HK1124321B - Substituted 4-phenyltetrahydroisoquinolines, method of producing them, their use as medicament, and also medicament containing them - Google Patents

Description

Substituted 4-phenyltetrahydroisoquinolines, process for their production, their use as medicaments and medicaments comprising them

The present invention relates to substituted 4-phenyltetrahydroisoquinolines. Medicaments comprising such compounds are useful in the prevention or treatment of various disorders. For example, these compounds are particularly useful in renal disorders such as acute or chronic renal failure, biliary dysfunction (disorders of biliary function) and respiratory disorders such as snoring or sleep apnea.

The present invention relates to compounds of formula I:

wherein:

r1, R2, R3 and R4

Each independently hydrogen, F, Cl, Br, I, CN, NO₂Or R11- (C)_mH_2m)-A_n-；

m is 0, 1, 2, 3 or 4;

n is 0 or 1;

r11 is hydrogen, methyl or C_pF_2p+1；

A is oxygen, NH, N (CH)₃) Or S (O)_q；

p is 1, 2 or 3;

q is 0, 1 or 2;

r5 is hydrogen; alkyl having 1, 2, 3, 4, 5, or 6 carbon atoms; or cycloalkyl having 3, 4, 5, or 6 carbon atoms;

r6 is hydrogen; OH; f; CF (compact flash)₃(ii) a Alkyl having 1, 2, 3, or 4 carbon atoms; or cycloalkyl having 3, 4, 5, or 6 carbon atoms;

r7 and R8

Each independently hydrogen, F, Cl, Br, CN, CO₂R12, NR13R14 or

R16-(C_mmH_2mm)-E_nn-；

R12 is hydrogen; alkyl having 1, 2, 3, or 4 carbon atoms; or cycloalkyl having 3, 4, 5, or 6 carbon atoms;

r13 and R14 are each independently hydrogen; alkyl having 1, 2, 3, or 4 carbon atoms; or

Cycloalkyl having 3, 4, 5, or 6 carbon atoms; or

R13 and R14 form together with the nitrogen atom to which they are bound a 4-, 5-, 6-or 7-membered ring in which one CH is₂The group may be replaced by NR15, S or oxygen;

r15 is hydrogen; alkyl having 1, 2, 3, or 4 carbon atoms; or cycloalkyl having 3, 4, 5, or 6 carbon atoms;

mm is 0, 1, 2, 3 or 4;

nn is 0 or 1;

r16 is hydrogen, methyl or C_ppF_2pp+1；

E is oxygen or S (O)_qq；

pp is 1, 2 or 3;

qq is 0, 1 or 2.

In one embodiment, compounds of formula I, and pharmaceutically acceptable salts and trifluoroacetates thereof, are preferred, as defined below, wherein:

r1, R2, R3 and R4

Each independently hydrogen, F, Cl, Br, CN or R11- (C)_mH_2m)-A_n-；

m is 0 or 1;

n is 0 or 1;

r11 is hydrogen, methyl or C_pF_2p+1；

A is oxygen, NCH₃Or S (O)_q；

p is 1 or 2;

q is 0, 1 or 2;

r5 is hydrogen, methyl, ethyl or cyclopropyl;

r6 is hydrogen or methyl;

r7 and R8

Each independently hydrogen, F, Cl, CN, CO₂R12, NR13R14 or

R16-(C_mmH_2mm)-E_nn-；

R12 is hydrogen, methyl or ethyl;

r13 and R14 are each independently hydrogen; alkyl having 1, 2, 3, or 4 carbon atoms; or

Cycloalkyl having 3, 4, 5, or 6 carbon atoms; or

R13 and R14 form together with the nitrogen atom to which they are bound a 5-, 6-or 7-membered ring in which one CH is₂The group may be replaced by NR15, S or oxygen;

r15 is hydrogen; alkyl having 1, 2, 3, or 4 carbon atoms; or cycloalkyl having 3, 4, 5, or 6 carbon atoms;

mm is 0, 1 or 2;

nn is 0 or 1;

r16 is hydrogen, methyl or C_ppF_2pp+1；

E is oxygen or S (O)_qq；

pp is 1 or 2;

qq is 0, 1 or 2.

Particular preference is given to compounds of the formula I and their pharmaceutically acceptable salts and trifluoroacetates, in which

R1 and R3 are each hydrogen;

r2 and R4 are each independently hydrogen, F, Cl, NH₂、NHCH₃Or N (CH)₃)₂；

R5 is hydrogen, methyl, ethyl or cyclopropyl;

r6 is hydrogen or methyl;

r7 and R8 are each hydrogen.

N-diaminomethylene-4- (6, 8-dichloro-2-methyl-1, 2, 3, 4-tetrahydroisoquinolin-4-yl) benzenesulfonamide and its pharmaceutically acceptable salts and trifluoroacetates are particularly preferred.

In one embodiment, compounds of formula I are preferably defined as follows: wherein the radicals R1, R2, R3 and R4 are each independently described as hydrogen, F, Cl, Br, CN or R11- (C)_mH_2m)-A_n-, where m and n are each independently 0 or 1, R11 is hydrogen, methyl or C_pF_2p+1And A is oxygen, NCH₃Or S (O)_qWherein p is 1 or 2 and q is 0, 1 or 2; particular preference is given to compounds of the formula I as defined below: wherein R1 and R3 are each hydrogen, and R2 and R4 are each independently hydrogen, F, Cl, NH₂、NHCH₃Or N (CH)₃)₂Such as Cl. In one embodiment, compounds of formula I are preferably defined as follows: wherein R2 and R4 are not hydrogen.

In another embodiment, compounds of formula I, defined as follows, are preferred: wherein R5 is described as hydrogen, methyl, ethyl or cyclopropyl, e.g. methyl.

In another embodiment, compounds of formula I, defined as follows, are preferred: wherein R6 is described as hydrogen or methyl.

In another embodiment, compounds of formula I, defined as follows, are preferred: wherein the radicals R7 and R8 are each independently described as hydrogen, F, Cl, CN, CO₂R12, NR13R14 or R16- (C)_mmH_2mm)-E_nn-, wherein R12 is hydrogen, methyl or ethyl, R13 and R14 are each independently hydrogen, alkyl having 1, 2, 3 or 4 carbon atoms, or cycloalkyl having 3, 4, 5 or 6 carbon atoms; or R13 and R14 together with the nitrogen atom to which they are bound form a 5-, 6-or 7-membered ring in which one CH is₂The group may be replaced by NR15, S or oxygen and wherein R15 is hydrogen, alkyl having 1, 2, 3 or 4 carbon atoms, or cycloalkyl having 3, 4, 5 or 6 carbon atoms; and wherein mm is 0, 1 or 2, nn is 0 or 1, and R16 is hydrogen, methyl or C_ppF_2pp+1Wherein E is oxygen or S (O)_qqWherein pp is 1 or 2 and qq is 0, 1 or 2; particular preference is given to compounds of the formula I as defined below: wherein R7 and R8 are each hydrogen.

If the compounds of formula I contain one or more asymmetric centers, they may each independently have the S and R configuration. These compounds may be in the form of optical isomers, diastereomers, racemates or mixtures thereof in all ratios.

The present invention encompasses all possible tautomeric forms of the compounds of formula I.

The invention also encompasses derivatives of the compounds of formula I, such as solvates (e.g., hydrates and alcohol adducts), esters, prodrugs and other physiologically acceptable derivatives of the compounds of formula I, as well as active metabolites of the compounds of formula I. The invention likewise encompasses all crystal modifications of the compounds of the formula I.

The alkyl group may be linear or branched. This also applies when they bear substituents or occur as substituents for other groups, for example in fluoroalkyl or alkoxy groups. Examples of alkyl groups are methyl, ethyl, n-propyl, isopropyl (═ 1-methylethyl), n-butyl, isobutyl (═ 2-methylpropyl), sec-butyl (═ 1-methylpropyl), tert-butyl (═ 1, 1-dimethylethyl), n-pentyl, isopentyl, tert-pentyl, neopentyl and hexyl. Preferred alkyl groups are methyl, ethyl, n-propyl, isopropyl and n-butyl. One or more, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14, hydrogen atoms in the alkyl group may be replaced by fluorine atoms. Examples of such fluoroalkyl groups are trifluoromethyl, 2, 2, 2-trifluoroethyl, pentafluoroethyl, heptafluoroisopropyl. The substituted alkyl group may be substituted at any position.

Alkylene radicals such as C_mH_2m、C_mmH_2mmOr C_rH_2rMay be straight chain or branched. When they bear substituents or substituents as other groups (e.g. in fluoroalkylene radicals, e.g. at C)_pF_2pAnd C_ppF_2ppIn (d), this also applies. Examples of alkylene are methylene, ethylene, 1-methylmethylene, propylene, 1-methylethylene, butylene, 1-propylmethylene, 1-ethyl-1-methylmethylene, 1, 2-dimethylethylene, 1-dimethylmethylene, 1-ethylethylene, 1-methylpropylene, 2-methylpropylene, pentylene, 1-butylmethylene, 1-propylethylene, 1-methyl-2-ethylethylene, 1, 2-dimethylpropylene, 1, 3-dimethylpropylene, 2-dimethylpropylene, hexylene and 1-methylpentylene. One or more of alkylene groups, e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9,10. 11 or 12 hydrogen atoms may be replaced by fluorine atoms. The substituted alkylene group may be substituted at any position. Having one or more CH groups in alkylene₂The groups may be replaced by oxygen, S, NH, N-alkyl or N-cycloalkyl.

Examples of cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. One or more, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12, hydrogen atoms in the cycloalkyl radical may be replaced by fluorine atoms. Substituted cycloalkyl groups may be substituted at any position. Cycloalkyl groups may also be branched, such as present as alkylcycloalkyl or cycloalkylalkyl, for example methylcyclohexyl or cyclohexylmethyl.

Derived from a compound in which R13 and R14 form a 4-, 5-, 6-or 7-membered ring with the nitrogen atom to which they are bonded, one CH₂Examples of rings of NR13R14, which groups may be replaced by NR15, S or oxygen, are morpholine, pyrrolidine, piperidine, piperazine and N-methylpiperazine.

If a variable appears more than once as a constituent, the definitions of the variables are independent of each other at each occurrence.

If the compounds of the formula I contain one or more acidic or basic groups or one or more basic heterocycles, the corresponding physiologically or toxicologically acceptable salts are also included in the scope of the invention, in particular the pharmaceutically acceptable salts. For example, the compounds of the formula I can be deprotonated at the acidic group and used, for example, in the form of the alkali metal salts, preferably the sodium or potassium salts, or in the form of the ammonium salts, for example with ammonia or organic amines or amino acids. Since the compounds of the formula I always contain at least one basic group, they can also be prepared in the form of physiologically acceptable acid addition salts, for example those formed with the following acids: inorganic acids such as hydrochloric acid, sulfuric acid, or phosphoric acid; or organic acids such as acetic acid, citric acid, tartaric acid, lactic acid, malonic acid, methanesulfonic acid, fumaric acid. Useful acid addition salts include those with all pharmacologically acceptable acids, for example halides, especially the hydrochloride, lactate, sulfate, citrate, tartrate, acetate, phosphate, methanesulfonate, p-toluenesulfonate, adipate, fumarate, gluconate, glutamate, glycerophosphate, maleate and pamoate (which groups also correspond to physiologically acceptable anions); also trifluoroacetic acid salts.

The invention also provides the following process for preparing the compound of formula I.

The compounds of formula I described herein can be prepared as follows: the compound of formula VIII is chlorosulfonated by methods known to those skilled in the art, and subsequently reacted with guanidine by methods known to those skilled in the art (e.g., as described in Synthetic Communications, 33(7), 1073; 2003).

It is not necessary to isolate the intermediate of formula XII obtained after chlorosulfonation, but rather a further reaction with guanidine can be carried out directly.

The starting compound of formula VIII can be prepared as follows:

tetrahydroisoquinolines of the formula VIIIa, wherein R1-R8 are each as defined above, can be prepared by methods known to the person skilled in the art by reduction of the carbonyl moiety in a compound of the formula VI, for example with sodium borohydride and subsequent acid-or base-catalyzed cyclization of the resulting alcohol of the formula VII (cf. Tetrahedron Lett.1989, 30, 5837; org. Prep. proced. int.1995, 27, 513).

To prepare alkyl-branched compounds of formula I wherein R6 is other than hydrogen, the corresponding diphenylacetate of formula IX is alkylated at the alpha position with R6 by known methods. Compounds of formula X can be converted by standard methods to the corresponding amides of formula XI which are converted in a Pictet-Schpengler (Pictet-Spengler) -like reaction to the desired tetrahydroisoquinolines of formula VIIIb (see Tetrahedron 1987, 43, 439; chem. pharm. Bull.1985, 33, 340), wherein R1-R8 are each as defined above and LG corresponds to a leaving group commonly used for alkylation, e.g. bromo, chloro, tosylate or mesylate.

The compounds of formula VI used above are preferably prepared in a manner known to the skilled person from benzylamines of formula IV and appropriate amino-substituted α -bromoacetophenone compounds of formula V, wherein R1-R8 are each as defined above.

The α -bromoacetophenone compounds of formula V can be obtained by bromination from the corresponding acetophenone precursors according to literature procedures.

If the benzylamine precursor of formula IV is not commercially available, it can be prepared by standard methods known to those skilled in the art from the corresponding benzyl halide of formula III, e.g., benzyl chloride or bromide, and the corresponding amine R5-NH₂Wherein R1-R5 are each as defined above and X is F, Cl, Br or I, especially Cl or Br.

Alternatively, the compounds of formula IV can also be obtained by reductive amination by aldehydes of formula IIIa by standard methods known to those skilled in the art, wherein R1-R5 are each as defined above.

Formulae III and IIIa, IX and R6-LG and R5-NH₂The compounds of (a) can be obtained commercially or can be prepared by literature methods known to the skilled worker or analogously thereto.

The products and/or intermediates are worked up and, if desired, purified by customary methods, such as extraction, chromatography or crystallization and customary drying.

The compounds of formula I have been shown to be significant inhibitors of the sodium hydrogen exchange pump (NHE), especially subtype 3 sodium hydrogen exchange pump (NHE 3). In addition, the compounds of formula I are also significant inhibitors of the subtype 5 sodium hydrogen exchange pump (NHE 5).

The NHE3 inhibitors known to date are, for example, compounds derived from acylguanidines (EP825178), norbornylamines (WO0144164), 2-guanidinoquinazolines (WO0179186) or benzamidines (WO0121582, WO 0172742). According to the current state of knowledge, squalamine, which has likewise been described as an inhibitor of NHE3 (M.Donowitz et al, am.J.Physiol.276(Cell Physiol.45): C136-C144), does not act as immediately as the compound of formula I, but rather acts by an indirect mechanism, thus reaching its maximum strength of action only after 1 hour.

Tetrahydroisoquinolines as inhibitors of the subtype 3 sodium hydrogen exchange pump (NHE3) have been described in patent applications WO03048129, WO2004085404 and german applications 102004046492.8 and 102005001411.9. Patent application WO03055880 describes tetrahydroisoquinolines as NHE3 inhibitorsRelated compound classes of salts. Has been surprisingIt was found that the compounds of formula I as described herein likewise constitute potent inhibitors of NHE3 and potent inhibitors of NHE5, and have advantageous pharmacological and pharmacokinetic properties.

NHE3 was found in vivo in different species, preferably in the gallbladder, intestine and kidney (Larry Fliegel et al, biochem. cell. biol. 76: 735-Asn 741, 1998), and also in the brain (E. Ma et al, Neuroscience 79: 591-Asn 603). NHE5 is expressed only in neurons and is therefore brain-specific (am.J.Physiol.cell.Physiol.281: C1146-C1157, 2001).

The compounds of the formula I are suitable for the prophylaxis and treatment of disorders which are caused by NHE activated or activated NHE, as well as disorders which have NHE-related damage as a secondary cause, owing to their NHE-inhibiting properties.

The compounds of formula I may also be useful in the treatment and prevention of conditions in which NHE is only partially inhibited, for example by using lower doses.

The use of the compounds of the present invention relates to the prevention and treatment of acute and chronic disorders in veterinary and human medicine.

As a result of their pharmacological action, the compounds of the formula I are particularly suitable for improving respiratory drive. They are therefore useful for treating disturbed respiratory states, which occur, for example, in the event of the following clinical states and conditions: disturbed central respiratory drive (e.g. central sleep apnea, sudden infant death, post-operative hypoxia), muscle-related respiratory disorders, respiratory disorders after prolonged ventilation, respiratory disorders during adaptation in mountains, obstructive and mixed forms of sleep apnea, chronic lung disease with hypoxia and hyperpnea.

Furthermore, the compounds increase the muscle tone of the upper respiratory tract in order to suppress snoring. The mentioned compounds are therefore advantageously used for the preparation of medicaments for the prophylaxis and treatment of sleep apnoea and muscle-related respiratory disorders and for the preparation of medicaments for the prophylaxis and treatment of snoring.

It was found that the combination of the NHE inhibitor of formula I with a carbonic anhydrase inhibitor (e.g. acetazolamide) can be advantageous, the latter causing metabolic acidosis and thereby increasing respiratory activity by itself, so that the effect of the active ingredient can be enhanced and its use reduced.

As a result of their NHE 3-inhibiting effect, the compounds of the invention protect cellular energy stores that are rapidly depleted in toxic and pathogenic events, thus leading to cell damage or cell death. Under the influence of NHE3 inhibitors, sodium absorption in the proximal tubule, which consumes energy-intensive ATP, is temporarily stopped, whereby the cells can survive acute pathogenic, ischemic or toxic conditions. These compounds are therefore suitable, for example, as medicaments for the treatment of ischemic etiologies, such as acute renal failure. Furthermore, these compounds are also suitable for the treatment of all chronic renal disorders and forms of nephritis, which lead to chronic renal failure due to increased protein deposition. The compounds of the formula I are therefore suitable for the preparation of medicaments for the treatment of late-stage diabetic lesions, diabetic nephropathy and chronic renal disorders, in particular all nephritis associated with protein/albumin deposits.

It has been found that the compounds used according to the invention have a mild laxative effect and can therefore also be used advantageously as laxatives or in the event of an urgent constipation (acute constipation).

Furthermore, the compounds of the invention can be advantageously used for the prevention and treatment of acute and chronic disorders of the intestinal tract which are induced, for example, by ischemic states in the intestinal region and/or by subsequent reperfusion or by inflammatory states and events. Such complications may arise, for example, as a result of inappropriate bowel movement, as observed in the event of constipation or a substantial decrease in bowel activity, such as is commonly observed following surgical intervention.

It is possible to prevent gallstone formation using the compounds of the present invention.

The NHE inhibitors of the invention are generally suitable for the treatment of conditions caused by ischemia and by reperfusion.

As a result of their pharmacological properties, the compounds of the invention are suitable as antiarrhythmic agents.

As a result of their cardioprotective component, NHE inhibitors are outstandingly suitable for the prophylaxis of infarcts and for the treatment of infarcts, and for the treatment of angina pectoris, in which case they also inhibit or significantly reduce the development of ischemia-induced states, in particular trigger pathophysiological processes in ischemia-induced arrhythmias. In particular their protective action against pathological hypoxic and ischemic conditions, the compounds of the formula I used according to the invention as cellular Na⁺/H⁺The outcome of the exchange mechanism can be used as a treatment for all acute or chronic injuries induced by ischemia or diseases induced primarily or secondarily therefrom.

The invention also relates to their use as medicaments for surgical interventions. For example, the compounds of the invention may be used in organ transplantation, in which case the compounds may be used to protect organs in donors prior to and during extirpation, to protect extirpated organs, for example during treatment with physiological baths or storage therein, and during transfer to recipient organisms pretreated with a compound of formula I.

These compounds are also valuable protective drugs in the manipulation of angioplasty interventions, such as angioplasty interventions on the heart and also on peripheral organs and blood vessels.

Furthermore, the compounds of the present invention may be used in the practice of bypass surgery, such as bypass surgery on coronary vessels and Coronary Artery Bypass Grafting (CABG).

The compounds of formula I according to the invention are also useful for resuscitation after cardiac arrest, depending on their effect against ischemia-induced damage.

Depending on their effect against ischemia-induced damage, these compounds are suitable as medicaments for the treatment of ischemia of the nervous system, in particular of the CNS, in which case they are suitable, for example, for the treatment of stroke or cerebral edema.

Since NHE inhibitors effectively protect human tissues and organs not only by combating the damage caused by ischemia and reperfusion, but also by combating the cytotoxic effects of drugs used in particular in cancer therapy and in the treatment of autoimmune diseases, their combined administration with compounds of formula I is suitable for reducing or inhibiting the cytotoxic effects of the therapy. The reduction in cytotoxicity, particularly cardiotoxicity, as a result of co-administration with NHE inhibitors also allows for increased doses of cytotoxic therapeutic agents and/or prolonged administration of such drugs. The therapeutic benefit of such cytotoxic therapies can be substantially increased by combination with NHE inhibitors. The compounds of formula I are particularly suitable for improving therapy with drugs containing undesired cardiotoxic components.

In general, the NHE inhibitors described herein may advantageously be combined with other compounds that also modulate intracellular pH, in which case possible combination components are inhibitors of the enzyme pool of carbonic anhydrase, inhibitors of systems that transport bicarbonate ions, such as sodium bicarbonate cotransporter (NBC) or sodium-dependent chlorine-bicarbonate exchange pump (NCBE), and may also advantageously be combined with wherein NHE inhibitors have an inhibitory effect on other NHE subtypes, as they are capable of enhancing or modulating the pharmacologically relevant pH-modulating effects of the NHE inhibitors described herein.

In view of their protective effect against ischemia-induced damage, the compounds of the invention are also suitable for use as medicaments for the treatment of ischemia of the nervous system, in particular of the central nervous system, in which case they are suitable, for example, for the treatment of stroke or cerebral edema.

The compounds of the formula I are also suitable for the treatment and prophylaxis of diseases and disorders which are caused by an excessive excitation of the central nervous system, in particular for the treatment of epileptic disorders, centrally induced paroxysmal and tonic spasms, psychological depression (depression) states, anxiety disorders and psychoses. In these cases, the NHE inhibitors of the invention may be used alone or in combination with other substances having anti-epileptic activity or substances having anti-psychotic activity or carbonic anhydrase inhibitors such as acetazolamide, and may also be used in combination with NHE or other inhibitors of the sodium-dependent chloro-bicarbonate exchanger pump (NCBE).

Furthermore, the compounds of the formula I according to the invention are likewise suitable for the treatment of shock types such as anaphylactic shock, cardiogenic shock, hypovolemic shock and bacterial shock.

The compounds of formula I are likewise useful for the prophylaxis and treatment of thrombotic disorders, since they can also inhibit the aggregation of platelets themselves as NHE inhibitors. They can also inhibit or prevent the excessive release of inflammatory and clotting mediators, especially von willebrand (von willebrand) factor and thromboselectin, that occurs after ischemia and reperfusion. This allows to reduce and eliminate the pathogenic effects of thrombosis and inflammatory-related factors. Thus, the NHE inhibitors of the invention can be combined with other anticoagulant and/or thrombolytic active ingredients such as recombinant or native tissue plasminogen activator, streptokinase, urokinase, acetylsalicylic acid, thrombin antagonists, factor Xa antagonists, drugs with fibrinolytic activity, thromboxane receptor antagonists, phosphodiesterase inhibitors, factor vila antagonists, clopidogrel, ticlopidine, and the like. The use of the NHE inhibitors of the invention in combination with NCBE inhibitors and/or with carbonic anhydrase inhibitors such as acetazolamide is particularly advantageous.

In addition, the NHE inhibitors of the present invention are characterized by a strong inhibitory effect on cell proliferation, such as fibroblast proliferation and vascular smooth muscle cell proliferation. The compounds of formula I can therefore be used as valuable therapeutic agents for conditions in which cell proliferation constitutes a primary or secondary cause, and therefore as antiatherosclerotic agents, substances against chronic renal failure, cancer. They are thus useful in the treatment of organ hypertrophy and hyperplasia, such as cardiac and prostate hypertrophy and hyperplasia. The compounds of the formula I are therefore suitable for the prophylaxis and treatment of heart failure (congestive heart failure ═ CHF) and also for the treatment and prophylaxis of prostatic hyperplasia or prostatic hypertrophy.

NHE inhibitors are also characterized by delaying or preventing fibrotic disorders. Thus, they are suitable as prominent substances for the treatment of cardiac fibrosis, pulmonary fibrosis, hepatic fibrosis, renal fibrosis and other fibrotic disorders.

Because of the significant elevation of NHE in essential hypertension, the compounds of formula I are suitable for the prevention and treatment of hypertension and cardiovascular disorders. In these cases, they can be used alone or in combination with suitable combination partners for the treatment of hypertension and for the treatment of cardiovascular disorders. For example, one or more diuretics having a thiazide-like action, loop diuretics, aldosterone and pseudoaldosterone antagonists such as hydrochlorothiazide, indapamide, polythiazide, furosemide, piretanide, torasemide, bumetanide, amiloride, triamterene, spironolactone, or eplerenone (eplerone) can be combined with a compound of formula I. In addition, the NHE inhibitor of the present invention may be used in combination with: calcium antagonists, such as verapamil, diltiazem, amlodipine or nifedipine; and ACE inhibitors such as ramipril, enalapril, lisinopril, fosinopril, or captopril. Other advantageous combination partners are also beta-blockers, such as metoprolol, salbutamol and the like; antagonists of the angiotensin receptor and its receptor subtypes, such as losartan, irbesartan, valsartan, omapatrilat, gemopatrilat (gemopatra); an endothelin antagonist; a renin inhibitor; an adenosine receptor agonist; potassium channel inhibitors and activators, such as glyburide, glimepiride, diazoxide, chromaffin, minoxidil, and derivatives thereof; mitochondrial ATP-sensitive potassium channel (mitok (ATP) channel) activators; other potassium channel inhibitors, such as Kv1.5 and the like.

Due to their anti-inflammatory effects, the NHE inhibitors of the present invention may be useful as anti-inflammatory agents. Inhibition of the release of inflammatory mediators is noteworthy in this respect as regards mechanism. Thus, these compounds may be used alone or in combination with anti-inflammatory agents to prevent or treat chronic and acute inflammatory conditions. The combination partners used are advantageously steroidal and non-steroidal anti-inflammatory drugs.

It has also been found that NHE inhibitors show beneficial effects on serum lipoproteins. Thus, they are useful for preventing and resolving atherosclerotic lesions by eliminating pathogenic risk factors. They include not only primary hyperlipidemia, but also certain secondary hyperlipidemia that occurs, for example, in the event of diabetes. Furthermore, NHE inhibitors result in a significant reduction in infarctions induced by metabolic abnormalities, and in particular in a significant reduction in the size and severity of the induced infarctions. Therefore, the NHE inhibitors of formula I are advantageously applied in the preparation of a medicament for the treatment of hypercholesterolemia; for the preparation of a medicament for the prevention of atherogenesis; for the preparation of a medicament for the prevention and treatment of atherosclerosis; for the preparation of a medicament for the prevention and treatment of diseases induced by elevated cholesterol levels; for the preparation of a medicament for the prevention and treatment of diseases induced by endothelial dysfunction; for the preparation of a medicament for the prevention and treatment of atherosclerosis-induced hypertension; for the preparation of a medicament for the prevention and treatment of atherosclerosis-induced thrombosis; for the preparation of a medicament for the prevention and treatment of hypercholesterolemia-induced and endothelial dysfunction-induced ischemic injury and post-ischemic reperfusion injury; for the preparation of a medicament for the prevention and treatment of cardiac hypertrophy and cardiomyopathy and Congestive Heart Failure (CHF); for the preparation of a medicament for the prevention and treatment of hypercholesterolemia-induced and endothelial dysfunction-induced coronary spasm and myocardial infarction; for the preparation of a medicament for use in combination with a hypotensive substance, preferably with an Angiotensin Converting Enzyme (ACE) inhibitor and an angiotensin receptor antagonist, for the treatment of said disorders. The combination of the NHE inhibitor of formula I with a lipid-lowering active ingredient, preferably with an HMG-CoA reductase inhibitor (e.g. lovastatin or pravastatin), constitutes an advantageous combination of enhanced action and reduced use of the active ingredient, wherein the latter produces a serum lipid-reducing effect and thus increases the serum lipid-reducing properties of the NHE inhibitor of formula I.

Therefore, NHE inhibitors result in effective protection against endothelial damage of different origins. This vascular protective effect against endothelial dysfunction syndrome means that NHE inhibitors are valuable drugs for the prevention and treatment of coronary artery spasm, peripheral vascular diseases, in particular intermittent claudication, atherogenesis and atherosclerosis, left ventricular hypertrophy and dilated cardiomyopathy and thrombotic disorders.

It has also been found that NHE inhibitors are suitable for the treatment of non-insulin dependent diabetes mellitus (NIDDM), in which case e.g. insulin resistance is limited. In this case, it is advantageous to enhance the antidiabetic activity and the effector qualities of the compounds of the invention by combining them with: biguanides, such as metformin; antidiabetic sulfonylureas such as glibenclamide, glimepiride, tolbutamide, and the like; a glucosidase inhibitor; PPAR agonists such as rosiglitazone, pioglitazone and the like; insulin products in different administration forms; a DB4 inhibitor; an insulin sensitizer; or meglitinide (meglitinide).

In addition to the acute antidiabetic effect, NHE inhibitors counteract the development of late complications of diabetes and are therefore useful as agents for the prevention and treatment of late damage caused by diabetes, such as diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, diabetic cardiomyopathy and other conditions which occur as a consequence of diabetes. They may advantageously be combined with the antidiabetic drugs mentioned above in the treatment of NIDDM. The combination with a beneficial dosage form of insulin may be particularly important in this regard.

In addition to protection against acute ischemic events and subsequent also acute stress reperfusion events, NHE inhibitors have also shown therapeutically useful direct effects against diseases and conditions of the mammalian organism as a whole, which are associated with manifestations of a chronic progressive aging process and which can also occur independently of acute ischemic states and under normal non-ischemic conditions. These pathologically age-related manifestations induced during long-term aging, such as disease, weakness and death (which can now be treated with NHE inhibitors), are diseases and disorders which are essentially caused by age-related changes in vital organs and their functions and which become increasingly important in aging organisms

Disorders associated with age-related dysfunction or with the manifestation of age-related organ wasting are, for example, inadequate vascular response and reactivity to contractile and relaxing responses. This age-related reduction in the vascular responsiveness to contraction and relaxation stimuli, which are important processes of the cardiovascular system and thus of life and health, can be significantly eliminated or reduced by NHE inhibitors. An important function and measure of maintenance of vascular reactivity is to block and or block age-related progression in endothelial dysfunction, which can be abolished with high significance by NHE inhibitors. Therefore, NHE inhibitors are notably suitable for the treatment and prevention of age-related processes in endothelial dysfunction, in particular for the treatment and prevention of intermittent claudication. Therefore, NHE inhibitors are also notably suitable for the prevention and treatment of myocardial infarction, Congestive Heart Failure (CHF), and also for the treatment, in particular for the prevention of age-related forms of cancer.

In this connection, useful combinations are with hypotensive agents, for example with ACE inhibitors, angiotensin receptor antagonists, diuretics, Ca²⁺Antagonists, etc. or with drugs that normalize metabolism, such as cholesterol-lowering substances. Thus, the compounds of formula I are also suitable for preventing age-related tissue changes and for maintaining health and prolonging life while maintaining a high quality of life.

The compounds of the invention are potent inhibitors of cellular sodium-proton antiporters (Na/H exchanger pumps) which are elevated in many diseases (essential hypertension, atherosclerosis, etc.) and even in those cells which are easy to measure, for example in erythrocytes, thrombocytes or leukocytes. The compounds used according to the invention are therefore suitable as outstanding and simple scientific tools, for example for their use as diagnostic substances for determining and distinguishing different forms of hypertension as well as atherosclerosis, diabetes and diabetic late complications, proliferative disorders, etc.

Furthermore, NHE inhibitors are suitable for the treatment of disorders (human and veterinary) induced by bacteria and by protozoa. Diseases induced by protozoa are in particular malarial disorders in humans and coccidiosis in livestock.

These compounds are also suitable for use as substances for controlling sucking parasites in human and veterinary medicine and in crop protection. Preferably as a substance against hematophagous parasites in human and veterinary medicine.

The mentioned compounds are therefore advantageously used, alone or in combination with other drugs or active ingredients, for the preparation of a medicament for the treatment or prophylaxis of respiratory drive disorders, respiratory disorders, sleep-related respiratory disorders, sleep apneas, snoring, acute and chronic renal disorders, acute and chronic renal failure, bowel dysfunction, hypertension, essential hypertension, central nervous system disorders, disorders due to CNS hyperexcitability, epilepsy and centrally induced convulsive or anxiety states, depressive states and psychotic states, ischemic states of the peripheral or central nervous system or of a stroke, acute and chronic injuries and disorders of the peripheral organs or limbs resulting from ischemic or reperfusion events, atherosclerosis, disorders of lipid metabolism, thrombosis, biliary dysfunction, ectoparasitic infections, disorders due to endothelial dysfunction, Protozoal disorders, malaria; and for preservation and storage of grafts for surgical procedures, surgical procedures and organ transplantation; or for the treatment of states of shock and diabetes and late-stage damage to diabetes or diseases in which cellular proliferation constitutes a primary or secondary cause; and for maintaining health and prolonging life.

The term "dementia" refers to a decrease in mental capacity. It is understood to mean in particular a decline in memory and thinking ability. Dementia in the elderly or senile dementia refers to a progressive, acquired mental decline in the elderly that may be attributed to structural and/or metabolic abnormalities of the central nervous system. Approximately 7% of the population over age 65 have dementia of varying degrees of severity. The causes of dementia are different. Alzheimer's disease is the most common form, accounting for up to 50%, followed by vascular dementia, such as multi-infarct dementia, and combinations of the two forms. Very rare causes are Tau mutations, prion diseases, polyglutamine expansion disorders such as Huntington's chorea and spinocerebellar ataxia, and Parkinson's syndrome. Also known are secondary dementias following and/or associated with infection (e.g. HIV infection), brain trauma, brain tumours or intoxication (e.g. alcoholism).

The concept of memory consolidation is based on the ability of new memory to stabilize over time and thereby become less susceptible to interference with new information and brain dysfunction. The basic aspects and mechanisms of memory formation and consolidation can be studied with the aid of a popular long-term potentiation (LTP) cell model (neuroscientist.9: 463-474.2003; Brain Res Rev.45: 30-37, 2004; Physiol Rev.84: 87-136, 2004).

One of the most important brain regions for storing and processing information is the hippocampal structure. It has long been known that certain patterns of electrical stimulation (tetanic) in the hippocampus result in altered synaptic efficacy (Bliss and Lomo, J physiol.232: 331-356, 1973), which is currently referred to as "long-term potentiation" or "LTP" and has subsequently been demonstrated in other areas of the brain of various mammals, both in vitro and in vivo. LTP is currently considered to be an important component of the neural mechanisms of learning and memory. It is also known that weak LTP is associated with short-term memory, whereas strong LTP is associated with long-term memory (J Neurosci.20: 7631-.

The hippocampus plays an important role in the situational, spatial and declarative learning and memory processes, which are essential for spatial localization and recall of spatial structures, and plays an important role in controlling autonomic and nutritional functions (McEwen 1999, "Stress and hippocampal plasticity" (Stress and hippopalamic plasticity), Annual Review of Neuroscience 22: 105-. In human dementing disorders, there are often impaired learning and memory processes in which the hippocampus is involved. Animal experiments on other mammals have shown similar results.

Thus, it could be confirmed that old mice had a deletion in spatial memory and LTP compared to young mice, and that the substance improving LTP simultaneously reduced the memory deletion (Bach et al, 1999, "Age-related spatial memory deletion in vitro was associated with a defect in the advanced stage of hippocampal long-term potentiation and was attenuated by drugs that enhance the cAMP signaling pathway" (Age-related defects in spatial memory corrected with defects in the late stage of hippocampal long-term potentiation; Proc Natl Acad USA 27; 96: 5280-5; Fujii & Sumikawa, "Acute and long-term contact nicotine conversion in rats reversible long-term potentiation induced decline of hippocampal-related diseases" (Age-related diseases-treatment), brain Res.894: 347-53, Clayton et al, 2002, "hippocampal NR2B deficiency can mimic the age-related changes in long-term potentiation and spatial learning in Fischer 344 rats" (Ahippopampal NR2B specific peptide-related changes in long-term evolution and spatial learning in the Fischer 344 rat), J neurosci.22: 3628-37).

Can be demonstrated in transgenic animals and in vivo and in vitro by administration of β -amyloid peptide: these peptides adversely affect LTP or interfere with its maintenance (Ye & Qiao 1999, "the inhibitory activity on the long-term potentiation by the β -amyloid peptide fragment 31-35 in rat hippocampus is N-methyl-D-aspartate receptor dependent: it is N-methyl-D-aspartate receptor dependent by (-) huperzine A-offset" (superior activity produced by beta-amyloid peptide fragment 31-35 on long-term potentination ratio hippompus is N-methyl-D-aspartate receptor-index: it's activity of beta-amyloid peptide A (-), Neurosci Lett.275: 187-90; Rowan et al, 2003, "Synaptic plasticity in early Alzheimer's disease animal model" cement gene, R821; cement modulus gene R8: cement gene R358, 2004, "Normal induced but accelerated decline of LTP in APP + PS1transgenic mice" (Normal induced butyl accessed decay of LTP in APP + PS1transgenic mice), Neurobiol Dis 15: 188-95). It is possible to correct impaired LTP and impaired memory function with rolipram and cholinesterase inhibitors such as those also used in the treatment of human Alzheimer's disease (Ye & Qiao 1999, Gong et al, 2004, "permanent improvement of synaptic and cognitive function after treatment with rolipram in a mouse model of Alzheimer's disease" (persistent improvement in synthetic and cognitive functions in an Alzheimer's disease mouse model), J Clin invest.114: 1624-34.).

It is thus expected that LTP-improving substances will also have beneficial effects on conditions associated with cognitive impairment and dementia.

It has been surprisingly found that inhibitors of cellular NHE5 can increase LTP. Thus, the inhibitor is expected to have a memory improving effect in dementing disorders such as alzheimer's form and dementia in alzheimer's-like form. The use of NHE5 inhibitors has advantages over active ingredients such as acetylcholinesterase inhibitors used to date for these disorders, i.e. a mild or absent systemic effect is expected, since NHE5 is expressed only in neurons and is therefore brain-specific (am.j. physiol.cell.physiol.281: C1146-C1157, 2001).

Therefore, NHE5 inhibitors are suitable for the treatment of neurodegenerative disorders, memory deficits and dementing disorders, such as dementia in elderly, alzheimer's disease, vascular dementia such as multi-infarct dementia, a combination of alzheimer's disease and cerebrovascular disorders, Tau mutations, prion diseases, polyglutamine-expansion disorders such as huntington's disease and spinocerebellar ataxia, and parkinson's syndrome; and is suitable for improving memory. NHE5 inhibitors are also suitable for the treatment of secondary dementia following and/or associated with infections such as HIV infection, brain trauma, brain tumors or intoxications such as alcoholism.

The invention further relates to the use of compounds of formula I and their pharmaceutically acceptable salts as medicaments.

The invention also relates to a human, veterinary or plant-protective medicament comprising an effective amount of a compound of the formula I and/or a pharmaceutically acceptable salt thereof, and to a human, veterinary or plant-protective medicament comprising an effective amount of a compound of the formula I and/or a pharmaceutically acceptable salt thereof, alone or in combination with one or more other pharmacologically active ingredients or medicaments.

Medicaments comprising a compound of formula I or a pharmaceutically acceptable salt thereof may be administered, for example, orally, parenterally, intramuscularly, intravenously, rectally, nasally, by inhalation, subcutaneously or by suitable transdermal dosage forms, the preferred administration depending on the particular manifestation of the condition. In veterinary and human medicine and in crop protection, the compounds of the formula I can be used alone or together with pharmaceutically acceptable excipients. The medicament comprises the active ingredient of formula I and/or a pharmaceutically acceptable salt thereof in an amount which is typically 0.01mg to 1g per dosage unit.

Those skilled in the art are familiar, from their expert knowledge, with which excipients are suitable for the desired pharmaceutical preparation. In addition to solvents, gel formers (gel formers), suppository bases, tablet excipients and other active ingredient carriers, it is also possible to use, for example, antioxidants, dispersants, emulsifiers, antifoams, flavoring agents, preservatives, solubilizers or colorants.

For oral administration, the active compounds are mixed with additives suitable for this purpose, such as carriers, stabilizers or inert diluents, and converted by customary methods into suitable dosage forms, such as tablets, coated tablets, hard gelatin capsules, aqueous, alcoholic or oily solutions. Examples of useful inert carriers are gum arabic, magnesium oxide, magnesium carbonate, potassium phosphate, lactose, glucose or starch, in particular corn starch. The preparation can take the form of dry granules or wet granules. Examples of useful oily carriers or useful solvents are vegetable or animal oils, such as sunflower oil or cod liver oil.

For subcutaneous, transdermal or intravenous administration, the active compounds used are converted into solutions, suspensions or emulsions, if desired together with substances customary for this purpose, such as solubilizers, emulsifiers or other excipients, into solutions, suspensions or emulsions. Examples of useful solvents are: water, physiological saline or alcohols (e.g. ethanol, propanol, glycerol) and sugar solutions (e.g. glucose or mannitol solutions), and mixtures of the different solvents mentioned.

Examples of pharmaceutical preparations suitable for administration in the form of an aerosol or spray are solutions, suspensions or emulsions of the active ingredient of formula I in a pharmaceutically acceptable solvent, in particular ethanol or water or mixtures of such solvents. The formulation may also contain other pharmaceutically acceptable excipients, such as surfactants, emulsifiers and stabilizers, and propellant gases, if desired. Such formulations typically comprise the active ingredient in a concentration of about 0.1 to 10% by weight, especially about 0.3 to 3% by weight.

The dose and frequency of administration of the active ingredient of the formula I administered depend on the efficacy and duration of action of the compounds used; in addition, it depends on the nature and severity of the disease to be treated, on the sex, age, weight and individual response of the mammal to be treated.

On average, for a patient weighing about 75kg, a daily dose of at least 0.001mg/kg, preferably 0.1mg/kg, up to 30mg/kg, preferably 1mg/kg body weight of a compound of formula I may be necessary in acute situations, for example immediately after an apneic state at high altitude, even higher doses. Up to 300 mg/kg/day may be necessary, especially for intravenous administration, e.g. for intravenous administration to heart attack patients in intensive care units. The daily dose may be divided into one or more, e.g. up to 4, single doses.

Description of the experiments and examples

List of abbreviations used:

AMPA may be substituted with alpha-amino-3-hydroxy-5-methylisoAzole-4-propionate activated receptor coupled channels

CA1 CA ═ hippocampal horn (armonhorn), CA1 region in hippocampus

EA Ethyl acetate

EPSP excitatory postsynaptic potential

ES⁺Electrospray ionization

HEP heptane

Conc.NH₃Saturated NH₃Aqueous solution

LTP long term enhancement

LTP1 early LTP (stage of LTP)

MeOH methanol

mp melting point

MS mass spectrometry

NMDA receptor-coupled channels activatable by N-methyl-D-aspartate

RT Room temperature

STP short-time range enhancement (stage of LTP)

THF tetrahydrofuran

Example 1: n-diaminomethylene-4- (6, 8-dichloro-2-methyl-1, 2, 3, 4-tetrahydro-isoquinolin-4-yl) benzenesulfonamide dihydrochloride

In argon atmosphere, 0.36g guanidine suspension in 30ml anhydrous THF, add 0.40g 4- (6, 8-dichloro-2-methyl-1, 2, 3, 4-four hydrogen isoquinoline-4-Yl) benzenesulfonyl chloride (WO 2003048129). The mixture was stirred at RT for 24 hours, then THF was distilled off. 10ml of water were added to the residue and the precipitate was filtered off. Washed with 10ml of water and dried in vacuo. The solid was then suspended in 10ml of EA and 10ml of a saturated solution of HCl in ether was added. The volatile constituents were removed in vacuo and the residue was suspended in 10ml EA and stirred at RT for 5 hours. The precipitate was then filtered off and dried in vacuo. 0.45g was obtained, mp 140 deg.C (decomposition). Rf (EA/HEP/CH)₂Cl₂/MeOH/concentrated NH₃＝10∶5∶5∶5∶1)＝0.30MS(ES⁺)：412

Pharmacological data:

NHE3 and NHE5 trials describe:

in this assay, the recovery of intracellular ph (phi) of LAP1 cells after acidification was determined, wherein LAP1 cells stably expressed different subtypes of the sodium-proton exchange pump (NHE). In the case of a functioning NHE, this recovery starts even in bicarbonate free conditions. For this purpose, the pHi was determined using the pH-sensitive fluorescent dye BCECF (Molecular Probes, Inc., Eugene, OR, USA; using the precursor BCECF-AM). First, cells were incubated with BCECF (5. mu. MBCECF-AM) in NH₄Cl buffer (NH)₄Cl buffer solution: 115mM choline hydrochloride, 20mM NH₄Cl，5mM KCl，1mM CaCl₂，1mM MgCl₂20mM Hepes, 5mM glucose; pH 7.4 was established using 1M KOH). By adding at NH₄NH-free cells incubated in Cl buffer₄Cl buffer (133.8mM choline hydrochloride, 4.7mM KCl, 1.25mM CaCl)₂，1.25mM MgCl₂，0.97mM K₂HPO₄，0.23mM KH₂PO₄5mM Hepes, 5mM glucose; pH 7.4 established using 1M KOH), thereby inducing intracellular acidification. After the washing operation, 90. mu.l were freed of NH₄The buffer of Cl remains on the cells. In an analytical instrument (FLIPR, "fluorescence imaging plate reader", Molecular Devices, Inc., Sunnyvale, Ca., USA) by adding 90. mu.l of Na-containing solution⁺Buffer (133.8mM NaCl, 4.7mM KC)l，1.25mM CaCl₂，1.25mM MgCl₂，0.97mM Na₂HPO₄，0.23mMNaH₂PO₄10mM Hepes, 5 mM-glucose; establishing a pH of 7.4 using 1M NaOH) to initiate pH recovery. BCECF fluorescence was measured at an excitation wavelength of 498nm and a FLIPR emission filter 1 (band gap 510-570 nm). Subsequent changes in fluorescence were recorded for 2 minutes for NHE3 and NHE5 as measures of pH recovery. For the calculation of the NHE-inhibitory potential of the test substance, cells were first tested in a buffer where complete pH recovery occurred or no recovery at all occurred. For complete recovery of pH (100%), cells were incubated in the presence of Na⁺In a buffer (see above) and Na-free⁺To determine the 0% value (see above). Test substances are added in the presence of Na⁺The buffer solution of (4). The intracellular pH recovery at each tested concentration of the substance was expressed as a percentage of the maximum recovery. IC of specific substances for each NHE subtype calculated from percent pH recovery by the XLFit program (idbs, Surrey, UK)₅₀The value is obtained.

	NHE3 IC50[μM]	NHE5 IC50[μM]
			Example 1	0.035	0.37

Description of the test: hippocampus slices long term test (in vitro)

Means of experiment

The LTP in the CA1 region is one that has been best characterized in vitro. The layered and input structure of this region allows field potential measurements to be performed in vitro over several hours. In the NHE study, a weak strong one based on the theta rhythm and inducing early LTP which returns to the initial value within 3 hours was used (Journal of neuroscience, 18(16), 6071 (1998); Eur J Pharmacol.502: 99-104, 2004). It has recently been demonstrated that an increased number of theta onset exercises induces PLTs of increased intensity and persistence (jneurophysiol.88: 249-255, 2002), i.e., a single weak stimulus induces unsaturated LTPs, rather than the most available saturated type of LTPs. The intensity (Behnisch, Reymann et al, Neurosci. Lett.1998, 253 (2): 91-94) and persistence (e.g.Neuroppeptides 26: 421-. The early LTP produced in our study was also unsaturated. Thus, substance-induced improvement or worsening of early-stage LTP can be determined. The early LTP studied consisted of an STP module known to last for about 30 minutes (Nature 335: 820-824, 1988) and an LTP1 module which usually lasted for the first 1-2 hours after LTP induction (LearnMem.3: 1-24, 1996).

Short-term (30-60 min) recording of the initial value of the strong pre-cisterns allowed an early effect of the substance of interest on the normal unstimulated synaptic transmission of interest. Since the major excitatory synapses are glutamatergic (J Clin neurophysiol.9: 252-263, 1992), i.e., the monosynaptic field EPSP is measured very predominantly by AMPA and only to a relatively low extent by NMDA receptors, the effect on glutamatergic transmission is tested simultaneously and indirectly.

The method comprises the following steps: hippocampus slices long term test (in vitro)

Animal type: rat

Age: 7-8 weeks old

Strain: weister (Wistar) (Shoe Wist, Shoe)

Sex: male sex

And (3) breeders: helen Winkelmann corporation, Artificial light (6-18.00h) and daily circadian rhythm

Preparing:

vibrating the corona: striking the back of the neck with a iron rod

And (3) killing: operation of broken head

Exposing the brain: the cranium is opened by incision along the sagittal suture of the skull from the dorsal to the ventral side.

Exposure of hippocampus: the brain was dissected between hemispheres, the right hemisphere was used as the starting material, and the hippocampus was pulled out using a blunt instrument.

Preparation of slices: the exposed hippocampus was transferred to a cooling block with wet filter paper and excess water was aspirated through another piece of filter paper. The hippocampus, which was fixed on a cooling block in this manner, was placed on a cutter and rotated in the horizontal direction until the hippocampus was at an appropriate angle to the slicing knife.

Cutting angle: in order to maintain the sheet-like structure of the hippocampus, it is necessary to slice the hippocampus at an angle of about 70 degrees with respect to a slicing knife (a cutter).

Slicing: hippocampus was sectioned at 400 μm intervals. The sections were removed from the blade by means of a very soft, fully moist brush (mink hair) and transferred to a cutter with a 95% O charge₂/5％CO₂In a glass container for cooling the nutrient solution. The total preparation duration does not exceed 5 minutes.

Storage of slices：

Dipping and slicing: the slices were placed 1-3mm below the liquid surface in a temperature-controlled chamber (33 ℃). The flow rate was 2.5 ml/min. The pre-gas treatment was carried out in a pre-heating chamber at slightly elevated pressure (about 1atm) and through a micro-needle. The slicing chamber is connected to a preheating chamber so that the micro-circulation can be maintained. Passing 95% O₂/5％CO₂The microcirculation is initiated by the outflow of the micromanipulation needle.

Slice adaptation: freshly prepared hippocampal slices were acclimatized in a slicing chamber at 33 ℃ for at least 1 hour.

Determination of test stimulus levels：

Stimulus level: fEPSP: maximum EPSP of 30%

Determination of focal potential：

Stimulation: monopolar stimulation electrodes were used to locally stimulate Schaffer collatoral (voltage: 1-5V, one polarity pulse width 0.1ms, total pulse 0.2ms), which included painted stainless steel and a constant current electrical biphasic stimulation generator (WPI A365).

And (3) determination: a glass electrode (borosilicate glass with fibers, 1-5MOhm, diameter: 1.5mm, diameter of the top circle: 3-20 μm) filled with normal nutrient solution was used to record the excitatory postsynaptic potential (fEPSP) from the irradiated layer. The field potential was measured using a DC voltage amplifier against a silver chloride reference electrode located at the edge of the chamber. The field potential is filtered by a low pass filter (5 kHz).

Measurement of field potential: the slope of fEPSP (fEPSP slope) was determined for statistical analysis of the experiment. The recording, analysis and control of the experiments were carried out with the aid of a software Program (PWIN) developed by the neurophysiology department. The formation and construction of the mean fEPSP slope was performed at each time point with the help of Excel software and automated data recording was performed macroscopically as appropriate.

Nutrient medium (ringer solution):

substance(s)	In mM	Grams per liter
			NaCl	124	7.248
KCl	4.9	0.356
			MgSO4*7H2O	1.3	0.321
CaCl2+ no water	2.5	0.368
			KH2PO4	1.2	0.164
NaHCO3	25.6	2.152
			Glucose	10	1.802
Osmolarity, in mOsm	330
			PH	7.4

Example 1 was dissolved in DMSO and diluted with ringer's solution to the final concentration of the experiment (final concentration 0.01% DMSO).

Summary of the experiment:

in control experiments, baseline synaptic transmission was initially recorded for 60-120 minutes. Subsequently, two dipulses were given 4 times at 200ms intervals, with a 10ms pulse interval for the dipulses and a 0.2ms (weak rigidity) width for the single pulse. The resulting EPSP enhancement was recorded for at least 60 minutes.

In experiments testing the effect of NHE5 inhibitors, baseline was again recorded initially for 60-120 minutes. NHE5 inhibitor (10 μ M) was infused 20 minutes prior to stimulation. Two dipulses were given 4 times at 200ms intervals as a control experiment, with the dipulses having a pulse interval of 10ms and a single pulse width of 0.2 ms. The material was washed away for 20 minutes after stimulation and EPSP enhancement was recorded for at least 60 minutes.

As a result:

the compound of example 1 had no intrinsic (inrinsic) effect on synaptic transmission at the concentrations used.

The enhancement after administration of example 1 was still 137% of baseline at 80 minutes post-stimulation, while the enhancement under control conditions returned almost to baseline levels, 113% of baseline. This clearly shows that even 10 μ M of the compound of example 1 improves the maintenance of weak LTP.

Claims (21)

1. A compound of formula I:

wherein:

r1 and R3 are each hydrogen;

r2 and R4 are each independently hydrogen, F, Cl, NH₂、NHCH₃Or N (CH)₃)₂；

R5 is hydrogen, methyl, ethyl or cyclopropyl;

r6 is hydrogen or methyl;

r7 and R8 are each hydrogen.

2. A compound of formula I according to claim 1, wherein the pharmaceutically acceptable salt is the trifluoroacetate salt, and the pharmaceutically acceptable salts thereof.

3. The use of a compound of formula I as claimed in claims 1 or 2 and pharmaceutically acceptable salts thereof for the manufacture of a medicament for the inhibition of NHE.

4. The use of a compound of formula I as claimed in claims 1 or 2 and pharmaceutically acceptable salts thereof for the manufacture of a medicament for inhibiting NHE 3.

5. The use of a compound of formula I as claimed in claims 1 or 2 and pharmaceutically acceptable salts thereof for the manufacture of a medicament for inhibiting NHE 5.

6. The use of a compound of formula I as claimed in claims 1 or 2 and pharmaceutically acceptable salts thereof for the preparation of a medicament for the treatment or prevention of respiratory disorders, acute and chronic renal disorders, intestinal dysfunction, hypertension, central nervous system disorders, ischemic states of the peripheral or central nervous system or of stroke, acute and chronic injuries and disorders of peripheral organs or limbs resulting from ischemic or reperfusion events, atherosclerosis, disorders of lipid metabolism, thrombosis, biliary dysfunction, ectoparasitic infections, disorders of endothelial function which are associated with NHE3/NHE 5; and for preservation and storage of grafts for surgical procedures, surgical procedures and organ transplantation; used for resuscitation after bypass surgery and cardiac arrest; or for the treatment of shock states associated with NHE3/NHE5 and diabetes and late-stage damage to diabetes or diseases in which cell proliferation constitutes a primary or secondary cause.

7. The use according to claim 6, wherein the medicament is for the treatment or prevention of respiratory dyskinesia associated with NHE3/NHE5, acute renal failure and chronic renal failure, essential hypertension, disorders due to CNS hyperexcitability, epilepsy and centrally induced convulsive or anxious states, depressive and psychotic states, protozoal disorders.

8. The use according to claim 6, wherein the medicament is for the treatment or prevention of sleep-related respiratory disorders associated with NHE3/NHE 5.

9. The use of claim 6, wherein the medicament is for the treatment or prevention of sleep apnea, snoring, malaria associated with NHE3/NHE 5.

10. The use of a compound of the formula I as claimed in claim 1 or 2 and of pharmaceutically acceptable salts thereof in combination with other medicaments or active ingredients for the preparation of medicaments for the treatment or prophylaxis of respiratory disorders, acute and chronic renal disorders, bowel dysfunction, hypertension, disorders of the central nervous system, ischemic states of the peripheral or central nervous system or of stroke, acute and chronic injuries and disorders of peripheral organs or limbs caused by ischemic or reperfusion events, atherosclerosis, disorders of lipid metabolism, thrombosis, biliary dysfunction, ectoparasite infections, disorders caused by endothelial dysfunction, which are associated with NHE3/NHE 5; and for preservation and storage of grafts for surgical procedures, surgical procedures and organ transplantation; used for resuscitation after bypass surgery and cardiac arrest; or for the treatment of shock states associated with NHE3/NHE5 and diabetes and late-stage damage to diabetes or diseases in which cell proliferation constitutes a primary or secondary cause.

11. The use according to claim 10, wherein the medicament is for the treatment or prevention of respiratory dyskinesia associated with NHE3/NHE5, acute renal failure and chronic renal failure, essential hypertension, disorders due to CNS hyperexcitability, epilepsy and centrally induced convulsive or anxious states, depressive and psychotic states, protozoal disorders.

12. The use according to claim 10, wherein the medicament is for the treatment or prevention of sleep-related respiratory disorders associated with NHE3/NHE 5.

13. The use of claim 10, wherein the medicament is for the treatment or prevention of sleep apnea, snoring, malaria associated with NHE3/NHE 5.

14. The use of a compound of the formula I according to claim 1 or 2 and/or of a pharmaceutically acceptable salt thereof, alone or in combination with other medicaments or active ingredients, for the preparation of a medicament for the treatment or prophylaxis of respiratory drive disorders and/or sleep-related respiratory disorders associated with NHE3/NHE 5.

15. The use of claim 14, wherein the medicament is for the treatment or prevention of sleep apnea associated with NHE3/NHE 5.

16. The use of a compound of the formula I according to claim 1 or 2 and/or of a pharmaceutically acceptable salt thereof, alone or in combination with other medicaments or active ingredients, for the preparation of a medicament for the treatment or prophylaxis of snoring associated with NHE3/NHE 5.

17. The use of a compound of the formula I as claimed in claim 1 or 2 and/or of a pharmaceutically acceptable salt thereof, alone or in combination with other medicaments or active ingredients, for the preparation of a medicament for the treatment or prophylaxis of acute or chronic renal disorders which are associated with NHE3/NHE 5.

18. The use of claim 17, wherein the medicament is for the treatment or prevention of acute renal failure or chronic renal failure associated with NHE3/NHE 5.

19. The use of a compound of the formula I according to claim 1 or 2 and/or of a pharmaceutically acceptable salt thereof, alone or in combination with other medicaments or active ingredients, for the preparation of a medicament for the treatment or prevention of bowel dysfunction associated with NHE3/NHE 5.

20. A pharmaceutical formulation for human or veterinary use comprising an effective amount of a compound of formula I according to claim 1 or 2 and/or a pharmaceutically acceptable salt thereof.

21. A pharmaceutical preparation for human or veterinary use comprising an effective amount of a compound of the formula I as claimed in claim 1 or 2 and/or a pharmaceutically acceptable salt thereof and a further pharmacologically active ingredient or drug.