inward rectifier potassium channel (Kir) belongs to a structural family containing two transmembrane domains, and the eighth functionally diverse family (TP or "2-pore") is the two tandems of this inward rectifier motif. It contains repetitions.

The potassium channel is usually formed of four alpha subunits, and can be homogeneous (consisting of the same alpha subunit) or heterologous (consisting of two or more different forms of alpha subunits). In addition, potassium channels consisting of Kv, KQT and Slo or BK subunits are often found to contain additional, structurally different adjuvants or beta, subunits. These subunits do not form the potassium channel itself, but instead act as auxiliary subunits to alter the functional properties of the channel formed by the alpha subunit. For example, Kv beta subunits are cytoplasmic and are known to increase surface expression of Kv channels and / or alter inactivation kinetics of channels (Heinemann et al., J. Physiol. 493: 625-633 (1996 Shi et al., Neuron 16 (4): 843-852 (1996)). In another example, the KQT family beta subunit, minK, primarily alters activation kinetics (Sanguinetti et al., Nature 384: 80-83 (1996)).

Slo or BK potassium channels are large conductance potassium channels found in a wide range of tissues in both the central and peripheral nervous systems. They play an important role in regulating processes such as neuronal integration, muscle contraction, and hormone secretion. They may also be involved in such processes as lymphocyte differentiation and cell proliferation, spermatogenic differentiation and sperm motility. Three alpha subunits of the Slo family, namely Slol, Slo2 and Slo3, were cloned (Butler et al., Science 261: 221-224 (1993); Schreiber et al., J. Biol. Chem., 273: 3509-16 (1998); and Joiner et al., Nature Neurosci. 1: 462-469 (1998). These Slo family members have been found to be voltage-formed and / or calcium gated and / or regulated by internal cell pH.

Certain members of the Kv family of potassium channels have recently been renamed (see Biervert, et al., Science 279: 403-406 (1998)). KvLQT1 was renamed KCNQ1 and KvLQT1-related channels (KvLR1 and KvLR2) were renamed KCNQ2 and KCNQ3, respectively. Recently, additional members of the KCNQ subfamily have been identified. For example, KCNQ4 has been identified as a channel expressed in perceptual outer hair cells (Kubisch, et al., Cell 96 (3): 437-446 (1999)). KCNQ5 (Kananura et al., Neuroreport 11 (9): 2063 (2000)), KCNQ2 / 3 (Main et al. Mol. Pharmacol. 58: 253-62 (2000), KCNQ3 / 5 (Wickenden et al. Br. J. Phanna 132: 381 (2001)) and KCNQ6 have also been described recently.

Benign family neonatal convulsions ("BFNC"), a type of neurological-specific potassium channel associated with idiopathic generalized epilepsy, have been shown (Leppert, et al., Nature 337: 647-648 (1989)). These channels are connected with M-current channels (see Wang, et al., Science 282: 1890-1893 (1998)). The discovery and characterization of these channels and currents provide clues to how these voltage-dependent (Kv) potassium channels work in different environments and how they respond to different activation mechanisms. This information will identify modulators of KCNQ2 and KCNQ3 potassium channels or M-currents and their use as therapeutic agents.

A potassium channel opener of great interest is retigabine (N- (2-amino-4- (4-fluorobenzylamino) -phenyl) carbamic acid ethyl ester). Retigabine was first described in European Patent No. 554,543. Compounds associated with retigabine have also been proposed for use as potassium channel modulators (see US patent application 10 / 022,579).

Retigabine has very high selectivity for potassium channels consisting of subunits KCNQ2 and KCNQ3. Retigabine also activates homologous multichannels containing only subunit KCNQ2. Only the minimum voltage-dependent current can be measured in the cell, which expresses only homologous channels from the KCNQ3 subunit (see US Pat. No. 6,472,165).

US patent application Ser. No. 10 / 075,521 describes 2,4-disubstituted pyrimidine-5-carboxamide derivatives as KCNQ potassium channel modulators.

US Patent Application No. 10 / 160,582 describes cinnamid derivatives as KCNQ potassium channel modulators.

U.S. Patents 5,565,483 and U.S. Patent Applications 10 / 312,123, 10 / 075,703 and 10 / 075,522 describe 3-substituted auxindol as KCNQ potassium channel modulator.

U.S. Patent 5,384,330 describes 1,2,4-triamino-benzene derivatives as KCNQ potassium channel regulators.

US 6,593,349 describes bisarylamine derivatives as KCNQ potassium channel modulators. The 2 aryl groups of the compounds described in US Pat. No. 6,593,349 are pyridine derivatives and 5-membered heterocycles.

A serious disadvantage of known KCNQ potassium channel modulators is that it is generally difficult to make complex multistage synthesis where they are required, and in some cases these modulators are nonspecific or even toxic.

Therefore, this need is widely recognized and it is very advantageous to have new and effective potassium channel regulators that are readily available and easy to synthesize.

The present invention is described in detail below with reference to the accompanying drawings, but is only an example. DETAILED DESCRIPTION With reference to the drawings in detail, the specific details shown are by way of example only, and merely illustrate by way of example only, and provide a description of the principles and concepts of the present invention in the most useful and easily understood. In this regard, the present invention has not been described in detail as necessary to fundamentally understand the present invention, and the results shown in the drawings, which are obvious to those skilled in the art, indicate how the present invention is actually implemented.

In the drawing,

1A-1D show the leftward conversion of the activation curve induced by meclofennamic acid (Compound 1) and 1-EBIO (Compound 10),

2A-2C show the results of an increase in KCNQ2 / 3 current induced by meclofennamic acid (Compound 1) and 1-EBIO (Compound 10),

3A-3E show the effect of meclofennamic acid (Compound 1) and 1-EBIO (Compound 10) on the deactivation process of KCNQ2 / 3 channel,

4A-4B show inhibition of neuronal action by meclofennamic acid (Compound 1),

5 shows the results demonstrating the neuroprotective effect of meclofennamic acid (Compound 1) from electric shock-induced fainting in adult rats,

6A-6C show an increase in KCNQ2 / 3 current by Diclofenac (Compound 2),

7A-7C show the effect of compound 6 on KCNQ2 / 3 current,

8A-8B show inhibition of neuronal action induced by compound 6,

9A-9C show the inhibitory effect of different concentrations of Compound 6 on spontaneous neuronal action,

10A-10C show the effect of compound 5 on neuronal action and KCNQ2 / 3 current,

11A-11B show an increase in KCNQ2 / 3 current induced by compound 3,

12A-12C show the effect of compound 4 on neuronal action and KCNQ2 / 3 current,

13 shows the effect of compound 9 on spontaneous neuronal action,

14A-14D show the effect of compound 7 on KCNQ2 / 3 channel and neuronal action,

15A-15B show the effect of compound 8 on induced and spontaneous neuronal action,

16A-16D show the selectivity of meclofennamic acid (Compound 1) for KCNQ2 and KCNQ3 homologous channels expressed in CHO cells, and

17 shows the chemical structural formula of compound 1-10.

The present invention generally provides compounds that are effective potassium channel modulators, particularly voltage-dependent potassium channels such as KCNQ2 channels, KCNQ3 channels and KCNQ2 / 3 channels. The present invention also provides compounds that are generally effective at inhibiting dermal neuronal action. Compounds of the present invention are derivatives of N-phenylanthranilic acid or 2-benzimidazolone. Some of the compounds of the present invention are known and readily available. Some of the compounds of the present invention are synthesized in several (usually one or two) high yield steps from novel but readily available starting materials.

The subject matter of the present invention is a method of regulating (preferably opening) a voltage-dependent potassium channel, N-phenylanthranilic acid, N-phenylanthranilic acid derivatives, 2-benzimidazolone and 2-benzimidazolone derivatives, Or administering to a patient in need thereof a therapeutically effective amount of a compound selected from the group consisting of pharmaceutically acceptable salts thereof.

Preferably, the compound has general formula (I) or (II) described later. Also preferably, the regulated voltage-dependent potassium channel is a KCNQ2 channel, a KCNQ3 channel and / or a KCNQ2 / 3 channel.

Another aspect of the present invention is a method for inhibiting dermal neuronal action, N-phenylanthranilic acid, N-phenylanthranilic acid derivatives, 2-benzimidazolone and 2-benzimidazolone derivatives, or pharmaceutical To provide a method comprising administering to a patient in need thereof a therapeutically effective amount of a compound selected from the group consisting of salts thereof.

Preferably, this compound has general formula (I) or (II) mentioned later.

Another aspect of the present invention is to provide a pharmaceutical composition for treating or preventing diseases or diseases in the central or peripheral nervous system, which is advantageous for regulating voltage-dependent potassium channels and / or inhibiting cortical neuronal action. Wherein the pharmaceutical composition is selected from the group consisting of N-phenylanthranilic acid, N-phenylanthranilic acid derivatives, 2-benzimidazolone and 2-benzimidazolone derivatives, or pharmaceutically acceptable salts thereof, as active ingredients Compound.

Preferably, this compound has general formula (I) or (II) mentioned later. Also preferably, the regulated voltage-dependent potassium channel is a KCNQ2 channel, a KCNQ3 channel and / or a KCNQ2 / 3 channel.

Formulas (I) and (II) according to the invention are as follows and include salts thereof:

(I) (II)

Where

Z is an A-G (= K) -X-Y group,

A is alkyl or absent,

G is selected from the group consisting of carbon, sulfur and substituted or unsubstituted phosphorus,

K is selected from the group consisting of oxygen and sulfur,

X is selected or absent from the group consisting of substituted or unsubstituted nitrogen, oxygen and sulfur,

Y is selected from the group consisting of hydrogen, alkyl, hydroxyalkyl, cycloalkyl, aryl and polyalkylene glycol moieties,

Each of Q and W is independently selected from the group consisting of substituted or unsubstituted nitrogen, oxygen, sulfur and carbon,

D is selected from the group consisting of oxygen and sulfur,

R ¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl and aryl,

R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are each independently hydrogen, alkyl, hydroxyalkyl, Trihaloalkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, Cyano, nitro, amino and -NR ¹⁵ R ¹⁶ , or at least two of R ² , R ³ , R ⁴ , R ⁵ and R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ At least two of and / or at least two of R ¹¹ , R ¹² , R ¹³ and R ^{14 form a} 5- or 6-membered aromatic, heteroaromatic, cycloaliphatic or heteroalicyclic ring,

R ¹⁵ and R ¹⁶ are each independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, carbonyl and sulfonyl, or R ¹⁵ and R ¹⁶ represent a 5- or 6-membered heteroalicyclic ring. Forming,

Here, when phosphorus and / or nitrogen are substituted, the substituent is selected from the group consisting of alkyl, cycloalkyl and aryl.

According to the invention, the compounds of the invention have the general formula (I). When the compound of the present invention has the general formula (I), Y is preferably selected from hydroxyalkyl and polyalkylene glycol residues. Preferably, the polyalkylene glycol residues have the general formula (III) :

[O- (CH ₂ ) m] n-OR ¹⁷ (III)

Wherein m and n are each independently an integer from 1 to 10 and R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl. According to a feature of the invention, R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are each independently selected from the group consisting of hydrogen, alkyl, halo and trihaloalkyl, R ⁷ , R ⁸ , R ⁹ And R ¹⁰ are each hydrogen.

According to another embodiment of the present invention, the compound of the present invention has general formula (II). When the compound of the present invention has the general formula (II), preferably Q and W are substituted or unsubstituted nitrogen, and D is oxygen, and more preferably Q is nitrogen substituted.

According to a preferred embodiment of the invention, the compound is selected from the group consisting of the following general formulas 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 and includes salts thereof:

In accordance with a feature of the invention, regulating voltage-dependent potassium channels and / or inhibiting cortical neuronal action is epilepsy, ischemic seizures, migraine, ataxia, muscle spasms, neuralgia, Alzheimer's disease, Parkinson's disease, old age For the treatment of diseases or disorders selected from the group consisting of amnesia, learning deficiency, bipolar disorder, trigeminal neuralgia, cramps, mood disorders, mental illness, brain tumors, hearing and vision loss, anxiety and motor neuron disease.

According to a feature of the invention, the compounds of the invention may be administered intranasally, subcutaneously, intravenously, intramuscularly, parenterally, oral, topical, intradermal, bronchial, oral, buccal, sublingual, anal and mucosal.

Accordingly, the compounds of the present invention are part of a pharmaceutical composition further comprising a pharmaceutically acceptable salt.

According to a feature of the invention, the pharmaceutical composition of the invention comprises an agent selected from the group consisting of antibacterial agents, antioxidants, buffers, bulking agents, anti-inflammatory agents, antiviral agents, chemotherapeutic agents and antihistamines.

In accordance with a feature of the invention, the pharmaceutical composition is packaged and printed on the packaging for use in the treatment or prevention of a disease or condition associated with the altered action of voltage-dependent potassium channels. Preferably, the disease or condition is epilepsy, ischemic attack, migraine, ataxia, muscle cramps, neuralgia, Alzheimer's disease, Parkinson's disease, old-fashioned amnesia, learning deficiency, bipolar disorder, trigeminal neuralgia, cramps, mood Disorders, mental illness, brain tumors, hearing and vision loss, anxiety and motor neuron disorders.

Another aspect of the invention provides a novel compound of formula (IV) or a pharmaceutically acceptable salt thereof:

(IV)

Where

Z is an A-G (= K) -X-Y group,

A is alkyl or absent,

G is selected from the group consisting of carbon, sulfur and substituted or unsubstituted phosphorus,

K is selected from the group consisting of oxygen and sulfur,

X is selected or absent from the group consisting of substituted or unsubstituted nitrogen, oxygen and sulfur,

Y is selected from the group consisting of hydroxyalkyl and polyalkylene glycol moieties,

R ¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl and aryl,

R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are independently hydrogen, alkyl, hydroxyalkyl, trihaloalkyl, cycloalkyl, alkenyl, alkynyl , Aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro, amino and -NR ¹⁵ R ¹⁶ At least two of R ² , R ³ , R ⁴ , R ⁵ and R ⁶ and / or at least two of R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are 5-membered or 6- To form an aromatic, heteroaromatic, cycloaliphatic or heteroalicyclic ring,

R ¹⁵ and R ¹⁶ are independently of each other selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, carbonyl and sulfonyl, or R ¹⁵ and R ^{16 form} a 5- or 6-membered heteroalicyclic ring and,

Where phosphorus and / or nitrogen are substituted, the substituent is selected from the group consisting of alkyl, cycloalkyl and aryl.

According to a feature of the invention, the polyalkylene glycol moiety of the mycobacterial compound of the invention has the following general formula (V):

[O- (CH ₂ ) m] n-OR ¹⁷ (V)

Wherein m and n are each independently an integer from 1 to 10 and R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl.

According to another feature of the novel compounds of the invention, G is carbon, K is oxygen, and R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are independently hydrogen, alkyl, halo and trihaloalkyl And R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each hydrogen.

According to a preferred embodiment of the invention, the novel compounds of the invention are selected from the group consisting of the following compounds 3, 4, 5, 6, 7, 8 and 9 or pharmaceutically acceptable salts thereof:

Another aspect of the present invention is to provide a pharmaceutical composition comprising a compound of the present invention having the general formula (IV) as an active ingredient.

It is a feature of the present invention to provide a pharmaceutical composition having any one of compounds 3, 4, 5, 6, 7, 8 and / or 9.

Another aspect of the present invention is to provide a method for synthesizing a compound of formula (IV). The method comprises obtaining N-phenylanthranilic acid or a derivative thereof and reacting N-phenylanthranilic acid or a derivative thereof with hydroxyalkyl or polyalkylene glycol having a reactive group at the end, wherein the reactive group is N- An ester bond can be formed with phenylanthranilic acid or a derivative thereof.

The ester bond is preferably selected from the group consisting of carboxy ester bonds, carboxy amide bonds, carboxy thioester bonds, S-carboxy thioester bonds and S-carboxy amide bonds, wherein the reactive groups are hydroxy, amine and thiohydroxy It is preferably selected from the group consisting of.

N-phenylanthranilic acid or derivatives thereof preferably have the following general formula (VI):

(VI)

Wherein A, G, K, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are as defined above.

Preferably, G is carbon, K is oxygen, R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are each independently selected from the group consisting of hydrogen, alkyl, halo and trihaloalkyl, R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each hydrogen.

Polyalkylene glycols having reactive groups at the end have the general formula (VII)

V- [O- (CH ₂ ) m] n-OR ¹⁷ (VII)

Where

V is hydroxy, amine or thiohydroxy,

m and n are each independently an integer of 1 to 10,

R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl.

The present invention successfully solves the disadvantages of known configurations by providing compounds that act to modulate potassium channels and / or inhibit cortical action, and are generally commercially available, and / or synthesized relatively easily.

Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. The same or similar methods and materials as described herein can be used in the testing or practical use of the present invention, suitable methods and materials are described below. In case of dispute, the patent specification, including patent definitions, is for reference. In addition, the materials, methods, and examples are illustrative and not intended to limit the invention.

Description of the preferred embodiment

The present invention provides compounds that are generally useful for the regulation of potassium ion flux and / or inhibit cortical neuronal action via voltage-dependent potassium channels, particularly KCNQ2 channels, KCNQ3 channels and KCNQ2 / 3 channels.

The principles and uses of the invention may be more readily understood by reference to the examples and the following description. Before describing at least one embodiment of the invention in detail, the invention is not limited by the embodiment. The invention is capable of other embodiments and of being practiced in various ways. In addition, the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting.

The subject matter of the present invention is a method of regulating (preferably opening) a voltage-dependent potassium channel, N-phenylanthranilic acid, N-phenylanthranilic acid derivatives, 2-benzimidazolone and 2-benzimidazolone derivatives, Or administering to a patient in need thereof a therapeutically effective amount of a pharmaceutically acceptable salt thereof.

As used herein, the term "derivative" refers to the chemical conversion, improvement, or alteration of a molecule or part thereof while maintaining its original function in at least one aspect.

The compound has general formula (I) or (II). The regulated voltage-dependent potassium channel is preferably a KCNQ2 channel, a KCNQ3 channel and / or a KCNQ2 / 3 channel.

Another aspect of the present invention is a method for inhibiting the action of dermal neuronal, N-phenylanthranilic acid, N-phenylanthranilic acid derivative, 2-benzimidazolone, 2-benzimidazolone derivative or pharmaceutically acceptable It is to provide a method comprising administering to a patient in need thereof a therapeutically effective amount of the salt.

Preferably, this compound has formula (I) or (II).

It is yet another aspect of the present invention to provide a pharmaceutical composition for treating or preventing a disease or condition in which it is advantageous to regulate voltage-dependent potassium channels and / or inhibit cortical neuronal action, wherein the pharmaceutical composition is active Components include N-phenylanthranilic acid, N-phenylanthranilic acid derivatives, 2-benzimidazolone, 2-benzimidazolone derivatives or pharmaceutically acceptable salts thereof.

Preferably, this compound has formula (I) or (II). Also preferably, the regulated voltage-dependent potassium channel is a KCNQ2 channel, a KCNQ3 channel and / or a KCNQ2 / 3 channel.

Formulas (I) and (II) according to the invention are as follows and include salts thereof:

(I) (II)

Where

Z is an A-G (= K) -X-Y group,

A is alkyl or absent,

G is selected from the group consisting of carbon, sulfur and substituted or unsubstituted phosphorus,

K is selected from the group consisting of oxygen and sulfur,

X is selected or absent from the group consisting of substituted or unsubstituted nitrogen, oxygen and sulfur,

Y is selected from the group consisting of hydrogen, alkyl, hydroxyalkyl, cycloalkyl, aryl and polyalkylene glycol moieties,

Each of Q and W is independently selected from the group consisting of substituted or unsubstituted nitrogen, oxygen, sulfur and carbon,

D is selected from the group consisting of oxygen and sulfur,

R ¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl and aryl,

R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are independently hydrogen, alkyl, hydroxyalkyl, tri Haloalkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, sia No, nitro, amino and —NR ¹⁵ R ¹⁶ , while at least two of R ² , R ³ , R ⁴ , R ⁵ and R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ At least two and / or at least two of R ¹¹ , R ¹² , R ¹³ and R ^{14 form a} 5- or 6-membered aromatic, heteroaromatic, cycloaliphatic or heteroalicyclic ring,

R ¹⁵ and R ¹⁶ are independently of each other selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, carbonyl and sulfonyl, or R ¹⁵ and R ^{16 form} a 5- or 6-membered heteroalicyclic ring and,

Here, when phosphorus and / or nitrogen are substituted, the substituent is alkyl, cycloalkyl or aryl.

According to an embodiment of the present invention, the compound of the present invention has general formula (I). When the compound of the present invention has general formula (I), Y is preferably selected from hydroxyalkyl and polyalkylene glycol moieties.

Preferably, the polyalkylene glycol moiety has the general formula (III)

[O- (CH ₂ ) m] n-OR ¹⁷ (III)

Wherein m and n are each independently an integer from 1 to 10 and R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl. According to a feature of the invention, R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are each independently selected from the group consisting of hydrogen, alkyl, halo and trihaloalkyl, R ⁷ , R ⁸ , R ⁹ And R ¹⁰ are each hydrogen.

According to another embodiment of the present invention, the compound of the present invention has general formula (II). When the compound of the present invention has the general formula (II), preferably Q and W are substituted or unsubstituted nitrogen, and D is oxygen, and more preferably Q is nitrogen substituted.

According to a preferred embodiment of the invention, the compound is selected from the group consisting of the following general formulas 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 and includes salts thereof:

The term "method" means a method for achieving a given task, including methods, means, techniques and processes readily developed or known from methods, means, techniques and processes known by the practitioners of chemistry, pharmacy, biology, biochemistry and medical technology. Means, means, techniques and processes, but is not limited to such.

"Treatment" or "prophylaxis" means that the compounds of the present invention are used as a therapeutic, prophylactic or ameliorating agent in connection with a pathology, illness or disease, signs or effects thereof.

There are numerous pathologies, ailments and diseases caused by defective potassium channel function. As with other potassium channel modulating compounds, the compounds of the present invention are in situ for the treatment of pathologies, diseases and conditions associated with defective potassium regulation to treat, alleviate, prevent, inhibit or limit the effects of diseases or pathologies in animals, including humans. It is for use in.

In particular, the present invention relates to diseases of the central or peripheral nervous system (local anemia, migraine, ataxia, Parkinson's disease, bipolar disorder, trigeminal neuralgia, cramps, mood disorders, brain tumors, mental disorders, muscle cramps, neuralgia, hearing and Provided are compounds, compositions, and methods useful for the treatment of blindness, Alzheimer's disease, Parkinson's disease, senile memory loss, learning deficiency, fear and motor neuron disease, and neuroprotective agents (eg, prevention of seizures, etc.). The compounds of the present invention have a use as a therapeutic agent for convulsive states such as the following epileptic epilepsy, epileptic seizure, psychomotor epilepsy or focal epilepsy. The compounds of the present invention are also useful for treating disease states such as gastroesophageal reflux disease and gastrointestinal dyskinesia. Other pathologies and diseases in which the compounds of the invention are useful for treating are described in, eg, US Pat. No. 6,348,486; 6,117,900; 6,589,986 and 6,593,349 and US patent applications 10 / 022,579; 10 / 075,703; 10 / 075,522; 10 / 114,148; 10 / 160,582 and 10 / 312,123, which are incorporated herein by reference.

As voltage dependent potassium channels are found in all animals, the compounds of the present invention are pharmacologically effective when administered to monkeys, dogs, cats, mice, rabbits, livestock, fish and most importantly human patients.

"Open" and "activation" refer to partial or full activation of a KCNQ channel by a compound that results in an increase in ion flux entering and exiting the cell in which the KCNQ channel is found and is used interchangeably.

Techniques for compounding and administering compounds as medicaments are described in "Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, latest edition," which is hereby incorporated by reference.

The compounds of the present invention are configured to cross the blood brain barrier to enable numerous forms of dosage forms. Nevertheless, the pharmaceutical compositions of the present invention are subcutaneous, including various preferred routes such as topical, intraarterial, intramuscular, subcutaneous and intramedullary injections as well as intramedullary space, direct intraventricular, venous, intraperitoneal, nasal or ocular injections. Need for treatment (treatment, prophylactic, improvement) by injection, topical, oral, intraperitoneal, intradermal, intravenous, intranasal, bronchial, oral, sublingual, anal, suppository, muscle, rectal, mucosal, intestinal or parenteral release It can be provided to an individual.

On the other hand, the composition of the present invention can be administered locally rather than systemically, such as by injecting the composition directly into the organ in a depo or sustained release formulation as described below.

By "therapeutically (or pharmaceutically) effective amount" is meant the amount of active ingredient necessary to achieve the desired result of preventing, alleviating or ameliorating the disease or manifestation of the disease. Determination of a therapeutically effective amount is within the determination of one skilled in the art, particularly in light of the description herein.

As used herein, a “compound of the invention” means a compound of Formula (I) or (II) or a combination thereof, and a pharmaceutically acceptable salt or combination of compounds of Formula (I) or (II) It refers to dissolved forms including hydrate forms such as monohydrate, dihydrate, trihydrate, hemihydrate, tetrahydrate and the like. This compound may be a true solvate or simply an accidental solvent or may be a mixture of solvates and accidental solvents.

"Pharmaceutically acceptable salt" means a salt of an active compound that is prepared with a relatively nontoxic acid or base, depending on the particular substituents found on the compounds described herein. When the compounds of the present invention have a relatively acidic action, base addition salts can be obtained by contacting a neutral (ie, non-ionized) form of the compound with a sufficient amount of the desired base in pure form or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts are the sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or similar salts. When the compound of the present invention contains a relatively basic function, acid addition salts can be obtained by contacting the compound in the neutral (ie non-ionized) form with a sufficient amount of the desired base in pure form or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include acetic acid, as well as salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrocarbonate, phosphoric acid, dihydrogen phosphate, sulfuric acid, monohydrosulfuric acid, hydroiodic acid, phosphorous acid, and the like. From relatively non-toxic organic acids such as propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. There is an induced salt. Also included are salts of amino acids such as alkylates, salts of organic acids such as glucuronic acid or galacturnoic acid (see Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain compounds of the present invention have both basic and acidic functions that allow them to be converted to base or acid addition salts.

The neutral form of the compound is preferably regenerated by contacting the salt with a base or an acid and separating the parent compound by known methods. Although the parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, the salt is the same as the parent form of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compounds in the form of prodrugs. Prodrugs of the compounds described herein are those compounds which readily undergo chemical changes under physiological conditions in vivo to provide the compounds of the present invention. Prodrugs can also be converted to compounds of the invention in an ex vivo environment by chemical or biochemical methods. For example, prodrugs can be slowly converted to the compounds of the present invention when placed in transdermal patch stores with suitable enzymes or chemicals.

Certain compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are the same as the unsolvated forms and are included within the scope of the present invention. Certain compounds of the present invention may exist in polycrystalline or amorphous form. In general, all physical forms are the same as those intended by the present invention and are within the scope of the present invention.

Certain compounds of the invention have asymmetric carbon atoms (optical centers) or double bonds; Racemates, diastereomers, geometric isomers and individual isomers are included within the scope of the present invention.

Compounds of the invention may also have an unnatural ratio of isotopes at one or more atoms that make up such compounds. For example, the compound may be radiolabeled with radioactive isotopes such as tritium ( ³ H), iodine-125 ( ¹²⁵ I) or carbon-14 ( ¹⁴ C). All isotopic variations of the compounds of the invention are included within the scope of the invention regardless of radioactivity.

As used herein below and in the claims, the term "alkyl" refers to saturated aliphatic hydrocarbons, including straight and branched groups. Preferably, the alkyl group has 1-20 carbon atoms. When the numerical range is referred to herein as "1 to 20", it means that the alkyl group contains 20 or less carbon atoms such as 1, 2, 3 and the like. More preferably, it is a medium sized alkyl having 1 to 10 carbon atoms. Most preferably, it is a lower alkyl group which has 1-4 carbon atoms. Alkyl groups may be substituted or unsubstituted. When substituted, examples of substituents include cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, halo, carbonyl, thiocarbonyl , O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy, nitro, sulfonamido, trihalo Methanesulfonamido, silyl, guanyl, guanidino, ureido, amino or NR _a R _b wherein R _a and R _b are independently hydrogen, alkyl, cycloalkyl, aryl, carbonyl, sulfonyl , Trihalomethylsulfonyl and their combined 5- or 6-membered heterocyclic rings.

By "cycloalkyl group" is meant any carbon monocyclic or fused ring (ie, rings which share adjacent pairs of carbon atoms) in which at least one ring does not have a fully conjugated pi electronic system. Examples of cycloalkyl groups include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexadiene, cycloheptane, cycloheptatriene, and adamantane. Cycloalkyl groups can be substituted or unsubstituted. When substituted, examples of substituents include alkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, halo, carbonyl, thiocarbonyl, C-carboxy, O-carboxy, O-carbamyl, N-carbamyl, C-amido, N-amido, nitro, amino and NR _a R _b as defined above.

"Alkenyl group" means an alkyl group consisting of two or more carbon atoms and one or more carbon-carbon double bonds.

By "aryl group" is meant any carbon monocyclic or mixed ring polycyclic group (ie, rings which share adjacent pairs of carbon atoms) with a fully conjugated pi-electron system. Examples of aryl groups include, but are not limited to, phenyl, naphthalenyl and anthracenyl. The aryl group can be substituted or unsubstituted. When substituted, examples of substituents include halo, trihalomethyl, alkyl, hydroxy, alkoxy, aryloxy, thiohydroxy, thiocarbonyl, C-carboxy, O-carboxy, O-carbamyl, N-carbamyl , O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, sulfinyl, sulfonyl, amino and NR _a R _b as defined above.

A "heteroaryl group" means a monocyclic or mixed ring having one or more atoms such as nitrogen, oxygen and sulfur in the ring, and also having a fully conjugated pi electron system (ie, rings which share adjacent pairs of carbon atoms) ). Examples of heteroaryl groups are pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline and purine. Heteroaryl groups can be substituted or unsubstituted. When substituted, examples of substituents include alkyl, cycloalkyl, halo, trihalomethyl, hydroxy, alkoxy, aryloxy, thiohydroxy, thiocarbonyl, sulfonamido, C-carboxy, O-carboxy, sulfinyl , Sulfonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, amino or NR _a R _b as defined above.

"Heterocycloaliphatic group" means a monocyclic or mixed ring having one or more atoms in the ring, such as nitrogen, oxygen, and sulfur. This ring may have one or more double bonds. Heteroalicyclic groups may be substituted or unsubstituted. When substituted, examples of substituents include alkyl, cycloalkyl, aryl, heteroaryl, halo, trihalomethyl, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, carr Carbonyl, thiocarbonyl, C-carboxy, O-carboxy, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, sulfinyl, sulfonyl, C-amido, N-amimi In addition, there is amino or NR _a R _b as defined above.

"Hydroxy group" means an -OH group.

"Azido group" means a -N = N group.

"Alkoxy group" means both an -O-alkyl and an -O-cycloalkyl group as defined herein.

"Aryloxy group" means both an -O-aryl group and an -O-heteroaryl group, as defined herein.

"Thiohydroxy group" means a -SH group.

"Thioalkoxy group" means a -S-alkyl group and -S-cycloalkyl group as defined herein.

"Thioaryloxy group" means both an -S-aryl group and -S-heteroaryl group, as defined herein.

"Carbonyl group" means -C (= 0), wherein R "is hydrogen, alkyl, cycloalkyl, aryl, heteroaryl (bonded via ring carbon) or heteroalicyclic (bonded through ring carbon) as defined herein; -R "group.

"Aldehyde group" means a carbonyl group in which R "is hydrogen.

"Tiocarbonyl group" means a group -C (= S) -R ", wherein R" is defined herein.

“C-carboxy group” means a group —C (═O) —O—R ″, wherein R ″ is defined herein.

"O-carboxy group" means R "C (= O) -O- group as defined herein.

"Carboxylic acid group" means a C-carboxy group wherein R "is defined herein.

"Halo" is fluorine, chlorine, bromine or iodine.

"Trihalomethyl group" means a -CX group where X is as defined herein.

“Trihalomethanesulfonyl group” means an X ₃ CS (═O) ₂ — group where X is as defined herein.

"Sulfinyl group" refers to the group -S (= 0) -R ", wherein R" is defined herein.

"Sulfonyl group" means a group -S (= 0) ₂ -R ", wherein R" is defined herein.

"S-sulfonamido group" means _a -S (= 0) ₂ -NR _a R _b group wherein R _a and R _b are defined herein.

"N-sulfonamido group" means R _a S (= 0) ₂ -NR _b in which R _a and R _b are defined herein.

"Trihaloalkyl methanesulfonamide amido" is R, and _a X ₃ CS (= O) X is as defined herein ₂ NR _a - means a group.

“O-carbamyl group” means —OC (═O) —NR _a R _b in which R _a and R _b are defined herein.

"N-carbamyl group" means _a group R _b OC (= 0) -NR _a -where R _a and R _b are defined herein.

"O-thiocarbamyl group" means _a group -OC (= S) -NR _a R _b where R _a and R _b are defined herein.

"N-thiocarbamyl group" means _a group R _b OC (= S) NR _a -where R _a and R _b are defined herein.

"Amino group" means a -NH ₂ group.

“C-amido group” means _a —C (═O) —NR _a R _b group in which R _a and R _b are defined herein.

“N-amido group” means _a group R _b C (═O) —NR _a , wherein R _a and R _b are defined herein.

"Quarter ammonium group" means an -NHR _a R _b group wherein R _a and R _b are independently alkyl, cycloalkyl, aryl or heteroaryl.

"Ureido group" means _a -NRaC (= 0) -NR _b R _c group, wherein R _a and R _b are defined herein and R _c is defined for R _a or R _b .

"Guanidino group" means _a -R _a NC (= N) -NR _b R _c group in which R _a , R _b and R _c are defined herein.

"Guanyl group" means _a group R _a R _b NC (= N)-, wherein R _a and R _b are defined herein.

"Nitro group" means a -NO ₂ group.

"Cyano group" means a -C≡N group.

"Silyl group" means a -Si (R ") ₃ group in which R" is defined herein.

According to a preferred embodiment, the compounds of the present invention have general formula (I). In a preferred embodiment, Y is selected from hydroxyalkyl and polyalkylene glycol moieties.

Preferably, the polyalkylene glycol moiety has the general formula (III)

[O- (CH ₂ ) m] n-OR ¹⁷ (III)

Wherein m and n are each independently an integer from 1 to 10 and R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl. Preferably, G is carbon, K is oxygen, R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are independently selected from the group consisting of hydrogen, alkyl, halo and trihaloalkyl, R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each hydrogen.

According to another embodiment of the present invention, the compound of the present invention has general formula (II). When the compound of the present invention has the general formula (II), preferably Q and W are substituted or unsubstituted nitrogen, and D is oxygen, and more preferably Q is nitrogen substituted.

Preferred compounds of the present invention include the following compounds 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 and salts thereof:

One 2

Meclofennam Diclofenac

Another aspect of the invention provides novel compounds useful herein. These novel compounds are generally derivatives of N-phenylanthranilic acid with covalently bonded hydroxyalkyl and polyalkylene glycol moieties. Hydroxyalkyl and polyalkylene glycol moieties generally increase the ability to cross the blood brain barrier.

Preferred novel compounds of the invention have the general formula (IV) and include salts thereof:

(IV)

Where

Z is an A-G (= K) -X-Y group,

A is alkyl or absent,

G is selected from the group consisting of carbon, sulfur and substituted or unsubstituted phosphorus,

K is selected from the group consisting of oxygen and sulfur,

X is selected or absent from the group consisting of substituted or unsubstituted nitrogen, oxygen and sulfur,

Y is selected from the group consisting of hydroxyalkyl and polyalkylene glycol moieties,

R ¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl and aryl,

R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are independently hydrogen, alkyl, hydroxyalkyl, trihaloalkyl, cycloalkyl, alkenyl, alkynyl , Aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro, amino and -NR ¹⁵ R ¹⁶ At least two of R ² , R ³ , R ⁴ , R ⁵ and R ⁶ and / or at least two of R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are 5-membered or 6- To form an aromatic, heteroaromatic, cycloaliphatic or heteroalicyclic ring,

R ¹⁵ and R ¹⁶ are independently of each other selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, carbonyl and sulfonyl, or R ¹⁵ and R ^{16 form} a 5- or 6-membered heteroalicyclic ring and,

Here, when phosphorus and / or nitrogen are substituted, the substituent is alkyl, cycloalkyl or aryl.

According to a feature of the invention, the residues of the polyalkylene glycols of the novel compounds of the invention have the general formula (V)

[O- (CH ₂ ) m] n-OR ¹⁷ (V)

Wherein m and n are each independently an integer from 1 to 10 and R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl.

According to another feature of the novel compounds of the invention, G is carbon, K is oxygen, and R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are each independently hydrogen, alkyl, halo and trihalo Selected from the group consisting of alkyl, each of R ⁷ , R ⁸ , R ⁹ and R ¹⁰ is hydrogen.

According to a preferred embodiment, the novel compounds of the invention are selected from the following compounds 3, 4, 5, 6, 7, 8 and 9, and pharmaceutically acceptable salts thereof:

Another aspect of the invention provides a pharmaceutical composition comprising a compound of the invention having the general formula (IV) as an active ingredient.

In accordance with a feature of the invention, there is provided a pharmaceutical composition having any one of compounds 3, 4, 5, 6, 7, 8 and / or 9.

It is yet another aspect of the present invention to provide a method for synthesizing the novel compounds described above. This method is carried out by reacting N-phenylanthranilic acid or a derivative thereof with hydroxyalkyl or polyalkylene glycol having a reactive group at the terminal. The reactive group is selected to be capable of forming an ester bond with N-phenylanthranilic acid or a derivative thereof.

As used herein, "ester bond" means J (where J is carbon, sulfur or phosphorus, preferably carbon, L is oxygen or sulfur, and M is oxygen, sulfur or nitrogen (substituted or unsubstituted as described above); = L) -M.

Preferred ester bonds according to the invention are carboxy ester bonds [-C (= O) -O-], carboxy amide bonds [-C (= O) -NR'-], carboxy thioester bonds [-C (= O) -S-], thiocarbonyl ester bond [-C (= S) -O-], thiocarbonyl thioester bond [-C (= S) -S-] and thiocarbonyl amide bond [-C (= S) -NR'-].

Thus, the reactive group is preferably hydroxy, amino or thiohydroxy, such as the fire protection described above, and the polyalkylene glycols having the reactive group at the end have the general formula (VII):

V- [O- (CH ₂ ) m] n-OR ¹⁷ (VII)

Wherein V is hydroxy, amine or thiohydroxy, each of m and n is independently an integer from 1 to 10 and R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl.

The starting material, N-phenylanthranilic acid or derivatives thereof preferably have the following general formula (VI):

(VI)

Wherein A, G, K, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are as described above.

Preferably, in formula (VI), G is carbon, K is oxygen, R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are independently hydrogen, alkyl, halo or trihaloalkyl, R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each hydrogen.

If the ester linkage is an amide linkage, the method according to the present invention further comprises converting N-phenylanthranilic acid or a derivative thereof to the corresponding ester before reacting the hydroxyalkyl or polyalkylene glycol. desirable.

As demonstrated in the examples below, the novel compounds of the present invention are synthesized easily and effectively by the methods of this gist of the present invention.

Compounds of the present invention include known compounds such as meclofennamic acid (compound 1), diclofenac (compound 2) and 1-EBIO (compound 10). 1-EBIO (Compound 10) has been shown to increase the open rate and openability of the channel (Po) in a single channel study conducted on the open rate and intermediate (IK) and small conductance (SK) Ca2 ⁺ -active K + channels (Syme). et al. Am. J. Physiol. 278: C570-C581 (2000).

Compounds of the present invention also include derivatives of meclofennamic acid, mainly derivatives of meclofennamic acid (compound 1) and novel compounds that are diclofenac (compound 2). Examples of novel compounds of the invention are compounds 3, 4, 5, 6, 7, 8 and 9.

These compounds are the openers of the KCNQ2 / 3 channel complex heterologously expressed in CHO cells. These compounds have also been found to reduce both induced and spontaneous action potentials in cortical neurons.

The compounds of the present invention have two main actions, the voltage polarization of KCNQ2 / 3 channel activation for slow polarization potential and slow channel deactivation. Similar to the effect on recombinant KCNQ2 / 3 channels, compounds 4, 5, 6 and 7 result in a threshold of M-current activation in cortical neurons, about 20 mV negative conversion from -50 mV to -70 mV. As a result of this leftward conversion of the KCNQ2 / 3 threshold of activation, there is a gradual hyperpolarization of the resting membrane potential. Without being bound by a particular theory, the data presented in the examples below suggest that compounds of the present invention stabilize KCNQ2 / 3 channels in the open state or destabilize closed channel structures. In addition, exposure of the channels of the compounds described herein will result in deactivation that contributes to stabilization of KCNQ2 / 3 channels in the open state. In particular, without being bound by any theory, the compounds of the present invention can improve channel gating by converting the voltage dependence of the voltage sensor S4 motion in the hyperpolarization direction.

From the functional point of view, the voltage-dependent leftward conversion and slow deactivation caused by the compounds of the present invention result in substantial M-current activation at normal restoring and subthreshold potentials. In the case of meclofennamic acid (Compound 1) at a potential of -60 / -50 mV (more than a tenfold increase of KCNQ2 / 3 width), particularly the activation of KCNQ2 / 3 channels, the compounds of the present invention To cause it. In addition, since the M-current (KCNQ2 / 3) is not inactivated, the significant activation made by the compounds of the present invention causes a large constant state of potassium conductance at sub-threshold and threshold potentials, resulting in braking against neuron firing. Act as. Indeed, the compounds of the present invention have been found to inhibit elicited and spontaneous cortical neuronal action.

The voltage range over which the compounds of the present invention activate KCNQ2 / 3 channels makes these compounds exceptionally useful in the treatment of ischemic attacks.

There is a similarity between the compounds of the present invention and the properties represented by retigabine.

First, both compounds and retigabine of the present invention shift the voltage dependence of KCNQ2 / 3 channel activation to the left, slow deactivation kinetics and hyperpolarize the resting membrane potential (Tatulian et al. J. Neurosci. 21: 5535-5545 (2001).

Second, as the compounds of the present invention act on the KCQN2 / 3 channel, retigabine exhibits a second inhibitory action on KCNQ at positive potentials (see Tatulian et al. J. Neurosci. 21: 5535-5545 (2001)). .

An interesting difference between retigabine and the compounds of the present invention is related to the selectivity towards the KCQN2 channel and KCQN3 subunit. On the other hand, while retigabine has the strongest opener action on KCQN3 homologous channels (see Tatulian et al. J. Neurosci. 21: 5535-5545 (2001)), the compounds of the present invention are stronger in KCQN2 homologous channels. .

From a functional point of view, the left shift and slow deactivation of the activation curve caused by the compounds of the present invention results in substantial M-current activation at normal resting and subthreshold potentials. In addition, since M-current (KCNQ2 / 3) is not inactivated, activation made by the compounds of the present invention results in a large constant state of potassium conductance at sub-threshold and threshold potentials, thus preventing neuronal firing. It is expected to. Indeed, compounds 4, 5, 6, 7 and 8 are induced and inhibit spontaneous cortical neuronal action.

The voltage range at which the compounds of the invention act is indicated to be exceptionally suitable for treating epilepsy, ischemic seizures and neuralgia.

As pointed out above, according to a feature of the invention, the compounds of the invention form part of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

According to another feature of the invention, the pharmaceutical composition of the invention is packaged in a packaging for use in the treatment or prophylaxis of diseases or diseases associated with the altered action of voltage-dependent potassium channels in the peripheral or central nervous system, for example. Printed on the packaging. The disease or disease so specified is one or more of the diseases or diseases for which the special pharmaceutical composition is suitable.

As used herein, the term "pharmaceutical composition" refers to a physiologically suitable carrier, excipient, lubricant, buffer, antimicrobial agent, elongate drug (e.g. mannitol), antioxidant (e.g. ascorbic acid or sodium bisulfite), anti-inflammatory, anti Means one or more compounds of the present invention (active ingredient), or physiologically acceptable salts or preparations thereof, together with other chemical components, including but not limited to viral agents, chemotherapeutic agents, antihistamines, and the like. do. The purpose of the pharmaceutical composition is to facilitate administration of a compound to a patient. By "active ingredient" is meant a compound capable of measuring a biological effect.

As used herein, "physiologically acceptable carrier" and "pharmaceutically acceptable carrier" refer to a diluent or carrier that does not cause significant irritation to the organism and does not destroy the properties and biological effects of the administered compound. "Excipient" means an inert substance added to a pharmaceutical composition to make administration of the compound easier. Examples include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and starches, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.

As mentioned above, techniques for administering and combining compounds as drugs are shown in "Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, latest edition", herein incorporated by reference.

The pharmaceutical composition of the present invention can be prepared by known methods, such as conventional mixing, dissolving, granulating, dragging, powdering, emulsifying, encapsulating, entrapping or lysifying.

Pharmaceutical compositions for use in the present invention may be formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate the processing of the active compounds in preparations which can be used pharmaceutically. . Proper formulation is dependent upon the route of administration chosen.

For injection, the compounds of the present invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution or physiological saline buffer. For transmucosal administration, a penetrant suitable for the barrier to penetrate is used in the formulation. Such penetrants are generally known in the art.

For oral administration, the compounds can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers known in the art. Such carriers allow the compound of the invention to be formulated into tablets, pills, powders, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for oral ingestion by a patient. Pharmaceutical preparations for oral use obtain solid tablets or powdered cores by using solid excipients, optionally by crushing the resulting mixture, adding the appropriate auxiliaries if necessary and then processing the granule mixture. Particularly suitable excipients are fillers such as sugars, including lactose, sucrose, mannitol or sorbitol, such as corn starch, flour starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium Cellulose preparations such as carbomethylcellulose; And / or physiologically acceptable polymers such as polyvinylpyrrolidone (PVP). If desired, disintegrating agents such as crosslinked polyvinyl pyrrolidone, agar, or alginic acid or salts thereof (eg sodium alginate) may be added.

Appropriate coating is provided to the drag core. For this purpose, concentrated sugar solutions can be used, which optionally contain gumarvik, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. can do. Dye pigments may also be added to the drag coating or tablet for characterization or to characterize different combinations of active compound dosage forms.

Pharmaceutical compositions that can be used orally include push-fit capsules made of gelatin, as well as soft sealed capsules made of gelatin and plasticizers such as glycerol or sorbitol. Push-fit capsules may contain the active ingredient together with fillers such as lactose, binders such as starch, lubricants such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. All combinations for oral administration should be in dosage forms suitable for the route of administration chosen.

For oral administration, the compositions may take the form of tablets or lozenges formulated in a conventional manner.

For inhalation administration, the compounds for use according to the invention are aerosols from packs or nebulizers compressed with suitable propellants, for example dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane or titanium dioxide. It is conveniently dispensed in a divided form. In the case of a compressed aerosol, the dosage unit can be measured by providing a valve to dispense the measured amount. Capsules and cartridges of gelatin for use in an inhaler or blower may be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch.

The formulations described herein may be formulated for parenteral administration, such as by large pill infusion or continuous infusion. Formulations for injection may be presented in unit dosage form, such as with preservatives optionally added to an ampoule or multi-dose container. The composition may be a suspension, solution or emulsion in an oily or aqueous medium and may contain blending agents such as suspending, stabilizing and / or dispersing agents.

Pharmaceutical compositions for parenteral administration include aqueous solutions of the active agent in the form of aqueous solutions. In addition, suspensions of the active compounds can be prepared as suitable injection suspensions in oil. Lipophilic solvents or media include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate, triglycerides, or liposomes. Aqueous injection suspensions may contain substances that enhance the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol or dextran. Optionally, the suspension may contain suitable stabilizers or agents that increase the solubility of the compound to produce a highly concentrated solution.

On the other hand, the active ingredient may be in powder form for use with a suitable medium, such as sterile, pyrogen-free water, before use.

The formulations of the present invention may also be formulated into rectal compositions such as suppositories or retention enemas using conventional suppository bases such as cocoa butter or other glycerides.

In addition to the combinations described above, the formulations of the invention may also be formulated for topical administration such as depot preparations. Such sustained combinations may be administered by implantation (eg, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the formulation may be combined with a suitable polymer or hydrophobic material (eg an emulsion in an acceptable oil) or an ion exchange resin, or may be formulated as a poorly soluble derivative such as a salt that hardly dissolves. Formulations for topical administration may also include, but are not limited to, lotions, suspensions, ointment gels, creams, drops, liquids, spray emulsions and powders.

Pharmaceutical compositions suitable for use in the present invention include compositions in which the active ingredient is contained in an amount effective to achieve the intended purpose. In particular, “therapeutically effective amount” means an amount of active ingredient effective to prevent, alleviate or ameliorate the disease and / or its symptoms and / or its effects.

Determination of a therapeutically effective amount is within the scope of those skilled in the art in light of the teachings herein.

For any agent used in the methods of the invention, the therapeutically effective amount or dosage can be initially predicted from cell culture assays. For example, the dosage form can be formulated into an animal model to achieve a range of circulating concentrations comprising an IC ₅₀ as determined in cell culture. This information can be used to accurately determine dosages useful for humans.

Toxicity and therapeutic effects of the compounds described herein can be determined by standard pharmaceutical procedures in cell culture or experimental animals, such as by measuring IC ₅₀ and LD ₅₀ (the lethal dose at which 50% of test animals die). Data obtained from these cell culture assays and animal studies can be used to calculate dosage ranges for use in humans. Dosage may vary depending on the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the physician depending on the patient's condition (Fingl, et al., 1975, in "The Pharmacological Basis of Therapeutics", Ch. 1P. 1).

Dosage and dosing intervals may be individually adjusted to provide plasma levels of the active moiety sufficient to maintain a kinase modulating effect called minimum effective concentration (MEC). MEC varies for each formulation but can be predicted from in vitro data. For example, the concentrations required to achieve 50-90% inhibition of kinases can be identified using the assays described herein. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. HPLC analysis or biological analysis can be used to determine the plasma concentration. MEC values can be used to determine dosing intervals. The formulation should be administered using a regimen that maintains plasma levels above the MEC for 10-90%, preferably 30-90%, most preferably 50-90% of the time.

In the case of topical administration or selective ingestion, the effective local concentration of the drug may not be related to the plasma concentration. In such cases, other procedures known in the art can be used to determine effective local concentrations.

Depending on the responsiveness and severity of the disease to be treated, the administration can be in the course of treatment lasting from days to weeks, or as a single dose of the sustained release composition until treatment is performed or a reduction in disease state is achieved.

The amount of composition to be administered will of course depend on the patient being treated, the extent of pain, the method of administration and the judgment of the prescribing physician.

The compositions of the present invention may be provided in a pack or dispensing device, such as an FDA approved kit, which may contain one or more unit dosage forms containing the active ingredient, if desired. Dosage instructions may be provided on the pack or dispensing device. The pack or dispenser is also provided with a warning in combination with the container in a form suggested by the governmental authority regulating the manufacture, use or sale of the drug, which reflects the authority's approval of the human or livestock dosage form of the composition. . Such warnings may be, for example, a label approved by the US Food and Drug Administration for the insertion of an approved product or prescription drug. Compositions comprising the formulations of the present invention formulated in compatible pharmaceutical carriers may be prepared and placed in appropriate containers and labeled for the treatment of the indicated conditions.

Pharmaceutical compositions of the present invention generally relate to a disease associated with the modified action of a voltage-dependent potassium channel or cortical neuron comprising as an active ingredient a compound of the present invention which acts by regulating each potassium channel or inhibiting cortical neuronal action. Or for the treatment, prevention or amelioration of the disease. In general, the regulated potassium channels are KCNQ2 channels and / or KCNQ3 channels and / or KCNQ2 / 3 channels. Diseases or diseases of the voltage-dependent potassium channel central nervous system or peripheral nervous system that are preferably treated or prevented by the pharmaceutical composition of the present invention include epilepsy, ischemic seizures, migraine, ataxia, muscle cramps, neuralgia, Alzheimer's disease, Parkinson's disease , Old age amnesia, learning deficiency, bipolar disorder, trigeminal neuralgia, convulsions, mood disorders, brain tumors, mental disorders, hearing and vision loss, fear and motor neuron disease. Preferably the composition is packaged and printed on the packaging to treat or prevent peripheral or central nervous system diseases or diseases associated with the altered action of voltage-dependent potassium channels.

Further objects, advantages and novel features of the invention will become apparent to those skilled in the art from the following examples, which do not limit the invention. In addition, several aspects of the invention shown in the following examples and claims are experimentally supported in the following examples.

The following examples are incorporated by reference with the above description and illustrate but do not limit the invention.

Materials and Experimental Methods

Chemical synthesis:

Meclofennamic acid (Compound 1), Diclofenac (Compound 2) and 1-ethylbenzimidazolone (1-EBIO, Compound 10) are commercially available and are available from Sigma-Aldrich (St. Louis, MO, USA). Can be purchased from).

The chemical structures of the compounds 1 to 10 according to the present invention are shown in FIG.

Synthesis of Compound 7:

Diclofenac (50 mg, 0.17 mmol) was dissolved in the catalytic amount of anhydrous dichloromethane (DCM) and N-dimethylaminopyridine (DMAP), followed by addition of diethylene glycol (0.8 ml, 0.85 mmol). The mixture was cooled to 0 ° C. with stirring and a solution of dicyclohexyl carbodiimide (DCC, 52.6 mg, 0.255 mmol) dissolved in DCM was added dropwise. The resulting suspension was monitored by TLC (using a 1: 1 mixture of EtOAc: hexane as eluent) with stirring at 0 ° C. for 30 minutes. The solid was then filtered off and washed with DCM. The filtrate was concentrated under reduced pressure and then chromatographed with silica gel to give the pure product (43 mg, 66%).

¹ H-NMR (200 MHz, CDC1 ₃ ): δ = 7.35 (2H, d, J = 8); 7.19-7.06 (2H, m); 6.99 (1H, d, J = 8); 6.96 (1H, doublet, J = 8.); 6.56 (1H, doublet, J = 8); 4.35 (2H, m); 3.8 (2H, s); 3.65-3.75 (4H, m); 3.53-3.57 (2H, m) ppm.

Synthesis of Compound 8

Compound 8 was prepared according to the method of compound 7 using diethylene glycol methyl ether instead of diethylene glycol. The product was obtained in 53% yield.

¹ H-NMR (200 MHz, CDC1 ₃ ): δ = 7.35 (2H, d, J = 8); 7.19-7.06 (2H, m); 6.99 (1H, d, J = 8); 6.96 (1H, doublet, J = 8.); 6.56 (1H, doublet, J = 8); 4.34-4.29 (2H, m); 3.85 (2H, s); 3.75-3.70 (2H, m); 3.61-3.59 (2H, m); 3.58-3.50 (2H, m) ppm.

Synthesis of Compounds 3, 4, 5, 6 and 9-General Methods

N-hydroxy succinimide (0.76 mmol) and DCC (0.76 mmol) were added to the corresponding acid (meclofennamic acid, diclothenac or derivatives) dissolved in dichloromethane. The mixture was stirred for 1 hour while monitored by TLC (using a 1: 1 mixture of EtOAc: hexane as eluent). After the reaction was completed, the mixture was filtered and the solvent was evaporated. The crude product was separated by column chromatography to give pure N-hydroxy succinimide ether intermediates of the following compounds 3, 4, 5, 6 and 9.

Intermediate of compound 3: yield 90%. ¹ H-NMR (200 MHz, CDC1 ₃ ): δ = 8.69 (1H, s); 8.09-8.14 (1H, doublet of doublets, J = 1.7, J = 8.6); 7.32-7.37 (1H, doublet of doublets, J = 1.8, J = 8.5); 7.14-7.28 (3H, m); 6.72-6.80 (2H, m); 2.9 (4H, s); 2.29 (3H, s) ppm.

Intermediate of compound 4: yield 90%. ¹ H-NMR (200 MHz, CDC1 ₃ ): δ = 8.91 (1H, s); 8.10-8.15 (1H, doublet of doublets, J = 1.6, J = 8.18); 7.33-7.48 (5H, m); 7.24 (1H, doublet, J = 7.8); 6.80-6.88 (1H, dt, J = 1.06, J = 7.1); 2.9 (4H, s) ppm.

Intermediate of Compound 5: Yield 100%. ¹ H-NMR (200 MHz, CDC1 ₃ ): δ = 8.6 (1H, s); 8.12 (1H, doublet of doublets, J = 1.53, J = 7.4); 7.32 (1 H, M); 7.06-7.13 (3H, m); 6.66-6.74 (2H, m); 2.9 (4H, s); 2.32 (3H, s); 2.14 (3H, s) ppm.

Intermediate of Compounds 6 and 9: Yield 89%. ¹ H-NMR (200 MHZ, CDC1 ₃ ): δ = 7.35 (3H, d, J = 8); 7.18-7. 26 (1 H, m); 6.93-7.07 (2H, m); 6.6-6.63 (1H, d, J = 7.9); 6.2 (1H, s); 4.13 (2H, s); 2.84 (4H, s) ppm.

For Compound 9 (1 equiv), the corresponding intermediate is dissolved in 1 ml of dimethylformamide (DMF), diethylene glycol amine or 2-hydroxyethyl) methylamine, and the mixture is added, and the mixture is TLC ( Stir for 30 minutes, monitoring with 1: 1 mixture of EtOAc: hexane as eluent. After the reaction was completed, the solvent was removed under reduced pressure. The product was purified by column chromatography to afford compounds 3, 4, 5, 6 and 9 as follows:

Compound 3: yield 55%. ¹ H-NMR (200 MHz, CDC1 ₃ ): δ = 9.31 (1H, s); 7.45-7.50 (1H, doublet of doublets, J = 0.6, J = 8); 6.99-7. 22 (5H, m); 6.67-6.94 (1H, dt, J = 1.8, J = 6.5); 3.54-3.74 (8H, m); 2.33 (3H, s) ppm.

Compound 4: yield 70%. ¹ H-NMR (200 MHz, CDC1 ₃ ): δ = 9.44 (1H, s); 7.46-7.50 (1H, doublet of doublets, J = 0.6, J = 8); 7.4 (2H, doublet, J = 9); 7.16-7. 32 (5H, m); 7.09 (1 H, bs); 6.79-6.84 (1H, doublet of doublets, J = 1.8, J = 6.5); 3.54-3.73 (8H, m) ppm.

Compound 5: yield 88%. ¹ H-NMR (200 MHz, CDC1 ₃ ): δ = 9.19 (1H, s); 7.41-7.45 (1H, doublet of doublets, J = 1.53, J = 7.4); 7.14-7.21 (2H, m); 7.06-7.13 (1H, t, J =); 6.93-6.96 (2H, m); 6.65-6.69 (2H, m); 3.6-3.8 (8H, m); 2.32 (3H, s); 2.20 (3H, s) ppm.

Compound 6: yield 72%. ¹ H-NMR (200MHZ, CDC1 ₃ ): δ = 7.35 (2H, d, J = 8); 7.19-7.06 (2H, m); 6.99 (1H, d, J = 8); 6.96 (1H, doublet, J = 8.); 6.56 (1H, doublet, J = 8); 3.69 (2H, s); 3.55-3.49 (4H, m) ppm.

Compound 9: yield 63%. ¹ H-NMR (200 MHz, CDC1 ₃ ): δ = 7.35 (2H, d, J = 8); 7.19-7.06 (2H, m); 6.99 (1H, d, J = 8); 6.96 (1H, doublet, J = 8.); 6.56 (1H, doublet, J = 8); 3.78-3.74 (2H, m); 3.64-3.57 (2H, m); 3.23 (2H, s); 2.95 (3H, s) ppm.

biology

CHO Cell Culture and Transfection:

CHO (Chinese Hamster Ovary) cells were grown in Dulbecco's improved Eagles culture supplemented with 2 mM glutamine, 10% fetal calf serum and antibiotics. In brief, 40,000 cells seeded on poly-D-lysine-coated glass coverslips (13 mm in diameter) in 24-multiwell plates were used as markers for transfection with pIRES-CD8 (0.5 μg) and KCNQ2 (0.5 μg) and And / or transfected with KCNQ3 (0.5 μg). For electrophysiological experiments, transfected cells were visualized using anti-CD8 antibody-coated beads (Jurman et al., 1994) about 40 hours after transfection. Transfection was performed using 3.5 μl of lipofectamine (Gibco-BRL) according to the manufacturer's experimental specifications.

Neuronal Cortex Culture:

Sprague Dawley rat fetus (E18) was removed by a caesarian section and the cortical cells were excised. Tissues were digested with papain (100 U; Sigma, St. Louis, Mo.) for 20 minutes, crushed into single-cell suspensions and then bovine collagen type IV on 13 mm diameter glass coverslips of 24-multiwell plates. Cultured at a density of 40,000 cells / ml on a substrate of 100 μg / ml poly-L-lysine. Cultures were modified Eagles culture containing 5% horse serum (Biological Industries, Beit HaEmek, Israel), B-27 neuron supplement (Invitrogen, Carlsbad, Calif.), 100 U / ml penicillin, 100 μg / ml streptomycin and 2 mM glutamine. D-glucose was corrected to a final concentration of 6 g / l. Cytosine-1-D-aribinofuranoside (5 μΜ) was added after 5 days to arrest rapid cell proliferation. All cultures were maintained in 37 ° C. wet air containing 5% CO ₂ .

Electrophysiology Experiments:

For current measurement in CHO cells, recording was made 40 hours after transfection using the whole cell configuration of the patch-clamp technique (Hamill et al., Nature 294: 462-464 (1981)). The signal was amplified using an Axopatch 200B patch-clamp amplifier (Axon Instruments, Foster City, California, USA), sampled at 2 kHz, and then filtered at 800 Hz with a 4-pole Bessel lowpass filter. Data was obtained using the pClamp 8.1 software (Axon Instruments, Foster City, California, USA) and the Elonex Pentium III computer with the DigiData 1322A interface (Axon Instruments, Foster City, California, USA). The patch pipette is pulled from a boron silicate glass (Warner Instrument. Corp., Hamden, Connecticut, USA) with a resistance of 2-5 MΩ, then charged with 130 KCl, 1 MgCl ₂ , 5 K ₂ ATP, 5 EGTA, 10 HEPES ( mM) and then adjusted to pH 7.4 (290 mOsm) with KOH. The external solution contained 140 NaCl, 4 KCl, 1.8 CaCl ₂ , 1.2 MgCl ₂ , 11 glucose, 5.5 HEPES in an amount of mM and adjusted to pH 7.4 (310 mOsm) with NaOH. Series resistance (3-13 MΩ) was compensated (75-90%) and monitored periodically. After incubation for current-clamp measurement of rat cortical neurons, recordings were made after 10-14 days. The patch pipette was charged with 135 KCl, 1 K ₂ ATP, 1 MgATP, 2 EGTA, 1.1 CaCl ₂ , 5 glucose, 10 HEPES in an amount of mM and then adjusted to pH 7.4 (315 mOsm) with KOH. The external solution contained 140 NaCl, 4 KCl, 2 CaCl ₂ , 2 MgCl ₂ , 5 glucose, 10 HEPES in an amount of mM and adjusted to pH 7.4 (325 mOsm) with NaOH. To induce neuronal action potentials, 50-300 pA currents were injected into the cells for 800 ms (square pulses). Records were sampled at 5 kHz and filtered with a 4-pole Bessel lowpass filter at 2 KHz. To measure the voltage-clamps of rat cortical neurons, patch pipettes were charged with 90 K-acetate, 40 KCl, 3 MgCl ₂ , 2 K ₂ ATP, 20 HEPES in an amount of mM, followed by pH 7.4 (310 to 310 to KEP). 315 mOsm). The external solution contains 120 NaCl, 23 NaHCO ₃ , 3 KCl, 2.5 CaCl ₂ , 1.2 MgCl ₂ , 11 glucose, 0.0005 Tetrodotoxin (TTX), 5 HEPES in an amount of mM, adjusted to pH 7.4 (325 mOsm) with NaOH. It was.

Current measurements in Xenopus oocytes were performed as described in "Peretz et al. J. Physiol 545: 751-766 (2002)". That is, two-electrode voltage-clamp measurements were performed after 3-5 days after cRNA microinjection into oocytes. Oocytes were wetted with an improved ND96 solution containing 96 NaCl, 2 KCl, 1 MgCl ₂ , 0.1 CaCl ₂ and 5 HEPES in an amount of mM and titrated to pH 7.4 with NaOH. Total cell currents were recorded at room temperature (20-25 ° C.) using a Gene Clamp 500 amplifier (Axon Instruments, Foster City, California, USA). Glass microelectrodes (AM Systems, Inc., Carlsborg, Wisconsin, USA) were filled with 3M KCl and had a tip resistance of 0.5-1.5 MΩ. Stimulation, data validation and analysis of the formulations were performed using a 586 personal computer Pentium 4 connected with pCLAMP 6.02 software (Axon Instruments, Foster City, California, USA) and Digidata 1200 interface (Axon Instruments, Foster City, California, USA). It became. The current signal was filtered at 0.5 kHz and digitized at 2 kHz.

Data analysis:

Clampfit program (pClamp 8.1, Axon Instruments, Foster City, California, USA), Microsoft Excel 98 (Microsoft Corp., Redmond, Washington, USA), Axograph 4.6 (Axon Instruments, Foster City, California, USA), and Prism 2.0 (GraphPad , San Diego, California, USA). Leak subtraction was performed off-line using the Clampfit program of pClamp 8.1 software. To analyze KCNQ2 / 3 channel deactivation, a single exponential pit was applied to the tail current. The code conductance (G) was calculated by the following equation:

G = I / (VV _rev )

I is the current strength measured at the end of the pulse,

V _rev is a calculated reverse potential that is assumed to be -90 mV in CHO cells and -98 mV in Zenof oocytes,

G was measured at several test voltages V and then normalized to the maximum conductance value G _max calculated at +40 mV. The activation curve was obtained by Boltzmann equation:

G / G _max = 1 / {1 + exp [(V ₅₀ -V) / s]}

Where V ₅₀ is the voltage at which the current is half activated and s is the slope element. All data are presented as mean ± SEM. Statistically significant differences were assessed by the student's t-test.

Experiment result

Analysis conducted with meclofennamic acid (compound 1), diclofenac (compound 2) and 1-EBIO (compound 10)

Example 1

Effect of Meclofennamic Acid (Compound 1) and 1-EBIO (Compound 10) on KCNQ2 / 3 Current

The leftward conversion of the activation voltage-dependence of the CKNQ2 / 3 current induced by meclofennamic acid (Compound 1) and 1-EBIO (Compound 10) is described with reference to FIGS. 1A-1D.

When KCNQ2 and KCNQ3 subunits are individually expressed as homologous channels in several expression systems, they generate relatively low potassium currents, particularly for KCNQ3 (Wang et al. Science 282: 1890-1893 (1998) and Yang et al. J. Biol. Chem. 273: 19419-19423 (1998). However, KCNQ2 co-expressed with KCNQ3 generates an electric current whose intensity is about 10 times greater than the sum of the two homologous channels, and its biological and pharmaceutical properties are very similar to the original M-current (Main et. Mol. Pharmacol. 58: 253-262 (2000), Wang et al. Science 282: 1890-1893 (1998) and Yang et al. J. Biol. Chem. 273: 19419-19423 (1998).

CHO cells were cotransfected in equal proportions with cDNAs of two corresponding KCNQ2 and KCNQ3 and then exposed to meclofennamic acid (Compound 1) and 1-EBIO (Compound 10) to effect these compounds on M-current It was confirmed.

Referring to FIGS. 1A and 1B, before (left panel) and after (right panel) external use of 100 μM meclofennamic acid (Compound 1, FIG. 1A) and 100 μM 1-EBIO (Compound 10, FIG. 1B) externally Representative traces from the same cells were recorded. The membrane potential was raised for 15 second pulses from -70 mV to +40 mV in increments of 10 mV and then raised to -60 mV with 0.75 second pulses to form a tail current. The holding potential of all experiments was -85 mV.

Normalized conductance (G / s) as a function of voltage step for cells treated with control (open square) and meclofennamic acid (compound 1, FIG. 1C) or 1-EBIO (compound 10, FIG. 1D) (solid square) Gmax) was obtained. The activation curve was obtained by Boltzmann's equation.

1A (left panel) shows a representative trace of KCNQ2 / 3 current activated above a voltage threshold of about −50 mV by step polarization. Figure 1A as 100 μM methoxy claw pen by the addition of acid in the external _{V 50 = -19.65 ± 1.85 mV (} n = 24) V 50 = -42.34 ± 2.08 mV (n = 14) in shown in (right panel) KCNQ2 / 3 A significant 22.7 mV leftward transformation was formed in the voltage-dependence of current activation. The slope of the Boltzmann fitting curve did not change significantly with s = -9.46 ± 0.41 mV / e and s = -10.50 ± 0.93 mV / e for control and meclofennamic acid (Compound 1), respectively.

Figure 1B (left panel) shows a leftward conversion of 7.9 mV (p <0.005) caused by 100 μM 1-EBIO (Compound 10) in the voltage-dependent nature of KCNQ2 / 3 current activation from V ₅₀ = -22.9 ± 1.8 mV. V ₅₀ = -30.8 ± 2.8 mV (n = 4). The slope of the Boltzmann fitting curve did not vary significantly with s = -10.2 ± 1.1 mV / e and s = -9.5 ± 0.7 mV / e for control and 1-EBIO (Compound 10), respectively.

In conclusion, when exposed to meclofennamic acid (Compound 1) or 1-EBIO (Compound 10), the KCNQ2 / 3 current is about -60 mV vs. -50 mV for the control (Figures 1C and 1D). It was activated at more superpolarized potentials above.

Example 2

Increase in KCNQ2 / 3 Current Intensity by Meclofennamic Acid (Compound 1) and 1-EBIO (Compound 10)

An increase in KCNQ2 / 3 current intensity by meclofennamic acid (Compound 1) and 1-EBIO (Compound 10) is described with reference to FIGS. 1A-1D and 2A-2C.

As shown in Figure 2A, KCNQ2 / 3 current increases in the presence of meclofennamic acid (Compound 1) or 1-EBIO (Compound 10). Traces in the presence or absence (control) of meclofennamic acid (Compound 1, left panel) and 1-EBIO (Compound 10, right panel) were recorded. Cells were staged up to -20 mV for 1.5 sec pulses. In this train specification, the interval between pulses was 30 seconds.

Figure 2B shows the percentage of current measured in the presence (+) or absence (-) of meclofennamic acid (Compound 1, left panel) or 1-EBIO (Compound 10, right panel), where the control is 100 %to be.

In FIG. 2C, the current intensity (nA) is the superpolarized potential to emphasize the negative conversion of the channel activity threshold in response to application of meclofennamic acid (Compound 1, left panel) or 1-EBIO (Compound 10, right panel). Obtained for step voltages in the range (-70 mV to -20 mV).

Within about 1 minute of external application of meclofennamic acid (Compound 1), it was observed that the KCNQ2 / 3 current intensity increased significantly beyond the test potential range between -50 and 0 mV (FIGS. 1C and 2A, left panel). . The effect of meclofennamic acid (Compound 1) can be fully reversed (data not shown). In the train regulation, when cells were staged up to -20 mV, applying meclofennamic acid (Compound 1) to 1451 ± 164 pA (n from 844 ± 130 pA for control and meclofennamic acid (Compound 1), respectively) Current intensity was increased by 72% width (Fig. 2B, left panel).

From the normalized conductance-voltage relationship (G / G _max ) and the normalized current-voltage relationship (I / I _max ) shown in FIGS. 1C and 2C, meclofennamic acid (Compound 1) is determined at the voltage-dependence of channel activation. It can be seen that the leftward conversion mainly increased the KCNQ2 / 3 potassium current. As the test potential became more positive and approached the saturation value of the activity curve (ie +20 mV), the effect of meclofennamic acid (Compound 1) on KCNQ2 / 3 current intensity was very small. The most prominent action of meclofennamic acid (Compound 1) was exerted at negative physiologically related potentials. At -50 mV, -40 mV and -30 mV, meclofennamic acid (Compound 1) increased KCNQ2 / 3 current intensity by 10, 5 and 2.5 times, respectively (Figures 1A, 1C and 2C, left panel). .

Likewise, although less pronounced than the effect of meclofennamic acid (Compound 1), addition of 100 μM 1-EBIO (Compound 10) immediately results in KCNQ2 / 3 current intensity across the test potential range between -50 mV and -10 mV. Increased (Figures 1B, 2A and 2B, right panel). The effect of 1-EBIO (Compound 10) can be fully reversed (data not shown). In this train regulation, when cells were staged up to -30 mV, application of 1-EBIO (Compound 10) to 1483 ± from 945 ± 135 pA (n = 14) for Control and 1-EBIO (Compound 10), respectively. The current intensity was increased by 57% width to 194 pA (n = 14).

From the normalized conductance-voltage relationship (G / G _max ) and the normalized current-voltage relationship (I / I _max ) shown in FIGS. 1D and 2C (right panel), 1-EBIO (Compound 10) is the voltage of channel activation. It can be seen that the KCNQ2 / 3 potassium current was mainly increased through the leftward transformation in dependence. As the test potential became more positive and approached the saturation value of the activity curve, the effect of 1-EBIO (Compound 10) on KCNQ2 / 3 current intensity was weaker. 1D shows the leftward transformation of the threshold for channel activation of 10 mV. In addition, FIG. 2C (right panel) shows that the most prominent action of 1-EBIO (Compound 10) was at the negative physiologically relevant potential. 1-EBIO (Compound 10) increased KCNQ2 / 3 current intensity by 10 and 3 times at -50 mV and -40 mV, respectively.

Example 3

Effect of Meclofennamic Acid (Compound 1) and 1-EBIO (Compound 10) on KCNQ2 / 3 Deactivation Kinetics

Reduction of KCNQ2 / 3 deactivation kinetics by meclofennamic acid (Compound 1) and 1-EBIO (Compound 10) is described with reference to FIGS. 3A-3E.

In Fig. 3A, the tail currents of the cells before and after the use of meclofennamic acid (Compound 1) are shown. The prepulse was -20 mV, while the tail potential was -60 mV.

3B shows the time constants obtained from the _exponential pits of tail current decay (τ _deact ) in the presence (+) or absence (-) of _{meclofennamic} acid (Compound 1).

Although meclofennamic acid (Compound 1) and 1-EBIO (Compound 10) did not affect KCNQ2 / 3 activation kinetics, they significantly reduced the deactivation process. The decrease in tail current or deactivation reflected the transition of the channel from the open state to the closed state. As shown in FIG. 3A, the cells were depolarized to −20 mV and then repolarized to −60 mV. The reduction in tail current was fitted using one exponential function. In response to the addition of 100 μM _{meclofennamic} acid (Compound 1), the time constant for deactivation increased about 2 times from τ _deact = 79.6 ± 4.5 msec to τ _deact = 167.5 ± 11.6 msec (n = 10). The result was very large as shown in Figures 3B and 3C.

As described above, 1-EBIO (Compound 10) had a significant (p <0.001) effect on KCNQ2 / 3 activation kinetics. The reduction in tail current was fitted using one exponential function. In response to the addition of 100 μM 1-EBIO (Compound 10), the time constant for deactivation increased from τ _deact = 91.2 ± 8.4 msec to τ _deact = 110.1 ± 9.5 msec (n = 14, Figure 3E). 3D shows that 1-EBIO (Compound 10) reduces the deactivation kinetics of KCNQ2 / 3 channel.

Example 4

Inhibition of Stimulation and Spontaneous Neuronal Action by Meclofennamic Acid (Compound 1) and 1-EBIO (Compound 10)

Inhibition of induction and spontaneous neuronal action by meclofennamic acid (Compound 1) and 1-EBIO (Compound 10) is described with reference to FIGS. 4A-4B.

4A-4B show inhibition of neuronal action by meclofenamic acid (Compound 1). In FIG. 4A, induced before (first row), during (second and third row) and after (fourth and fifth row) application of different concentrations of meclofennamic acid (Compound 1). Rat cortical neuronal action. Each column corresponds to a different neuron.

In FIG. 4B, the spontaneous action recorded before, during and after application of 10 μM meclofennamic acid (Compound 1) is shown.

As mentioned above, one of the main functions of M-current is to monitor the excitability of neurons in the brain. From these results, it is clear that meclofennamic acid (Compound 1) and 1-EBIO (Compound 10) enhance the heterologously expressed M-current. It remains questionable how meclofennamic acid (Compound 1) and 1-EBIO (Compound 10) regulate the excitability of neurons expressing M-current. The answer to this question was that primary cultures of the current clamp configuration of rat cortical neurons and patch clamp technology were used.

First, how meclofennamic acid (Compound 1) and 1-EBIO (Compound 10) influence the induced action potential action of rat cortical neurons. 4A shows that the induced potential is reversibly suppressed by meclofenamic acid (Compound 1) in the range of 5 to 20 µM. Each lane of Figure 4A is recorded from different neurons using the current-clamp configuration of the patch clamp technique.

Second, the spontaneous action of rat cortical neurons is shown to be completely but reversibly inhibited by 10 μM meclofennamic acid (Compound 1).

Example 5

Protection of Mice from Electric Shock Induced by Electric Shock with Meclofennamic Acid (Compound 1)

The effect of meclofennamic acid (Compound 1) on protecting mice from electroshock induced fainting will be described with reference to FIG.

Each group of 10 ICR mice received an intraperitoneal administration of saline or meclofennamic acid (Compound 1) at a rate of 25 mg / kg, 50 mg / kg, 100 mg / kg and 150 mg / kg after 30 minutes (50 mA, for 0.2 s, 60 Hz) was added. Relative proportions of mice not stunned were calculated in FIG. 5 for each dose.

In terms of the strong inhibition enteric of meclofennamic acid (Compound 1) against cortical neurons, the anticonvulsive action was tested in mice stunned by electric shock. Meclofennamic acid (Compound 1) dissolved in saline was injected intraperitoneally (in a volume of 10 ml / kg) into ICR adult mice in an amount of 25 mg / kg to 150 mg / kg, and in case of saline and convulsions Preventive action was compared. Thirty minutes after drug administration, the patient was stunned by electric shock (50 ma, 0.2 sec, 60 Hz). FIG. 5 shows that 50 mg / kg of meclofennamic acid (Compound 1) significantly protected 50% of mice from electric shock and sufficiently prevented fainting in an amount of 100 mg / kg. Meclofennamic acid (Compound 1) at 150 mg / kg stabilized the mice.

Example 6

Effect of Diclomenac (Compound 2) on Voltage-dependent Activation of KCNQ2 / 3 Current and Deactivation of KCNQ2 / 3 Current

The leftward conversion of the activation voltage dependence of the CKNQ2 / 3 current caused by Diclofenac (Compound 2) is described with reference to FIGS. 6A-6C.

6A-6C show an increase in KCNQ2 / 3 current induced by Diclofenac (Compound 2). 6A shows the total cell current of KCNQ2 / 3 heterologously expressed in CHO cells recorded before and after injection of 50 μM Diclofenac. In Figure 6A, the cell membrane was stepped from -90 mV to -50 mV (1 second) to -60 mV (0.75 second) wide. Recordings were made every 30 seconds. FIG. 6B shows the percentage of the current provided (control is 100%) in the presence (+) or absence (-) of diclofenac obtained from the experiment provided in FIG. 6A. In FIG. 6C, normalized conductance (G / G _max ) is shown as a function of voltage step in control (open square) and diclofenac (closed square) for KCNQ2 / 3 current.

In Figure 6C, 50μM diclofenac (Compound 2) was added causing a leftward when converted to _{V 50 = -30.9 ± 4.1 mV V} 50 = -45.4 ± 2.7 mV (n = 7, p <0.01) from the in the KCNQ2 / 3 active It shows.

6C shows that, like meclofennamic acid (Compound 1), treatment of CHO cells with Diclofenac (Compound 2) reduces the deactivation kinetics of KCNQ2 / 3 channels.

6A and 6B show that KCNQ2 / 3 current intensity is increased by Diclofenac 2 at physiologically relevant potential. In this train regulation, when cells stepped up from -85 mV to -50 mV, diclofenac (Compound 2) increased the current intensity by 262 ± 26% (n = 6).

The effect of Diclofenac (Compound 2) can be fully reversed (data not shown).

In general, meclofennamic acid (Compound 1) and Diclofenac (Compound 2) are shown to have a stronger effect of opener on KCNQ2 than on KCNQ3 channel subunits, as shown in Example 7 below.

Example 7

Selectivity of Meclofenacic Acid (Compound 1) Action

The opener properties of the meclofenacic acid (compound 1) and diclofenac for the heterologous KCNQ2 / 3 channel raise questions about whether these compounds work more selectively or equally well for subunits. To address this problem, the effects of 50 μM on homologous KCNQ2 and homologous KCNQ3 channels individually expressed in CHO cells were tested. The results are shown in Figs. 16A-D.

As shown in FIG. 16C, meclofennamic acid ranges from V ₅₀ = −23.6 ± 2.2 mV (n = 8) to V ₅₀ = −50.5 ± in the activation curve of KCNQ2 channel in control and meclofennamic acid-treated cells. It was found to exhibit a generally stronger action on CKNQ2 than on the CKNQ3 channel, as it caused an actual leftward conversion of -26.9 mV width to 1.4 mV (n = 5) (FIG. 16C, left panel, p <0.01). For the activation curves of the claw KCNQ3 channel methoxy leftward transformation caused by the pen acid, respectively, and methoxy-verified claw pen acid - V ₅₀ = -54.0 ± in _{V 50 = -39.0 ± 3.5 mV (} n = 11) in the treated cells It became weaker at 2.0 mV (n = 6) (-15 mV) (FIG. 16C, right panel, p <0.01).

_{Meclofennamic} acid (Compound 1) is a KCNQ2 channel closure with a time constant of deactivation increasing from τ _deact = 92.6 ± 3.9 msec to τ _deact = 152.7 ± 4.9 msec (Figures 16A and 16D, n = 8, p <0.001). Significantly reduced the speed. In contrast, there was no effect on deactivation kinetics of KCNQ3 channels (τ _deact = 319.6 ± 40.8 msec and τ _deact = 317.2 ± 30.2 msec, n = 8 for control and _{meclofennamic} acid-treated cells).

When cells step up from -85 mV to -40 mV (FIG. 16A and B; n = 8, p <0.001), 50 μM meclofennamic acid is reflected, reflecting the stronger action of the opener on KCNQ2 versus KCNQ3 channels. When applied, the KCNQ2 and KCNQ3 current intensities were increased to 240 ± 26% and 120 ± 4%, respectively.

Interestingly, homologous KCNQ1 and heterologous KCNQ1 / KCNE1 currents were not enhanced by 50 μM diclofenac or 50 μM meclofennamic acid over a range of test potentials of −50 to 0 mV (data not shown). Instead, both openers reduced the current intensity at positive potential (from 0 mV to 40 mV).

Assays conducted with compound 3-9

As described below, KCNQ2 / 3 open action was tested for novel compounds 3, 4, 5, 6, 7, 8 and 9. It is important that compound 3-9 was tested, did not affect KCNQ1 / KCNE1 heart channel and showed selective brain specificity. The result is not shown.

Example 8

Effect of Compound 6 on KCNQ2 / 3 Channels and Rat Cortical Neurons

The effect of Compound 6 on heterologously expressed recombinant KCNQ2 / 3 potassium channels and rat cortical neurons heterologously in CHO cells is described with reference to FIGS. 7A-7C, 8A-8B and 9A-9C.

In Figures 7A-7C, the effect of compound 6 on the KCNQ2 / 3 current is shown. 7A shows the total-cell currents recorded before and after injection of 25 μM Compound 6. FIG. 7B shows the percentage of the current provided (control is 100%) in the presence (+) or absence (−) of Compound 6 obtained from the experiment provided in FIG. 7A. In FIG. 7C, normalized conductance (G / G _max ) is obtained as a function of voltage step for control (open square) and compound 6 (closed square) for KCNQ2 / 3 current.

In FIGS. 8A-8B, inhibition of neuronal action induced by compound 6 is shown. In FIG. 8A, the neuronal action induced by square polarization amplification current, inhibited by 10 μM compound 6 and recovered after washing is shown. In Figure 8A, the polarization erase current was 50 pA for 800 msec. In FIG. 8B, the induced neuronal action is recorded before and after injecting Compound 6 externally using a lamp regime and restored after washing. In the ramp specification shown in Fig. 8B, the polarization cancellation current increased from 0 pA to 300 pA within 800 msec.

9A-9C show inhibition of spontaneous neuronal action by different concentrations of compound 6. The spontaneous action recorded before and after the addition of Compound 6 is shown for 20 μM in FIG. 9A, 10 μM in FIG. 9B and 5 μM in FIG. 9C.

7C shows the effect of 25 μM Compound 6 on recombinant KCNQ2 / 3 channels expressed in CHO cells. Methoxy claw pen acid (Compound 1) and diclofenac (2) and similarly, when applied externally, compound 6 is the voltage of the KCNQ2 / 3 activation - from V ₅₀ = -30.9 ± 4.1 mV in dependence V ₅₀ = -43.45 ± 2.3 mV A large leftward transformation of about 13 mV occurred until (n = 7, p <0.01).

7A shows that Compound 6 reduced the deactivation kinetics of KCNQ2 / 3 channel.

7A and 7B show that KCNQ2 / 3 current intensity was increased by compound 6 at physiologically relevant potentials. In this train regulation, when cells step up from -85 mV to -50 mV, using 25 μM compound 6 increased the current intensity to 220% (n = 10).

The effect of compound 6 may be reversed (data not shown).

One of the main functions of the M-current is to attenuate neuronal spiking discharge, thus testing how Compound 6 affects the induction and spontaneous action potential of cultured rat cortical neurons. By using the current-clamp configuration of the patch clamp technique, the neuron action induced by a squared pulse (FIG. 8A; 50-300 pA, 800 msec) or a ramp of polarized current (FIG. 8B; 0-300 pA, 800 msec) The effect of compound 6 on the potential was tested. The resting membrane potential was close to -60 mV and maintained at this level by applying a DC current if necessary. Superfusion of 10 μM Compound 6 drastically and reversibly reduced the number of action potentials induced in cortical neurons. After 10 μM Compound 6 externally fused within 1 minute, cortical neurons excited the action potential by widening the interspike interval (FIG. 8A, second row). After 2 minutes of open exposure, only one spike could be caused by the same polarization clearance current (Figure 8A, row 3). Therefore, compound 6 consistently reduced the number of action potentials induced. The neurons recovered their initial spiking action after washing the compounds for less than 2 minutes (Figure 8A, row 5).

Similar results were obtained with the lamp current shown in FIG. 8B.

Spontaneous spiking action was recorded using higher density cultures of rat cortical neurons (FIG. 9). Compound 6 significantly inhibited spontaneous action potentials within 2 minutes dose-dependently (5 μM-20 μM). The inhibitory action of compound 6 can be reversed by washing compound 6 for all 3 concentrations.

Example 9

Effect of Compound 5 on KCNQ2 / 3 Channels and Rat Cortical Neurons

The effect of Compound 6 on heterologously expressed recombinant KCNQ2 / 3 potassium channels and rat cortical neurons heterologously in CHO cells is described with reference to FIGS. 10A-10C.

10A-10C, the effect of compound 6 on neuronal action and KCNQ2 / 3 current is shown. 10A shows the induced rat cortical neuronal action recorded before, after and after washing with 25 μM Compound 5. 10B shows the KCNQ2 / 3 currents recorded before (top panel) and after (bottom panel) use of 25 μM Compound 5. In FIG. 10B the cell membrane was stepped up to a width of 10 mV from -80 mV to -40 mV (holding potential = -90 mV). In FIG. 10C, normalized conductance (G / G _max ) is shown as a function of voltage step for control (open square) and compound 5 (closed square).

10A-10C, it can be seen that Compound 5 has KCNQ2 / 3 potassium current openness similar to that of Compound 6.

In Figure 10C, 25 μM Compound 5 KCNQ2 / 3 voltage in the activation-dependent in V ₅₀ = -30.9 ± from _{4.1 mV V 50 = -38.3 ± 1.9} mV (n = 5, p <0.05) up to a large approximately 8 mV Indicates a leftward conversion.

In FIG. 10B, compound 5 shows reduced deactivation kinetics of KCNQ2 / 3 channel. 15 μM Compound 5 also shows an increase in current intensity at physiologically relevant potentials of −40 and −50 mV.

In FIG. 10A, it is also shown that superfusion of 25 μM Compound 5 reversibly inhibited the number of action potentials induced in cortical neurons.

Example 10

Effect of Compound 3 on KCNQ2 / 3 Channel

The effect of compound 3 on recombinant KCNQ2 / 3 potassium channels heterologously expressed in CHO cells is described with reference to FIGS. 11A-11B.

11A-11B show an increase in KCNQ2 / 3 current in the presence of compound 3. 11A shows the current in the presence and absence (control) of 25 μM Compound 3. In FIG. 11A, the cells went up to -50 mV for 1.5 seconds pulses and the interval between pulses was 30 seconds. In FIG. 11B, the percentage of current provided (with control is 100%) in the presence (+) or absence (−) of compound 3.

11A-11B show that Compound 3 has strong KCNQ2 / 3 potassium channel openness at 25 μM. In this train regime, when cells step up from -85 mV to -50 mV, application of 25 μM compound 3 increased the current intensity to 299 ± 47% (n = 6, p <0.002).

The effect of compound 3 could be fully reversed (data not shown).

Example 11

Effect of Compound 4 on KCNQ2 / 3 Channels and Rat Cortical Neurons

The effect of compound 4 on recombinant KCNQ2 / 3 potassium channels and murine cortical neurons heterologously expressed in CHO cells is described with reference to FIGS. 12A-12C.

12A-12C show the effect of compound 4 on neuronal action and KCNQ2 / 3 current. 12A shows the 12A KCNQ2 / 3 current recorded before (left panel) and after (right panel) injection of 50 μM Compound 4. The cell membrane was stepped up to a width of 10 mV from -80 mV to -40 mV (holding potential = -90 mV). 12B shows the spontaneous cortical neuronal action recorded before and after addition of 20 μM Compound 4 and after washing. In FIG. 12D, normalized conductance (G / G _max ) is obtained as a function of voltage step for control (open square) and compound 4 (closed square).

12A-12C, it can be seen that Compound 4 is a strong KCNQ2 / 3 channel opener.

As shown in FIG. 12A, when CHO cells step up from -85 mV to -50 mV and -40 mV, 50 μM Compound 4 is current intensities more than four and 1.5 times greater as described above for Compound 3, respectively. Increased.

12B, it can be seen that when applied to rat cortical neurons, 25 μM Compound 4 significantly inhibited spontaneous spiking action. As shown in FIG. 12B, the effect of compound 4 could be fully reversed.

Example 12

Effect of Compound 9 on KCNQ2 / 3 Channels and Rat Cortical Neurons

The effect of compound 9 on rat cortical neurons is described with reference to FIG. In FIG. 13, spontaneous neuronal action (action potential) as shown by 20 μM Compound 9 is shown.

As shown in FIG. 13, and in contrast to other molecules of the present invention, 20 μM of compound 9 showed an inhibitory effect on the induction and spontaneous spiking action of cortical neurons. However, in contrast to other molecules of the invention, compound 9 exhibited only one week opener action on recombinant KCNQ2 / 3 channels heterologously expressed in CHO cells. These results indicate that Compound 9 is neuronal inhibitory through a mechanism that does not include KCNQ2 / 3 channels.

Example 13

Effect of Compound 7 on KCNQ2 / 3 Channels and Rat Cortical Neurons

The effect of compound 7 on recombinant KCNQ2 / 3 potassium channels heterologously expressed in CHO cells is described with reference to Figures 14A-14D.

14A-14D show the effect of 20 μM compound 7 on KCNQ2 / 3 channel and neuronal action. In FIG. 14A, KCNQ2 / 3 total cell current recorded before and after injection of 20 μM Compound 7 is shown. In FIG. 14B, the percentage of current provided (control is 100%) in the presence (+) or absence (-) of compound 7 obtained from the experiment provided in FIG. 14A. In FIG. 14C, normalized conductance (G / G _max ) of KCNQ2 / 3 current is obtained as a function of voltage step for control (open square) and compound 7 (closed square). In FIG. 14D, regulation of spontaneous neuronal action (action potential) by 20 μM compound 7 is shown.

14 shows the effect of compound 7 on KCNQ2 / 3 channel. Like compounds 6 and 5 described above, compound 7 is a large KCNQ2 / 3 channel opener with a significant leftward transformation in the voltage dependent activation curve. This effect resulted in a large increase in channel strength as determined by train regulation (FIGS. 14A and 14B). Likewise, 20 μM compound 7 showed a very strong inhibitory effect on the induction and spontaneous spiking action of cortical neurons (FIG. 14D). This effect could be reversed sufficiently.

Example 14

Effect of Compound 7 on KCNQ2 / 3 Channels and Rat Cortical Neurons

The effect of Compound 8 on recombinant KCNQ2 / 3 potassium channels heterologously expressed in CHO cells and murine cortical neurons is described with reference to FIGS. 15A-15B.

15A-15B show the induced and spontaneous neuronal action regulated by compound 8. 15A shows the induced rat cortical neuronal action recorded before, during and after applying 10 μM Compound 8. In 15B, spontaneous neuronal action (action potential) is regulated by 5 μM Compound 8.

15A-15B show that low concentrations of compound 8 have a strong inhibitory effect on the induction and spontaneous spiking action of cortical neurons. However, like compound 9 described above, compound 8 exhibited only weak opener action of recombinant KCNQ2 / 3 potassium channels heterologously expressed in CHO cells. This result implies that compound 8 has neuronal inhibitory action through a mechanism that does not include KCNQ2 / 3 channels.

Features of the invention described in separate embodiments may be provided in combination with a single embodiment. In other words, the various features of the invention described in a single embodiment may also be provided individually or in any suitable combination.

Although the present invention has been described in connection with particular embodiments, numerous alternatives, improvements, and variations will occur to those skilled in the art. Accordingly, it is intended to embrace all such alterations, modifications and variations that fall within the spirit and broad scope of the claims. All publications, patents, and patent applications mentioned in this specification are incorporated into the specification to the extent that individual publications, patents, or patent applications refer to the invention. In addition, citation or identification of references mentioned in this application is not considered to be a known technology for the present invention.

Claims (64)

A therapeutically effective amount of a compound selected from the group consisting of N-phenylanthranilic acid, N-phenylanthranilic acid derivatives, 2-benzimidazolone and 2-benzimidazolone derivatives, or pharmaceutically acceptable salts thereof, is administered to a patient in need thereof. A method of regulating a voltage-dependent potassium channel comprising: The method according to claim 1, wherein the compound is the following general formula (I) or (II), or a pharmaceutically acceptable salt thereof: (I) (II) Where Z is an A-G (= K) -X-Y group, A is alkyl or absent, G is selected from the group consisting of carbon, sulfur and substituted or unsubstituted phosphorus, K is selected from the group consisting of oxygen and sulfur, X is selected or absent from the group consisting of substituted or unsubstituted nitrogen, oxygen and sulfur, Y is selected from the group consisting of hydrogen, alkyl, hydroxyalkyl, cycloalkyl, aryl and polyalkylene glycol moieties, Q and W are each independently selected from the group consisting of substituted or unsubstituted nitrogen, oxygen, sulfur and carbon, D is selected from the group consisting of oxygen and sulfur, R ¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl and aryl, R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are each independently hydrogen, alkyl, hydroxyalkyl, Trihaloalkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, Cyano, nitro, amino and -NR ¹⁵ R ¹⁶ , or at least two of R ² , R ³ , R ⁴ , R ⁵ and R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ At least two of and / or at least two of R ¹¹ , R ¹² , R ¹³ and R ^{14 form a} 5- or 6-membered aromatic, heteroaromatic, cycloaliphatic or heteroalicyclic ring, R ¹⁵ and R ¹⁶ are each independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, carbonyl and sulfonyl, or R ¹⁵ and R ¹⁶ represent a 5- or 6-membered heteroalicyclic ring. Forming, Here, when phosphorus and / or nitrogen are substituted, the substituent is selected from the group consisting of alkyl, cycloalkyl and aryl. The method of claim 2 wherein said compound has formula (I). 4. The process of claim 3 wherein Y is selected from the group consisting of hydroxyalkyl and polyalkylene glycol moieties. The process of claim 4 wherein the polyalkylene glycol moiety has the formula (III) [O- (CH ₂ ) m] n-OR ¹⁷ (III) Wherein m and n are each independently an integer from 1 to 10 and R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl. 4. The compound of claim 3 wherein G is carbon, K is oxygen, and R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are each independently selected from the group consisting of hydrogen, alkyl, halo and trihaloalkyl , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each hydrogen. The method of claim 2 wherein said compound has formula (II). 8. The method of claim 7, wherein Q and W are each substituted or unsubstituted nitrogen, and D is oxygen. 9. The method of claim 8, wherein Q is each substituted nitrogen. The method of claim 2, wherein the compound is selected from the group consisting of the following compounds 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 or pharmaceutically acceptable salts thereof: The method of claim 1, wherein the regulated voltage-dependent potassium channel comprises a KCNQ2 channel and / or a KCNQ3 channel and / or a KCNQ2 / 3 channel. 2. The method of claim 1, wherein the regulation of voltage-dependent potassium channels comprises epilepsy, ischemic seizures, migraine, ataxia, muscle cramps, neuralgia, Alzheimer's disease, Parkinson's disease, old age memory loss, learning deficiency, bipolar disorder, trigeminal neuralgia , Spasm, mood disorders, mental disorders, brain tumors, hearing and blindness, anxiety and motor neuron disease. The method of claim 1, wherein said administration consists of intranasal, subcutaneous, intravenous, intramuscular, parenteral, oral, topical, intradermal, bronchial, oral, buccal, sublingual, anal and mucosal. The method of claim 1, wherein the compound forms part of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier. 15. The method of claim 14, wherein said pharmaceutical composition further comprises an agent selected from the group consisting of antimicrobial agents, antioxidants, buffers, intestinal expandables, anti-inflammatory agents, antiviral agents, chemotherapeutic agents and antihistamines. A method for inhibiting cortical neuronal action by administering to a patient in need thereof a therapeutically effective amount of a compound selected from the group consisting of N-phenylanthranilic acid, N-phenylanthranilic acid derivatives, 2-benzimidazolone and 2-benzimidazolone derivatives. . The method of claim 16, wherein the compound is of formula (I) or (II), or a pharmaceutically acceptable salt thereof: (I) (II) Where Z is an A-G (= K) -X-Y group, A is alkyl or absent, G is selected from the group consisting of carbon, sulfur and substituted or unsubstituted phosphorus, K is selected from the group consisting of oxygen and sulfur, X is selected or absent from the group consisting of substituted or unsubstituted nitrogen, oxygen and sulfur, Y is selected from the group consisting of hydrogen, alkyl, hydroxyalkyl, cycloalkyl, aryl and polyalkylene glycol moieties, Q and W are each independently selected from the group consisting of substituted or unsubstituted nitrogen, oxygen, sulfur and carbon, D is selected from the group consisting of oxygen and sulfur, R ¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl and aryl, R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are independently hydrogen, alkyl, hydroxyalkyl, tri Haloalkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, sia No, nitro, amino and —NR ¹⁵ R ¹⁶ , while at least two of R ² , R ³ , R ⁴ , R ⁵ and R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ At least two and / or at least two of R ¹¹ , R ¹² , R ¹³ and R ^{14 form a} 5- or 6-membered aromatic, heteroaromatic, cycloaliphatic or heteroalicyclic ring, R ¹⁵ and R ¹⁶ are independently of each other selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, carbonyl and sulfonyl, or R ¹⁵ and R ^{16 form} a 5- or 6-membered heteroalicyclic ring and, Here, when phosphorus and / or nitrogen are substituted, the substituent is selected from the group consisting of alkyl, cycloalkyl and aryl. 18. The method of claim 17, wherein said compound has formula (I). 19. The method of claim 18, wherein Y is selected from the group consisting of hydroxyalkyl and polyalkylene glycol moieties. 20. The process of claim 19, wherein the polyalkylene glycol moiety has the formula [O- (CH ₂ ) m] n-OR ¹⁷ (III) Wherein m and n are each independently an integer from 1 to 10 and R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl. 19. The compound of claim 18, wherein G is carbon, K is oxygen, and R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are each independently selected from the group consisting of hydrogen, alkyl, halo and trihaloalkyl , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each hydrogen. 18. The method of claim 17, wherein said compound has formula (II). The method of claim 22, wherein Q and W are each substituted or unsubstituted nitrogen, and D is oxygen. 23. The method of claim 22, wherein each Q is substituted nitrogen. 18. The method of claim 17, wherein the compound is selected from the group consisting of the following compounds: 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 or pharmaceutically acceptable salts thereof. 17. The method of claim 16, wherein the inhibition of cortical neuronal action is epilepsy, ischemic attack, migraine, ataxia, muscle cramps, neuralgia, Alzheimer's disease, Parkinson's disease, old age memory loss, learning deficiency, bipolar disorder, trigeminal neuralgia, A method for treating a disease or condition selected from the group consisting of convulsions, mood disorders, mental disorders, brain tumors, hearing and vision loss, anxiety and motor neuron disease. The method of claim 16, wherein said administration consists of intranasal, subcutaneous, intravenous, intramuscular, parenteral, oral, topical, intradermal, bronchial, oral, buccal, sublingual, anal and mucosal. The method of claim 16, wherein the compound forms part of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier. 29. The method of claim 28, wherein said pharmaceutical composition further comprises an agent selected from the group consisting of antimicrobial agents, antioxidants, buffers, enteric drugs, anti-inflammatory agents, antiviral agents, chemotherapeutic agents and antihistamines. An active ingredient comprising a compound selected from the group consisting of N-phenylanthranilic acid, N-phenylanthranilic acid derivatives, 2-benzimidazolone and 2-benzimidazolone derivatives, and pharmaceutically acceptable salts thereof; A pharmaceutical composition for the treatment or prevention of an illness or disease, wherein it is advantageous to modulate dependent potassium channels and / or to inhibit cortical neuronal action. The pharmaceutical composition according to claim 30, further comprising a compound of formula (I) or (II), or a pharmaceutically acceptable salt thereof: (I) (II) Where Z is an A-G (= K) -X-Y group, A is alkyl or absent, G is selected from the group consisting of carbon, sulfur and substituted or unsubstituted phosphorus, K is selected from the group consisting of oxygen and sulfur, X is selected or absent from the group consisting of substituted or unsubstituted nitrogen, oxygen and sulfur, Y is selected from the group consisting of hydrogen, alkyl, hydroxyalkyl, cycloalkyl, aryl and polyalkylene glycol moieties, Each of Q and W is independently selected from the group consisting of substituted or unsubstituted nitrogen, oxygen, sulfur and carbon, D is selected from the group consisting of oxygen and sulfur, R ¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl and aryl, R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are each independently hydrogen, alkyl, hydroxyalkyl, Trihaloalkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, Cyano, nitro, amino and -NR ¹⁵ R ¹⁶ , or at least two of R ² , R ³ , R ⁴ , R ⁵ and R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ At least two of and / or at least two of R ¹¹ , R ¹² , R ¹³ and R ^{14 form a} 5- or 6-membered aromatic, heteroaromatic, cycloaliphatic or heteroalicyclic ring, R ¹⁵ and R ¹⁶ are each independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, carbonyl and sulfonyl, or R ¹⁵ and R ¹⁶ represent a 5- or 6-membered heteroalicyclic ring. Forming, Here, when phosphorus and / or nitrogen are substituted, the substituent is selected from the group consisting of alkyl, cycloalkyl and aryl. The pharmaceutical composition according to claim 31, wherein the compound has general formula (I). The pharmaceutical composition according to claim 32, wherein Y is selected from the group consisting of hydroxyalkyl and polyalkylene glycol moieties. The pharmaceutical composition according to claim 33, wherein the polyalkylene glycol moiety has the following general formula (III): [O- (CH ₂ ) m] n-OR ¹⁷ (III) Wherein m and n are each independently an integer from 1 to 10 and R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl. 33. The compound of claim 32, wherein G is carbon, K is oxygen, and R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are each independently selected from the group consisting of hydrogen, alkyl, halo and trihaloalkyl , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each a pharmaceutical composition. The pharmaceutical composition according to claim 31, wherein the compound has general formula (II). The pharmaceutical composition according to claim 36, wherein Q and W are each substituted or unsubstituted nitrogen, and D is oxygen. The pharmaceutical composition according to claim 37, wherein Q is each substituted nitrogen. 32. A pharmaceutical composition according to claim 31, wherein said compound is selected from the group consisting of compounds 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 or pharmaceutically acceptable salts thereof. The pharmaceutical composition according to claim 30, wherein the voltage-dependent potassium channel comprises a KCNQ2 channel and / or a KCNQ3 channel and / or a KCNQ2 / 3 channel. 31. The method of claim 30, wherein the disease or condition is epilepsy, ischemic attack, migraine, ataxia, muscle cramps, neuralgia, Alzheimer's disease, Parkinson's disease, old age memory loss, learning deficiency, bipolar disorder, trigeminal neuralgia, cramps, A pharmaceutical composition selected from the group consisting of mood disorders, mental disorders, brain tumors, hearing and vision loss, anxiety and motor neuron disease. 31. A pharmaceutical composition according to claim 30, printed on or packaged in or on said packaging for use for the treatment or prevention of diseases or diseases associated with the altered action of voltage-dependent potassium channels and / or cortical neurons. 43. The method of claim 42, wherein the disease or condition is epilepsy, ischemic attack, migraine, ataxia, muscle cramps, neuralgia, Alzheimer's disease, Parkinson's disease, old age memory loss, learning deficiency, bipolar disorder, trigeminal neuralgia, cramps, A pharmaceutical composition selected from the group consisting of mood disorders, mental disorders, brain tumors, hearing and vision loss, anxiety and motor neuron disease. 44. The pharmaceutical composition of claim 43, further comprising a medicament selected from the group consisting of antimicrobial agents, antioxidants, buffers, extendible drugs, anti-inflammatory agents, antiviral agents, chemotherapeutic agents and antihistamines. A compound comprising N-phenylanthranilic acid or a derivative thereof and a hydroxyalkyl moiety or a polyalkylene glycol moiety covalently attached thereto, or a pharmaceutically acceptable salt thereof. 46. A compound according to claim 45 or a pharmaceutically acceptable salt thereof: (IV) Where Z is an A-G (= K) -X-Y group, A is alkyl or absent, G is selected from the group consisting of carbon, sulfur and substituted or unsubstituted phosphorus, K is selected from the group consisting of oxygen and sulfur, X is selected or absent from the group consisting of substituted or unsubstituted nitrogen, oxygen and sulfur, Y is selected from the group consisting of hydroxyalkyl and polyalkylene glycol moieties, R ¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl and aryl, R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each independently hydrogen, alkyl, hydroxyalkyl, trihaloalkyl, cycloalkyl, alkenyl, alky Neyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro, amino and -NR ¹⁵ R ¹⁶ At least two of R ² , R ³ , R ⁴ , R ⁵ and R ⁶ and / or at least two of R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are 5-membered or 6 -To form an aromatic, heteroaromatic, cycloaliphatic or heteroalicyclic ring, R ¹⁵ and R ¹⁶ are independently of each other selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, carbonyl and sulfonyl, or R ¹⁵ and R ^{16 form} a 5- or 6-membered heteroalicyclic ring and, Where phosphorus and / or nitrogen are substituted, the substituent is selected from the group consisting of alkyl, cycloalkyl and aryl. 47. The compound of claim 46, wherein the polyalkylene glycol moiety has the formula (V) [O- (CH ₂ ) m] n-OR ¹⁷ (V) Wherein m and n are each independently an integer from 1 to 10 and R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl. 48. The compound of claim 47, wherein G is carbon, K is oxygen, and R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are independently selected from the group consisting of hydrogen, alkyl, halo and trihaloalkyl, And each of R ⁷ , R ⁸ , R ⁹ and R ¹⁰ is hydrogen. A compound selected from the group consisting of the following compounds 3, 4, 5, 6, 7, 8 and 9 or pharmaceutically acceptable salts thereof: A pharmaceutical composition comprising the compound of claim 45 as an active ingredient. A pharmaceutical composition comprising the compound of claim 46 as an active ingredient. 52. A pharmaceutical composition according to claim 51 comprising as an active ingredient Compound 3 or a pharmaceutically acceptable salt thereof: 52. A pharmaceutical composition according to claim 51 comprising as an active ingredient Compound 4 or a pharmaceutically acceptable salt thereof: 52. A pharmaceutical composition according to claim 51 comprising as an active ingredient Compound 5 or a pharmaceutically acceptable salt thereof: 52. A pharmaceutical composition according to claim 51 comprising as an active ingredient Compound 6 or a pharmaceutically acceptable salt thereof: 52. A pharmaceutical composition according to claim 51 comprising as an active ingredient Compound 7 or a pharmaceutically acceptable salt thereof: 52. A pharmaceutical composition according to claim 51 comprising as an active ingredient Compound 8 or a pharmaceutically acceptable salt thereof: 52. A pharmaceutical composition according to claim 51 comprising as an active ingredient Compound 9 or a pharmaceutically acceptable salt thereof: Obtaining N-phenylanthranilic acid or a derivative thereof; And Reacting N-phenylanthranilic acid or a derivative thereof with hydroxyalkyl or polyalkylene glycol having a reactive group at the end, wherein the reactive group can form an ester bond with N-phenylanthranilic acid or a derivative thereof, A method of synthesizing the compound of claim 45. 60. The method of claim 59, wherein said ester bond is selected from the group consisting of carboxy ester bonds, carboxy amide bonds, carboxy thioester bonds, S-carboxy ester bonds, S-carboxy thioester bonds and S-carboxy amide bonds. 60. The method of claim 59, wherein said reactive group is selected from the group consisting of hydroxy, amine and thiohydroxy. 60. The method of claim 59, wherein the N-phenylanthranilic acid or derivative thereof has the general formula: (VI) Where A is alkyl or absent, G is selected from the group consisting of carbon, sulfur and substituted or unsubstituted phosphorus, wherein when said phosphorus is substituted, the substituent is selected from the group consisting of alkyl, cycloalkyl and aryl; K is selected from the group consisting of oxygen and sulfur, R ¹ is selected from the group consisting of hydrogen, alkyl, cycloalkyl and aryl, R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each independently hydrogen, alkyl, hydroxyalkyl, trihaloalkyl, cycloalkyl, alkenyl, alky Neyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro, amino and -NR ¹⁵ R ¹⁶ At least two of R ² , R ³ , R ⁴ , R ⁵ and R ⁶ and / or at least two of R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are 5-membered or 6 -To form an aromatic, heteroaromatic, cycloaliphatic or heteroalicyclic ring, R ¹⁵ and R ¹⁶ are independently of each other selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, carbonyl and sulfonyl, or R ¹⁵ and R ^{16 form} a 5- or 6-membered heteroalicyclic ring do. 63. The compound of claim 62, wherein G is carbon, K is oxygen, and R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are each independently selected from the group consisting of hydrogen, alkyl, halo and trihaloalkyl And R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each hydrogen. 60. The process of claim 59, wherein the polyalkylene glycol having a reactive group at the end has the general formula (VII): V- [O- (CH ₂ ) m] n-OR ¹⁷ (VII) Where V is hydroxy, amine or thiohydroxy, m and n are each independently an integer of 1 to 10, R ¹⁷ is hydrogen, alkyl, cycloalkyl or aryl.