HK1076276B - 1-amino-alkylcyclohexanes as 5-ht3 and neuronal nicotinicreceptor antagonists - Google Patents

Description

As 5-HT3And 1-amino-alkylcyclohexanes of neuronal nicotinic receptor antagonists

no marking

Technical Field

New use of 1-amino-alkylcyclohexane is provided.

Prior Art

Representative of the prior art is our prior patent USP6,034,134 on 7/3/2000 and our published applications WO 99/01416, PCT/EP98/04026 and Parsons et al Neuropharmacology 38, 85-108(1999) in which the active compounds used in the present invention are disclosed and which are NMDA receptor antagonists and anticonvulsants.

The invention

The present invention relates to a novel use of a 1-amino-alkylcyclohexane compound selected from the group consisting of compounds of the following formula and their enantiomers, optical isomers, hydrates and pharmaceutically acceptable salts and their pharmaceutical compositions,

wherein R is- (CH)₂)_n-(CR⁶R⁷)_m-NR⁸R⁹；

Wherein n + m is 0, 1 or 2;

wherein R is¹To R⁷Independently selected from hydrogen and lower alkyl (1-6C); and

wherein R is⁸And R⁹Each represents hydrogen and lower alkyl (1-6C) or together represents lower alkylene- (CH)₂)_x-, where x is 2 to 5, including 2 and 5; and the use of these compounds and compositions as 5HT₃And neuronal nicotinic receptor antagonists and neuroprotective agents for the treatment of living animals to alleviate conditions responsive thereto.

Representative of these compounds are as follows:

MRZ 2/579: 1-amino-1, 3, 3, 5, 5-pentamethylcyclohexane, HCl

601: 1-amino-1-propyl-3, 3, 5, 5-tetramethylcyclohexane, HCl

607: 1-amino-1, 3, 3, 5 (trans) -tetramethylcyclohexane (axial amino), HCl

615: 1-amino-1, 3, 5, 5-tetramethyl-3-ethylcyclohexane (mixture of diastereomers), HCl

616: 1-amino-1, 3, 5-trimethylcyclohexane (mixture of diastereomers), HCl

617: 1-amino-1, 3-dimethyl-3-propylcyclohexane (mixture of diastereomers), HCl

618: 1-amino-1, 3 (trans), 5 (trans) -trimethyl-3 (cis) propylcyclohexane, HCl

620: 1-amino-1, 3-dimethyl-3-ethylcyclohexane, HCl

621: 1-amino-1, 3, 3-trimethylcyclohexane, HCl

625: 1-amino-1, 3 (trans) -dimethylcyclohexane, HCl

627: 1-amino-1-methyl-3 (trans) propylcyclohexane, HCl

629: 1-amino-1-methyl-3 (trans) ethylcyclohexane, HCl

632: 1-amino-1, 3, 3-trimethyl-5 (cis) ethylcyclohexane, HCl

633: 1-amino-1, 3, 3-trimethyl-5 (trans) ethylcyclohexane, HCl

640: n-methyl-1-amino-1, 3, 3, 5-pentamethylcyclohexane, HCl

641: 1-amino-1-methylcyclohexane, HCl

642: n, N-dimethyl-1-amino-1, 3, 3, 5, 5-pentamethylcyclohexane, HCl₂O

705: n- (1, 3, 3, 5, 5-pentamethylcyclohexyl) pyrrolidine, HCl

680: 1-amino-1, 3 (trans), 5 (trans) -trimethylcyclohexane, HCl

681: 1-amino-1, 3 (cis), 5 (cis) -trimethylcyclohexane, HCl₂O，

682: 1-amino- (1R, 5S) trans-5-ethyl-1, 3, 3-trimethylcyclohexane, HCl

683: 1-amino- (1S, 5S) cis-5-ethyl-1, 3, 3-trimethylcyclohexane, HCl₂O，

1-amino-1, 5, 5-trimethyl-3 (cis) -isopropyl-cyclohexane HCl,

1-amino-1, 5, 5-trimethyl-3 (trans) -isopropyl-cyclohexane HCl,

1-amino-1-methyl-3 (cis) -ethyl-cyclohexane HCl,

1-amino-1-methyl-3 (cis) -methyl-cyclohexane HCl,

1-amino-5, 5-diethyl-1, 3, 3-trimethyl-cyclohexane HCl, and

also, 1-amino-1, 3, 3, 5, 5-pentamethylcyclohexane,

1-amino-1, 5, 5-trimethyl-3, 3-diethylcyclohexane,

1-amino-1-ethyl-3, 3, 5, 5-tetramethylcyclohexane,

n-ethyl-1-amino-1, 3, 3, 5, 5-pentamethylcyclohexane,

n- (1, 3, 5-trimethylcyclohexyl) pyrrolidine or piperidine,

n- [1, 3 (trans), 5 (trans) -trimethylcyclohexyl ] pyrrolidine or piperidine,

n- [1, 3 (cis), 5 (cis) -trimethylcyclohexyl ] pyrrolidine or piperidine,

n- (1, 3, 3, 5-tetramethylcyclohexyl) pyrrolidine or piperidine,

n- (1, 3, 3, 5, 5-pentamethylcyclohexyl) pyrrolidine or piperidine,

n- (1, 3, 5, 5-tetramethyl-3-ethylcyclohexyl) pyrrolidine or piperidine,

n- (1, 5, 5-trimethyl-3, 3-diethylcyclohexyl) pyrrolidine or piperidine,

n- (1, 3, 3-trimethyl-cis-5-ethylcyclohexyl) pyrrolidine or piperidine,

n- [ (1S, 5S) cis-5-ethyl-1, 3, 3-trimethylcyclohexyl ] pyrrolidine or piperidine,

n- (1, 3, 3-trimethyl-trans-5-ethylcyclohexyl) pyrrolidine or piperidine,

n- [ (1R, 5S) trans-5-ethyl-1, 3, 3-trimethylcyclohexyl ] pyrrolidine or piperidine,

n- (1-ethyl-3, 3, 5, 5-tetramethylcyclohexyl) pyrrolidine or piperidine, and

n- (1-propyl-3, 3, 5, 5-tetramethylcyclohexyl) pyrrolidine or piperidine,

and their optical isomers, enantiomers, as well as hydrochloride, hydrobromide, hydrochloride hydrate or other pharmaceutically acceptable salts of any of the foregoing compounds.

Of particular interest are those in which at least R¹、R⁴And R⁵A compound of the above formula which is lower alkyl, and wherein R¹To R⁵Compounds which are methyl, in which x is 4 or 5, especially the compound N- (1, 3, 3, 5, 5-pentamethylcyclohexyl)) Pyrrolidine, and optical isomers, enantiomers, hydrates and pharmaceutically acceptable salts thereof.

In our USP6,034,134 of 3/7/2000, we disclose compounds of the above formula, their pharmaceutical compositions and their use as NMDA-receptor antagonists and anticonvulsants. It has been found that the compounds of the above formula and their optical isomers, enantiomers, hydrates and pharmaceutically acceptable salts, in addition to their NMDA antagonist and anticonvulsant properties, have a high degree of 5HT which is quite unpredictable₃And neuronal nicotinic receptor antagonism, making them useful in the treatment of diseases or conditions in which it is important to block these receptors.

Summary of The Invention

We therefore believe that the invention comprises what can be summarized in particular as the following text:

a method of live animal therapy for inhibiting the progression of and alleviating a disorder mediated by 5HT₃Or a neuronal nicotinic receptor antagonist, comprising the steps of: administering to said living animal an amount of a 1-aminoalkylcyclohexane compound selected from the group consisting of compounds of the formula

Wherein R is- (CH)₂)_n-(CR⁶R⁷)_m-NR⁸R⁹；

Wherein n + m is 0, 1 or 2;

wherein R is¹To R⁷Independently selected from hydrogen and lower alkyl (1-6C);

wherein R is⁸And R⁹Independently selected from hydrogen and lower alkyl (1-6C) or together represent lower alkylene- (CH)₂)_x-, where x is 2 to 5, including 2 and 5; and their optical isomers, enantiomers, hydrates andpharmaceutically acceptable salts, which compounds are effective for said purpose;

such a method: wherein at least R¹、R⁴And R⁵Is lower alkyl;

such a method: wherein R is¹To R⁵Is methyl;

such a method: wherein R is¹Is ethyl;

such a method: wherein R is²Is ethyl;

such a method: wherein R is³Is ethyl;

such a method: wherein R is⁴Is ethyl;

such a method: wherein R is⁵Is ethyl;

such a method: wherein R is⁵Is propyl;

such a method: wherein R is⁶Or R⁷Is methyl;

such a method: wherein R is⁶Or R⁷Is ethyl;

such a method: wherein X is 4 or 5;

such a method: wherein the condition being treated or inhibited is selected from emesis, anxiety, schizophrenia, drug and alcohol abuse disorders, depression, cognitive disorders, Alzheimer's disease, cerebellar tremor, Parkinson's disease, Tourette's syndrome, pain, and appetite disorders;

such a method: wherein the compound is selected from:

1-amino-1, 3, 3, 5, 5-pentamethylcyclohexane,

1-amino-1-propyl-3, 3, 5, 5-tetramethylcyclohexane,

1-amino-1, 3, 3, 5 (trans) -tetramethylcyclohexane (axial amino),

1-amino-1, 3, 5, 5-tetramethyl-3-ethylcyclohexane (mixture of diastereomers),

1-amino-1, 3, 5-trimethylcyclohexane (mixture of diastereomers),

1-amino-1, 3-dimethyl-3-propylcyclohexane (mixture of diastereomers),

1-amino-1, 3 (trans), 5 (trans) -trimethyl-3 (cis) -propylcyclohexane,

1-amino-1, 3-dimethyl-3-ethylcyclohexane,

1-amino-1, 3, 3-trimethylcyclohexane,

1-amino-1, 3 (trans) -dimethylcyclohexane,

1-amino-1-methyl-3 (trans) propylcyclohexane,

1-amino-1-methyl-3 (trans) ethylcyclohexane,

1-amino-1, 3, 3-trimethyl-5 (cis) ethylcyclohexane,

1-amino-1, 3, 3-trimethyl-5 (trans) ethylcyclohexane,

n-methyl-1-amino-1, 3, 3, 5-pentamethylcyclohexane,

1-amino-1-methylcyclohexane,

n, N-dimethyl-1-amino-1, 3, 3, 5, 5-pentamethylcyclohexane,

1-amino-1, 5, 5-trimethyl-3 (cis) -isopropyl-cyclohexane,

1-amino-1, 5, 5-trimethyl-3 (trans) -isopropyl-cyclohexane,

1-amino-1-methyl-3 (cis) -ethyl-cyclohexane,

1-amino-1-methyl-3 (cis) -methyl-cyclohexane,

1-amino-5, 5-diethyl-1, 3, 3-trimethyl-cyclohexane, and

n- (1, 3, 3, 5, 5-pentamethylcyclohexyl) pyrrolidine, and

optical isomers, enantiomers, hydrates and pharmaceutically acceptable salts of any of the foregoing; and

such a method: wherein said compound is administered in the form of a pharmaceutical composition thereof comprising said compound in combination with one or more pharmaceutically acceptable diluents, excipients or carriers.

Furthermore, 1-aminoalkylcyclohexanes selected from the compounds of the formula

Wherein R is- (CH)₂)_n-(CR⁶R⁷)_m-NR⁸R⁹；

Wherein n + m is 0, 1 or 2;

wherein R is¹To R⁷Independently selected from hydrogen and lower alkyl (1-6C);

wherein R is⁸And R⁹Independently selected from hydrogen and lower alkyl or together represent lower alkylene- (CH)₂)_x-, where x is 2-5, including 2 and 5, and their optical isomers, enantiomers, hydrates and pharmaceutically acceptable salts, for the preparation of a medicament for the treatment of living animals to alleviate the passage of 5HT₃Use in medicine of a condition alleviated by a receptor antagonist,

use in which at least R¹、R⁴And R⁵Is a lower alkyl group;

use of R in which¹To R⁵Is methyl;

use wherein x is 4 or 5;

use wherein the compound is selected from

1-amino-1, 3, 3, 5, 5-pentamethylcyclohexane,

1-amino-1-propyl-3, 3, 5, 5-tetramethylcyclohexane,

1-amino-1, 3, 3, 5 (trans) -tetramethylcyclohexane (axial amino),

1-amino-1, 3, 5, 5-tetramethyl-3-ethylcyclohexane (mixture of diastereomers),

1-amino-1, 3, 5-trimethylcyclohexane (mixture of diastereomers),

1-amino-1, 3-dimethyl-3-propylcyclohexane (mixture of diastereomers),

1-amino-1, 3 (trans), 5 (trans) -trimethyl-3 (cis) propylcyclohexane,

1-amino-1, 3-dimethyl-3-ethylcyclohexane,

1-amino-1, 3, 3-trimethylcyclohexane,

1-amino-1, 3 (trans) -dimethylcyclohexane,

1-amino-1-methyl-3 (trans) propylcyclohexane,

1-amino-1-methyl-3 (trans) ethylcyclohexane,

1-amino-1, 3, 3-trimethyl-5 (cis) ethylcyclohexane,

1-amino-1, 3, 3-trimethyl-5 (trans) ethylcyclohexane,

n-methyl-1-amino-1, 3, 3, 5-pentamethylcyclohexane,

1-amino-1-methylcyclohexane,

n, N-dimethyl-1-amino-1, 3, 3, 5, 5-pentamethylcyclohexane,

1-amino-1, 5, 5-trimethyl-3 (cis) -isopropyl-cyclohexane,

1-amino-1, 5, 5-trimethyl-3 (trans) -isopropyl-cyclohexane,

1-amino-1-methyl-3 (cis) -ethyl-cyclohexane,

1-amino-1-methyl-3 (cis) -methyl-cyclohexane,

1-amino-5, 5-diethyl-1, 3, 3-trimethyl-cyclohexane, and

n- (1, 3, 3, 5, 5-pentamethylcyclohexyl) pyrrolidine, and

optical isomers, enantiomers, hydrates and pharmaceutically acceptable salts of any of the foregoing; and finally

Such a use, wherein the condition to be treated is selected from emesis, anxiety, schizophrenia, drug and alcohol abuse disorders, depression, cognitive disorders, Alzheimer's disease, cerebellar tremor, Parkinson's disease, Tourette's syndrome, pain, and appetite disorders.

Detailed Description

Background and pharmacology

5-HT₃Receptor antagonists

5-HT₃The receptor is a cation permeable ligand-gated ion-tropic receptor. In humans, 5-HT₃Receptors exhibit the highest density on enterochromaffin cells in the gastrointestinal mucosa, which is dominated by the afferent vagus nerve and the rearmost region of the brainstem, which forms the trigger zone for chemical receptors.

Due to 5-HT₃Receptors not only have high density in the posterior region, but also in the hippocampal and amygdala regions of the limbic system, and thus 5-HT has been suggested₃Selective antagonists may have a psychotropic effect (Greenshaw)& Silverstone，1997)。

Indeed, early animal studies indicate 5-HT₃Receptor antagonists other than they are knownIn addition to the antiemetic use of (1), it may have clinical use in various fields. These include anxiety disorders, schizophrenia, drug and alcohol abuse disorders, depression, cognitive disorders, Alzheimer's disease, cerebellar tremor, Parkinson's disease treatment-related psychosis, pain (migraine and irritable bowel syndrome), and appetite disorders.

Neuronal nicotinic receptors

Nine alpha subunits (. alpha.1-. alpha.9) and four beta (. beta.1-. beta.4) subunits of nicotinic receptors are known at present. The α 4 β 2 receptor is probably most prevalent in the CNS, particularly in the hippocampus and striatum. They form non-selective cation channels (type II) with slow, incomplete desensitization currents. Homomeric α 7 receptors are presynaptic and postsynaptic receptors found in the hippocampus, motor cortex and limbic system, and peripheral autonomic nervous system. These receptors are characterized by their high Ca content²⁺Permeability and rapid, strong desensitization (form 1A).

Changes in nicotinic receptors are implicated in a number of diseases. These diseases include Alzheimer's disease, Parkinson's disease, Tourette's syndrome, schizophrenia, drug abuse, and pain.

Based on the observation that nicotinic agonists nicotine itself appears to have beneficial effects, current drug development targets the discovery of selective nicotinic agonists.

On the other hand, it is unclear whether the role of nicotinic agonists in e.g. tourette's syndrome and schizophrenia is due to activation or inactivation/desensitization of neuronal nicotinic receptors.

The effect of an agonist on a neuronal nicotinic receptor depends mainly on the contact time. Rapid reversible desensitization occurs within milliseconds, attenuation occurs within seconds, irreversible inactivation of α 4 β 2 and α 7-containing receptors occurs within hours, while their upregulation occurs within days.

In other words, the effect of the nicotinic "agonist" may actually be due to partial agonism, inactivation, and/or desensitization of neuronal nicotinic receptors. In turn, moderate concentrations of neuronal nicotinic receptor channel blockers may produce the same effects reported for nicotinic agonists in the above indications.

Amino-alkylcyclohexanes as 5-HT₃And neuronal nicotinic receptor antagonists

We hypothesized whether a novel amino-alkylcyclohexane organism (USP 6,034,134), described as an uncompetitive NMDA receptor antagonist and anticonvulsant, might also be useful as a 5HT receptor antagonist₃And neuronal nicotinic antagonists. These properties will allow blocking of 5HT to be of all importance₃Or nicotinic receptors, using amino-alkylcyclohexanes. Our findings are positive.

Method

Synthesis of

The synthesis of the novel amino-alkylcyclohexanes used in the present invention has been disclosed in USP6,034,134, 3 months and 7 days, 2000.

Alternative methods

The 1-cyclic amino compound can also be prepared by reacting the corresponding 1-free amino-alkylcyclohexane with a selected α, ω -dihaloalkyl compound, such as 1, 3-dibromopropane, 1, 4-dibromobutane or 1, 5-dibromopentane, according to the following representative examples:

n- (1, 3, 3, 5, 5-pentamethylcyclohexyl) pyrrolidine hydrochloride

1, 3, 3, 5, 5-pentamethylcyclohexylamine hydrochloride (12g, 58.3mmol), potassium carbonate (48.4g, 350mmol) and 1, 4-dibromobutane (7.32ml, 61.3mmol) were reheated in acetonitrile (250ml) for 60 hours. After cooling to room temperature, the mixture was filtered and the precipitate was washed with diethyl ether (600 ml). The filtrate was concentrated by rotary evaporation under vacuum and the residue was fractionated under reduced pressure (11 mm/Hg). The fractions at 129 ℃ were collected to give a colourless oil (8.95 g). This oil was dissolved in diethyl ether (120ml) and a 2.7M HCl solution in diethyl ether (30ml) was added. The resulting precipitate was filtered off, washed with diethyl ether (3 x 30ml) and washed withNaOH was dried under vacuum to give N- (1, 3, 3, 5, 5-pentamethylcyclohexyl) pyrrolidine hydrochloride hydrate (12.9g, 68%) at m.p.158 ℃. PMR spectra: (DMSO-d6, TMS) d: 0.97(6H, s, 3, 5-CH)₃)；1.11(6H，s，3，5-CH₃) (ii) a 0.8-1.4(2H, cyclohexane 4-CH)₂)1.41(3H，s，1-CH₃) (ii) a 1.69(4H, m, cyclohexane 2, 6-CH)₂) (ii) a 1.84(4H, m, pyrrolidine 3, 4-CH)₂) (ii) a 3.20(4H, m, pyrrolidine 2, 5-CH)₂)；10.9ppm(1H，brs，NH+)。

Elemental analysis (C)₁₅H₂₉n*HCl*H₂O) found (%) C65.0; h11.7; n5.0 calcd (%) C64.8; h11.6; and (5.0) N.

Electrophysiology

Hippocampus was obtained from rat embryos (E20-E21) and then transferred to Ca-free on ice²⁺And Mg²⁺Hank's buffered saline solution (Gibco). Cells were mechanically dissociated in 0.05% DNase/0.3% ovomucoid (Sigma) and then preincubated with 0.66% trypsin/0.1% DNase (Sigma) for 8 minutes. The dissociated cells were then centrifuged at 18G for 10 minutes, resuspended in minimal essential medium (Gibco), and grown at 150,000 cell cm^-2Density plates were plated on plastic petri dishes (Falcon) pre-coated with poly-DL-ornithine (Sigma)/laminin (Gibco). With NaHCO supplemented with 5% fetal bovine serum and 5% horse serum (Gibco)₃The cells were fed in a HEPES buffered minimal essential medium (Gibco) and 5% CO at 37 ℃₂And cultured at 95% humidity. After approximately 5 days of in vitro culture, further glial mitosis was inhibited with cytosine- β -D-arabinofuranoside (ARAC, 5 μ M Sigma), and the medium was then replaced in its entirety.

After 15-21 days in vitro culture, patch-clamp recordings were performed from these neurons using a polished glass electrode (2-3 M.OMEGA.) in whole cell mode with the aid of EPC-7 amplifiers (List) at room temperature (20-22 ℃). Improved rapid application system using theta glass with 100 μ M opening diameter (Clark TGC 200-10) pulled with Zeiss DMZ (Augsburg, Munich) horizontal pullerThe test substances were applied to the system (SF-77B Fast Step, Warner Instruments). The content of the intracellular solution is generally as follows (mM): CsCl (95), TEACl (20), EGTA (10), HEPES (10), MgCl₂(1)、CaCl₂(0.2), glucose (10), Tris-ATP (5), Di-Tris-phosphocreatinine (20), creatine phosphokinase (50U); the pH was adjusted to 7.3 with CsOH or HCl. The extracellular solution had the following basic composition (mM): NaCl (140), KCl (3), CaCl₂(0.2), glucose (10), HEPES (10), sucrose (4.5), tetrodotoxin (TTX 3 x 10)^-4)。

N1E-115 cells were purchased from the European Collection of cell cultures (ECACC, Salisbury, UK) and stored at-80 ℃ until later use. The cells were measured at 100,000 cell cm^-2Was plated on plastic petri dishes (Falcon) and supplemented with 15% fetal bovine serum (Gibco) using NaHCO₃The cells were fed in HEPES buffered Minimal Essential Medium (MEM) and 5% CO at 37 deg.C₂And cultured at 95% humidity. The medium was replaced all daily. Cells were re-seeded into fresh petri dishes after treatment with trypsin-EDTA (1% in PBS), resuspended in MEM, and centrifuged at 1000 rpm for 4 minutes once every three days.

2-3 days after seeding, patch-clamp recordings were performed from lifted cells (liftedcells) using EPC-7 amplifiers (List) in whole cell mode using a polished glass electrode (2-3 M.OMEGA.) at room temperature (20-22 ℃). The test substances were applied as for the hippocampal cells. The content of the intracellular solution was as follows (mM): CsCl (130), HEPES (10), EGTA (10), MgCl₂(2)、CaCl₂(2) K-ATP (2), Tris-GTP (0.2), D-glucose (10); the pH was adjusted to 7.3 with CsOH or HCl. The extracellular solution had the following basic composition (mM): NaCl (124), KCl (2.8), HEPES (10), adjusted to pH7.3 with NaOH or HCl.

Results received only from stable cells were included in the final assay, i.e., showed at least 75% recovery of response to agonist (5-hydroxytryptamine or Ach) after removal of the test antagonist. Nevertheless, due to the attenuation of certain cells (10% in 10 minutes), the recovery of the drug effect is not always 100%. When present, it is always compensated by basing the% antagonism at various concentrations on control and recovery basis and assuming that this decay is a linear time course. All antagonists were evaluated in the case of steady state blockade using 3-6 concentrations on at least 5 cells. Equilibrium blockade is achieved within 2-5 agonist applications depending on the concentration of antagonist.

Results

Table 1 shows the general structure of the amino-alkylcyclohexanes selected for this study.

Basic structure of amino-alkylcyclohexanes

MRZ	R1	R2	R3	R4	R5	R*
							579	CH3	CH3	CH3	CH3	CH3	NH2
601	CH3	CH3	CH3	CH3	C3H7	NH2
							607	CH3	CH3	H	CH3	C3H7	NH2
615	CH3	CH3	C2H5(CH3)	CH3(C2H5)	CH3	NH2
							616	CH3(H	H(CH3	H(CH3)	CH3(H)	CH3	NH2
617	H	H	CH3(C3H7)	C3H7(CH3)	CH3	NH2
							618	CH3	H	C3H7	CH3	CH3	NH2
620	H	H	C2H5(CH3)	CH3(C2H5)	CH3	NH2
							621	H	H	CH3	CH3	CH3	NH2
625	H	H	H	CH3	CH3	NH2
							627	H	H	H	C3H7	CH3	NH2
629	H	H	H	C2H5	CH3	NH2
							632	CH3	CH3	C2H5	H	CH3	NH2
633	CH3	CH3	H	C2H5	CH3	NH2
							640	CH3	CH3	CH3	CH3	CH3	NHCH3
641	H	H	H	H	CH3	NH2
							642	CH3	CH3	CH3	CH3	CH3	NH(CH3)2
705	CH3	CH3	CH3	CH3	CH3	NH(CH2)4

TABLE 1

Substitution in parenthesesAlternatives in the racemic mixture, e.g. CH₃(C₃H₇) Meaning CH₃Or C₃H₇。

*****

Brief description of the drawings

FIGS. 1A and 1B show 5HT caused by MRZ2/633 in N1E-115 cultured cells₃Concentration-dependent blocking of receptors. 5-hydroxytryptamine (10. mu.M) was applied every 30 seconds for 2 seconds in the continuous presence of various concentrations of MRZ2/633 (1-10. mu.M).

A: as shown in the bar, the raw data of 5-hydroxytryptamine using a single N1E-115 cell was used. The left and right panels represent control and recovery responses, respectively. The middle three panels show equilibrium reactions in the continuous presence of MRZ 2/6331, 3 and 10. mu.M, respectively.

B: peak and steady state (plateau) 5-hydroxytryptamine current responses were normalized to control levels and mean (± SEM) was plotted against MRZ2/633 concentration (n-8). IC according to the 4 parameter logarithmic equation (Grafit, Erithocus Software)₅₀Estimation of s and curve fitting.

Figures 2A and 2B show that nicotine acts as a functional antagonist of neuronal nicotinic (Ia ═ α 7) receptors in hippocampal neurons by inducing receptor desensitization. Ach (1mM) was applied every 30 seconds for 2 seconds in the continuous presence of various concentrations (-) nicotine (1-10. mu.M).

A: raw data for single hippocampal neuron-Ach was applied as shown in bars. The left and right panels represent control and recovery responses, respectively. The middle three panels show the equilibrium reactions in the continuous presence of (-) nicotine 1, 3 and 10. mu.M, respectively.

B: peak Ach current responses were normalized to control levels and mean (± SEM) versus (-) nicotine concentration (n-12/concentration) were plotted. IC according to the 4 parameter logarithmic equation (GraFit, Erithocus Software)₅₀Estimation of s and curve fitting.

Fig. 3A and 3B show concentration-dependent blockade of neuronal nicotinic (Ia ═ α 7) receptors by MRZ2/616 in hippocampal neurons. Ach (1mM) was applied every 30 seconds for 2 seconds in the continuous presence of various concentrations of MRZ2/616 (1-100. mu.M).

A: raw data for single hippocampal neuron-Ach was applied as shown in bars. The left and right panels represent control and recovery responses, respectively. The middle three panels show the equilibrium reactions in the continuous presence of MRZ 2/61610, 30 and 100. mu.M, respectively.

B: peak Ach current responses were normalized to control levels and mean (± SEM) was plotted against MRZ2/616 concentration (n-11/concentration). IC according to the 4 parameter logarithmic equation (Grafit, Erithocus Software)₅₀Estimation of s and curve fitting.

Fig. 4A and 4B show concentration-dependent blockade of neuronal nicotinic (Ia ═ α 7) receptors by MRZ2/705 in hippocampal neurons. Ach (1mM) was applied every 30 seconds for 2 seconds in the continuous presence of various concentrations of MRZ2/705 (0.3-30. mu.M).

A: raw data for single hippocampal neuron-Ach was applied as shown in bars. The left and right panels represent control and recovery responses, respectively. The middle three panels show the equilibrium reactions in the continuous presence of MRZ 2/7050.3, 1.0 and 3.0. mu.M, respectively.

B: peak Ach current responses were normalized to control levels and mean (± SEM) was plotted against MRZ2/705 concentration (n-9/concentration). IC according to the 4 parameter logarithmic equation (GraFit, Erithocus Software)₅₀Estimation of s and curve fitting.

*******

Amino-alkylcyclohexane on 5-HT₃Action of receptor

All 10 tested amino-alkylcyclohexanes antagonize 5-hydroxytryptamine-induced inward currents in N1E-115 cells with similar potency to the NMDA-induced inward currents previously reported (fig. 1, see also Parsons et al, 1999). Against 5-HT permanently expressed in HEK-293 cells₃When receptors were tested, similarities were observed for the same compoundsThe function of (1). Thus, the amino-alkylcyclohexane tested was on 5-HT₃The receptor action is similar to that of many antidepressants previously reported (Fan, 1994), i.e. they antagonize the response by inducing desensitization.

MRZ27	[3H]MK-	PC NMDA	5HT3
				579	1.4	1.3	1.7
601	7.7	10.0	1.3
				607	7.7	13.8	22.3
615	2.29	1.30	2.5
				616	10.4	33.2	38.7
621	30.6	92.4	20.3
				632	2.8	6.4	2.4
633	4.7	13.9	7.7
				640	4.8	14.6	10.8
642	10.7	42.5	35.5

TABLE 2

For the amino-alkylcyclohexane pairs NMDA and 5-HT₃Summary of the receptor's potency. A reference to the replacement of [ 2 ] in rat cortex membranes was obtained from Parsons et al, 1999³H]MK-801 binding and antagonism of NMDA-induced inward current (at-70 mV) in cultured rat hippocampal neurons. anti-5-HT₃Efficacy of the receptor was assessed as the IC of the steady state "response to 2 sec 5-hydroxytryptamine (10. mu.M) against N1E-115 cells₅₀s(μM)。

Effect of amino-alkylcyclohexanes on neuronal nicotinic receptors

Ach (1mM) concentration-clamp applied to cultured hippocampal cell leadsResulting in a rapid, significant inward current that rapidly desensitizes to much lower plateau levels. Nicotine leads to concentration-dependent blockade of neuronal responses to Ach, and the concentrations reached in the CNS of smokers lead to substantial antagonism (figure 2, IC)₅₀＝1.17μM)。

We next also evaluated the efficacy of various amino-alkylcyclohexanes as α 7 neuronal nicotinic antagonists. In position R¹To R⁴Simple amino-alkylcyclohexanes with lower alkyl substitutions (see table 1) are potent α 7 neuronal nicotinic antagonists, and certain amino-alkylcyclohexanes are actually much more potent in this regard than previously reported for NMDA receptors, as exemplified by MRZ2/616 (see figure 3 and Parsons et al, 1999).

The N-pyrrolidine derivative MRZ2/705 was 16-fold more potent as an α 7 neuronal nicotinic antagonist than as an NMDA receptor antagonist (table 3 and figure 4).

MRZ	[3H]M	PC	PC Ach
				579	1.44	1.30	30.00
615	2.29	2.90	2.21
				616	9.94	33.20	3.40
617	36.08	63.90	1.16
				618	22.79	57.50	0.65
620	24.18	99.00	2.44
				621	30.56	92.40	0.65
625	48.98	244.90	3.29
				627	67.30	150.00	2.60
629	46.74	218.60	2.05
				641	135.86	＞100	2.40
642	10.73	42.50	1.00
				705	7.09	20.80	1.30

TABLE 3

Summary of the efficacy of amino-alkylcyclohexanes on NMDA and α 7 neuronal nicotinic receptors. A reference to the replacement of [ 2 ] in rat cortex membranes was obtained from Parsons et al, 1999³H]MK-801 binding and antagonism of NMDA-induced inward current (at-70 mV, PC NMDA) in cultured rat hippocampal neurons. Efficacy against the α 7 neuronal nicotinic receptor (PC ACh) was assessed as the IC against the peak response of cultured hippocampal neurons to application of 2 seconds ACh (1mM)₅₀s(μM)。

Conclusion

The present data show that amino-alkylcyclohexanes are 5-HT₃A receptor antagonist. These effects can be seen at concentrations similar to or even lower than those required for uncompetitive antagonism of the NMDA receptor as reported by Parsons et al, 1999. Thus these compounds are active against NMDA and 5-HT₃The combined antagonism of the receptors results in positive synergy, by increasing the target effects-cognitive enhancement and antidepressant effects, while further reducing possible side effects of NMDA receptor antagonism by e.g. decreasing mesolimbic (meso-limbic) dopamine hyperactivity, leading to its therapeutic safety and efficacy in alzheimer's disease. And, 5-HT₃Antagonism is itself used in the treatment of cognitive deficits, depression, alcohol abuse, anxiety, migraine, irritable bowel syndrome and emesis.

The present data also indicate that certain aminoalkylcyclohexanes as α 7 neuronal nicotinic receptor antagonists are actually more potent than their NMDA and/or 5-HT₃The receptor action is more effective. It is likely that many of these agents are also antagonists of the α 4 β 2 receptor, as reported by Buisson et al (1998) for agents such as memantine and amantadine. We propose that the positive effects of neuronal nicotinic agonists reported by others in animal models of various diseases are actually due to α 7 receptor desensitization and α 4/β 2 receptor inactivation/downregulation or other forms of functional antagonism resulting from, for example, partial antagonism. Thus, intermediate concentrations of neuronal nicotinic receptor antagonists are useful for neuroprotection or treatment of diseases associated with disorders of nicotinic transmission, such as Alzheimer's disease, Parkinson's disease, schizophrenia, Tourette's syndrome, drug abuse and pain.

Pharmaceutical composition

The active ingredients of the present invention may be incorporated in the form of pharmaceutical compositions and unit doses thereof with one or more conventional adjuvants, carriers or diluents, and may be employed as such in the form of: solids, such as coated or uncoated tablets or filled capsules, or liquids such as solutions, suspensions, emulsions, elixirs or elixir filled capsules, all for oral administration; suppository or capsule dosage forms for rectal administration or sterile injectable liquid dosage forms for parenteral (including intravenous or subcutaneous) application. Such pharmaceutical compositions and unit dosage forms thereof may thus contain conventional or novel ingredients in conventional or specific proportions, with or without additional active compounds or ingredients, and such dosage forms may contain any suitable effective amount of the active ingredient commensurate with the intended daily dosage range to be employed. Thus, tablets containing twenty (20) to one hundred (100) milligrams of active ingredient per tablet, or more broadly, ten (10) to two hundred and fifty (250) milligrams per tablet, are representative of unit dosage forms.

Method of treatment

Due to their high activity and low toxicity, and exhibiting a very favourable therapeutic index, the active ingredients of the invention can be administered to a subject in need thereof, such as the body of a living animal (including a human), for the treatment, alleviation or amelioration, palliation or elimination of the symptoms or conditions susceptible thereto, or representative thereof as described elsewhere in this application, preferably in parallel, simultaneously or together with one or more pharmaceutically acceptable excipients, carriers or diluents, particularly and preferably in the form of pharmaceutical compositions thereof, administered in an effective amount by oral, rectal or parenteral (including intravenous or subcutaneous) or in some cases even topical routes. The dosage range may be from 1 to 1000 mg per day, preferably from 10 to 500 mg per day, in particular from 50 to 500 mg per day, depending generally on the exact mode of administration, the form of administration, the indication for which the administration is to be made, the subject concerned and the body weight of the subject concerned, as well as the preference and experience of the attending physician or veterinarian.

Examples of representative pharmaceutical compositions

The reaction product may be formulated into tablets, coated tablets, capsules, drops, suppositories, injections, infusion preparations and the like with the aid of conventional solvents, adjuvants and carriers, and may be used therapeutically by oral, rectal, parenteral and other routes. Representative pharmaceutical compositions are as follows.

(a) Tablets suitable for oral administration containing the active ingredient may be prepared by conventional tableting techniques.

(b) With respect to suppositories, the active ingredient may be incorporated therein by conventional procedures using any conventional suppository base such as polyethylene glycol which is solid at normal room temperature but melts at or about body temperature.

(c) With regard to parenteral (including intravenous and subcutaneous) sterile solutions, conventional amounts of the active ingredient and conventional ingredients, such as sodium chloride and appropriate amounts of double distilled water, are used concurrently and aseptically by conventional procedures such as filtration, aseptic filling into ampoules or IV ampoules and autoclaving.

Other suitable pharmaceutical compositions will be readily apparent to those skilled in the art.

The following examples are given by way of illustration only and are not to be construed as limiting.

Example 1

Tablet formulation

A suitable formulation for a tablet containing 10 mg of active ingredient is as follows:

	Mg.
		active ingredient	10
Lactose	63
		Microcrystalline cellulose	21
Talc	4
		Magnesium stearate	1
Colloidal silica	1

Example 2

Tablet formulation

Another suitable formulation for a tablet containing 100 mg of active ingredient is as follows:

	Mg.
		active ingredient	100
Potato starch	20
		Polyvinylpyrrolidone	10
Film coating and coloring
		Composition of film coating material:
lactose	100
		Microcrystalline cellulose	80
Gelatin	10
		Polyvinylpyrrolidone, crosslinked	10
Talc	10
		Magnesium stearate	2
Colloidal silica	3
		Pigment	5

Example 3

Capsule formulation

A suitable formulation for a capsule containing 50 mg of active ingredient is as follows:

	Mg.
		active ingredient	50
Corn starch	20
		Dibasic calcium phosphate	50
Talc	2
		Colloidal silica	2

Filling in gel capsules.

Example 4

Injection solution

Suitable formulations for injectable solutions containing 1% active ingredient are as follows:

active ingredient mg	12
		Sodium chloride mg	8
Sterile water	To prepare 1ml

Example 5

Liquid oral formulation

A suitable formulation for a 1 litre liquid mixture containing 2 mg of active ingredient in 1ml of the mixture is as follows:

	G.
		active ingredient	2
Sucrose	250
		Glucose	300
Sorbitol	150
		Orange flavour	10
Sunset yellow.
		Purified water was added to prepare a total of 1000ml

Example 6

Liquid oral formulation

Another suitable formulation for a 1 litre liquid mixture containing 20 mg of active ingredient in 1ml of the mixture is as follows:

	G.
		active ingredient	20
Astragalus gum	7
		Glycerol	50
Sucrose	400
		Hydroxybenzoic acid methyl ester	0.5
Propyl p-hydroxybenzoate	0.05
		Red currant spice	10
Soluble red pigment	0.02
		Purified water was added to prepare a total of 1000ml

Example 7

Liquid oral formulation

Another suitable formulation for a 1 litre liquid mixture containing 2 mg of active ingredient in 1ml of the mixture is as follows:

	G.
		active ingredient	2
Sucrose	400
		Bitter orange peel tincture	20
Sweet orange peel tincture	15
		Purified water was added to prepare a total of 1000ml

Example 8

Aerosol formulation

180g of aerosol solution contained:

	G.
		active ingredient	10
Oleic acid	5
		Ethanol	81
Purified water	9
		Tetrafluoroethane	75

15ml of the solution are placed in an aluminium aerosol can, the metering valve is closed and 3.0bar purging (purge) is carried out.

Example 9

TDS formulation

100g of solution contained:

	G.
		active ingredient	10.0
Ethanol	57.5
		Propylene glycol	7.5
Dimethyl sulfoxide	5.0
		Hydroxyethyl cellulose	0.4
Purified water	19.6

1.8ml of the solution was placed on the fleece covered by an adhesive backing. The system is closed by a protective liner which is removed prior to use.

Example 10

Nanoparticle formulations

10g of polybutylcyanoacrylate nanoparticles comprise:

	G.
		active ingredient	1.0
Poloxamers	0.1
		Butyl cyanoacrylate	8.75
Mannitol	0.1
		Sodium chloride	0.05

The polybutylcyanoacrylates are prepared by emulsion polymerization in a water/0.1N HCl/ethanol mixture as polymerization medium. Finally the nanoparticles in the suspension are lyophilized under vacuum.

Thus, the compounds of the present invention find use in the treatment of diseases in a living animal body, particularly a human, at 5HT₃And nicotinic receptor indications for symptomatic and neuroprotective purposes.

As previously mentioned, methods of treating a living animal to inhibit the progression of, and alleviate, a selected disease using the compounds of the present invention can be carried out by any of the generally accepted pharmaceutical routes, using selected dosages effective to alleviate the particular disease to be alleviated.

Use of a compound of the invention in the preparation of a medicament for treating a living animal to inhibit a selected disease or condition, particularly susceptible to treatment with 5HT₃Or nicotinic receptor antagonists, comprising the step of admixing an effective amount of a compound of the present invention with a pharmaceutically acceptable diluent, excipient or carrier, and the methods of treatment, pharmaceutical compositions and use of the compounds of the present invention in the manufacture of medicaments are in accordance with the foregoing and our prior USP6,034,134 for the same 1-amino compound, and representative acid addition salts, enantiomers, isomers and hydrates and processes for their preparation are similarly disclosed in our prior USP and published WO applications for 1-amino-alkylcyclohexane compounds.

Representative pharmaceutical compositions prepared by mixing the active ingredient with a suitable pharmaceutically acceptable excipient, diluent or carrier include tablets, capsules, solutions for injection, liquid oral formulations, aerosol formulations, TDS formulations and nanoparticle formulations to produce a medicament for oral, injectable or dermal application are also consistent with the pharmaceutical composition examples previously described and given in U.S. patent 6,034,134 for these 1-amino-alkylcyclohexanes.

*****

It is to be understood that the invention is not limited to the specific details of operation or to specific compositions, methods, procedures or embodiments shown and described, since obvious modifications and equivalents will be apparent to those skilled in the art, and it is intended that the invention be limited only by the full scope of protection legally afforded by the appended claims.

Reference to the literature

Buisson, b., Bertrand, d., 1998, Open-channel blockers of the alpha4beta2 neuronal nicotinic acetylcholine receptor (Open channel blockers of the human α 4 β 2 neuronal nicotinic acetylcholine receptor), mol.

Fan，P.，1994，Effects of antidepressants on the inwardcurrent mediated by 5-HT₃receptors in rat nodose grandioneurones (antidepressant on 5-HT in rat ganglion neurons)₃The effect of receptor-mediated inward current), Br J Pharmacol 112, 741-744.

Greenshaw，A.J.，Silverstone，P.H.，1997，Thenonantiemetic uses of serotonin 5-HT₃Acceptor antadonosts (5-hydroxytryptamine 5-HT)₃Non-antiemetic use of receptor antagonists) drugs 53, 20-39.

Parsons, c.g., Danysz, w., Bartmann, a., Spielmanns, p., Frankiewicz, t., Hesselink, m., Eilbacher, b., quick, g., 1999, Amino-alkyl cyclic amine novel comprehensive NMDA recording agents with string voltage-dependency and fast blocking kinetics: in vitro and in vivo catalysis (amino-alkylcyclohexane is a novel uncompetitive NMDA receptor antagonist with strong voltage dependence and rapid blocking kinetics.) Neuropharmacology 38, 85-108.

Claims (9)

1. Use of a 1-aminoalkylcyclohexane compound of the formula or an optical isomer, enantiomer, hydrate or pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating a living animal to inhibit the progression of or alleviate a condition selected from emesis, cerebellar tremor, appetite disorders and irritable bowel syndrome:

wherein:

r is- (CH)₂)_n-(CR⁶R⁷)_m-NR⁸R⁹；

n + m is 0, 1 or 2;

R¹to R⁷Independently selected from hydrogen and C_1-6An alkyl group;

R⁸and R⁹Independently selected from hydrogen and C_1-6Alkyl or together represent C_2-5-an alkylene group.

2. The use of claim 1, wherein at least R¹、R⁴And R⁵Is C_1-6An alkyl group.

3. The use of claim 2, wherein R¹To R⁵Is methyl.

4. The use of claim 1, wherein R¹、R²、R³、R⁴、R⁵、R⁶And R⁷One of which is ethyl.

5. The use of claim 1, wherein R⁵Is propyl.

6. The use of any one of claims 1-3, wherein R⁶Or R⁷Is methyl.

7. The use of any one of claims 2-6, wherein R⁸And R⁹Together represent C₄Or C₅An alkylene group.

8. The use of claim 1, wherein the compound is selected from

1-amino-1, 3, 3, 5, 5-pentamethylcyclohexane,

1-amino-1-propyl-3, 3, 5, 5-tetramethylcyclohexane,

1-amino-1, 3, 3, 5 (trans) -tetramethylcyclohexane (axial amino),

1-amino-1, 3, 5, 5-tetramethyl-3-ethylcyclohexane (mixture of diastereomers),

1-amino-1, 3, 5-trimethylcyclohexane (mixture of diastereomers),

1-amino-1, 3-dimethyl-3-propylcyclohexane (mixture of diastereomers),

1-amino-1, 3 (trans), 5 (trans) -trimethyl-3 (cis) -propylcyclohexane,

1-amino-1, 3-dimethyl-3-ethylcyclohexane,

1-amino-1, 3, 3-trimethylcyclohexane,

1-amino-1, 3 (trans) -dimethylcyclohexane,

1-amino-1-methyl-3 (trans) propylcyclohexane,

1-amino-1-methyl-3 (trans) ethylcyclohexane,

1-amino-1, 3, 3-trimethyl-5 (cis) ethylcyclohexane,

1-amino-1, 3, 3-trimethyl-5 (trans) ethylcyclohexane,

n-methyl-1-amino-1, 3, 3, 5, 5-pentamethylcyclohexane,

1-amino-1-methylcyclohexane,

n, N-dimethyl-1-amino-1, 3, 3, 5, 5-pentamethylcyclohexane,

1-amino-1, 5, 5-trimethyl-3 (cis) -isopropyl-cyclohexane,

1-amino-1, 5, 5-trimethyl-3 (trans) -isopropyl-cyclohexane,

1-amino-1-methyl-3 (cis) -ethyl-cyclohexane,

1-amino-1-methyl-3 (cis) -methyl-cyclohexane,

1-amino-5, 5-diethyl-1, 3, 3-trimethyl-cyclohexane, and

n- (1, 3, 3, 5, 5-pentamethylcyclohexyl) pyrrolidine, and

optical isomers, enantiomers, hydrates and pharmaceutically acceptable salts of any of the foregoing compounds.

9. The use of any one of claims 1-8, wherein the medicament is a pharmaceutical composition comprising the compound in combination with one or more pharmaceutically acceptable diluents, excipients or carriers.