Spiraeoideae containing five-membered rings, the pharmaceutical composition

 

(57) Abstract:

Spiraeoideae General formula (1), their pharmaceutically acceptable salts, enantiomers, racemates and Quaternary compounds, representing derivatives of these compounds containing a tertiary nitrogen atom, which contain a five-membered group, in which spirostane is implemented by the carbon atom included as part of the five-membered groups, and the composition of the saturated ring system containing one nitrogen atom, the ring system a is a residue of formula (a-b), in the structure (a) bridge one end attached to the position 1 and the other end to position 4 or 5, m = 2, n and p = 1, and n+p= 2; R1- H1-6alkyl, C2-6quinil; specified the remainder of the five-membered 3-utilization, 1-acetylserotonin, 3-methylhydantoin and others specified in paragraph 1 of the claims. Compounds 1 are active in relation to the Central and peripheral nervous system, in particular agonistic activity towards muscarine. 3 S. and 7 C.p. f-crystals, 1 Il., 5 table.

< / BR>

Scope and background of the invention

The application is a continuation application of U.S. 08/094855, published July 20, 1993, for which priority is claimed.

At the HQ, as described below, spiroborate attached ring, with or without the jumper and containing one nitrogen atom, and pharmaceutical composition comprising the above spiraeoideae.

Known (US 4104397) spiraeoideae, in which 1,3-dioxolane and Hinkley associated normal spirogermanium atom. Describes the synthesis and characterization of (activity and selectivity of binding between sites M1and M2optical isomers of one such connection ("AF30") (Saunders J. et al., J. Med. Chem. 30: 969 (1987)); the published synthesis spirodiclofen/piperidinovyh analogues AF30 and their agonistic effect in relation to cholinergic muscarinic activity (J. Saunders et al., 3. Med. Chem. 31: 486 (1988)); also known spirooxazine/hinoklidina analogues exhibiting obvious muscarinic activity (Nordvall et al., J. Med. Chem. 35: 1541 (1992)).

Found spiraeoideae active at least in relation to the Central nervous system, in which oxathiolane ring spiroborate connected with hinoksalinovym rings (see EP-A-0205247, US 4855290, EP-A-0303391, US 4981858, which describes four different optical isomers of the same compound of this kind, as well as EP-A-0314444, US 4900830, US 4876260, which deals with the us (US 5053412) and spiritualiy (EP-A-0452101). It should be emphasized that the entire contents of the aforementioned patents and published patent applications, in which at least one of the applicants is among the applicants of the present invention, as well as all other patents, published patent applications and scientific articles cited in the present description, as well as the content of the application (WO 95/03303) (since it is not mentioned in the revised version of this application) is incorporated into the present application by reference. Conducted a large-scale study of biological activity of 2-methylspiro(1,3-oxathiolan-5,3')hinoklidina, which exists in the form of geometrical CIS - and TRANS-isomers in the matter is whether the 2-methyl group on the same side oxathiolane ring, and the nitrogen atom that is part of hinolinovogo ring (CIS-), or on the opposite side from the specified nitrogen atom (TRANS); on the basis of pre-clinical tests it is shown that the CIS-compound (conventionally denoted by AF102B) it seems particularly promising in terms of control senile dementia type Alzheimer's disease (SSTA). Each of the above CIS - and TRANS-isomers may be subjected to optical division, additionally, you can issledovat which the system is saturated N-heterocyclic ring spiroborate connected with oxalanilide, tiolovymi, oxathiolane, dioxolane or dithiolane rings are agonists cholinergic activity or exhibit therapeutic activity (EP-A-0311313, EP-A-0189370). Known compounds, in which the saturated N-heterocyclic ring spiroborate attached to 2-position of the 1,3-oxathiolan-4-(or 5-)ones, or dioxolane or dithiolane analogues exhibiting muscarinic agonistic activity against receptors M1(J-AND-02/247183); other compounds in which the system is saturated N-heterocyclic ring spiroborate attached to the oxo - or thiono-oxylidinum or oxo or thiono-isoxazolidine, have activity against Central nervous system or can be used in the treatment of mental illness (J-AND-S/208590, J-AND-02/164882).

Published more than 50 spiroperidol (or pyrrolidin)/hydantoins and some of their Tihonovich analogs exhibiting anti-inflammatory properties (Winters et al., Farmco, Ed. Sci. 25(9): 681-93 (1970)). There is a message group spiroperidol (or tetrahydropyran, or thiopyran)/hydantoins having inhibitory activity against the enzyme, but no significant antispasmodic action the doctrine of the new helical compounds. Another task that provide an acceptable pharmaceutical composition containing these compounds will be apparent from the following description.

Description of the invention

One aspect of the present invention provides spiraeoideae that meet the following formula (AA)

< / BR>
including their pharmaceutically acceptable salts, enantiomers, racemates and Quaternary compounds, representing derivatives of these compounds containing a tertiary nitrogen atom, which contain a five-membered group, in which spirostane is implemented by the carbon atom included as part of the five-membered groups, and the composition of the saturated ring system containing one nitrogen atom, and

the specified five-membered residue selected from the group comprising 3-utilization; 1-acetylserotonin; 3-methylhydantoin; 3-propargylglycine; 2,4-dirigeante; 2-thiohydantoin; oxazolidin-2 - tion; 3-ethyloxazole-2-he; oxazolidine,4-dione; 3-ethyloxazole-2,4-dione; 2-methyl-1,4-oxazolidin-3-one; 2-methyl-1,4-thiazolidin-3-one; 2,4-dimethyl-1,4-thiazolidin-3-one; 2-ethyl-1,4 - thiazolidin-3-one; 2-ethyl-4-methyl-1,4-thiazolidin-3-one; 3-methyl-1,4 - oxathiolan-2-he; 2-ethyl-1,4-thiazolidin-3-one; 5-methyl-1,3 - oxazolidin; 4-ethyl-1,3 is the as; 3-(4-pyrrolidino-2-butynyl)-as; 3-(2 - butynyl)-as; 2,5-bis(methylthio)-4H-imidazole; 3-ethyl-4 - thiohydantoin; 4-methylthiazole-2-tion; 3-ethyl-2,4 - dirigeante; 4 ethylthio-3-imidazolin-2-tion; 1-ethyl-2-ethylthio-2 - imidazolin-5-tion; 2-thio-4-hydroxyethylaminophenol; 2,5 - bis(aminomethyl)-4H-imidazole; 2-methyl-2-thiazoline; 2-methyl-2 - imidazoline; 2-methyl-2-oxazoline-4-one; 2-methyl-4H(5H)-imidazole-5(4) -; 2-methylthio-5-methoxy-4H-imidazole; 2-methylthio-5-amino-4H - imidazole; 2-methylthio-5-aminomethyl-4H-imidazole; 2 tion-3-utilization; 2 tion-3-tert-Butylimino; 2-methylthio-2-imidazolin-5(4) -; 1-ethyl-2-ethylthio-2-imidazolin-5-Oh and 1-ethyl-2-imidazolin-5-he;

specified saturated ring system containing one nitrogen atom, selected from a number of groups, including groups K, M, N:

< / BR>
< / BR>
in the structure To bridge one end attached to the position 1 and the other end to position 4 or 5;

m is 2;

n and p independently of one another are equal to 1, and n + p = 2;

R1denotes a group selected from among the following: a hydrogen atom, a C1-6-alkyl, C2-6-quinil.

One of the variants of the present invention provides spirochetemia in which the specified saturated ring system containing inertron and Hinkley.

Specific helical compounds according to the invention include the following:

1 methylpiperidin-4-Spiro-4'-(2',5'-bis(methylthio)-4H-imidazole);

1 methylpiperidin-4-Spiro-5'-(3'-ethyl-4'-thiohydantoin);

1 methylpiperidin-4-Spiro-5'-(4'-methylthioadenosine-2'-tion);

1 methylpiperidin-4-Spiro-5'-(3'-ethyl-2',4'-dirigeante);

1 methylpiperidin-4-Spiro-5'-(4'-ethylthio-3'-imidazolin-2'-tion);

1 methylpiperidin-4-Spiro-4'-(1'-ethyl-2'-ethylthio-2' -imidazolin-5'-tion);

1 methylpiperidin-4-Spiro-5'-(2'-thio-4' -hydroxy - etilaminopodandov);

1 methylpiperidin-4-Spiro-4'-(2',5'-bis(aminomethyl)-4'H - imidazol);

1 methylpiperidin-4-Spiro-5'-(3'-ethyloxazole-2'-he)

1 methylpiperidin-4'-Spiro-4'-(2'-methyl-2'-thiazolin);

1 methylpiperidin-4-Spiro-4'(5')-(2'-methyl-2'-imidazolin);

1 methylpiperidin-4-Spiro-5'-(2'-methyl-2'-oxazoline-4'-he);

1 methylpiperidin-4-Spiro-4'(5')-[2'-methyl-4'H(5 N)- imidazole-5'(4')-he];

1 methylpiperidin-4-Spiro-4'-(2'-methylthio-5'-methoxy-4'H-imidazol);

1 methylpiperidin-4-Spiro-4'-(2'-methylthio-5'-amino-4 N-imidazole);

1 methylpiperidin-4-Spiro-4'-(2'-methylthio-5'-aminomethyl-4 N-imidazole);

1 methylpiperidin-4-Spiro-5'-(3'-utilization);

1 methylpiperidin-4-Spiro-5'-(1'-acetylimino);

piperidine-4-Spiro-5'-(3'-ethyl) - Rev. BR>1 methylpiperidin-4-Spiro-5'-(3'-propargylglycine);

N-methylnitramine-3-Spiro-5'-as;

N-methylnitramine-3-Spiro-5'-(3'-methylhydantoin);

N-methylnitramine-3-Spiro-5'-(3'-methylhydantoin);

1 methylpiperidin-4-Spiro-5'-(2',4'-dirigeante);

1 methylpiperidin-4-Spiro-5'-(2'-thiohydantoin);

1 methylpiperidin-4-Spiro-5'-(oxazolidin-3'-tion);

1 methylpiperidin-4-Spiro-4'-(3'-ethyloxazole-2',4'-Oh);

1 methylpiperidin-4-Spiro-5'-(oxazolidin-2',4'-dione);

1 methylpiperidin-4-Spiro-5'-(3'-ethyloxazole-2',4'-dione);

1 methylpiperidin-4-Spiro-5'-(2'-methyl-1',4'-oxazolidin-3'-Oh);

1 methylpiperidin-4-Spiro-5'-(2'-methyl-1',4'-thiazolidine-3'-Oh);

1 methylpiperidin-4-Spiro-5'-(2',4'-dimethyl-1',4'-thiazolidine-3'-Oh);

1 methylpiperidin-4-Spiro-5'-(2'-ethyl-1',4'-thiazolidine-3'-Oh);

1 methylpiperidin-4-Spiro-5'-(2'-ethyl-4-methyl-1',4'- thiazolidine-3'-Oh);

piperidine-4-Spiro-5'-(3'-methyl-1',4'-oxathiolan-2'-he);

piperidine-4-Spiro-5'-(2'-methyl-1',4'-thiazolidine-3'-Oh);

1 methylpiperidin-4-Spiro-5'-(3'-methyl-1',4'- oxathiolan-2'-he);

1 methylpiperidin-4-Spiro-2'-(5'-methyl-1',3'-oxazolidin);

1 methylpiperidin-4-Spiro-2'-(4'-ethyl-1',3'-oxazolidin);

1 methylpiperidin-4-Spiro-5'-(3'-ethyl-1',4'-oxathiolan-2'-he);

1 methylpiperidin-4-Spiro-5'-(2'-AIRO-2'-(5'-methyl-1',3'-dioxolane-4'-he);

1 methylpiperidin-4-Spiro-5'-(2'-methyl-1',4'- thiazolidine-3'-tion);

2-N-methylspiro-(1,3-succinimide-4,3')Hinkley;

2-N-utilisera-(1,2-succinimide-4,3')Hinkley;

1 methylpiperidin-4-Spiro-5'-[3'-(4-pyrrolidino-2-butynyl) -as];

1 methylpiperidin-4-Spiro-5'-(3'-tert-Butylimino);

1-propolypeptide-4-Spiro-5'-(3'-utilization);

1 methylpiperidin-4-Spiro-5'-[3'-(2-butynyl)-as];

piperidine-4-Spiro-5'-(3'-propargylglycine);

2-methyl-1,4-thiazolidin-3-he-Spiro[5, 3']-Hinkley;

1 methylpiperidin-4-Spiro-5'-(2'-tion-3'-utilization);

1 methylpiperidin-4-Spiro-4'-(2'-tion-3'-tert-Butylimino);

1 methylpiperidin-4-Spiro-4'(5')-(2'-methylthio-2'-imidazolin-5'(4') -);

1 methylpiperidin-4-Spiro-4'-(1'-ethyl-2'-ethylthio-2'-imidazolin-5'-he);

1 methylpiperidin-4-Spiro-4'-(1'-ethyl-2'-imidazolin-5'-he).

It should be noted any (one or more) dedicated right - and left-torque enantiomers, selected from among the following:

d - and l-1 methylpiperidin-4-Spiro-5'-(2'-methyl-1',4'- thiazolidine-3'-Oh), and

d - and l-1 methylpiperidin-4-Spiro-5'-(2'-ethyl-1', 4'- thiazolidine-3'-Oh), including their pharmaceutically acceptable salts and Quaternary compounds.

Provided by the present invention the soy is their money for example, in therapy. More specifically, provided by the present invention spiraeoideae affect the functioning of the Central and/or peripheral nervous system. A common characteristic of these compounds is their activity in relation to the cholinergic system, and the considered compounds are ligands of muscarinic receptors. Thus, another aspect of the present invention provides spiraeoideae formula AA with muscarinic activity. Compounds according to the invention, as defined by the authors of the application, is additionally possess the biological activity of one of the following types:

necrotroph-like or synergistic activity when combined with NEF,

the activity that causes the secretion of the protein precursor of amyloid a (BPA) and reduce the toxicity of amyloid,

activity, increasing the share of dephosphorylating-proteins and NGF-like activity.

Such compounds constitute another aspect of the invention.

Tests used to assess the biological activity of the compounds provided by the present invention, are described below.

Another option vopl the molecular characteristics,

where r denotes the relative position determined by the location of the anion relative to the cationic form of the nitrogen atom (denoted N*) that is not a double bond and is composed of ring A (or A') in the most stable conformation of the compounds being considered;

X*is a heteroatom, which is part depicts the 5-membered ring, and said heteroatom is in the adjacent position relative to the corresponding spirotone carbon;

Z*denotes an atom in the following depicts the 5-membered ring through from 1 atom X*;

Q*is an atom or N, located at the end of the side chain that is attached to the atom in the structure depicted ring between atoms X*and Z*; the hydrogen atoms included in the side chain, are not taken into account;

these molecular characteristics can take the following specific values:

flat angle r-X*-Q*-Z*ranging from -54oto -170o;

molecular distances: r-N*= 3.0 angstroms (relative distance);

r-X*= 5,7-6.75 angstroms;

r-Q*= 7,9-8,90 of angstroms;

x is regular features, exhibit agonistic activity towards muscarine. The above pattern, derived on the basis of molecular characteristics, confirmed by the following biological tests (see Table 1).

In addition, a pharmaceutical composition intended for use in the treatment of diseases of the Central and peripheral nervous system in mammals; this composition includes effective for the treatment of the aforementioned diseases, the number of at least one compound corresponding to the formula (AA), including pharmaceutically acceptable salts, enantiomers and racemates of these compounds. The composition may contain at least one pharmaceutically acceptable diluent, carrier or additive. In the preferred case, the composition is either in a format suitable for oral, rectal, parenteral or subcutaneous injection (in this case, consider the composition may additionally include a low molecular weight fatty acids), either in the form intended for injection or injection into the nose, and may be in the form of a unit dosage. At least one connection provided on the to 100 mg, in the preferred case is approximately from 5 to 100 mg, more preferred is from about 10 to 50 mg.

In accordance with one alternative embodiment of the present invention, the pharmaceutical composition described in the preceding paragraph can optionally include at least one pharmaceutically active compound selected from among the following: physostigmine, tetrahydroaminoacridine, choline, lecithin, piracetam, aniracetam, pramiracetam, oksiratsetam, 4-aminopyridine, 3,4-diaminopyridine, somatostatin, pirenzepine, N-methylatropine, N-butylscopolamine, scopolamine, clonidine, kantaminen, propantheline, methantheline, glycopyrrolate, trapezium, nortriptyline, amitriptyline, imipramine, minupren, recoverin, AFDX-116, nicotine, alaproclate, zimelidine, deprenil, and Nerve Growth Factor.

Another variant of the embodiment of the present invention provides a pharmaceutical composition, which includes at least one compound of formula AA, and Nerve Growth Factor (NGF), and provided by the present invention, the connection (connection) is contained in such an amount that stimulates nerve growth caused by the activity of NGF. Most Sousa effect on the Central or peripheral nervous system, not able to cause the growth of nerves in the absence of NEF, which provides better control in that case, if therapy want to stimulate nerve growth. However, some of the most active compounds provided by the present invention, can cause nerve growth regardless of NGF.

Provided by the present invention compounds can be used for the treatment of diseases in mammals, if these diseases susceptible to treatment under the action of an effective amount of the compounds (compounds) expressing muscarinic activity; this connection can be used in the form of pharmaceutical compositions comprising an effective amount of these compounds and corresponding to other parameters outlined above.

In addition, the present invention provides the use of compounds corresponding to the above formula (AA), including pharmaceutically acceptable salts, Quaternary compounds, represents a structural derivatives of these compounds having a tertiary nitrogen atom, and enantiomers and racemates of these compounds, to obtain the drug, oblad the present invention, well-known specialists in the field of organic chemistry as methods of obtaining five-membered rings, substitution on the ring, changing the degree of saturation/unsaturation of the ring, internal rearrangements of salts and bases, the formation of Quaternary salts and so on. Thus, despite the fact that in the further description will list some methods of obtaining specific compounds provided by the present invention, for obtaining the compounds can also be used and other methods obvious to the expert.

In that case, if the desired five-membered ring represents, for example, as specified ring can be obtained by the reaction of the corresponding saturated N-heterocyclic ketone (for example 1 methylpiperidin-4-it) (NH4)2CO3+ CN-after which the nitrogen atom located in the 3'-position of the considered rings can be replaced in a known manner. These reactions may be illustrated as follows:

< / BR>
(AA') + R0- (tsepliaeva group)

< / BR>
including: AF160 (R0=Et); AF178 (R0=Me); AF185 (R0=propargyl); AF167 (R0= Me, similar*); AF168 (R0=Et, anavol jumper i.e. analogues of N-methylnitramine.

Tsepliaeva group, which is part of the specified reagent "R0-(tsepliaeva group)" may represent a bromine atom, a chlorine atom, or a residue of p-toluensulfonate acid, while the group R0corresponds to that used above designation with the exception of the hydrogen atom, alkoxy and alkanoyl. This substitution reaction can be carried out under well known conditions, including the interaction of 3'-unsubstituted as in the presence of alkali (such as KOH), or when using a solvent, such as ethanol. The corresponding 1',3'-disubstituted compound can be obtained in the above reaction using an excess of the specified reagent "R0-(tsepliaeva group) or in the interaction of the compound (B) with a reagent R0-(tsepliaeva group)",

1-methyl group, a subset of the patterns (B), can be removed by the reaction with demetrious agent such as CH3CH(Cl)OCOCl

< / BR>
including AF160(Des) (R0=Et); AF179 (R0=Me).

Structure (B) can also be obtained in the following way:

< / BR>
When replacing connection R0-N=C=S during the same rectilineal a halide, or alkanoyl anhydride carried out under standard conditions, causes the following substitution at the specified 1'position:

< / BR>
In that case, if the desired five-membered ring represents dirigeante, these compounds can be obtained, for example, by forming the above-mentioned ring as a result of interaction between the corresponding saturated N-heterocyclic ketone and CN-, NH4Cl and CS2. If the hydantoins substituted (for example by alkylation) on respective nitrogen atoms, based thiohydantoins receive the products, substituted by nitrogen atoms and/or sulfur. The following examples illustrate the reaction conditions conducive to the formation of different products or product mixtures that can be separated. These reactions may be illustrated as follows (example of a suitable N-heterocyclic ketone is 1-methylpiperidin-4-one):

< / BR>
< / BR>
and/or

< / BR>
and/or

< / BR>
Specified tsepliaeva group, and R0included in the reagent "R0-(tsepliaeva group)" may be the same groups described above in the case of hydantoins. The reaction is obtained with reactions between the corresponding hydantoins and R2S2resulting in

< / BR>
In that case, if the compound (D) result in interaction with 20% HCl, the corresponding link S-R0hydrolyses receive the following connections:

< / BR>
A similar reaction between the compound (G) and 20% HCl leads to the formation of compounds having the following structure (see in particular Example 12):

< / BR>
The specified connection is also formed by hydrolysis of the corresponding analogue carrying a balance =NR0instead remainder =O; this analogue can be obtained, resulting connection AF173 in contact with the reagent R0NH2(see in particular Example 13, where R=- hydroxyethyl).

Provided by the present invention compounds represent oxo or thiono-substituted oxazolidine, can be obtained, for example, as follows:

< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
As indicated tsepliaeva group, and R0included in the reagent "R0-(tsepliaeva group)" may be the same groups described above in the case of hydantoins. Substitution reactions can be performed under well known conditions.

Provided this is to be obtained, for example, as follows:

< / BR>
3 carboethoxy-3-carbomethoxybiphenyl

< / BR>
Provided by the present invention of thiazoline can be synthesized, for example, as follows:

< / BR>
< / BR>
It should be noted that the connection

< / BR>
while the compounds corresponding to the formula (E), have not been shown in this application as concrete examples. However, now unexpectedly found that the compounds provided by the formula (E), in particular the connection AF151(S), are much more promising from a pharmacological point of view compared to the class of compounds, which include the connection AF150(S).

Provided by the present invention imidazoline can be obtained, for example, in accordance with the following scheme:

< / BR>
< / BR>
< / BR>
In this case, the resulting product, in particular AF190 (R=Me), may be in the form of a tautomeric mixture of compounds (G) and (H).

Provided by the present invention oxazolidine and thiazolidine, such as compounds (I), in particular AF264 (R=Me, R'=H) and AF268 (R=H, R'=Et), as well as connections (K), in particular AF261 (R=Me, R0=H), AF267 (R=Et, R0=H) and AF266 (R=R0=Me), can be obtained, for example, the following method is icon in interaction with, accordingly, HOCHRCHR'NH2or HSCHRCO2H + R0NH2.

The following compounds in which R and R0correspond to the notation used above, however, are largely stands or ethyl, are also preferred compounds provided by the present invention.

< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
< / BR>
It should be emphasized that if in the description below, as examples of compounds provided by the present invention, discusses the connection includes piperidine, nortropinone and hinoklidina ring any nitrogen - containing heterocyclic ring, referred to in this description as convenient for the education of zeroconfiguration with listed spiroplatin ring, may be substituted. A similar clarification is true with respect to specific Examples, which are primarily illustrative, and not restrictive.

Provided by the present invention spiraeoideae potentially suitable primarily for the treatment of presenilny and senile dementia, senile SL (1990)), caused multiple infarct dementia together with Alzheimer's disease, age-related memory disorders (GRP), attacks of acute disorientation, mental disorders and attention, mania, slowness, hypermodernity, mixed Alzheimer's and Parkinson's disease, aphasia, hallucinatory-paranoid States, syndrome post brain amnesia symptoms of alcohol intoxication, Huntingtonbeach traucki, disease Peak, ataxia, Frederick, disease, Gilles de La Touré, and down syndrome, because all of these diseases are disorders in which there is a place at least some decrease in Central cholinergic activity. Also provided by the present invention compounds potentially suitable for the treatment of progressive nanolearning paralysis; these compounds are also potential analgesic agents, and therefore can be used in the treatment of severe painful conditions such as rheumatism, arthritis and diseases of the nervous endings.

As was already briefly mentioned above, provided by the present invention spiraeoideae can be used in combination with the smaller m is, is akimi as fototermin or tetrahydroaminoacridine; in combination with a precursor of acetylcholine, such as choline or lecithin; together with "nootropic" drugs such as piracetam, aniracetam, oksirazetam or pramiracetam; together with compounds affecting Ca2+-view channels, such as 4-aminopyridine or 3,4-diaminopyridine; or together with peptides, are able to exert a regulatory effect on the release of acetylcholine, such as somatostatin; together with peripheral agent exhibiting activity antimuskarinovoe act occurs (such as pirenzepine, N-methylatropine, N-butylscopolamine, propantheline, methantheline, glycopyrrolate or trapezoideum), to prevent unwanted peripheral effects such as salivation, diarrhea, the secretion of gastric juice or vomiting that may be caused by high doses of the specified agent or in combination with subcutaneously injected with scopolamine, such as Scopoderm(R)to prevent nausea and/or vomiting; in combination with antidepressants, such as nortriptyline, amitriptyline, imipramine, minupren, to facilitate as mental disorders, and depressive symptoms sometimes associated with SSTA, GRP and mixed disease (SSTA/Parkinso (Hammer et al. Life Sci. 38: 1653 (1986)) to prevent unwanted peripheral side effects that can be expected when using high doses respective agents, to prevent the inhibitory effects exerted considered agonists by inhibiting presynaptic and postsynaptic receptors M2-type in the Central nervous system, as well as to enhance the production of acetylcholine in the suppression of inhibiting autoreceptors M2-type in the intact nerve endings; in combination with nicotinic agonists, such as nicotine, for stimulation of nicotinic and muscarinic receptors of the brain; in combination with an agonist adrenergically activity (clonidine or kapamilya) to facilitate mental and other disorders associated with mixed cholinergic and noradrenergicheskoy failure when SSTA; in combination with inhibitors of reuptake of serotonin neurons (alaproclate, zimelidine) to facilitate mental and other emotional functions when SSTA; in combination with inhibitors monoamine oxidase, such as deprenyl, to facilitate mental and other locomotor disorders associated with mixed disease type STA/BP; in combination with the LASS="ptx2">

Provided by the present invention spiraeoideae, both in the presence and in the absence of the above-mentioned additional active agents can be used, for example, by injection in an acceptable diluent or carrier, orally, rectally in the form of suppositories, by injection or injection into the nose, through infusion or subcutaneously using an acceptable carrier in the presence or in the absence of fitotecnia or tetrahydroaminoacridine.

Provided by the present invention spiraeoideae can also be of interest in terms of treatment of diseases requiring the use of long-acting cholinergic agents having a mild local activity. This agent is required in the treatment of diseases, such as glaucoma; as specified spirochaetaceae not destroyed by the enzyme, inactivating acetylcholine (in particular, acetyl, butyryl-cholinesterase), it can also be used to treat peripheral cholinergic diseases, such as myasthenia pregnant, dysfunction of the bladder, disease ADI and disease Eaton-Lambert. These compounds can also be used in cases where smotriny the present invention spiraeoideae are agents, showing antiholinergicescuu activity (which can be easily determined by the expert), these compounds may find use in the treatment of diseases caused by the hyperactivity of the cholinergic activity, regardless of whether the hyperfunction of spontaneous or induced by drugs. Also provided by the present invention compounds can be used in the treatment of various diseases, such as BP, pseudo-PSU, mixed SS/BP, primary dystonia, spastic torticollis, cranial dystonia, depression, decreased mobility, akathisia (arising after neuroleptic intoxication), mediated by the Central nervous system, increased stress, traumatic brain injury, mixed mental deficiency and BP, manic-depressive syndrome; as an aid in surgery (instead of atropine, scopolamine, and the like); in case of poisoning due to excess acetylcholine arising, for example, the suppression of acetylcholinesterase. In addition, these compounds can be used in ophthalmology, if you need long-term or short-term mydriasis.

the surveillance of diseases, accompanied by an increased activity similar to the activity of the peripheral nervous system; among these diseases include asthma, chronic pulmonary insufficiency, peptic ulcer. In the case of listed diseases of the peripheral system is particularly recommended to use Quaternary salt compounds provided by the present invention.

Currently, the Quaternary ammonium salts are widely used in therapy. In particular, the number of such agonists cholinergic activity include acetylcholine chloride, beta-nehal chloride, and carbachol (Goodman & Gilman''s "The Pharmacological Basis of Therapeutics, Seventh Edition. Macmillan Publishing Co. (1985), p. 104). Quaternary agents exhibiting anticholinesterase activity, serve as neostigmine bromide, ambenonium chloride, pyridostigmine bromide, indoforum chloride, demecarium bromide, and hativat iodide; pralidoxime chloride is used as reactivate cholinesterase (Goodman & Gilman''s "The Pharmacological Basis of Therapeutics". Seventh Edition. Macmillan Publishing Co. (1985), p.122-123).

Quaternary derivatives of alkaloids of belladonna (including, methscopolamine bromide and gomatropin methyl bromide), and synthetic Quaternary compounds (h is Gilman''s "The Pharmacological Basis of Therapeutics", Seventh Edition. Macmillan Publishing Co. (1985), p.139-140).

On the basis of the chemical structure of Quaternary ammonium ions predicted that these ions can cause paralysis, similar to the paralysis caused by curare (A. Burger "Medicinal Chemistry". Second Edition. Interscience Publishers (1960), p.497). This feature Quaternary compounds used in anesthesia, for example, as additional measures during surgical anesthesia to relax skeletal muscles (Goodman & Gilman, "The Pharmacological Basis of Therapeutics, Seventh Edition. Macmillan Publishing Co. (1985), p.222-235).

The above-mentioned active Quaternary compounds, causing neuromuscular blockade did not prevent the development and clinical application of Quaternary compounds in subsequent years. The specialist must know that the choice of any connection (including Quaternary compound) for use in clinical therapy is influenced by many factors, including the effectiveness in terms of the desired destination, safety, possible side effects, and therapeutic index. Thus, an experienced specialist should understand, what does the expression "pharmaceutically acceptable Quaternary compounds, which is a structural derivative I provided in the present description and the claims in terms of relevant existing knowledge.

Hereinafter the present invention will be illustrated in the following Examples, without restrictive.

Example 1: 1 Methylpiperidin-4-Spiro-5'-(3'-utilization) AF160

a) 1-Methylpiperidin-4-Spiro-5'-as

The mixture of solutions 1 methylpiperidin-4-it (36,44 g, 0,322 M) in ethanol (150 ml), ammonium carbonate (93.0 g; 0,968 M) in water (400 ml) and potassium cyanide (25,8 g; 0,396 M) in water (82 ml) was kept at a temperature of 60oC for 2.5 hours, then left overnight at room temperature. Was filtered fallen in sediment 1 methylpiperidin-4-Spiro-5'-as and washed his small quantities of cold water, ethanol and ether to obtain crystalline powder (27,0 g). Concentrating the obtained filtrate and washing liquid were allocated second portion of the product (20,0 g). The isolated product was led from methanol, so pl. 265-276oC (decomposition).

IR (KBr) 3170 (NH); 1700 (C=O) cm-1.

Mass spectroscopy m/z 183 (M+, 38%); 71 (100%).

1H-NMR (D2O) 1,8 (2H); 1.06 (the sextet, 2H); 2.49 USD (s, -CH3); of 2.58 (t, 2H); 3,14 (t, 1H); 3,20 (t, 1H) ppm.

b) 4-Amino-1-methylpiperidin-4-carboxylic acid

A mixture of 1-methylpiperidin-4-Spiro-5'-as (9.75 g; 0,0533 M) and octahydrate hydro is keep four of these tanks was filtered and precipitated precipitated barium carbonate. The obtained filtrate was neutralized solid CO2and the resulting precipitate was collected by filtration. Concentrated highlighted the filtrate to obtain 4-amino-1-methylpiperidin-4-Spiro-carboxylic acid (32,0 g, 95%), so pl. 275-280oC (decomposition).

IR (KBr) 3300, 1655, 1580 cm-1.

Mass Spectroscopy m/z 183 (M+, 90%); 141 (98%, M-OH); 113 (12%, M-CO2H); 96 (100%); 71 (52%).

1H-NMR (C5D5N+D2O) 1,2 (m, 2H); of 1.46 (sextet, CH3N-); and 1.7 (m, 2H); 1,9 (m, 2H); 2,0 (m, 2H) ppm.

in 1 Methylpiperidin-4-Spiro-5'-(3'-utilization), AF160

To a mixture of 1-methylpiperidin-4-Spiro-5'-as (5 g, 27 mm) and potassium hydroxide (2,08 g; 37 mm) in 100 ml of absolute ethanol was added ethyl bromide (15 g; 137 mm). The resulting mixture was heated to a temperature of 80oC, every 0.5 h were sampled and compared them by GLC with an internal standard (difenilmetana). To maintain the alkalinity of the solution was performed titration (1 N. HCl), after which was added potassium hydroxide (2.1 g). Getting maximum output (after 2.5 h), evaporated above solution was added water (50 ml), and the resulting aqueous solution was extracted with chloroform and subjected to chromatography on silicagel column using tarali in the air and adding HCl in isopropanol, was besieged by the corresponding hydrochloric acid salt, so pl. 278-280oC.

Mass spectroscopy m/z 211 (M+, 45%); 71 (100%).

1H-NMR (free base, CDCl3) 1,2 (t, J=6 Hz, 2H); 1.6 to 1.7 (m, 2H); 1.9 to 1,95 (m, 2H); from 2.1 to 2.2 (m, 2H); of 2.34 (s, 3H); 2,85-2,95 (m, 2H); 3,5 (q, J=6 Hz, 2H);

1H-NMR (HCl salt, D2O) 1,1 (t, J=6 Hz, 3H); 1,95-2,05 (m, 2H); about 2.2-2.3 (m, 2H); 2,85 (s, 3H); to 3.0-3.2 (m, 2H); 3,4-3,5 (m, 2H); 3,5 (q, J=6 Hz, 2H) ppm.

13C-NMR (free base, CDCl3) 14,0; 33,1; 33,1; 46,0; 52,8; 59,9; 157,0; 177,0 ppm.

UV (free base, H2O)max208 nm ( 3500).

Example 2: 1 Methylpiperidin-4-Spiro-5'-(1'-acetylimino) AF164

a) AF164A. A mixture of 1-methylpiperidin-4-Spiro-5'- as (3.25 g) in acetic anhydride (50 ml), kept at the temperature of reflux distilled for 3 hours Under reduced pressure selected in excess of the specified reagent to obtain a solid substance, which was dispersible in ether and was filtered emitting white solid (3.75 g), crystallized from a mixture of methanol and dichloromethane, so pl. 250-254oC (decomposition) AF164A.

1H-NMR (D2O) 1,89 (m, 2H); 2,44 (s, CH3CO); 2,86 (s, CH3N); 2,98 (m, 2H); to 3.41 (m, 2H); to 3.67 (m, 2H) ppm.

13C-NMR (D2), dioxane as an internal one hundred); 166,0 (C2'); 173,8 (CH3CO); 189,2 (C4') ppm.

MC m/z 255 (M+); 210; 166; 155; 123; 95; 71 (100%); 70.

b) AF164B. The connection part AF164A (1.10 g) was podslushivaet saturated aqueous Na2CO3and were extracted with a mixture of methanol and dichloromethane, evaporated the resulting extract, and the resulting residue was extracted again with the same mixture of solvents were filtered extract, evaporated corresponding to the filtrate, and the selected residue (1.0 g) triturated with acetone to yield compounds AF164B in the form of a white solid substance, so pl. 225-230oC (decomposition) (crystallization from CH2Cl2-CH3-CH3CN).

1H-NMR (D2O) of 1.53 (m, 2H); of 2.21 (s, CH3CO); 2,39 (s, CH3N); 2,65 is 2.80 (m, 6H) ppm.

13C-NMR (D2O, dioxane as internal standard) 26,9 (CH3CO -), AND 28.3 (C3and C5); 45,0 (CH3N); 50,9 (C2and C6); 64,9 (C4); 67,3 (dioxane); 168, 8mm (C2'); 173,6 (CH3CO); 193,9 (C4') ppm.

MC m/z 225 (M+); 183 (M+-CH2=C=0); 166; 154; 123; 95; 71 (100%).

in) AF164 (hydrochloride). In order to obtain hydrochloride of the compound AF164, a solution of compound AF164A or AF164B in methanol was treated with HCl dissolved in isopropanol, to acidic pH (pH 1-2). Chere is. the HP 301-302oC (decomposition).

1H-NMR (D2O) 2,17 (m, 2H); 2.50 each (s, CH3CO-); at 2.93 (s, CH3N); 3,10 (m, 2H); 3,48 - 3,71 (m, 4H) ppm.

13C-NMR (D2O, dioxane as internal standard) 26,7 (C3and C5); 26,9 (CH3CO); 43,9 (CH3N); 51,1 (C2and C6); 61,8 (C4); 67,3 (dioxane); 154,8 (C2'); 173,4 (CH3CO); 176,0 (C4') ppm.

Hydrolysis of compounds AF164:

Connection AF164A and AF164B within 1-2 h hydrolyzed at a temperature of reflux distilled at 0.2 N. aqueous NaOH solution to obtain 1-methylpiperidin-4-Spiro-5'-as identified by matching it with TLC and1H-NMR spectrum with an authentic sample.

Example 3: piperidine-4-Spiro-5'-(3'-utilization) AF164(Des)

To a solution of dried connection AF160 (2.0 g; 9.5 mm) in dichloroethane (25 ml, dried over molecular sieve) at room temperature was added - chloroethylphosphonic (1.0 ml; 9.3 mm), and the resulting mixture was stirred at a temperature of 60oC for 1 h In vacuum was removed dichloroethane, selected solid was dissolved in 20 ml of methanol, and the resulting solution was kept at a temperature of 60oC for a further 30 minutes and Then vacuum was removed, methanol, and dedicated oily tuccio was extracted with chloroform, and the extract evaporated to yield roughly refined oil, which was further purified by chromatography on silicagel column. The elution was performed with a mixture of chloroform, methanol and ammonia in the ratio of 4:1:0,1 emitting compounds AF160 (Des) (1.12 g; 60% yield) as a white powder, so pl. 225-227oC.

MS m/z 197 (M+the main peak); 57.

1H-NMR (CDCl3) of 1.17 (t, J=6 Hz, 3H); of 1.65 to 1.7 (m, 2H); 1.85 to 2.0 a (m, 2H); of 2,75 2,85 (m, 2H); 3,05 is 3.15 (m, 2H); 3,5 (q, J=6 Hz, 2H) ppm.

Example 4: 1-Methylpiperidin-4-Spiro-5'-(3'-methylhydantoin) AF178

To a mixture of 1-methylpiperidin-4-Spiro-5'-as (5.0 g; 27,3 mm) and sodium hydroxide (2.0 g; 50 mm) in 120 ml of methanol was added to the methyl tosylate (11.2 g; 60 mm). The resulting reaction medium was stirred over night at room temperature by evaporation of methanol was removed, and the selected oily residue was dissolved in aqueous potassium carbonate solution and was extracted with chloroform. Evaporated appropriate organic extract, and the obtained crudely purified product was further purified by chromatography on silicagel column, elution with a mixture of chloroform, methanol and ammonia in the ratio of 9:1:0.1 s allocation of 1.2 g (22%) white donovania, CDCl3) to be 1.6-1.7 (m, 2H); to 2.1-2.3 (m, 4H); of 2.34 (s, 3H); 2,85-2,95 (m, 2H); to 3.02 (s, 3H) ppm.

Example 5: piperidine-4-Spiro-5'-(3-methylhydantoin) AF179

A white powdery product obtained in the same manner as described in Example 3, was dissolved in isopropanol and acidified with hydrochloric acid to yield a white precipitate. So pl. above 320oC (decomposition).

MS m/z 183 (M+the main peak); 57.

1H-NMR (hydrochloride, D2O) 2,0 (m, 2H); 2,2 (m, 2H); 2,96 (s, 3H); 3,3 (m, 2H); 3,6 (m, 2H).

Example 6: Synthesis of 1-methylpiperidin-4-Spiro-5'- (3'-propargylglycine) AF185

A suspension of KH (11 g; 0.1 M; 35% suspension in mineral oil) and 1 methylpiperidin-4-Spiro-5'-as (25 g; 0,13 M; dried over P2O5) was stirred at room temperature in dry DMF (500 ml). Added propargyl chloride and withstand the resulting reaction mixture at a temperature of 50oC for 20 minutes this mixture was cooled, acidified aqueous hydrochloric acid to pH 3, and then DMF was removed, extragere its oil mixture of ether and the ether in the ratio 1:1, and then ether. The corresponding aqueous phase was podslushivaet with sodium carbonate to pH 10 and was twice extracted with chloroform. United received a chloroform extracts, Vissi column. The elution was performed with a mixture of chloroform, methanol and ammonia in the ratio of 80:19:1 with the release of 3.0 g of purified compounds AF185, as well as several fractions (total 10 g) containing a small amount of impurities. The corresponding free base was besieged in the form of the hydrochloride; using crystallization from methanol was obtained 3.2 g of a white hygroscopic salt.

1H-NMR (D2O hydrochloride) of 2.0 to 2.35 (m, 4H); of 2.64 (t, 1H, J=3.5 Hz); 2.91 in (s, CH3N); 3,1-3,2 (m, 2H); of 3.5-3.7 (m, 2H); 4,27 (bs, 2H) ppm.

1H-NMR (D2), excess sodium carbonate) of 1.65 and 1.75 (m, 2H); 1.9 to 2.05 is (m, 2H); of 2.25 (s, CH3N); about 2.2-2.3 (m, 4H); 2,8-2,9 (m, 4H); 4.25 in (s, 2H) ppm.

1H-NMR (CDCl3- free basis) of 1.85-of 1.95 (m, 4H); 2,2 (m, 2H); 2.23 to (t, 1H, J=3.5 Hz) and 2.4 (s, CH3N); 2,9-2,95 (m, 2H); 4,3 (d, 2H, J=3.5 Hz); at 6.4 (bs, 1H) ppm.

MC m/z 221 (M+the main peak); 206 (M-15); 149.

Example 7: 1-Methylpiperidin-4-Spiro-4'-(2',5'-bis (methylthio)-4'H-imidazol) AF177

To a solution of 1-methylpiperidin-4-Spiro-5'-(2',4'-dirigeante (1,00 g; 4,65 mm; see Example 11) in methanol (15 ml) was added NaOH (0,30 g; 7.50 mm), and then gradually a solution of methyl iodide (1,00 g;? 7.04 baby mortality mm) in methanol (3.0 ml) and the resulting mixture was stirred at room temperature for 1.5 h deposition was Performed using NaBr, hoteltravel the STI, then solvent was removed; the selected residue was podslushivaet with K2CO3and was extracted with ether. The resulting organic extract was dried on Na2SO4and solvent was removed with the release of the residue, which was subjected to chromatography on silicagel column. The elution was performed with a mixture of ether, chloroform, methanol and ammonia in the ratio of 78:18:3:1 with the release of the connection AF177 (465 mg); after crystallization from hexane so pl. 101-102oC.

1H-NMR (CDCl3) of 1.33 (m, 2H); to 1.98 (m, 2H); 2,39 (s, CH3N); 2,56 (s, CH3S); 2,59 (s, CH3S); 2,48-of 2.64 (m, 2H); 2,82 (m, 2H) ppm.

13C-NMR (CDCl3) 14,1 (CH3S); 14,2 (CH3S); 34,9 (C3and C5); 46,1 (CH3N); 52,2 (C2and C6); 83,0 (C4); 171,6 (C2'-); 203,5 (C5') ppm.

MC m/z 244 (M++1); 185; 149; 93; 75.

IR (KBr) 2920; 2797; 1535; 1477; 1565; 1452; 1378; 1316; 1286; 1210; 1108; 1054; 1000; 965; 942; 900; 776; 696 cm-1.

UV (EtOH)max257 nm ( 16100).

Example 8: 1-Methylpiperidin-4-Spiro-5'-(3'-ethyl-4'-thiohydantoin) AF182

An aliquot of 1 methylpiperidin-4-Spiro-4'-(1'-ethyl-2'-ethylthio - 2'-imidazolin-5'-thione) (100 mg; compound AF170, see Example 12) was dissolved in 20% HCl (1 ml) and the resulting solution was subjected to reflux distilled for 1.5 hours With pooveli her dichloromethane. The resulting organic extract was dried over Na2SO4and solvent was removed with the release 1 methylpiperidin-4-Spiro-5'-(3'-ethyl-4'-thiohydantoin) as a white solid (74 mg), crystallized from a mixture of petroleum ether and dichloromethane, so pl. 176-178oC.

1H-NMR (CDCl3) 12,5 (t, J=7.2 Hz, CH3CH2-); and 1.54 (m, 2H); 2,10 (m, 2H); of 2.36 (s, CH3N); is 2.41 (m, 2H); 2,96 (m, 2H); of 3.94 (q, J=7.2 Hz, -CH2CH3); 7.23 percent (NH) ppm.

13C-NMR (CDCl3) 11,8 (CH3CH2-); 36,9 (C3and C5); 37,4 (-CH2CH3); 46,1 (CH3N); 51,2 (C2and C6); 68,1 (C4); 157,1 (C2'); 208,1 (C4') ppm

MC m/z 227 (M+); 211 (M+-OH); 194 (M+-SH); 170 (M+-C3H7N); 71; 70 (100%).

UV (EtOH)max280 nm ( 13600), 229 nm ( 4400).

Example 9: 1-Methylpiperidin-4-Spiro-5'-(4'-methylthio-3'- imidazolin-2'-tion) AF183

Reproducing the Example 7, but using an equivalent amount of methyl iodide, sodium hydroxide and litigiation, in addition to bis-(methylthio)-derived compounds AF177 received 1 methylpiperidin-4-Spiro-5'-(4'-methylthio-2'- imidazolin-2'-tion), the connection AF183, so pl. 218-220oC (decomposition) (from a mixture of CH2Cl2and acetone).

1H-NMR (CDCl3+-CH3S); 123; 122; 70.

UV (EtOH)max312 nm ( 12000), 280 nm ( 15300).

Example 10: 1-Methylpiperidin-4-Spiro-5'-(3'-ethyl-2',4'- dirigeante) AF 163

Thoroughly mixed powders 1 methylpiperidin-4-Spiro-5'-(3'- utilizaton) (0,570 g) and pentasulfide phosphorus (0,570 g) and the resulting mixture was subjected to reflux distilled in tetraline (15 ml) for 2 h Indicated the reaction mixture was allowed to cool to room temperature, after which formed thick brown sludge treatment under reduced pressure was removed, tetralin, getintegervalue selected precipitate and washed it ether. The resulting mixture was podslushivaet concentrated aqueous solution of NaOH and was extracted with dichloromethane. The corresponding extract was dried over Na2SO4and evaporated emitting residue (0,250 g), which was subjected to chromatography on a column of silica gel (Merck, 60,15 g). The elution was performed with a mixture of chloroform, ether, methanol and ammonia in the ratio of 78:18:4:1 with a yield of pure compounds AF163 (50 mg), crystallized from a mixture of CH2Cl2and ether, so pl. 223-225oC (decomposition).

1H-NMR (CDCl3) of 1.27 (t, J=7.2 Hz, CH3CH2-); 1,55-to 1.67 (m, 2H); 2,03-2,19 (m, 2H); 2.26 and is 2.46 (m, 2H); of 2.36 (s, CH3N); 2,89 totaling 3.04 (m, 2H); 4,27 (q, J= 7.2 Hz, -CH2-CH2-CH3; 45,9 (CH3N); A 49.9 (C2and C6); 72,9 (C4); 179,8 (C2'); 207,8 (C4') ppm.

MC m/z 243 (M+); 186 (M+-C3H7N); 149; 71; 70 (100%); 57.

IR (KBr) 3177 (NH); 2930; 2778; 1513; 1434; 1357; 1231; 1116; 1090; 1070; 1040; 961; 801; 780; 626; 546; 457 cm-1.

UV (EtOH)max302 nm ( 34200), 226 nm ( 7700).

Synthesis of tartrate salt of compound AF163

To a solution of the free base compounds AF163 (0,735 g; 3,025 mm) in a mixture of methylene chloride (15 ml) and methanol (5 ml) was added a solution of L(+)-tartaric acid (0,214 g; 1,427 mm) in methanol (2.0 ml). The resulting mixture was stirred at room temperature for 0.5 h, then evaporated these solvents, and the selected residue suspended in a mixture of ether and methylene chloride, filtered and washed with a mixture of the above solvents with the release of yellow solid, which represents a tartaric acid salt of the compound AF163 (0,923 g; 96% yield), so pl. 221-225oC.

1H-NMR (D2O) of 1.27 (t, J=7.2 Hz, CH3CH2-); of 2.08 (m, 2H); 2.49 USD (m, 2H); 3,01 (s, CH3N-); and 3.31 (m, 2H); 3,76 (m, 2H); 4.26 deaths (q, J=7.2 Hz, -CH2CH3); 4,37 (s, -CHCHOH) ppm.

Example 11: 1-Methylpiperidin-4-Spiro-5'-(2',4'- dirigeante) AF173

A solution of 1-methyl-4-piperidone (29,35 g; 0,260 M), KCN (26,57 g; 0,408 M), NH45oC) within 8 hours Specified reaction medium was left overnight at room temperature, was filtered, the resultant precipitate was washed with water and then ethanol to yield yellow solid (26,3 g; 47,0% yield), crystallized from methanol, so pl. 252-254oC (decomposition).

1H-NMR (DMSO-d6) 1,43-of 1.56 (m, 2H); 1,89-2,05 (m, 2H); (C3and C5); of 2.21 (s, CH3N-); and 2.26-2.40 a (m, 2H); 2,64-and 2.79 (m, 2H); 11,14 (brs., -NH-) ppm.

13C-NMR (DMSO-d6) 36,2 (C3and C5), WITH 45.5 (CH3N); A 49.9 (C2and C6); 74,8 (C4); 181,5 (C2'); 212,0 (C4') ppm.

MC m/Z 215 (M+); 183 (M+S); 182 (M+-SH); 181; 158 (M+-C3H7N); 123; 102; 77; 71; 70 (100%).

IR (KBr) 3130 (NH); 1484; 1449; 1350; 1259; 1231; 1198; 1176; 1145; 1083; 1062; 957; 721; 545; 504 cm-1.

< / BR>
Example 12: 1-Methylpiperidin-4-Spiro-5'-(3'-ethyl-2', 4'-dirigeante) AF163; 1 Methylpiperidin-4-Spiro-5'-(4' -ethylthio-3'-imidazolin-2'-tion) AF176; 1 Methylpiperidin-4-Spiro-4'- (1'-ethyl-2'-ethylthio-2'-imidazolin-5'-tion) F170

To a suspension of 1-methylpiperidin-4-Spiro-5'-(2', 4'-dirigeante) (connection AF173; 3,30 g; 15,3 mm) in dry DMF (30 ml) was added NaOH (0,760 g; 60% mixture in mineral oil; 19,0 mm) and stirred the mixture at a temperature of 50oC for the ur 75-80oC for 4 h, After considering the mixture was stirred over night at room temperature, the formed precipitate (NaBr), which was filtered and washed with ether. Combined corresponding to the filtrate and washing liquid and dried them to produce oil, which was separated insoluble in dichloromethane solid (1.5 g), caught the reference compound (identified using NMR and TECH). The obtained residue was subjected to chromatography on silicagel column. The elution was performed with a mixture of ether, chloroform, methanol and ammonia in the ratio of 68: 27:4:1 with the output of the first 1 - methylpiperidin-4-Spiro-5'-(3'-ethyl-2', 4'-dirigeante) (connection AF163; 0.95 g), identical to the product obtained previously in the reaction between the compound AF160 and P2S5. Continued elution with output 1 methylpiperidin-4-Spiro-5'-(4'-ethylthio-3'- imidazolin-2'-thione) (connection AF176; 0.15 g). Crystallizes from a mixture of hexane and CH2Cl2so pl. 212 - 215oC (decomposition).

1H-NMR (CDCl3) to 1.42 (t, J=7.2 Hz, CH3CH2-); at 1.73 (m, 2H); to 2.06 (m, 2H); 2,31 (m, 2H); of 2.38 (s, CH3N); 2,95 (m, 2H); to 3.33 (q, J=7.2 Hz, -CH2CH3); 10,0 (br. -NH-) ppm.

13C-NMR (DMSO-d6) 14,2 (CH3CH2'
); 198,1 (C4') ppm.

MC m/z 243 (M+); 182 (M+-EtS, 100%); 156 (M+-EtSCN); 123; 124; 71; 70; 57.

IR (KBr) 3135 (NH); 2930; 2788; 1476; 1463; 1446; 1281; 1257; 1232; 1158; 1141; 1121; 1096; 958; 711; 674; 535 cm-1.

UV (EtOH) max314 nm ( 5400), 282 nm ( 6700).

Repeating the above reaction with a 1.5 molar excess EtBr in relation to the connection AF173, in addition to the above monoethylene derivative gain diethylene derived: 1 methylpiperidin-4-Spiro-4'-(1'-ethyl-2'-ethylthio-2'- imidazolin-5'-tion) (connection AF170), so pl. 66-67oC (crystallized from hexane).

1H-NMR (CDCl3) to 1.24 (t, J=7.2 Hz, CH3CH2-); 1,24 (m, 2H); of 1.44 (t, J=7.2 Hz, CH3CH2-); and 2.27 (m, 2H); 2,39 (s, CH2N); 2,52 (m, 2H); of 2.81 (m, 2H); 3.24 in (q, J=7.2 Hz, -CH2CH3); 3,93 (q, J=7.2 Hz, -CH2CH3) ppm.

13C-NMR (DMSO-d6) 14,2 (CH3CH2-); 13,9 (CH3CH2-); 25,7 (-CH2CH3); 36,8 (C3and C5); 39,0 (-CH2CH3); 46,1 (CH3N); 51,5 (C2and C6); AND 82.2 (C4); 158,5 (C2'); 216,7 (C5') ppm.

MC m/z 271 (M+); 242 (M+-Et); 214 (M+-C3H7N); 185; 165; 75; 71; 70 (100%); 57.

IR (KBr) 1574 (C=N); 1446; 1372; 1354; 1212; 1071; 1060; 936 cm-1.

UV (EtOH)max296 ( 22000), 252 nm ( 17700).

Acid hydroly salsa smell of mercaptan. This solution was stirred at room temperature for 1 h, then evaporated at a temperature of 50oC and reduced pressure to obtain hydrochloride of 1-methylpiperidin-4-Spiro-5'-(2'-tingitania) (X).

1H-NMR (D2O) 2,10 is 2.43 (m, 4H); 2,94 (s, CH3N); 3,20 (m, 1H); 3,47-of 3.78 (m, 3H) ppm.

MC m/z 199 (M+); 181; 171 (M+-CO); 156 (M+-HNCO); 142 (M+-C3H7N); 111; 96; 75; 71; 70; 57.

UV (H2O)max264 nm ( 20400), 224 nm ( 8600).

Example 13: 1-Methylpiperidin-4-Spiro-5'-(2'-thiohydantoin) AF195

a) a Solution of 1-methylpiperidin-4-Spiro-5'-(2',4'-dirigeante) (connection AF173; 10.0 g) in a mixture of ethanolamine (40 ml) and water (75 ml) was subjected to reflux distilled for 1.5 hours Under reduced pressure to remove the used solvent and excess reagent. As a result of consistent kristallizatsii selected balance:

- a mixture of acetonitrile and dichloromethane,

- acetone and

- mixture of ethanol and acetonitrile were obtained pure 1-methylpiperidin - 4-Spiro-5'-(2'-thio-4'- (8 - hydroxyethylaminophenol), so pl. 230-231oC (decomposition).

1H-NMR (D2O) 11,74 (m, 2H) was 1.94 (m, 2H); and 2.26 (s, CH3N); 2,17-of 2.34 (m, 2H); 2,92 (m, 2H); of 3.54 (q, J=5.4 Hz, -CH2OH); 3,74 (q, J=5.4 Hz, =NCH2-) ppm.

13C-I IS(C4); 182,5 (-C=S); 195,0 (-C=N ) ppm.

MC m/z 242 (M+); 224 (M+-H2O); 199; 185; 172 (M+-70); 154 (M+-H2O); 71; 70.

< / BR>
The specified derived aminoguanine (1.40 g) was dissolved in water 50% HCl (5.0 ml) and subjected to reflux distilled for 1 h the reaction mixture is evaporated under reduced pressure, and the selected balance was led from methanol to yield the hydrochloride of 1-methylpiperidin-4-Spiro-5'-(2'-thiohydantoin) (0,756 g), identical to the product that was obtained by hydrolysis of compounds AF176 (see Example 12).

b) a Solution of 1-methylpiperidin-4-Spiro-5'-(2',4'-dirigeante) (connection AF173; 10.0 g; 0,0465 M) and n-butylamine (17.0 g; 0,233 M) in ethanol (80 ml) was kept at the temperature of reflux distilled within 1.5 hours From the cooled reaction mixture was filtered precipitated precipitated crystalline solid, washed it with a small amount of ethanol, then ether, and then with petroleum ether to obtain 1-methylpiperidin-4-Spiro-5'-(2'-thio-4'-Ethylenediamine) (connection AF189; 11.1 g; 0,0437 M; 94% yield). The isolated product was led from a mixture of dichloromethane and methanol to yield needle-like crystals so pl. 236-239oC (decomposition).

1H-NMR (CDClrepresented 33.47 per (t, J=7.2 Hz, -CH2-N) ppm.

MC m/z 254 (M+); 197 (M+-C4H9); 184 (M+-70); 149; 128; 71; 70; 57 (C4H9+).

The specified derived aminoguanine (connection AF189; 10,45 g) was dissolved in water 16% HCl (15.0 ml) and subjected to reflux distilled for 1 h the reaction mixture was distilled as long as there was a small amount of fluid. To the selected residue was added ethanol (50 ml) and cooled the resulting mixture, which resulted in precipitation, which was filtered and washed with a small amount of ethanol and then ether to obtain the hydrochloride of 1-methylpiperidin-4-Spiro-5'- (2'-thiohydantoin) (connection AF195; 8,42 g; 87% yield), so pl. > 295oC (with decomposition), is identical to the previously selected product.

Example 14: 1-Methylpiperidin-4-Spiro-5'-(oxazolidin-2'- tion) AF165

To stir anhydrous DMSO (10 ml) containing a small amount of powdered KOH was added 4-aminomethyl - 4-hydroxy-1-methylpiperidin (0,595 g; 4,13 mm), and then carbon disulfide (0,340 g; 4,47 mm). The resulting reaction mixture was stirred at a temperature of 50oC for 2 h Used the solvent was removed in vacuo, and the selected balance multiple RA is. 190-195oC (21 mg).

1H-NMR (CDCl3) 1,80-to 1.98 (m, 2H); 2,03-of 2.16 (m, 2H); 2,32 (s, CH3N); 2,44-of 2.72 (m, 2H); 3,51 (s, -CH2N-C=S) ppm.

13C-NMR (DMSO-d6) 35,1 (C3Cl5); 45,7 (CH3N); 51,5 (C2Cl C6); 53,1 (-CH2NH-); 86,4 (C4); 187,2 (-C=S) ppm.

Example 15: N-Methylnitramine-3-Spiro-5'-(3'-methylhydantoin) AF167 and N-Methylnitramine-3-Spiro-5'-(3'- utilization) AF168

a) N-methylnitramine-3-Spiro-5'-as

The mixture tropinona (45 g; 0,32 M) in ethanol (160 ml), ammonium carbonate (93 g; 0,96 M) in water (400 ml) and KCN (25,8 g, and 0.40 M) in water (84 ml) was kept at a temperature of 60oC for 2 h and then at room temperature for 16 hours Selected N-methylnitramine-3-Spiro-5'-as (61,33 g; 0,29 M; 92% yield) was dried in a desiccator, so pl. 330oC.

Mass spectroscopy m/z 209 (M+).

1H-NMR (CD3COOD) to 2.1 (m, 2H, H6=H7 ()); ); 2,3 (m, 2H, H2=H4 ()); 2,4 (m, 2H, H2=H4 ()); 2,7 (m, 2H, H6=H7 ()); 2,9 (s, 3H); 3,0 (bs, NH); to 4.1 (bs, 2H, H1=H5) ppm.

1H-NMR (DCl, D2O) to 2.3 (m, 4H, h6=H7( ) and H2=H4( )) ; 2,5 (m, 2H, H2=H4 ()); 2,7 (m, 2H, H6=H7 ()); 2,9 (s, 3H); 4,15 (bs, 2H, H1=H5) ppm.

13C-NMR (DCl, D2O) 25,5 (C6=C7, t); 33,5 (C2=C4, t); 39,7 (CH3, q); 59,3 (C3-C5', s); 63,0 (C1=C5, d); 159,3 (C2', s); 180,2 (C4', s) ppm.

b) Connection AF167 and AF168

M-Methylnitramine-3-Spiro-5'-g is n aqueous solution was added dropwise methyl iodide or ethyl bromide (2 parts) in methanol. Specified, the reaction mixture was extracted with chloroform and dried over magnesium sulfate, and then evaporated. Desired products (connection AF167 and AF168) received approximately 10% output.

Connection AF167.

1H-NMR (DCI, D2O) of 1.65 (m, 2H, H6=H7 ()); 1,8 (m, H2=H4 ()); 2,2 (m, 2H, H2= H4 ()); 2,4 (m, 2H, h6=H7 ()); to 2.55 (s, 3H); 2,85 (s, 3H); of 3.25 (bs, 2H, H1=H5) ppm.

1H-NMR (CDCl3) of 1.55 (m, 2H, H6=H7 ()); to 1.75 (m, 2H, H2=H4 ()); ; 2,2 (m, 2H, H2=H4 ()); 2,4 (s, 3H); of 2.45 (m, 2H, H6=H7 ()); 3,0 (s, 3H); 3,3 (bs, 2H, H1=H5); and 6.3 (bs, NH) ppm.

Mass spectroscopy m/z 223 (M+).

Connection AF168.

1H-NMR (CDCl3CD3OD) of 1.1 (t, 3H); 1,6 (m, 2H, H6-H7 ()); ; to 1.75 (m, H2= H4 ()); 2,2 (m, 2H, H2=H4 ()); to 2.35 (s, 3H); 2,4 (m, 2H, H6=H7 ()); 3,25 (bs, 2H, H1=H5); 3,55 (q, 2H) ppm.

1H-NMR (CDCl3) 1,2 (t, 3H); 1,6 (19, 2H, H6=H7 ()); to 1.75 (m, H2=H4 ()); 2,2 (m, 2H, H2=H4 ()); 2,4 (s, 3H); of 2.45 (m, 2H, h6=H7 ()); 3,3 (bs, 2H, H1=H5); 3,55 (q, 2H); 6,3 (bs, NH) ppm.

13C-NMR (CDCl3, ) 13 (CH3CH2), 25 (C6=C7); 35 (C2=C4); 40 (N-CH3); 40,5 (N-CH2); 59 (C3-C5'); 60 (C1=C5); 159,3 (C2'); 180 (C4') ppm.

Mass spectroscopy m/z 237 (M).

Example 16: 1-Methylpiperidin-4-Spiro-5'-(3'- ethyloxazole-2'-he) AF172

a) 4-Atsetamidometil-4-hydroxy-1-methylpiperidin

At room temperature for 2 h, mixed 4-aminomethyl - 4-hydroxy-1-methylpiperidin (2,95 g; 0,0 is ulali sodium hydroxide and was extracted with above solution with chloroform. Viparita the corresponding extract was allocated a yellow oil, which was identified as 4-atsetamidometil-4-hydroxy-1-methylpiperidin (3.4 g; 0,018 M; 91% yield).

b) 4-Ethylaminomethyl-4-hydroxy-1-methylpiperidin

A solution of 4-atsetamidometil-4-hydroxy-1-methylpiperidine (3.4 g; 0,018 M) in dry THF was subjected to reflux distilled in the presence of lithium hydride-aluminum (4 g). After 3 days the resulting mixture was poured into a mixture of water and ice and filtered through Celite. Evaporated the solvent, was added water, the resulting solution was extracted with chloroform, and the extract was dried over magnesium sulfate and evaporated with the release of 1.03 g (33% yield) roughly purified substances. Thus obtained 4-ethylaminomethyl-4 - hydroxy-1-methylpiperidin used without further purification.

in 1 Methylpiperidin-4-Spiro-5'-(3'-ethyloxazole-2'-he) AF172

4-Ethylaminomethyl-4-hydroxy-1-methylpiperidin (16,8 g; 0.1 M) under nitrogen atmosphere were mixed with N',N'-carbodiimide (32 g, 0.2 M) in 400 ml of chloroform. Viparita this mixture, received 50 g crudely purified substance, which was intensively washed with hexane and then evaporated to yield the desired compound AF172 - 1 methylpiperidin - 4-Spiro-5'-(3'-ethyloxazole-2'-o); 2,3 (s, 3H); to 2.55 (m, 2H); or 3.28 (s, 2H); of 3.32 (q, 2H); ppm.

13C-NMR (CDCl3) 13 (CH3CH2); 22 (CH3CH2); 37 (CH3CH2N); 39 (CH2NEt); 52 (NCH2CH2); 55 (C-O); 157,5 (C=O) ppm.

Mass spectroscopy m/z 198 (M+).

Example 17: 1 Methylpiperidin-4-Spiro-4'-(3'- ethyloxazole-2'-he) AF174

a) 4-Acetamido-4-hydroxymethyl-1-methylpiperidin

At room temperature for 2 h and was stirred 4-amino - 4-hydroxymethyl-1-methylpiperidine (2,95 g; 0.02 M), potassium carbonate (6.5 g; 0,047 M) and acetic anhydride (8.5 g; 0.08 M) in methanol. To neutralize received MESI was added sodium hydroxide and was extracted with above solution with chloroform. Viparis the corresponding extract, obtained white solid, which was led warm acetone to yield 4-acetamido-4 - hydroxymethyl-1-methylpiperidine (1,67 g; 0,009 M; 45% yield).

b) 4-Ethylamino-4-hydroxymethyl-1-methylpiperidin

A solution of 4-acetamido-4-hydroxymethyl-1-methylpiperidine (1.6 g; 8,6 mm) in dry THF was subjected to reflux distilled in the presence of alumoweld lithium (3 g). After 4 days the resulting mixture was poured into a mixture of water and ice and filtered through Celite. The solvent evaporated, and after evaporating most of the water was extracted with what hodom 1.13 g (77% yield) rather crudely purified substances. Thus obtained 4-ethylamino-4-hydroxymethyl-1-methylpiperidin used without further purification.

in 1 Methylpiperidin-4-Spiro-4'-(3'-ethyloxazole-2'-he) AF174

A mixture of 4-ethylamino-4-hydroxymethyl-1-methylpiperidine (172 mg, 1 mm) and N', N'-carbodiimide (486 mg, 3 mm) for 3 h were mixed in chloroform under nitrogen atmosphere. Viparita this mixture, received roughly purified substance, which was intensively washed with hexane and then evaporated to yield the desired compound AF172 - 1 methylpiperidin-4-Spiro-4'-(3'- ethyloxazole-2'-she) in the form of a white solid (140 mg; 0.71 mm; 71% yield).

1H-NMR (CDCl3) of 1.10 (t, 3H); 1,6 (m, 2H); 1,95 (m, 4H); of 2.25 (s, 3H); to 2.85 (m, 2H); 3,2 (q, 2H); to 4.1 (s, 2H) ppm.

Mass spectroscopy m/z 198 (M+).

Example 18: 2-N-Methylspiro-(1,3-succinimide-4,3')Hinkley AF133

a) Ethyl (3-finallydecided)-cyanoacetate

A mixture of 3-binucleation (30 g; 0.24 M), ethyl cyanoacetate (40 g; 0.35 M), ammonium acetate (3.8 g), acetic acid (11 g) and 120 ml of benzene was kept at the temperature of reflux distilled, and then using azeotropic distillation was removed, water (4 ml). Cooled obtained benzene solution, was added potassium carbonate (30 g) in 120 ml of water, and the resulting mixture three times the way the product was besieged in the form of the hydrochloride with a yield of 63 g (95% yield) of the crudely purified product.

TECH on silica gel Art 5735 (Merck), 2% mixture (volumetric ratio) of 25% aqueous ammonium hydroxide solution in methanol Rf0,67.

Specified product may be further purified by crystallization in ethanol or isopropanol.

1H-NMR (CDCl3-TMS) free base: of 1.29 (t, 3H, CH3); to 4.2 (q, 2H, CH2); 1,7-1,9; 2,8-3,2 (m, chinuklidinyl skeleton).

13C-NMR (CDCl3-TMS): 14 (CH3); 62 (CH2); 189 (C=O); 162 (C=N); 115 (C-CN); 100 (C=C); BY 33.7 (C-H).

b) 3-Carboethoxy-3-carbomethoxybiphenyl

A mixture of ethyl (3-genocidist)-cyanoacetate (64 g; 0.24 M) and potassium cyanide (17 g; 0,26 M) dissolved in 25 ml of water, was dissolved in 125 ml of ethanol. This mixture for 20 min was subjected to reflux distilled, cooled, filtered to remove potassium chloride and filtered potassium chloride twice washed with 50 ml ethanol. Were combined and evaporated corresponding alcohol solution, and the obtained oily residue was dissolved in 250 ml of concentrated hydrochloric acid and subjected to reflux distilled for 24 hours Then evaporated above solution, and the selected residue is repeatedly washed with acetone and dried. The dried solid was dissolved in ethanol saturated with hydrochloric kisk was carefully podslushivaet sodium carbonate and was extracted with chloroform. Was dried and evaporated corresponding to a chloroform solution, and the selected roughly purified fluids were additionally purified by chromatography on a column using 2% solution of methanol in chloroform as eluent.

MS m/z 2,69 (M+); the main peak of m/z 196 (M-C-OEt).

1H-NMR (CDCl3-TMS): 1,2 (dt, 6H, CH3); 4,2-4,3 (dt, 4H, CH2O); 1,3-1,6; 2,6-3,1 (m, chinuklidinyl skeleton).

a) 2-N-Methylspiro-(1,3-succinimide-4,3')Hinkley AF133

3 Carboethoxy-3-carbomethoxybiphenyl (3,35 g; 12 mm) was dissolved in 4.5 g of methylamine and within the 90 h, kept at a temperature of 190oC and high pressure. Cooled and evaporated specified the reaction mixture, and the resulting solid residue was purified using silicagel chromatography, elution of 2% solution of methanol in chloroform containing 0.2 ml of ammonia. Connection AF133 was allocated in the form of 1.2 g (5.7mm) of a white solid substance, so pl. 94-96oC.

MS 209 (M+).

1H-NMR (CDCl3-TMS) and 3.4 (d, 1H) (H2); 2,96 (s, 3H) (CH3); 2,5 (d, 1H) (H2); of 1.5-1.9 (m, chinuklidinyl skeleton).

Example 19: 2-N-Ethyl-Spiro-(1,3-succinimide-4,3')Hinkley AF134

Roughly purified 3-carboethoxy-3-carbomethoxybiphenyl (20 kω pressure. Over the course of the reaction was monitored by GC. Formed roughly purified product was extracted with chloroform, and the corresponding chloroform fraction was then dried and evaporated. The obtained oily residue was purified by chromatography on silica gel, elution with a mixture of chloroform, petroleum ether, ethanol and aqueous ammonia in the ratio 17:13:3:0,4. The corresponding free base was besieged in the form of the hydrochloride with a yield of 6.6 g of a white solid substance, so pl. 270-272oC. TECH on silica gel Art 5735 (Merck), 2% mixture (volumetric ratio) of 25% aqueous ammonium hydroxide solution in methanol Rf0,47.

MS 222 (M+).

1H-NMR (CDCl3-TMS) free base: 1,5 (dt, 3H, CH3); 3,5 (q, 2H, N-CH2); 1,6-3,3 (m, chinuklidinyl skeleton).

Example 20: 1-Methylpiperidin-4-Spiro-5'-(oxazolidin-2',4'-dione) AF169 and 1 Methylpiperidin-4-Spiro-5'-(3'-ethyloxazole-2',4'-dione) AF180

a) 4-Hydroxy-4-cyano-1-methylpiperidin

To fresh 1-methylpiperidin-4-ONU (81,72 g; 0,72 M) in water (200 ml) was added to approximately 100 ml of 37% HCl to pH 3. The specified reaction medium was cooled in an ice bath, and then was gradually added a solution of potassium cyanide (49 g; 0.75 M) in water (200 ml), t is the reaction mixture was stirred for 2 h, and then was filtered. The filtered product was washed with water and dried to obtain 4-hydroxy-4 - cyano-1-methylpiperidine (67 g; 0,48 M; 67% yield) as a white powder, so pl. 135oC.

1H-NMR (CDCl3) to 1.9 (m, 2H); 2,2 (m, 2H) and 2.4 (s, 3H); of 2.45 (m, 2H); to 2.75 (m, 2H); 2-3,5 (bm, 1H, OH) ppm.

b) 4-Hydroxy-4-carbarnoyl-1 methylpiperidin

To sulfuric acid (80 ml) under conditions of external cooling was slowly added 4-hydroxy-4-cyano-1-methylpiperidine (36,4 g; 0,26 M). This mixture over 41 h and kept at room temperature, then added to it crushed ice (30 g). The resulting solution was neutralized with barium carbonate (376 g) to pH 8-9, added water, was filtered precipitated precipitated barium sulfate and washed it with methanol. The corresponding filtrate was concentrated under reduced pressure. The desired product 4-hydroxy-4-carbarnoyl-1 methylpiperidin (28,16 g; 0.18 M; 69% yield) was led from ethanol in the form of a white solid substance, so pl. 180oC.

1H-NMR (CD3OD) is 1.51 (m, 2H); to 2.15 (m, 2H); of 2.25 (s, 3H); 2,4 (m, 2H); to 2.7 (m, 2H) ppm.

Mass spectroscopy m/z 158 (M+).

g) 1-Methylpiperidin-4-Spiro-5'-(oxazolidin-2',4'-dione) AF169

To a solution of potassium methoxide (9,8 g; 0,14 M) in the dry etano is in ethanol (300 ml). The formed mixture for 60 h were subjected to reflux distilled at a temperature of 80oC. Evaporated specified, the reaction mixture was allocated the residue was dissolved in cold water (70 ml) and neutralized 2 N. HCl to pH 7. The resulting solution was concentrated to 2 times was filtered and the resulting white precipitate. Rubbing the specified residue in ethanol, was obtained 1-methyl-piperidine-4-Spiro - 5'-(oxazolidin-2', 4'-dione) (compound AF169; 22 g; 0.12 M; 70-cent yield) as a white solid, so pl. 285oC (decomposition).

1H-NMR (D2O, pH 7) 2,07 (m, 2H); and 2.27 (m, 2H); 2.95 and (s, 3H); 3,30 (m, 2H); of 3.60 (m, 2H) ppm.

Mass spectroscopy (pH connection is equal to 7) m/z 185 and 184 (M++1 and M+).

in 1 Methylpiperidin-4-Spiro-5'-(3'-ethyloxazole-2',4'-dione) AF169

To a solution of 1-Methylpiperidin-4-Spiro-5'-(oxazolidin-2',4'-dione) (2.8 g; 0,015 M) in dry DMF (100 ml) was slowly added potassium hydride (65% oil mixture) until such time as the reaction medium did not cease to be heated (used 4.9g). The resulting white suspension for 1 h were subjected to reflux distilled, and then cooled. Then was added dropwise ethyl bromide (4.6 g, 0.04 M). Thus obtained, the reaction medium spontaneously warmed up. The specified reaction environment Univesrity was filtered white solid and washed it with ethanol. Evaporated solvents and allocated roughly purified product was subjected to chromatography on silicagel column, elution with a mixture of chloroform and methanol. Evaporated fraction containing the desired product. The specified product was isolated and identified in the form of free base. For the practical use of the above base was treated with a mixture of HCl, ether and ethanol into the hydrochloride, falling precipitated from this mixture in the form of hydrochloride of 1-methylpiperidin-4-Spiro-5'-(3'-ethyl-oxazolidin-2',4'-dione) (0,746 g; 0,003 M; 21% yield), so pl. 305oC (decomposition).

1H-NMR (CDCl3- free basis) of 1.2 (t, 3H); to 1.75 (m, 2H); to 2.15 (m, 2H); 2,3 (s, 3H); 2,32 (m, 2H); 2,80 (m, 2H); 3,55 (q, 2H) ppm.

13C-NMR (CDCl3, free base) 13 (CH3CH2); 32 (CH2CH3); 35 (CH2-CH2-C); 46 (N-CH3); 50 (CH2-N-CH3); 85 (Spiro); 155 (OC=O); 175 (O=C= O) ppm.

Mass spectroscopy (free base) m/z 212 (M+).

Example 21: 1 Methylpiperidin-4-Spiro-4'-(2'-methyl-2'-thiazolin) AF151(S)

a) a Mixture of 1-methylpiperidin-4-Spiro-4'-(2'-methyl-2'-oxazoline) (connection AF151; 1.85 g; br11.01 mm), pentasulfide phosphorus (1,83 g; 8,23 mm) and monohydrate p-toluensulfonate acid (4,20 g; 22,08 mm) in the used solvent, and the resulting residue was podslushivaet concentrated aqueous solution of NaOH and then was extracted with dichloromethane. Appropriate organic extract was dried over Na2SO4and solvent was removed to obtain a brown oil (2.10 g), which was subjected to chromatography on silicagel column (Kieselgel S; 0,032-0,063 mm; Riedel De-NAEP, 70 g). Elwira with a mixture of chloroform and methanol in the ratio of 97: 3, containing 10 M ammonium, received the fractions containing pure compound AF151(S) (0,90 g).

b) the Powders 4-acetamido-4-hydroxymethyl-1-methylpiperidine (8.00 g; 0,043 M) and pentasulfide phosphorus (6,10 g; 0,0275 M) for 6 h were thoroughly stirred in xylene (120 ml) on the magnetic rocking at reflux distilled. Specified, the reaction mixture was left overnight at room temperature, was filtered, the resulting precipitate and washed his oil ether (40-60oC) obtaining a gray powder (13,0 g). Cooled selected powder and podslushivaet its concentrated aqueous solution of NaOH and then was extracted several times with dichloromethane. Combined appropriate extracts were dried over Na2SO4and evaporated. The obtained residue was extracted with hexane, then hexane was removed with vyhoda C (0.4 mm RT. century), which in turn was subjected to chromatography on silicagel column (Merck, 100 g). The elution was performed with a mixture of chloroform, ether, methanol and ammonia in the ratio of 70:25:4:1 with a yield of pure compounds AF151(S) (1,71 g).

1H-NMR (CDCl3) 1,65-of 1.78 (m, 2H); 1,90-2,04 (m, 2H); 2,19 (s, CH3C=N-); 2,32 (s, CH3N); 2,30 is 2.43 (m, 2H); 2,60-of 2.75 (m, 2H); 3,11 (s, -CH2S) ppm.

13C-NMR (CDCl3) 19,9 (CH3C= N-); 35,8 (C3and C5); 43,1 (-CH2S); 45,6 (CH3N); 52,1 (C2and C6); 78,5 (C4); 161,0 (S-C=N) ppm.

MC m/Z 184 (M+, 100%); 110 (26%); 109 (56%); 72 (93%); 71 (24%).

Example 22: 1-Methylpiperidin-4-Spiro-4'(5)'-(2'-methyl-2' -imidazolin) AF190

a) 4-amino-4-cyano-1-methylpiperidin

A mixture of 1-methylpiperidin-4-it (33,0 g; 0,292 M), potassium cyanide (19.5 g; 0,299 M) and ammonium chloride (16.5 g; 0,308 M) suspended in a mixture of methanol (225 ml) and water (150 ml) and the resulting suspension was stirred at room temperature for 12 days. The precipitate was filtered, and the filtrate evaporated under reduced pressure. To remove the water possibly present in the selected residue to the specified residue was added ethanol, and then was carried out by azeotropic distillation. To the obtained residue, again to relax the inorganic compounds and washed them with ethanol. Combined corresponding to the filtrate and the washing liquid and the solvent was removed emitting a thick oil (35.5 g), which is being investigated using TIH (a mixture of chloroform, methanol and ammonia in the ratio of 17:2:1 on silica gel) resulted in the appearance of two spots. The above oil was led from ether to obtain a solid substance, which once again was subjected to crystallization from the specified solvent to yield 4-cyano-4-hydroxy-1-methylpiperidine in the form of crystals, so pl. 130-133oC. When the concentration of the remaining mother liquor was receiving another portion of the crystalline solid, which mainly consisted of 4-amino-4-cyano-1-methylpiperidin.

1H-NMR (CDCl3) 1,72-of 1.88 (m, 2H); for 2.01 (m, 2H); 2,25-is 2.37 (m, 2H); 2,32 (s, CH3N); 2,74-and 2.83 (m, 2H) ppm.

MC m/z 139 (M+); 112 (M+-HCN); 71; 70.

Exposing selected 4-amino-4-cyano-1-methylpiperidin acetylation using acetic anhydride and pyridine was obtained 4-acetamido-4-cyano-1-methylpiperidin, which was led from the oil mixture of ether and dichloromethane, so pl. 143-144oC.

1H-NMR (CDCl3) 1,78 is 1.96 (m, 2H); 2,04 (s, CH3CON-); 2,32 (s, CH3N); 2,35-of 2.50 (m, 4H); 2.66 per-2,84 (m, 2H); from 6.22 (s, -NHCO-) 40; 2804; 2242 (C=N); 1670 (amide) cm-1.

Exposing selected 4-amino-4-cyano-1-methylpiperidin acidic hydrolysis using H2SO4received 4-amino-4-carbarnoyl-1 methylpiperidin (so pl. 145-147oC), which was led from a mixture of ethyl acetate and dichloromethane.

1H-NMR (CDCl3) of 1.44 (m, 2H); 1,68 (br-NH2); 2,12-of 2.34 (m, 4H); 2,30 (s, CH3N); 2,70-2,82 (m, 2H); vs. 5.47 (br, -NH-); 7,39 (br,-NH) ppm.

MC m/z 158 (M++1); 157 (M+); 140 (M+-NH3, 100%); 113 (M+-CONH2); 96; 71.

b) 4-Amino-4-aminomethyl-1 methylpiperidin

A solution of 4-amino-4-cyano-1-methylpiperidine (of 3.60 g) in dry dimethoxyethane was added under nitrogen atmosphere to a mechanically stirred suspension of lithium hydride-aluminum (3.0 g) in diethoxyethane so that the temperature did not exceed 50oC. after the specified adding the resulting mixture was stirred at the temperature of reflux distilled for 6 hours in order to destroy the excess LiAlH4to the above mixture, cooled to 0oC, under stirring in nitrogen atmosphere was sequentially added 4M NaOH (10 ml), water (3 ml), a saturated solution of NaOH (10 ml) and again water (5 ml). Separated the organic solvent, and an appropriate aqueous phase is several times the extras the Wali over Na2SO4and removed the used solvents to yield the named compound as a viscous oil (3,17 g; so pl. 60-62oC), which was purified by distillation at 0.08 mm Hg).

1H-NMR (CDCl3) was 1.43 (m, 2H); was 1.58 (m, 2H); 2,15 (br. -NH2); 2,30 (s, CH3N); is 2.30 (m, 2H); 2,56 (s, -CH2NH2); of 2.56 (m, 2H) ppm.

Acetylurea specified diamine, got diacetamide in the form of a solid substance (so pl. 175-176oC), crystallized from acetonitrile.

1H-NMR (CDCl3) of 1.74 (m, 2H); 1,96 is 2.10 (m, 2H); 1,99 (s, CH3CON-); 2,02 (s, CH3CON-); of 2.21 (m, 2H); 2,28 (s, CH3N); of 2.51 - 2,61 (m, 2H); 3,50 (d, J= 5.7 Hz, -CH2NH-); 5,62 (s, -NHCO-); 7,18 (t, -CH2NHCO-) ppm.

MC m/z 227 (M+); 184 (M+-CH3CO); 169 (M+-CH3CONH); 168 (M+-CH3CONH2); 167; 155; 112; 109 (100%); 96; 71; 70.

The specified diacetamide also received, Giriraja 4-amino-4-cyano - 1-methylpiperidin hydrogen (50 psi) when using as a catalyst of Raney Nickel in a hot (60oC) acetic anhydride containing sodium acetate.

in 1 Methylpiperidin-4-Spiro-4'(5)'-(2'-methyl-2'-imidazolin) AF190

To a solution of 4-amino-4-aminomethyl-1 methylpiperidine (0,248 g; 1,734 mm) in dichloromethane (5 ml) were added hydrochloride ethyl acetimidate remove the used solvent, the selected residue was podslushivaet using a concentrated solution of Na2CO3and then was extracted with dichloromethane. The corresponding extract was dried over Na2CO3and solvent was removed to obtain a colorless oil (191 mg), which was subjected to chromatography on silicagel column. The elution was carried out with a mixture of methanol, chloroform, and 1% ammonia; with increasing methanol concentration from 10 to 99% received net connection AF190 in the form of oil, solidified by freezing.

1H-NMR (CDCl3) 1,57-of 1.84 (m, 4H); 1.93 and (s, CH3C=N-); 2,17 is 2.33 (m, 2H); to 2.29 (s, CH3N); of 2.56 (m, 2H); 3,37 (s, -CH2-N) ppm.

13C-NMR (CDCl3) 15,2 (CH3C=N-); 37,3 (C3and C5); 46,1 (CH3N); 52,5 (C2and C6); 59,7 (C4); 63,7 (-CH2N); 162,1 (-C=N) ppm.

MC m/z 167 (M+); 152 (M-CH3); 138; 109 (100%); 97; 96; 72; 71; 70.

IR (net) 3260; 2933; 2852; 2800; 1620 (-C=N) cm-1.

Example 23: 1-Methylpiperidin-4-Spiro-4'(5')-[2-methyl-4'H(5 N)- imidazole-5'(4')'-he] AF230

4-Acetamido-4-cyano-1-methylpiperidine (of 1.03 g) was dissolved in concentrated sulfuric acid (4.0 ml) and kept at room temperature for 4 days. The resulting reaction medium was made in the cold is not observed. Precipitated precipitated barium sulphate was filtered and washed with water and ethanol. United received the filtrate and washing liquid, using concentrated NaOH solution was brought to pH to 13 and remove the solvents under reduced pressure. The selected residue was extracted with ethanol, evaporated the corresponding extract, the resulting residue was extracted with dichloromethane, and the extract evaporated to yield an oil (0.75 g) which was subjected to chromatography on a column of silica gel 60 (Merck 0,040-0,063 mm, 32 g). The elution was performed with a mixture of methanol (containing 15 weight percent ammonia) and chloroform in a ratio of 1: 9 to obtain pure compounds AF230 (0,185 g; so pl. 231-234oC) crystallizing from a mixture of dichloromethane and ether.

1H-NMR (CDCl3) 1,49 (m, 2H); of 1.97 (m, 2H); 2,20 (s, CH3-); of 2.34 (s, CH3-); 2,48 (m, 2H); 2,80 (m, 2H); 9,83 (br. -NH-) ppm.

1H-NMR (D2O) of 1.55 (m, 2H) a 1.88 (m, 2H); 2,22 (s, CH3-); to 2.29 (s, CH3-); of 2.33 (m, 2H); 2,90 (m, 2H) ppm.

MC m/z 181 (M+); 111 (M+-70); 104; 94; 77; 71 (100%); 70.

UV (EtOH) max224 nm ( 4650), 248 nm ( 2450).

IR (KBr) 3140; 2795; 2540; 1665; 1540 cm-1.

In addition, in the basic environment, you can get the cyclization of 4-acetamido-4-cyano-1-methylpiperid NaOH in water, you can also get a connection AF230. In the latter case, additionally formed 4-acetamido - 4-carbarnoyl-1 methylpiperidin (so pl. 207-208oC, with decomposition), crystallized from a mixture of dichloromethane and methanol.

1H-NMR (CDCl3) 2,05 (s, CH3CO); 2,08-of 2.30 (m,6H); 2,28 (s, CH3N); a 2.36 (m, CH2-); to 5.40 (brs, -NH-); to 5.56 (brs, -NH -);? 7.04 baby mortality (brs, -NH) ppm.

MC m/z 199 (M+); 181 (M+-H2O); 155 (M+-CONH2); 140 (M+-CH2CO-NH2); 122; 112; 111; 96; 71 (100%); 70.

Connection AF230 can exist in the form of tautomers, as shown in its name.

Example 24: 1-Methylpiperidin-4-Spiro-5'-(2-methyl-2'- oxazoline-4'-he) AF238

a) 4-Acetamidomethyl-4-acetoxy-1-methylpiperidin

To a mixture of 4-cyano-4-hydroxy-1-methylpiperidine (2.55 g; 18 mm) and acetic anhydride (11 ml; 108 mm) in the three-neck flask was added dropwise to 5.1 g (2 equivalents) of a 60% aqueous solution of perchloric acid (HClO4). Was carried out exothermic reaction, however, after 20 min the temperature of the mixture was decreased. This solution was stirred for 2 h, then kept at room temperature during the night. Was filtered, the resulting white precipitate was washed its ester oil and ether. As a result, the on.

1H-NMR [perchlorate] (D2O) 2,2-2,4 (m, 2H); and 2.26 (m, 3H); and 2.27 (s, 3H); 2,5 (m, 2H); 2.95 and (s, 3H); of 3.32 (m, 2H); of 2.58 (m, H) ppm.

1H-NMR [free base] (CDCl3) 2,05-of 2.27 (m, 4H); to 2.13 (s, 3H); of 2.28 (s, 3H); 2,67 is 2.75 (m, 4H); and 8.4 (bs, 1H, NH) ppm.

MC m/z 242 (M+); 182; 167; 139; 123 (100%); 114; 96; 82; 70; 60.

UV (H2O)max206 nm ( 20400).

IR (KBr) 3380; 3020; 1740; 1670 (sh); 1550; 1400; 1380; 1250 cm-1.

b) 1-Methylpiperidin-4-Spiro-5'-(2-methyl-2'-oxazoline-4'-he) AF238

The mixture perchlorates salt 4-acetamidomethyl-4-acetoxy-1 - methylpiperidine (70 mg, 0.25 mm) in xylene kept at a temperature of 172oC (bath temperature silicone oil) up until white suspended solid matter does not become yellow. During the reaction there was a strong smell of acetic acid. Using TECH showed complete conversion of starting compound 1-methylpiperidin-4-Spiro-5'-(2 - methyl-2'-oxazoline-4'-he] (connection AF238), characterized as the corresponding salt of perchloric acid.

1H-NMR (D2O) 2,2 (m, 3H); to 2.25 (m, 2H); of 2.45 (m, 2H); 2,8 (s, 3H); of 3.25 (m, 2H); 2,5 (m, H) ppm.

MS m/z 182 (M+); 140; 123; 112; 104; 96; 77; 70 (100%).

Example 25: 1 Methylpiperidin-4-Spiro-5'-(1'-methyl-3'- utilization) AF161

To a mixture of compound AF160 (100 mg; 0.47 mm) and the PRC was stirred for 10 min at room temperature and acidified with oxalic acid in ether. The precipitate was dissolved in water, podslushivaet and was extracted with petroleum ether, and the extracts were concentrated and subjected to chromatography on silica gel, elution with a mixture of chloroform, methanol and ammonia in the ratio of 90:10:1. Were combined and evaporated pure fractions and selected the residue was dissolved in ether and precipitated as hydrochloride (85 mg; 72% yield).

1H-NMR (free base, CDCl3) 1,2 (t, J=6 Hz, 3H); 1,6-1,65 (m, 2H); 2,0-2,1 (m, 2H); 2,39 (s, 3H); of 2,75 2,85 (m, 4H); of 2.86 (s, 3H); 3,55 (q, J=6 Hz, 2H) ppm.

1H-NMR (hydrochloride, D2O) 1,2 (t, J=6 Hz, 3H); from 2.1 to 2.15 (m, 2H); 2.3 to 2.4 (m, 2H); 2.95 and (s, 3H); 3,0 (s, 3H); 3,5 (q, J=6 Hz, 2H) ppm.

MS m/z 225 (M+18%); 71 (100%).

Example 26: 1 Methylpiperidin-4-Spiro-5'-(1',3'- diarylheptanoid) AF162

To a mixture of 1-methylpiperidin-4-Spiro-5'-as (110 mg, 0.6 mm) and KH (0.2 g; 35% by weight in mineral oil) in 3 ml DMF was added ethyl bromide (0.5 g, 4.6 mm), the resulting solution was stirred for 30 min at room temperature, diluted with ether, and acidified with an excess of oxalic acid. The precipitate was dissolved in water, podslushivaet and was extracted several times with chloroform, the combined relevant extracts are concentrated and their powerglove alcohol in the ratio of 90:10:1, with the release of net connection AF162 (60 mg, 42% yield).1H-NMR (free base, CDCl3) 1,2 (t, J=6 Hz, 6N); of 1.65 to 1.7 (m, 2H); 1,96-of 2.15 (m, 2H) and 2.4 (s, 3H); 2,5-2,6 (m, 4H); 3,3 (q, J=6 Hz, 2H) ppm.

1H-NMR (hydrochloride, D2O) to 1.15 (t, J=6 Hz, 6N); 2,05-of 2.15 (m, 2H); of 2.25 to 2.35 (m, 2H); 2,9 (s, 3H); 3,3 (q, J=6 Hz, 3H) ppm.

13C-NMR (hydrochloride, D2O) 14,0; 28,6; 35,0; 44,0; 52,7; 59,2; 157,0; 177,0 ppm.

MC m/z 239 (M+75%); 71 (100%).

Example 27 ; 1 Methylpiperidin-4-Spiro-4'-(2',5'-dimethyldi - 4'H-imidazol) AF191

A mixture of 1-methylpiperidin-4-Spiro-4'-(2', 5'-dimethyldi-4 N - imidazole (0,700 g; 2,881 mm) and sodium methoxide (0,360 g; 6,667 mm) was kept at the temperature of reflux distilled in methanol (15 ml) for 3.5 hours was observed gas evolution. The resulting reaction medium solvent was removed and the resulting residue was extracted with dichloromethane. Appropriate organic extract is evaporated with the separation of the solid residue (0,613 g), which were extracted oil ether. From the above extract, the solvent evaporated with the release of the balance (to 0.263 g), which was led from the oil ether to obtain pure compounds AF191, so pl. 84-85oC.

1H-NMR (CDCl3) of 1.40 (m, 2H); of 1.97 (m, 2H); of 2.36 (s, CH3N); 2,52 (s, CH3S); 2,53 (m, 2H); 2.70 height is 2.80 (m, 2H); 4,10 (s9 (CH3O-); 74,2 (C4); 171,6 (-N=C-SCH3); 196,0 (-N=C-OCH3) ppm.

MC m/z 228 (M++1).

Example 28: 1-Methylpiperidin-4-Spiro-4'-(2'-methylthio-5'- amino-4'H-imidazol) AF192

A solution of compound AF177 (0,411 g) reagent (15 ml) obtained by dissolving ammonia (15% by weight) in liquid ammonia was stirred for 3 days at room temperature. Evaporated the resulting reaction mixture was allocated to the residue was added 15 ml of fresh specified reagent and repeating the above reaction two more times. Finally, consider the reaction medium is evaporated under reduced pressure and the obtained residue was washed with acetone to yield compounds AF192 (0,266 g) as a white solid (so pl. > 240oC, with decomposition), crystallized from ethanol.

1H-NMR (D2O) to 1.48 (m, 2H) 1,89 (m, 2H); to 2.29 (s, CH3N); the 2.46 (m, 2H); 2,48 (s, CH3S); of 2.86 (m, 2H) ppm.

MC m/z 212 (M+); 142 (M+-70); 70.

Example 29: 1-Methylpiperidin-4-Spiro-4'-(2'-methylthio-5'- aminomethyl-4'H-imidazol) AF193 and 1 methylpiperidin-4-Spiro-4'- (2',5'-bis(aminomethyl)-4'H-imidazol) AF194

A solution of compound AF177 (0,338 g) in an aqueous solution of methylamine (35%; 5 ml) was kept at a temperature of 80oC for 2 h using viparea on silicagel column (Merck 60; 0,040-0,063 mm). The elution was performed with a mixture of chloroform, methanol and ammonia in the ratio of 80:20:1 with obtaining pure compounds AF193 (0,061 g; so pl. 193-194oC), crystallized from acetonitrile.

1H-NMR (CDCl3) of 1.44 (m, 2H); of 1.85 (m, 2H); of 2.38 (s, CH3N); 2,52 (s, CH3S); to 2.66 (m, 2H); of 2.81 (m, 2H); to 3.06 (s, CH3NH-); 6,47 (-NH-) ppm.

MC m/z 226 (M+); 179 (M+-CH3S); 170 (M+-CH3NHCN); 169; 156 (M+-70).

By elution with a mixture of chloroform, methanol and ammonia in the ratio of 50: 50:1 has received the connection AF194 (0,130 g; so pl. 113-114oC in the form of a white substance crystallized from acetonitrile.

1H-NMR (CDCl3+ CO3OD) of 1.44 (m, 2H); is 1.81 (m, 2H); is 2.37 (s, CH3N); to 2.57 (m, 2H); 2,80 (m, 2H); 2.95 and (s, CH3NH-); 2,96 (m, CH3N) ppm.

MC m/z 209 (M+); 152; 139 (M+-70).

Repeating the above reaction using 2 M methylamine in each M connection AF194, the primary formed product was a connection AF193 in the presence of only traces of connection AF194.

Example 30: 1 Methylpiperidin-4-Spiro-4'-(2'-methyl-2'- oxazoline) AF150

a) Hydrochloride of 1-methyl-4-nitromethylene-4-ol

The specified parent compound was synthesized using natantia (1,28 M) in ethanol, the temperature of which was maintained at a level of 5-8oC, was added a mixture of N-methylpiperidine (142 g; 1,28 M) and nitromethane (78,1 g; 1,28 M). Formed white precipitate, mixing was continued for 20 min, and then for a further 40 min at room temperature. The resulting solution was acidified with 500 ml of 7.2 N. HCl in isopropyl alcohol. The corresponding hydrochloride, and inorganic salts were extracted with three portions of methanol (300 ml), and the used solvent was removed in vacuum to yield the named compound, so pl. 180-182oC (non-hygroscopic), m/z 174 (M+free base, 100%); 157 (M-OH, 20%); 127 (M-H-NO2, 25%); 113 (M-NO2-CH3, 40%).

b) To a solution of 1-methyl-4-nitromethylene-2-ol (133,5 g) in methanol (1500 ml) in parts) was added palladium-carbon catalyst (10%, 4 g). The specified connection within 48 h were subjected to hydrogenation in a Parr apparatus at a pressure of 55 psi and room temperature. The resulting solution was carefully filtered, treated with activated charcoal, the solvent was removed, and the selected residue is triturated with ethanol (200 ml) to yield the named compound, so pl. 177-179oC. m/z 144 (M+free base, 15%); 127 (M-OH, 25%); 114 (M-CH2-NH2, 100%).

2CO3in 50 ml of water and then concentrating the resulting mixture under vacuum until dry. Selected white solid was twice extracted with 50 ml of chloroform, treated with activated carbon, dried over Na2SO4and solvent was removed to yield the named compound (62,5% yield; so pl. 45oC; sublimation at a temperature of 40oC and 0.05 mm Hg), forming a single spot in TECH on silica gel, elution of 2% NH3in CH3HE, Rf=0,4.

m/z 168 (M free base, 100% at 7.5 EV).

1H-NMR (300 MHz, CDCl3): of 3.56 (2H, q, J=1.5 Hz); 2,53 (4H, m); of 2.34 (3H, s); a 1.96 (3H, t, J=1.5 Hz); is 1.82 (4H, m).

Replacing in the above Example, the KONA on the equivalent amount of NaOH or et 3 n, obtained similar results.

d) Dibenzoyl-D-tartrate soedinenii Dibenzoyl-D-tartaric acid. The resulting precipitate was allowed to settle and then decanted supernatant. The remaining solid is washed three times with 100 ml of dry toluene and dried under reduced pressure to get a 8.4 g (80% yield) of a white slightly hygroscopic solid.

TECH, a mixture of chloroform and aluminum oxide (Merck Art 5581) Rf=0,4. m/z: 168 (M+).1H-NMR (300 MHz; D2O, containing 1.5 mg of Na2CO3/0.5 ml of D2O)): 1,95 (s, 6H, CH3-With); to 2.35 (s, 6N, CH3N); 3,5 (s, 4H, CH2); to 5.7 (s, 2H); 7,5-8,2 (m, 10H, aromatic hydrogens).

Example 31: 1 Methylpiperidin-4-Spiro-4'-(2'-ethyl-2 - oxazoline) (2'-ethyl analogue connection AF150)

The specified connection was obtained in a similar manner to the compound of Example 31, using an equivalent quantity of the hydrochloride of ethyl propionamide instead of the hydrochloride of ethyl acetimidate. The corresponding product was isolated in the form of liquid, so Kip. 53oC/0.03 mm Hg (65% yield).

1H-NMR (300 MHz; CDCl3): to 3.52 (2H, t, J=1.5 Hz); 2,47 (4H, m); of 2.30 (3H, s); 2.26 and [2H, Quartet, J=7 Hz), triplet (J=1.5 Hz)]; to 1.86 (2H, m); 1,72 (2H, m); of 1.18 (3H, t).

Example 32: 1 Methylpiperidin-4-Spiro-5'-(2'-methyl-2'-oxazoline) AF151

a) 1-Methylpiperidin-4-Spiro-5'-as

The mixture rustia (25,8 g; 0,396 M) in water (82 ml) was kept at a temperature of 60oC for 2.5 h and then left overnight at room temperature, resulting in the deposition occurred 1 methylpiperidin-4-Spiro-5'-as. Was specified filtered the precipitate off, washed with small quantities of cold water, ethanol and ether to yield crystalline powder (27,0 g). Concentrating corresponding to the filtrate and washing liquid were receiving another portion of the product in question (20,0 g). The isolated product was led from methanol, so pl. 265-276oC (decomposition).

IR (KBr) 3170 (NH); 1700 (C=0) cm-1.

m/z 183 (M+, 38%); 71 (100%).

1H-NMR (300 MHz; D2O): 1,8 (2H); 2.06 to (sextet, 2H); 2.49 USD (s, CH3); of 2.58 (t, 2H); 3,14 (t, 1H); 3,20 (t, 1H).

b) 4-Amino-1-methylpiperidin-4-carboxylic acid

A mixture of 1-methylpiperidin-4-Spiro-5'-as (9.75 g; 0,0533 M) and exagerate of barium hydroxide (28.8 g; 0,00913 M) in water (150 ml) was kept in an autoclave at a temperature of 160oC for 3 hours Combined contents of four of these tanks was filtered and precipitated precipitated barium carbonate. The corresponding filtrate was neutralized with solid carbon dioxide, and the resultant precipitate was removed through the n-4-carboxylic acid (32,0 g; 95% yield), so pl. 275-280oC (decomposition).

IR (KBr) 3300; 1655; 1580 cm-1.

m/z 158 (M+, 90%); 141 (90%, M-OH); 113 (12%, M-CO2H); 96 (100%); 71 (52%).

1H-NMR (300 MHz; C5D5N + D2O): 1,2 (m, 2H); to 1.48 (s, CH3N-); and 1.7 (m, 2H); 1,9 (m, 2H); 2,0 (m, 2H).

b) 4-Amino-4-hydroxymethyl-1-methylpiperidin

Powder of lithium aluminum hydride (15.62 wide g; 0,412 M) in dry tet rageragerage (THF; 600 ml) was kept at the temperature of reflux distilled for 15 min, then in a nitrogen atmosphere with efficient stirring in parts) was added 4-amino-1-methylpiperidin-4 - carboxylic acid (31.6; 0,196 M) in the form of a dry powder. When you are finished adding the resulting reaction mixture was stirred at the temperature of reflux distilled for 4 h, then cooled to a temperature of 0oC with efficient stirring in a nitrogen atmosphere and the reaction was stopped, carefully adding water (20 ml), 15% ammonia (20 ml) and again water (10 ml). Filtered by the specified reaction medium, and the selected residue was extracted three times with 150 ml of boiling THF. Combined appropriate THF-filtrate and extracts, and the solvent was removed at 25 mm Hg with a yellow thick oil (28,0 g; that 98.9% yield).

IR (net who.

1H-NMR (300 MHz; CDCl3): of 1.41 (m, 2H) and 1.60 (m, 2H); 2,24 (s, CH3N); 2,9 (m, 2H); 2,48 (m, 2H); 2.50 each (br., -NH2); 3,29 (s, -CH2OH).

g) 1-Methylpiperidin-4-Spiro-5'-(2'-methyl-2'-oxazoline) F151

A mixture of 4-amino-4-hydroxymethyl-1-methylpiperidine (1,80 g), acetic acid (20 ml) and xylene (20 ml) was subjected to azeotropic distillation in the next 28 hours Remaining acetic acid and xylene was removed under reduced pressure (25 mm Hg) to obtain the remainder in the form of a thick oil, which was podslushivaet to pH 11 with aqueous solution of Na2CO3. Extragere the specified residue with chloroform, and viparita the corresponding extract was allocated a small amount of residue in the form of a brown oil (0.27 g). To remove the water evaporated aqueous solution remaining after extraction with chloroform, the resulting solid was extracted with chloroform, and the extract was dried over Na2SO4and evaporated to yield the remainder in the form of a very hygroscopic solid (3.0 g). Using TEH showed that the solid gives mainly one spot, the more polar compared to the original linesperson. When heated portions of this hygroscopic solid (so pl. 155oC and a pressure of 0.15 mm Hg. When storing consider oil in the freezer it was formed crystalline needles, melting at room temperature. Product distillation was acetic acid salt of the named compounds.

IR (pure) 1664 (C=N); 1565 and 1398 (CO2--); 1256 (C-O) cm-1.

m/z 168 (M+free base); 109; 70.

1H-NMR (300 MHz, CDCl3) to 1.77 (m, 2H); to 1.96 (m, 2H); to 1.98 (s, CH3-); 2,0 (s, CH3-); 2,49 (s, CH3N); only 2.91 (m, 4H); 3,95 (s, CH2O-); of 9.30 (br.s, -CO2H).

13C-NMR (300 MHz, CDCl3) 14,0 (CH3CO2-); 22,9 (CH3C=N-); 35,6 (C3and C5); 44,4 (CH3N+); 51,1 (C2and C6); 67,0 (C4); 77,4 (C5'); 164,3 (C=N); 176,7 (-CO2-).

MC m/z 167 (M+); 152 (M-CH3); 138; 109 (100%); 97; 96; 72; 71; 70.

1H-NMR of free base (300 MHz; CDCl3): of 1.64 (m, 2H); of 1.84 (m, 2H); to 1.98 (s, CH3-); and 2.26 (m, 2H); 2,30 (s, CH3-); 2,69 (m, 2H); 3,94 (s, -CH2-).

Example 33: 1 Methylpiperidin-4-Spiro-5'-(2'-ethyl-2 - oxazoline) (2'-ethyl analogue connection AF151)

A mixture of 4-amino-4-hydroxymethyl-1-methylpiperidine (3.0 g), propionic acid (50 ml) and xylene (90 ml) was subjected to azeotropic distillation for 5 h Formed is OK chloroform and viparita the corresponding extract, allocated a mixture of non-polar compounds (0,80 g). To remove the water evaporated aqueous solution remaining after extraction with chloroform, the resulting solid was extracted with chloroform, and the extract was dried over Na2SO4and evaporated to yield the remainder in the form of a hygroscopic solid (3.6 g). Using TIH (silica gel, elution with a mixture of methanol, chloroform and ammonia in the ratio of 40:58:2), showed that the solid gives mainly one spot, the more polar compared to the original amino alcohol. When heated portions of this hygroscopic solid (1.5 g) in vacuum it almost immediately began to perekanatisya as a thick colorless oil at a temperature of 50oC and a pressure of 0.1 mm Hg. Product distillation was propionato acid salt of the named compounds.

m/z 182 (M+free base, 14%); 167 (5%); 154 (71%); 125 (9%); 109 (100%); 96 (45%); 81 (30%); 74 (57%); 70 (89%); 57 (64%).

1H-NMR (300 MHz, CDCl3) of 1.12 (t, J=7.5 Hz, CHCH2-); of 1.17 (t, J=7,6 Hz, CH3CH2-); to 1.75 (m, 2H); 2,00 (m, 2H); to 2.29 (q, J=7.5 Hz, CH3CH2-); 2,30 (q, J= 7,6 Hz, CH3CH2-); 2,56 (s, CH3N); to 3.02 (m, 2-CH2-); 3,95 (s, -CH2O-); to 7.25 (br) was added a saturated aqueous solution of K2CO3until then, until it stopped production of CO2. Then the resulting mixture was stirred for 0.5 h, and share in the resulting phase. The corresponding aqueous phase was extracted with CHCl3merged the previously selected CHCl3-phase and the extracts were dried over Na2SO4and evaporated with the release of the named compound (free base) as a residue colorless oil (550 mg), which analyses using TECH gave a single spot.

1H-NMR (300 MHz, CDCl3) of 1.17 (t, J=7,6 Hz, CHCH2-); to 1.61 (m, -CH2-); to 1.86 (m, -CH2-); to 2.18 (m, -CH2-); to 2.29 (q, J=7,6 Hz, CH3CH2-); 2,30 (s, CH3N); a 2.71 (m, -CH2-); of 3.94 (s, -CH2O-).

m/z 182 (M+25%); 167 (9%); 154 (78%); 125 (17%); 109 (100%); 96 (65%); 81 (54%); 74 (96%); 57 (77%).

An alternative method of preparing compounds AF150 and AF151 is cyclodehydration corresponding amides. As dehydrating agents in the considered reactions can be used such compounds as P2O5sulfuric acid , BF3-apirat, CaCl2and molecular sieves. Obtaining respective thiazolino instead of oxazolines can be done using similar reactions using P2S5.

m/z 186 (M+)

1H-NMR (300 MHz, CDCl3) 1,60 (multi, 4H, H3 and H4); 2,01 (singlet, 3H, CH3-With); to 2.29 (singlet, 3H, CH3N); 2,38 (multi, 2H, H1); 2,55 (multi, 2H, H2); 2,98 (plural, 1H, NH); 3,26 (double, 2H, H5) ppm.

1H-NMR (300 MHz, D2O) 1,42 (multi, 4H, H3 and H4); 1,81 (singlet, 3H, CH3-C); 2,08 (singlet, 3H, CH3N); 2,27 (multi, 2H, H1); 2,46 (multi, 2H, H2); 3,03 (singlet, 2H, M) ppm.

Impurities give a peak at 3,44 ppm.

b) 1-Methylpiperidin-4-Spiro-5'-(2'-methyl-2'-thiazolin) AF150(S)

A mixture of 4-atsetamidometil-4-hydroxy-1-methylpiperidine (6.5 g; 35 mm) and pentasulfide phosphorus (10 g, 22 mm) was kept at a temperature of 220oC for 30 min, then cooled and dissolved in 30 ml of concentrated aqueous HCl. The resulting solution was mixed with 100 ml holodnie extracts, was dried and evaporated emitting 5 g of black oily residue, which was purified by distillation at a temperature of 75oC and a pressure of 1 mm Hg to yield 1.8 g of a transparent liquid.

m/z 184 (M+)

1H-NMR (300 MHz, CDCl3) of 1.8-2.0 (m, 4H); 2,17 (t, 3H, CH3-C); 2,2 (s, 3H, CH3N); 3,2 (q, 2H, CH2-thiazoline ring).

Example 35: 1-Methylpiperidin-4-Spiro-5'-(2'-tion-3'-utilization) AF181

To a solution of 4-amino-1-methylpiperidin-4-carboxylic acid (1.78 g; 0,0113 M) and sodium hydroxide (0,47 g; 0,0118 M) in water (3.0 ml) was added ethyl isocyanate (1,00 g; 0,0111 M), and the resulting mixture was stirred at the temperature of reflux distilled for 6,5 hours With concentrated hydrochloric acid indicated the reaction mixture was acidified to pH 1 and continued dephlegmation for another 1.5 hours In the future, consider the reaction mixture was stirred at room temperature, resulting in the formed solid substance (0,61 g), which was led from methanol to obtain crystals (83 mg), so pl. > 260oC. spectrum Analysis1H-NMR indicated the solids showed that it is a hydrochloride connection AF181.

1H-NMR (D2O) to 1.16 (t, J=7.2 Hz, CH3CHOcnye solutions remaining after separation and crystallization of the above-mentioned salts with concentrated aqueous sodium hydroxide solution was podslushivaet them to pH 13, and then was extracted with dichloromethane. The corresponding extract was dried over Na2CO3and evaporated to obtain yellow solid, which was subjected to chromatography on silicagel column (Merck 60; 0.04 to 0.06 mm). The elution was carried out with a mixture of chloroform, methanol and ammonia in the ratio of 96:3:1 with the output of the first 1,3-diethyl-2-urea (0,287 g; so pl. 76-78oC; crystallization from ether), and then - 8-methyl-3-ethyl-1,3,8 - diazaspiro[4,5]Decan-4-one-2-thione (AF181; 135 mg) in the form of free base. The specified connection was led from a mixture of ether and dichloromethane to obtain needle-like crystals so pl. 180oC

1H-NMR (CDCl3) to 1.24 (t, J=7.2 Hz, CH3CH2-); 1.69 in (m, 2H); 2.05 is-is 2.30 (m, 4H); 2,35 (s, CH3N); 2,90 (m, 2H); a 3.87 (q, J=7.2 Hz, CH3CH2-) ppm.

MS m/z 227 (M+); 71; 70.

Example 36: 1 Methylpiperidin-4-Spiro-5'-(2'-tion-3'- tert-butyl-as) AF184

A mixture of 4-amino-1-methylpiperidin-4-carboxylic acid (1.80 g; 0,0114 M) t-utilizationof (1,00 g; 0,0087 M) in water (3.0 ml) was kept at a temperature of 80oC for 5 h T the C was added ethanol (4.0 ml) and kept it at the temperature of reflux distilled for a further 1 h Using concentrated hydrochloric acid indicated the reaction mixture was acidified to pH 1 and continued dephlegmation for a further 1 hour In the future consider the reaction mixture during the night kept at room temperature, resulting in a formed solid precipitate, which was filtered to obtain the original aminokisloty (0,60 g). Evaporated the remaining mother liquor was podslushivaet using concentrated sodium hydroxide solution, and then was extracted with dichloromethane. The corresponding extract was dried over Na2SO4and solvent was removed to obtain crystalline solid (0,101), forming needle-like crystals during crystallization from a mixture of ether and dichloromethane, so pl. 200-201oC.

1H-NMR (CDCl3) and 1.63 (m, 2H); 1,81 [s, (CH3)3-]; 2,00-of 2.21 (m, 4H); 2,33 (s, CH3N); 2,87 (m, 2H); of 7.82 (bs, -NH-) ppm.

MC m/z 255 (M+); 199 (M+-SN); 96; 71; 70; 57.

Example 37: 1-Methylpiperidin-4-Spiro-5'-(3'-tert-Butylimino) AF213

Named compound was synthesized in the same manner as the connection AF181. The corresponding product was obtained as hydrochloride 1 methylpiperidin-4-Spiro-5'-(3'-t-butylhydrazine) (connection AF213), W
)3-]; a 2.01 (m, 2H); 2,22 (m, 2H); 2,92 (s, CH3N); 2,97 to 3.5 (m, 2H); 3,62 (m, 2H) ppm.

Selected free base was led from a mixture of ether and dichloromethane, so pl. 221-223oC.

1H-NMR (CDCl3) of 1.57 (m, 2H); 1,60 [S, (CH3)3-]; 2,01 - to 2.18 (m, 4H); 2,31 (s, CH3N); is 2.88 (m, 2H); of 4.77 (bs, -NH-) ppm.

13C-NMR (CDCl3) 28,6 [(CH3)3C]; 33,5 (C6 and C10); 46,2 (CH3N); 51,0 (C7 and C9); 57,5 58,1 and [(C5 and (CH3)3C-]; to 158.4 (C2); 177,4 (C4) ppm.

MC m/z 239 (M+); 237; 224 (M-CH3); 194; 181; 155; 110; 104; 71; 56; 43.

Example 38: 1-Propolypeptide-4-Spiro-5'-(3'-utilization) AF196

A mixture of piperidine-4-Spiro-5'-(3'-utilizaton) (connection AF160-DES; 40,9 r; 0.21 mm) and propargylamine (24,7 mg; 0.21 mm; 80% by weight solution in toluene) was stirred in methanol (1.0 ml) at a temperature of 25oC for 24 h using a stream of nitrogen was removed used methanol, and the remaining mass was podslushivaet aqueous solution of sodium carbonate and was extracted with chloroform. Evaporated corresponding chloroformic extracts and selected the residue was purified on a plate of silica gel for preparative chromatography (elution with a mixture of chloroform, methanol and ammonia in the ratio of 90: 10: 1) and precipitated with an excess of oxalic acid to obtain the; ,1-2,35 (m, 6N); of 3.12 (t, 2H, J=3.5 Hz); 3,5 (q, J=7.2 Hz, CH3CH2-); 3,7-of 3.85 (m, 2H); 4,12 (d, J=3.5 Hz) ppm.

MC m/z 235 (M+); 196 (M+-C3H3); 95; 80; 67; 56.

Example 39: 1 Methylpiperidin-4-Spiro-5'-[3'-(4-pyrrolidino-2-butynyl) as] AF197

A mixture of 1-methylpiperidin-4-Spiro-5'-(3'-propargylglycine) (connection AF185; 204 mg; 0,92 mm), pyrrolidine (88 mg; 1,23 mm) and copper chloride (4.2 mg) was stirred in dioxane (10 ml) at room temperature for 70 hours using a stream of nitrogen was removed used dioxane, and the remaining mass was podslushivaet aqueous solution of sodium carbonate and was extracted with chloroform. Evaporated appropriate chloroform extracts, and 50% (by weight) of the selected residue was purified on a plate of silica gel for preparative chromatography (elution with a mixture of chloroform, methanol, ether and ammonia in the ratio of 150: 20: 100: 6) and has twice led from ether to obtain crystalline solid (25 mg).

1H-NMR (CDCl3- free basis) of 1.12 (m); 2,1-2,2 (m); to 2.35 (s, 3H); 2,6 (m); 2,9-2,95 (m); 3,4 (t, 2H, J=3.5 Hz); 4,15 (t, 2H, J=3.5 Hz); at 5.9 (bs, 1H) ppm.

MC m/z 304 (M+); 234 (M+-C4H8N); 71 (C4H9N+).

Example 40: 1 Methylpiperidin-4-Spiro-5'-(2'-methyl-1', 4'- ACS is g) and p-toluensulfonate acid (2.1 g) in toluene (100 ml) were introduced into a steel vessel, designed for high-pressure, and kept at a temperature of 107oC and stirring for 24 h formed From the reaction medium by distillation of the toluene was removed, and the obtained residue was subjected to chromatography on silicagel column (Merck 60). The elution was carried out with a mixture of chloroform, methanol and ammonia in the ratio of 92:7:1 with obtaining fractions (1.01 g), which was further separated using chromatography on silicagel column. Elwira mixture of chloroform, ether, methanol and ammonia in the ratio of 54:37:7:2, received roughly purified product (0,441 g), which was led from a mixture of ether and petroleum ether and then from toluene to yield 1-methylpiperidin-4-Spiro-5'-(2'-methyl - 1',4'-oxazolidin-3'-one) (compound AF260) in the form of needle crystals (85 mg), so pl. 148-150oC.

1H-NMR (CDCl3) to 1.42 (d, J=6,6 Hz, CH3CH-); of 1.85 (m, 4H); 2,31 (s, CH3N); 2,52 (m, 4H); however, 4.40 (q, J=6.6 Hz, CH3CH-); to 7.64 (bs, -NH-) ppm.

MC m/z 184 (M+); 156; 126; 114; 84; 71; 70; 43 (100%).

Example 41: 1 Methylpiperidin-4-Spiro-5'-(2'- methyl-1',4'-thiazolidine-3'-Oh) AF261

Example 42: 1 Methylpiperidin-4-Spiro-5'-(2', 4'-dimethyl-1', 4'-thiazolidine-3'-Oh) AF266

A mixture of 1-methyl-4-piperidone (5,65 g; 0,050 M), timelocal acid (6,37 is e which sublimated solid (ammonium carbonate), settling out of the reaction flask. From time to time subliminable solid was again added to the reaction flask along with a new extension portion of ammonium carbonate (a total of 6.2 g). Finally, using distillation removed used benzene, and the resulting residue was washed with ether splitting soluble in ether substances (0,76 g) and insoluble in ether substances (12.9 g), which was dissolved in a small volume of methanol and podslushivaet with a solution of ammonia in methanol. Evaporated the resulting solution was subjected him chromatography on silicagel column (Merck 60). Elwira mixture of chloroform, methanol and ammonia in the ratio of 89:10:1, was obtained fraction (0,61 g) which was identified as 1-methylpiperidin-4-Spiro-5'-(2', 4'-dimethyl - 1', 4'-thiazolidine-3'-one) (compound AF266). The specified connection is a hygroscopic solid substance with a low melting point.

1H-NMR (CDCl3) of 1.53 (d, J=7 Hz, CH3CH-); 1.69 in (m, 2H); 2,16 is 2.44 (m, 4H); 2,32 (s, CH3N); 2,87 (m, 2H); 2,90 (s, CH3NCO-); with 3.79 (q, J=7 Hz, CH3CH) ppm.

13C-NMR (CDCl3) 19,7 (CH3CH-); 27,2 (CH3NCO-); 36,0 and 37.3 (C6 and C10); and 39.7 (C2); 45,3 (CH3N); 51.8, 52,2 (C7 and C9); 68,5 (C5); to 172.8 (C3) ppm.

the thief of hydrochloric acid in isopropyl alcohol, in the result of which was allocated hygroscopic solid, which was led from isopropyl alcohol, so pl. 238-240oC (decomposition).

1H-NMR (D2O hydrochloride) to 1.48 (d, J=7.2 Hz, CH3CH-); was 2.05 (m, 2H); 2.57 m (m, 2H); 2,89 and only 2.91 (2s, CH3N+and CH3NCO-); and 3.31 (m, 2H); 3,61 (m, 2H); Android 4.04 (q, J=7.2 Hz, CH3CH) ppm.

MS m/z 214 (M+); 181; 156; 125; 124; 96; 71; 70; 57 (100%); 43.

Continuing the elution of the specified column with a mixture of chloroform, methanol and ammonia in the ratio of 85:14:1, received a fraction (3.50 g), which was led from a mixture of ether and dichloromethane to yield 1-methylpiperidin-4-Spiro-5'-(2'-methyl-1', 4'-thiazolidine-3') (connection AF261; to 1.83 g) in the form of needle-shaped crystals, so pl. 133-134oC.

1H-NMR (CDCl3) of 1.53 (d, J=7 Hz, CH3CH-); 1,9-of 2.20 (m, 4H); 2,20-to 2.42 (m, 2H); 2,30 (s, CH3N); in 2.68 (m, 2H); of 3.85 (q, J=7 Hz, CH3CH-); make 6.90 (bs, -NH-) ppm.

13C-NMR (CDCl3) 19,9 (CH3CH-); 41,0 and 41,3 (C6 and C10); 41,3 (C2); 45,7 (CH3N); 52,5 and 52.8 (C7 And C9); 63,6 (C5); 176,3 (C3) ppm.

Hydrochloride connection AF261 (so pl. 285-287oC) has been received, the processing solution of the corresponding free base in ether solution of hydrochloric acid in isopropyl alcohol.

1H-NMR (hydrochloride in D2O) a 1.50 (d, J=7,1 Hz,-piperidone (5,65 g; 0,050 M), timelocal acid (6,37 g; to 0.060 M) and ammonium carbonate (7.20 g) in benzene (100 ml) were introduced into a steel vessel designed for high-pressure, and kept at a temperature of 95oC and stirring with a magnet. The resulting reaction medium by distillation of the solvent was removed and the resulting residue was dissolved in a small volume of methanol and applied on a dry silicagel column (Merck 60; 200 g). The elution was carried out with a mixture of chloroform, methanol and ammonia in the ratio of 94:5:1 with the first output connection AF266 (0.71 g), and then connect AF261 (3,02 g).

C) a Mixture of 1-methyl-4-piperidone (2.50 g; 22,1 mm), timelocal acid (2,81 g; 26.5 mm) and methylamine (0,90 g; 29,0 mm) in toluene (60 ml) were introduced into a steel vessel designed for high-pressure, and kept at a temperature of 96oC and stirring for 24 h the Resulting reaction medium was a thick oil, which was divided into the sediment and in toluene solution. Chose this solution and using distillation solvent was removed. The obtained residue (1.68 g) was subjected to distillation under reduced pressure to yield pure 1-methylpiperidin-4-Spiro-5'- (2',4'-dimethyl-1', 4'-thiazolidine-3') (connection AF266) as Tverdov ethylpiperidine-4-Spiro-5'-(2'-ethyl-1',4'- thiazolidine-3'-Oh) AF267

Example 44: 1 Methylpiperidin-4-Spiro-5'-(2'-ethyl-4'- methyl-1',4'-thiazolidine-3'-Oh) AF272

Named connection AF267 was obtained in the same manner as in the case of compound AF261 using 2-mercaptoethanol acid instead of 2-mercaptopropionate (timelocal) acid. The specified connection was led from acetone, so pl. 140-141oC.

1H-NMR (CDCl3) of 1.02 (t, J=7.2 Hz, CH3CH2-); 1,75 and to 2.06 (m, CH3CH2-); to 1.98 (m, 4H); 2,30 (s, CH3N); 2,32 (m, 2H); to 2.67 (m, 2H); 3,81 (dd, -CHS-); 6,35 (bs, -NH-) ppm.

13C-NMR (CDCl3) 11,4 (CH3CH2-); 27,1 (CH3CH2-); 41,5 (C6 and C10); 45,8 (CH3N); 49,0 (C2); 52,7 and 52.9 (C7 and C9); 63,7 (C5); 175,5 (C3) ppm.

MS m/z 214 (M+); 181; 140; 71; 57.

Hydrochloride connection AF267 was characterized by so pl. 267-269oC (decomposition). As a side product was formed 1 methylpiperidin-4-Spiro-5'-(2'-ethyl-4'-methyl-1',4'- thiazolidine-3'-one) (compound AF272) as a thick oil.

1H-NMR (CDCl3) and 1.00 (t, J=7,4 Hz, CH3CH2-); to 1.67 (m, 2H); 1,58-1,78 (m) and 2.13 (m, CH3CH2-); 2,20-to 2.42 (m, 4H); 2,32 (s, CH3N); 2,89 (s, CH3NCO-); 2,90 (m, 2H); 3,76 (dd, J1=3,6 Hz, J2=9,0 Hz, CH3CH2CH) ppm.

MC m/z 228 (M+); 195; 170; 138; 125; 96; 71; 70; 57.

Example 45: 1 Methylpiperidin-4) (connection AF261; 48,4 mg; 0.24 mm) in 2.0 ml of acetic acid and 0.5 ml of water 32% hydrogen peroxide is stirred at a temperature of 25oC within 20 minutes after the Specified reaction medium was acidified with excess hydrochloric acid and diluted with 20 ml of a mixture of petroleum ether and ether in the ratio of 4:1. The resulting residue was dissolved in aqueous sodium carbonate solution, and the resulting solution was extracted with chloroform. Evaporated corresponding chloroformic extract, and the resulting mass was subjected to preparative chromatography on a plate of silica gel (elution with a mixture of chloroform, methanol and ammonia in the ratio of 80:20:1) with the release of 3.3 g of oil, which, by means of oxalic acid was besieged from the ether with the output of one isomer (more than 90% purity) as a solid.

1H-NMR (CDCl3- free basis) of 1.52 (d, 3H, J=7 Hz, CH3CH-); 1,85-2,7 [m, 8H] ; a 2.36 (s, CH3N); 3,55 (q, J=6 Hz, CH3CH-); 7,0 (bs, 1H) ppm.

MC m/z 216 (M+); 199; 167; 149; 125; 111.

Example 46: piperidine-4-Spiro-5'-(2'-methyl-1',4'- thiazolidine-3'-Oh) AF263

A mixture of 1-methylpiperidin-4-Spiro-5'-(2'-methyl-1', 4'- thiazolidine-3') (connection AF261; 250 mg; 1.25 mm) and chloroethyl of chloroformate (0.15 ml; 1.4 mm) in 5 ml of dichloroethane was kept at a temperature of 60ooC for 1 h Evaporated methanol, highlighted the residue was dissolved in aqueous sodium carbonate solution, was extracted with chloroform and separated on a plate of silica gel for preparative chromatography (elution with a mixture of chloroform, methanol and ammonia in the ratio of 80:20:1) to yield 20 mg of pure reason, which was besieged in the form of crystalline oxalic salt.

1H-NMR (D2O, oxalic salt) is 1.51 (d, J=6 Hz, CH3CH-a); 2,2-to 2.35 (m, 4H); 3.1 to 3.6V (m, 4H); to 4.1 (q, J=6 Hz, CH3CH) ppm.

MC m/z 186 (M+); 153; 126; 57.

Example 47: 1 Methylpiperidin-4-Spiro-5'-(3'-methyl-1',4'- oxathiolan-2'-he) AF265

A solution of 1-methyl-4-piperidone (11.3 g; 0.1 M), timelocal acid (15 ml) and p-toluensulfonate acid in acetonitrile was subjected to reflux distilled for 24 h, and then TECH on silica gel (elution of 20% methanol in chloroform). Evaporated above solution, was dissolved in chloroform and washed with sodium bicarbonate solution and then with water. Appropriate organic phase was dried over sodium sulfate and evaporated. The isolated product was purified on silicagel column (Merck 60). The elution was carried out with a mixture of chloroform, methanol and ammonia with access 10,27 g butter, crystallized sickened-1', 4'-oxathiolan-2'-one) (compound AF265; so pl. 92oC).

1H-NMR (CDCl3) to 11.6 (d, 3H); 1,98-2,12 (m, 2H); 2,13-2,3 (m, 2H); 2,34 is 2.55 (m, 2H); 2,55-of 2.72 (m, 2H); 2,30 (s, 2H); 4,06 (q, 1H) ppm.

13C-NMR (CDCl3) 18 (CH3CH, Quartet); 39 (C6 and C10, one triplet); 40 (CH3CH, doublet), with 45.5 (CH3-N, Quartet); 51.8, 52 (C7 and C9, two different triplet); 87 (C5, singlet); 175,5 (C=O) ppm.

MS m/z 201 (M+).

Dedicated connection AF265 (free base) was dissolved in acetone and was added oxalic acid (1 M to 2 M of free base). After a few seconds, then formed a white precipitate. Was specified filtered the precipitate off, washed it with acetone, chloroform and ether, then dried in a desiccator. Consider the precipitate was identified as oxalic salt compounds AF265. The stability of this salt is investigated in the present. Of a specified salt was decomposed in the water, possibly due to hydrolysis of the corresponding lactone; evidence of the availability of the products resulting from the disclosure of the ring, can be detected in the spectrum of13C-NMR, recorded in D2O for at least 3 h Stanovoy accumulation. In the corresponding spectrum1H-NMR, measured directly after RELCOL connection AF265, dissolved in water, characterized by open ring; so pl. 120oC.

1H-NMR (D2O) of 1.55 (d, 3H); 2,3-2,6 (m, 4H); 2,9 (s, 3H); 3,2-3,4 (m, 2H); 3,4-3,7 (m, 2H); 4,4 (q, 1H) ppm.

13C-NMR (D2O) 17,5 (CH3CH); 36 and 36.5 (C6 And C10); 41,5 (CH3CH); 43 (CH3N); 51 (C7 and C9); 84 (C5); 167 (CO in the composition of oxalic acid); 177 (C=O) ppm.

Example 48: piperidine-4-Spiro-5'-(3'-methyl-1',4'- oxathiolan-2'-he) AF269

Used as educt 1 methylpiperidin - 4-Spiro-5'-(3'-methyl-1', 4'-oxathiolan-2'-one) (compound AF265; 80 mg) was dissolved in benzene and the resulting solution was evaporated until dry to ensure the absence of moisture. In an atmosphere of nitrogen was added dichloromethane (approximately 4 ml), dried over molecular sieve, and then 1-chloroethylphosphonic (56 mg). The resulting reaction mixture was stirred at a temperature of 100oC for approximately 1 h, and then evaporated. Added chloroform, filtered, the resulting white solid was washed its carbon tetrachloride, and then dried. The isolated product was identified as the hydrochloride, piperidine-4 - Spiro-5'-(3'-methyl-1',4'-oxathiolan-2'-she) (connection AF265).

1H-NMR (D2O) of 1.55 (d, CH3CH); 2,25-to 2.55 (m, 4H); 3l-1', 3' -oxazolidin) AF264

A mixture of 1-methyl-4-piperidone (11,30 g; 0,10 M) and DL-1-amino-2-propanol (9.75 g; 0.3 M) was kept at the temperature of reflux distilled for 1.5 hours the reaction medium was subjected to distillation under reduced pressure. Removing faction prehoneymoon at a temperature of 30-110oC (pressure of 18 mm Hg) and containing water, an excess of the corresponding amerosport and the desired product, the distillation of pure 1-methylpiperidin-4-Spiro-2'- (5'-methyl-1', 3'-oxazolidine) (connection AF264; 10,70 g) is conducted at a temperature of 110-115oC (pressure of 18 mm Hg).

1H-NMR (CDCl3) to 1.22 (d, J= 7 Hz, CH3CH-); to 1.75 (m, 4H); 2,30 (s, CH3N); 2,49 (m, 4H); 2,70 (dd) 3.27 (dd) (J1=6,6 Hz, J2=12 Hz, -CH2NH-); Android 4.04 (m, J1=6,6 Hz, J2=6 Hz, CH3CHO-) ppm.

13C-NMR (CDCl3) 20,3 (CH3CHO-); 35,3 and 36.7 (C6 or C10), with 45.5 (CH3N); 51,7 (-CH2NH-); 52,8 (C7 and C9); 71,8 (CH3CHO-); 93,3 (C5) ppm.

MC m/z 170 (M+); 169 (M+-1); 123; 112; 85; 83 (100%); 71; 70; 58.

Example 50: 1 Methylpiperidin-4-Spiro-2'-(4'-ethyl-1', 3' -oxazolidin) AF268

A mixture of 1-methyl-4-piperidone (5,65 g; 0.05 M) and DL-2-amino-1 - butanol (6,24 g; of 0.07 M) was kept at the temperature of reflux distilled for 3 hours the reaction medium was subjected to distillation at reduced is containing a series of predominantly the desired product, as well as smaller amounts of water and reagents, the distillation of pure 1-methylpiperidin-4-Spiro-2' -(4'-ethyl-1', 3'-oxazolidine) (connection AF268; 4.5 g) is conducted at a temperature 129-130oC (pressure of 19 mm Hg).

1H-NMR (CDCl3) to 0.97 (t, J=7.5 Hz, CH3CH2-); 1,45 and 1.69 (m, CH3CH2-); 1.70 to 1,89 (m, 4H); 2,30 (s, CH3N); 2,32-2,62 (m, 4H); 3.25 and of 3.96 (m, -CH2O-); 3,29 (m, -CH2CH3) ppm.

13C-NMR (CDCl3) 10,7 (CH3CH2-); 26,1 (CH3CH2-); and 34,9 36,3 (C6 and C10); 45,3 (CH3N); 52,5 and 52.7 (C7 and C9); 58,5 (C3); 69,5 (C2); br93.1 (C5); ppm.

Example 51: 1 Methylpiperidin-4-Spiro-5' -(3'-ethyl-1',4'- oxathiolan-2'-he) AF271

A solution of 1-methylpiperidine (1,921 g; 0,017 M), 2-mercaptoethanol acid (2 g; 0,0167 M) and p-toluensulfonate acid (300 mg) in acetonitrile was subjected to reflux distilled within 36 hours Then evaporated above solution, was dissolved in methanol and provided the desired product in silicagel column, elution with a mixture of chloroform, methanol and ammonia in the ratio of 97: 2,5: 0,5. Additional purification was performed on silicagel column, elution with a mixture of chloroform and methanol in the ratio 95:5.

1H-NMR (CDCl3) 1,0 (t, CH3CH2); 1,6 (m, 3H); 2,1 (m, 1H, CH3CH2); 2,25-2,4 (m, 4H); 2,3 (s, - propargylglycine)and AF186

Connection AF185 has Demetrashvili in accordance with the method developed in relation to the connection AF160(Des) (see Example 3), and then was purified by chromatography on silicagel column (elution with a mixture of chloroform, methanol and ammonia in the ratio of 80:19:1). The corresponding free base precipitated as a white hygroscopic hydrochloride.

1H-NMR (D2O hydrochloride) a 2.0 to 2.15 (m, 2H); about 2.2 to 2.35 (m, 2H); in 2.68 (t, 1H, J= 3.5 Hz); 2.91 in (s, CH3N); of 3.25 to 3.35 (m, 2H); 3,55-the 3.65 (m, 2H); 4,3 (d, 2H, J=3.5 Hz) ppm.

MC m/z 207 (M+the main peak); 179; 151; 113.

Example 53: Synthesis of 1-methylpiperidin-4-Spiro-5'-[3'-(2- butynyl)as)]and AF199

Connection AF199 synthesized in accordance with the method developed in relation to the connection AF185 (see Example 50), and then was purified by chromatography on silicagel column (elution with a mixture of chloroform, methanol and ammonia in the ratio of 80:19:1). The corresponding free base precipitated as a white hygroscopic hydrochloride.

1H-NMR (CDCl3- free basis) of 1.65 to 1.8 (m, 2H); 1,78 (t, 3H, J= 1.7 Hz); 1,9-of 2.25 (m, 4H); 2,33 (s, CH3N); 2,8-2,95 (m, 2H); 4,22 (q, 2H, J=1.7 Hz); 6,7 (bs, 1H) ppm.

MC m/z 207 (M+); 165; 154; 71 (main peak).


In a flask with a capacity of 250 ml was made the connection AF261 in the form of the free base (3,23 g, 16 mm) and di-p-toluene-D-tartaric acid (5.8 g; 15 mm) was added isoamyl alcohol (28 g) and toluene (130 g) and the resulting mixture was subjected to reflux distilled within 5 minutes the Resulting clear solution is partially evaporated during reflux distilled in a stream of nitrogen to slight turbidity, after which he left for the night. Then the precipitation was filtered and recrystallized several times its from a mixture of isoamyl alcohol and toluene to more than 98% purity of the enantiomer according to the results of GC and NMR. Was podslushivaet selected solid product and besieged him in the form of the hydrochloride with a yield 207 mg of a white hygroscopic solid. []2D5= -54,8(hydrochloride in methanol).

Evaporated mother liquor was podslushivaet it was treated with di-p-toluene-L-tartaric acid and recrystallized several times from a mixture of isoamyl alcohol and toluene to more than 98% purity of the enantiomer according to the results of GC and NMR. Was podslushivaet selected solid product and besieged him in the form of the hydrochloride with a yield 207 mg of a white hygroscopic solid. []2D

The purity of each enantiomer was determined by GC using capillary column Chrompack XE-60-S-Val-S -- PEA; WCOT-attached silicon oxide 50 m; internal diameter of 0.26 mm; outer diameter 0.35 mm; flow 0.9 ml of N2; temperature 175o; the retention time of 64 minutes and 65 minutes is an NMR Spectrum of the above hydrochloride was identical to that in case (a) AF269.

Example 55: 1 Methylpiperidin-4-Spiro-5'-(2'-ethyl-1',4'-thiazolidine-3'- Oh) AF267 and its optical isomers (+)AF267 and (-)AF267

Connection AF267 in the form of a free base and di-p-toluene-D - tartaric acid salt was treated in accordance with the method developed in relation to the connection AF261 (see Example 53) to more than 90% purity of the enantiomer according to the results of GC. Was podslushivaet selected solid product and besieged him in the form of the hydrochloride with a yield 207 mg of a white hygroscopic solid. []2D5= -67,0(hydrochloride in methanol, > 90% optical purity). Evaporated mother liquor, Podlachia the product (characterized by more than 90% purity of the enantiomer), which was podslushivaet and precipitated as hydrochloride, []2D5= +70,8(hydrochloride in methanol, > 95% optical purity).

The purity of each enantiomer was determined by GC using capillary column Chrompack XE-60-S-Val-S -- PEA; WCOT-attached silicon oxide 50 m; internal diameter of 0.26 mm; outer diameter 0.35 mm; flow 0.9 ml of N2; temperature 175o; the retention time of 83 minutes and 88 minutes is an NMR Spectrum of the above hydrochloride was identical to that in case (a) AF267.

Example 56: Synthesis of 1-methylpiperidin-4-Spiro-2'-(5'- methyl-1',3'-dioxolane-4'-he) AF274

8 ml of lactic acid azeotrope dried in benzene. To a dry and hot lactic acid was added an additional amount of dry benzene, and then p-toluensulfonate acid (300 mg) and 1-methylpiperidin (5,00 g; 0,027 M). The resulting mixture was subjected to reflux distilled up until they ceased not to separate water. After that evaporated the specified solution, highlighted the residue was dissolved in chloroform and washed with saturated sodium bicarbonate solution (25 ml). The corresponding aqueous phase was washed with chloroform (25 ml), combined organic phase was twice washed with water (10 ml). Washed organicheskoi with the release of 8.7 g crudely purified oily product. Rubbed the specified oil in a mixture of hexane, ether and petroleum ether (40-60), the remaining oil was dissolved in acetone, and then to the resulting solution was added oxalic acid (1 EQ. ). After a while she rolled a white precipitate, which was identified as oxalic salt 1 methylpiperidin-4-Spiro-2'-(5'-methyl-1',3'-dioxolane-4'-she) (connection AF274).

1H-NMR (oxalic salt, CDCl3+DMSO-d6) and 1.5 (d, 3H, J=7 Hz); of 2.15 (m, 4H); 2.63 in (s, 3H); 2,9 (m, 2H); 3,05 (m, 2H); 4,55 (q, 1H, J=7 Hz) ppm.

1H-NMR (free base, CDCl3) and 1.5 (d, 3H, J=7 Hz); 1,95 (m, 4H); 2,32 (s, 3H); 2,5 (m, 2H); 2,6 (m, 2H); 4.5 m (q, 1H, J=7 Hz) ppm.

Mass spectrometry m/z 185 (M+).

Example 57: 2-Methyl-1,4-thiazolidin-3-he-Spiro[5,3']Hinkley AF273

A mixture of 3-binucleation (2.50 g; 20.0 mm), timelocal acid (2.55 g; 24.0 mm) and ammonium carbonate (2.9 g) in toluene (100 ml) were introduced into a steel vessel designed for high-pressure, and kept at a temperature of 108oC and stirring for 28 hours From the resulting reaction medium was removed emerged in the oil, and the remaining toluene solution was evaporated emitting 2.5 g of residue. The specified residue was led from acetone to yield crystalline solid, to which was led from acetonitrile, and then from dichloromethane with the release of one of the pure enantiomeric pair () connection AF273 (0,210 g; so pl. 201 - 202oC).1H-NMR (CDCl3) of 1.53 (d, J=7.2 Hz, CH3CH-); of 1.65 (m, 2H); of 1.92 (m, 2H); 2,03 (m, 1H); and 2.83 (m, 4H); 3,18 and of 3.32 (dd, J= 15 Hz, NCH2CNH-); to 3.89 (q, J=7.2 Hz, CH3CH-); 8,58 (bs, -NH-) ppm.

MC m/z 212 (M+); 155; 142 (M+-70); 123; 96; 71; 70 (100%); 58.

Mother liquor remaining after the first crystallization and enriched the other enantiomeric pair was subjected to chromatography on silicagel column (Merck 60). The elution was carried out with a mixture of chloroform, methanol and ammonia in the ratio of 96:3:1 with the output of the first, second enantiomeric pair connection () AF273B (0,106 g; so pl. 181-182oC), crystallization from acetonitrile.

1H-NMR (CDCl3) of 1.52 (d, J=7 Hz, CH3CH-); of 1.64 (m, 2H); 1.93 and (m,2H); 2,03 (m, 1H); 2,70 are 2.98 (m, 4H); 3,19 (s, NCH2CNH-); of 3.85 (q, J= 7 Hz, CH3CH-); at 8.60 (bs, -NH-) ppm.

MC is similar to that in the case of compound (a) AF273A.

Further received a mixture of all enantiomers and, finally, another pure enantiomeric pair. () AF273A.

Example 58: 1 Methylpiperidin-4-Spiro-5'-(2'-methyl-1',4'- thiazolidine-3'-tion) AF275

A suspension of 1-methylpiperidin-4-Spiro-5'-(2'-methyl-1', 4'- thiazolidine-3') (connection AF261; 1,00 g; 5.00 mm) and reagent Lisa reaction medium solvent was removed, and the resulting residue was purified on a dry silicagel column (Merck 60; 0,040-0,065). The elution was carried out with a mixture of chloroform, methanol and ammonia in a volume ratio of 96: 3: 1 with a yield of pure 1-methylpiperidin-4-Spiro-5'- (2'-methyl-1', 4'-thiazolidine-3'-thione) (connection AF275; 0,81 g) in the form of needle-like crystals formed by crystallization from acetone, so pl. 171-172oC (decomposition).

1H-NMR (CDCl3) was 1.69 (d, J=7,1 Hz, CH3CH-); 2,00 (m, 2H); from 2.00 to 2.35 (m, 4H); 2,30 (s, CH3N); 2,80 (m, 2H); 4.26 deaths (q, J=7,1 Hz, CH3CH-); a total of 8.74 (bs, -NH-) ppm.

MC m/z 216 (M+); 156; 98; 97; 96 (100%); 70.

Example 59: 1 methylpiperidin-4-Spiro-4'(5')-(2'-methylthio-2'- imidazolin-5'(4')-he) AF187

a) To a stirred solution of sodium hydroxide (0.36 g; 9,00 mm) in methanol (30 ml) were added hydrochloride 1 methylpiperidin-4 - Spiro-5'-(2'-thiohydantoin)and (hydrochloride of the compound AF195; 1,00 g; 4,25 mm). Formed precipitate (NaCl). To this mixture at room temperature was added methyliodide (0,664 g; 4,68 mm) and continued stirring for an additional 1.3 hours the reaction mixture is evaporated under reduced pressure, and the selected residue was extracted with dichloromethane. Removing the used dichloromethane, received the remainder (0,58 g) which was purified on sinikalliontie 89:10:1 to yield 1-methyl-piperidine - 4-Spiro-4'(5')-(2'-methylthio-2'-imidazolin-5'(4')-it) (connection AF187; to 0.23 g; so pl. 176-177oC), crystallization from acetone.

1H-NMR (CDCl3) of 1.57 (m, 2H); 2,00 (m, 2H); 2,35 (s, CH3N); 2,52 (m, 2H); 2,56 (s, CH3S); 2,84 (m, 2H); 6.73 x (bs, -NH-) ppm.

13C-NMR (CDCl3) 12,6 (s, CH3S); 32,7 (C3); 45,8 (CH3N); 50,8 (C2); 68,3 (C4); 163,8 (-C-SCH3); 188,1 (-C=O) ppm.

MC m/z 213 (M+); 198 (M+-CH3); 143 (M+-70); 71.

< / BR>
b) a Solution of 1-methylpiperidin-4-Spiro-4'-(2'-methylthio-5'- aminomethyl-4'-imidazole) (connection AF193; 30 mg) in water 16% hydrochloric acid (0.5 ml) was kept at room temperature for 3 days. Then under reduced pressure, the solvent was removed and the resulting residue was podslushivaet using concentrated aqueous sodium hydroxide solution, and then was extracted with a mixture of dichloromethane and methanol. The corresponding extract was dried over Na2SO4and solvent was removed with the release of the residue, which was purified by thin-layer plate of silica gel (Merck Kieselgel 60 F254), treated with a mixture of chloroform, methanol and ammonia in a volume ratio of 79: 20: 1 connection output AF187 (8 mg), similar to that of an authentic sample.

Example 60: 1-Methylpiperidin-4-Spiro-4'-(1'-ethyl-2'- ethylthio-5'-imidazolin-5'-he) is droxide potassium (3,85 g; 0,0686 M) in ethanol (100 ml) was added ethyl bromide (ceiling of 5.60 g; 0,0514 M), and the resulting mixture was subjected to reflux distilled under stirring. After 2 hours reflux distilled the main reaction product remained monoethylene derived the specified connection, therefore, to consider the reaction mixture was added an additional amount of potassium hydroxide (0.95 g; 0,0196 M) and ethyl bromide (2.00 g; 0,0184 M) and continued dephlegmation for another 3 hours and Then under reduced pressure, the solvent evaporated, to the residue was added water (20 ml) and was extracted with him ether. The corresponding extract was dried over Na2SO4and solvent was removed with the release of the balance (0,973 g), which was led from petroleum ether to obtain pure 1-methylpiperidin-4-Spiro-4'-(1'-ethyl-2'-ethylthio-2'-imidazolin-5'- she) (connection AF188; 0,513 mg; so pl. 95-96oC). The remaining mother liquor after removal of the solvent was subjected to chromatography on a dry silicagel column (20 g; Merck 60, 0,40-0,065 mm), elution with a mixture of chloroform, methanol and ammonia in a ratio of 84: 5:1 with the release of additional quantities of pure compounds AF188 (0,310 g).

1H-NMR (CDCl3) to 1.19 (t, J= 7.2 Hz, CH3CH2N); of 1.40 (t, J=7.2 Hz, CH3CHB>CH2N); ppm.

13C-NMR (CDCl3) 14,1 (2 CH3CH2-); 24,4 (CH3CH2S); 33,0 (C6); 35,1 (CH3CH2N); 46,2 (CH3N); 51,0 (C7); 68,6 (C5); 158,8 (-N=C-); 183,9 (-C=O) ppm.

MC m/z 255 (M+); 226 (M+-Et); 185 (M+-70); 71; 70.

Example 61: 1 Methylpiperidin-4-Spiro-4'-(1'-ethyl-2'- imidazolin-5'-he) AF220

To a solution of 1-methylpiperidin-4-Spiro-4'-(1'-ethyl - 2'-ethylthio-2'-imidazolin-5'-one) (compound AF188; 0,214 g) in ethanol (10 ml) was added Raney Nickel (0,30 g) and the resulting mixture was subjected to reflux distilled for 4 hours because after this time, an appropriate response has not yet ended, was added an additional amount of Raney Nickel (0,30 g) and continued dephlegmation for another 4 hours Filtered by the specified reaction medium, and the selected catalyst was washed with methanol and dichloromethane. Combined appropriate filtrate and washing liquid, and then evaporated to their exit 1-methylpiperidin-4 - Spiro-4'-(1'-ethyl-2'-imidazolin-5'-one) (compound AF220; 0,160 g) in the form of butter.

1H-NMR (CDCl3) of 1.27 (t, J=7,3 Hz, CH3CH2-); of 1.46 (m, 2H); 2,03 (m, 2H); of 2.36 (s, CH3N); 2,48 (m, 2H); 2,82 (m, 2H); of 3.53 (q, J=7,3 Hz, CH3CH2-); 7,72 (s, -CH=N) ppm.

13C-NMR (CDCl3) 13,9 (CH3

Trying to clear the specified product by chromatography on a dry silicagel column (20 g; Merck 60, 0,40-0,065 mm, elution with a mixture of chloroform, ether, methanol and ammonia in a volume ratio of 25:17:3: 1), as the primary connection received the product resulting from the hydrolysis of disclosure of the respective rings: 1-methylpiperidin-4-formylamino-4-N - ethylcarbodiimide (connection AF221; so pl. 108-109oC) crystallizing from a mixture of ether and dichloromethane.

1H-NMR (CDCl3) of 1.13 (t, J=7.2 Hz, CH3CH2-); 2,04 of-2.32 (m, 6N); 2,28 (s, CH3N); to 2.65 (m, 2H); 3,28 (dq, CH3CH2-); 5,73 (s, HCONH-); 6,97 (bs, EtNHCO-); 8,18 (s, HCON-) ppm.

The resulting hydrolysis product AF221 can be re-cycletour in connection AF220 when heated to a temperature of 150oC in vacuum.

The above agonistic or antagonistic ability of the considered compounds were investigated in a series of tests, including computational molecular analysis and biological testing of in vivo and in vitro.

Computational molecular analysis

For the study of agonistic activity towards muscarine was developed pharmacopoiea model based on nih agonists, required for the activity;

- analysis of their mutual spatial arrangement, and also

- (to some extent) the definition of the greatest number, in which the ligand is an agonist.

< / BR>
< / BR>
In the above formulas, the point r denotes a negative charge, which interacts with the cationic part of the agonist. This model determines the position of this point relative to the corresponding nitrogen atom. Significant distances (D) are as follows: D(r-N*) = 3,0; D(r-X*) = 6,50; D(r-Q*) = 8,70; D(x*-Q*) = 2,45. Optimal flat angle r-x*- Q*-z*= -85. Deviations from these optimal model parameters in a certain range does not lead to violation of the relevant activity (table 1).

Below are listed a specific of the significant features of the model:

1. The model allows to distinguish between partial and full agonists of muscarine. This distinction is based on the correlation between the distance r-X*and agonistic activity.

2. Within one class of muscarinic agonists atom in position 4 and the corresponding carbonyl atom color the tx2">

3. In that case, if the distance r-Q*becomes too large for the display of agonistic activity, are weakly bind agonists.

Based on this model, based on the structural stability of a specific agonist, the nature of the group Z*and the model parameters, it is possible to predict the specificity of the muscarinic agonist with respect to subtypes of receptor.

Described pharmacophoric model can be used as follows for screening new compounds, potentially having muscarinic activity:

a) optimization of the new structure to identify its low-energy conformers;

b) the study of these conformers in terms of correct positioning of pharmacophoric elements. Alternatively pharmacophoric conformation can be given a new structure using one of the traditional ways. Further, the resulting conformation compared with low-energy conformation, characteristic of the same structure. Using the described method gave real results in all investigated cases (table 1). Despite the fact that it is impossible to predict whether muscarinic activity appear what if, are active agonists.

Size limits cationic site in the structure of muscarinic agonists was investigated in experiments on shortening of the relevant structures (see Table 1) included in a molecular model of the transmembrane domain of muscarinic m1 receptor. Connections that are larger than the derivative of hinoklidina cannot be recognized macromolecular binding site.

The compounds listed in Table 1 (see end of description) divided into 4 groups. The first group includes several well-known agonists muscarine. Their structure is characterized by optimal pharmacophoric parameters. The second group includes agonists, different suboptimal pharmacophoric parameters and mainly because of partial agonists. The structure of the compounds included in the third group, characterized by altered pharmacophoric parameters compared to the above groups of compounds exhibiting muscarinic activity. These compounds are either antagonists muscarinic activity, or are deprived of it. In the fourth group are some potential agonists muscarine, preds the isolation of the ileum of the Guinea pig.

I provided by the present invention compounds were investigated for their agonistic and antagonistic activity shown on the preparations of the isolated ileum (Fisher et al. , J. Pharm. Exp. Therap. 257: 394-403 (1991). The data obtained for some compounds, are presented in Table 2 (see end of description).

Test N. 2. Binding to muscarinic receptors in the brain; competition with [3H] QNB, [3H] NMS and [3H]OXO-M on membranes isolated from cortex and cerebellum of rats.

For testing new compounds (Tables 3 and 4) (see the end of the description) used membrane preparations cortex and cerebellum of rats, as well as ligands [3HNMS, [3H]pirenzepine and [3H]oxotremorine-M.

The value of the ratios KiPZ/KiNMSor KiPZ/KiQNB(Table 3) is a typical measure of the selectivity of muscarinic ligands with respect to M1 receptors, while low values of these ratios indicate a greater selectivity. In addition, the magnitude of the ratios KiPZ/KiQNBdefined for membrane preparations cortex and cerebellum of rat brain, are also indicators selectivity of muscarinic ligands with respect to M1 receptors; the values of m which could demonstrate, some of the compounds provided by the present invention are characterized by a relatively high preference in relation to muscarinic receptors M1. Among such compounds include the following: AF133, AF134, AF160, AF160(Des), AF177, AF178, AF181-AF185, AF261, AF265, AF267.

In particular, the connection AF185 showed 3 times higher affinity for sites of competitive binding with [3H]PZ compared with sites of competitive binding with [3H]NMS (Ki = 1,4 0,15 µm 1 and 5 μm, respectively; table 3). These data contrast sharply with the results obtained for compounds AF102B (U.S. Patent), which was characterized by almost the same levels of affinity for sites of competitive binding of both ligands (Ki = 1,43 0.2 μm and 1.86 of 0.43 μm, respectively; table 3). As of [3H]PZ is considered a specific marker of receptor M1AChR on the membranes of the cerebral cortex of rats, we obtained new evidence suggests that the connection AF185 is more specific ligand relative to the receptor M1 than the connection AF102B.

Curves of binding of [3H]NMS, for certain membranes of the cerebral cortex using some new connections, specifies the interaction with dum connections. This fact may indicate high agonistic activity of the compounds in the above-mentioned drugs. Connections of this kind were the following: AF260, AF261, AF263 AND AF265.

Some of these new compounds unexpectedly characterized dvuhlistovymi curves of competitive binding [3H]PZ in the cerebral cortex of rats (table 3). Among such compounds include the following: AF160, AF160(Des), AF260, AF261, AF265. Dvuhlitrovye curves of competitive binding [3H] PZ obtained for these compounds were converted into simple curves "dose-effect" in the presence of the stable GTP analogue: GppNHp (data not shown). Similar observations sensitivity to GppNHp were carried out in the case of competition with [3H]NMS (data not shown). The results testified to the fact that these compounds behave as an effective agonists with respect to M1 receptors in the cerebral cortex of rats. This finding differs from the results obtained in the competitive tests using connection AF102B, which is characterized by typical curves "dose-effect". The data seems to indicate some differences between the connection AF102B, videopom AF267 was characterized by single-site competitive binding curves of [3H]NMS and [3H]PZ on the membranes of the cerebral cortex of rats. The following study of the hydrolysis of phosphoinositides (PI) and the release of arachidonic acid showed that this compound is a partial agonist. Therefore, the patterns of binding of type "dose-effect" were similar to those in the case of connection AF102B, which is a selective agonist with respect to M1 receptors, in relation to muscarinic receptors considered drug. We have previously shown that treatment of the membranes of the cerebral cortex of rats 0.1 mm CuSO4can detect hidden agonistic features linking connection AF102B (Fisher et al. , J. Pharm. Exp. Therap. 257: 392-403 (1991)); the published data is based on competition connection AF102B and [3H]QNB, however, similar observations obtained with [3H]NMS and [3H]PZ). In addition, this treatment can increase the proportion of sites having a high affinity with respect to certain agonists (for example the connection AF160), which reveal the presence of sites of high affinity and in the absence of processing ions Cu2+. Therefore, we investigated the features of the binding compounds AF267 after treatment of the ions Cu2+. The specified processing, e affinity connection AF267 (KH= 22 nm, i.e. approximately 100-fold higher affinity compared to the KL= 2.5 μm) without a significant change in the number of sites with low affinity. The data suggests that the connection AF267 can communicate with muscarinic receptors in the cerebral cortex of rats as well as the connection AF102B. The ability of ions of Ca2+to identify sites with high affinity to certain partial agonists may be due to stabilization of the interaction between receptor and G-protein due to coordination complexes between metal ions and essential sulfhydryl groups in the structure of these macromolecules (Gurwitz et al., BBRC 120: 271-277 (1984)). In the control membranes such interaction is definitely not enhanced in the presence of a partial agonist, which, thus, does not exhibit high affinity in the competitive tests using labeled antagonists.

The ability of compounds to modify the binding of [3H]oxo - tremorine-M ([3H] OXO-M) allows to measure the degree of affinity of the corresponding receptor in a state of high affinity for the agonist. The ratio of the Ki values of the substitution of [3H]NMS or [who is superior to 100, typical full agonists; antagonists characteristic ratio close to 1; the intermediate values typical for partial agonists (Orlek et al., J. Med. Chem. 34: 2726 (1991)).

The above new compounds were investigated in the test for competitive binding of labeled, non-selective muscarinic agonist [3H]OXO-M using membranes of the cerebral cortex of rats; high agonistic properties of the compounds AF179 and AF160 reflected in the relatively high values of KiNMS/KiOXO-Mtypical of these compounds (table 4).

Connection AF160(Des) and AF185 sudden I had only studied agents, characterized by a nonlinear dependence "dose-effect" with [3H]OXO-M and membranes of the cerebral cortex of rats (table 4). Extremely effective competition of these compounds with sbraccia receptors located on the membrane of the cerebral cortex of rats and labeled with [3H] OXO-M, apparently? suggests that these compounds are highly specific with respect to a specific receptor subtype. Thus, the calculated ratio KiNMS/KHOXO-M(Table 4) suggest that the connection AF160(Des) pentiu with other agents, related connection AF160, as well as connection AF102B. However, such data were not received: curves of competition as connections AF160(Des) and the connection AF185 with [3H]NMS show a simple linear dependence "dose-effect". Thus, the assumption of heterogeneity of the subfamily of receptors, which is revealed only by studying the binding of [3H]OXO-M, but not of [3H]NMS, does not seem convincing. An alternative interpretation may have some interaction with allosteric sites of muscarinic receptors, which are not implemented in the binding of the receptor with such antagonists as [3H]NMS, but is manifested when using the agonist [3H]-OXO-m. Another possible explanation for the weak performance of competition in the test for binding of [3H] OXO-M is that your system has not reached equilibrium. In tests on competition using connection AF160(Des) or AF185 equilibrium was not reached within 30 min, as the characteristics of the binding after 30 and 60 min at a temperature of 25oC was different. In that case, if the test was interrupted after 30 min, weak competition with [3 the act is what connections AF160(Des) and AF185 have a relatively slow kinetics (low coefficients of both the Association and dissociation) compared the connection AF102B. This fact can lead to very slow to establish equilibrium in the tests on competition, causing a clear identification dvuhshipovyh isotherm binding in non-equilibrium conditions. Note that the coefficients of the Association and dissociation are relatively low: in that case, if only the coefficients of the Association of compounds AF160(Des) and AF185 were low compared with the connection AF102B, these compounds were characterized by much lower affinity compared with the connection AF102B. On the other hand, if the below were the only factors dissociation of these compounds, the corresponding figures affinity would be significantly higher than in connection AF102B.

The rat cerebellum is relatively homogeneous in terms of expressed subtypes of mAChR receptors (especially M2). Connection AF102B shows similar effectiveness of competition with [3H]OXO-M as in the cortex and cerebellum of rats. On the contrary, the connection AF160(Des) and AF185 more active with respect to the membranes of the cortex, rather than the cerebellum; this fact was discovered, according to the presence of duhaieva the result may indicate different kinetics of the considered compounds in the cortex compared with the cerebellum.

Test N. 3. Activation of second messengers in the brain slices and cell cultures.

To conduct a test to measure the efficiency of the investigated compounds as agonists of muscarinic receptors M1 received slices of the cerebral cortex of rats (200 μm3). To test the metabolism of phosphoinositides (PI) these sections of the brain was treated with [3H]Inositol (4 µci/ml), incubare them into the buffer salt Krebs solution containing a labeled ligand for 1 h at a temperature of 37oC and oxygen flow. Then washed the above sections and added in aliquots of 50 ál in each tube, containing 10 mm LiCl in fresh Krebs solution in the presence or absence of the tested compound. Incubated for 20 min at a temperature of 37oC, and then stopped the reaction and was isolated labeled products on columns AG-1-X8 (Berridge, Biochem. J. 258: 849-858 (1983)). Depending on your connection, partial agonists caused less than 80% activation of PHOSPHOINOSITIDE hydrolysis in comparison with the churches of Christ (a full agonist). So, the connection AF160 resulted in a significant increase in the concentration of HF3(1.6 times). Compared with the churches of Christ, the connection AF160 was a partial agonist exhibiting 50% of the activity ukplease another agonist receptors M1, connection AF102B was less active than compound AF160, while the connection AF178 was inactive in this test.

In order to measure the increase in the activation of second messengers under the action of the investigated compounds, used cell culture enriched in one subpopulation of muscarinic receptors. To study the metabolism of PHOSPHATIDYLINOSITOL used the method of Berridge (Berridge, Biochem. J. 258: 849-858 (1983)); to study the mobilization of arachidonic acid cells for 16 h were labeled with [3H] arachidonic acid (or 0.2 µci/ml) in the original growth medium. Before the test cells 6 times washed with serum-free medium DME containing HEPES (20 mm) and albumin serum (1 mg/ml). After this rinse was added to 0.5 ml of the same medium with the addition of the studied ligand. To stop testing the above medium was transferred into Eppendorf tubes and centrifuged for 10 min at 6000 g. Determined the radioactivity of the supernatant was expressed in dpm released [3H]arachidonic acid per well. The accumulation of cyclic AMP in intact cells was evaluated in accordance with (Pinkas-Kramarski et al., Neurosci. Lett. 108: 335-340 (1990)), while the activity adenylyl cyclase activity in selected member the tx2">

Preferred are compounds corresponding to the formula AA and characterized by the highest percentage (over 25%) of PHOSPHOINOSITIDE metabolism and/or mobilization of arachidonic acid (but not significant activation adenylyl cyclase activity). The number of connections of this kind can be identified as follows: AF160, AF160(Des), AF179, AF180, AF185, AF260, AF261, AF263, AF265, AF266, AF267. These compounds are able to activate muscarinic receptors M1.

Unlike acetylcholine, the churches of Christ, oxotremorine-M and other classic full agonists, these compounds cause selective activation of the transmission of signals via muscarinic receptors M1 (or M3). In particular, selective activation of PHOSPHOINOSITIDE hydrolysis in the absence (or minimal level) the activation of accumulation of camp is a common characteristic of the activity inherent to consider new connections. The specified properties of the specified selective muscarinic ligands can be based on their ability to induce the binding of the muscarinic M1 receptor to different G-proteins. Thus, in addition to activation of M1 receptors, the compounds being considered can also show selectivity at the level of different second messengers. This hypothesis, pdin the same muscarinic receptor, recently described by the authors of the present invention in the form of the concept of signal transmission due to selective ligand using CHO cells, transfected with the gene m1AChR rats (Fisher et al. , Biorganic &Medicinal Chem. Lett. 2: 839 - 844 (1992)), and when using cell lines of neural type, in particular, cell lines RSM (Soc. Neuroci. Abs., November, 1993). Judging by the detection of elevated levels of G-proteins in patients suffering from Alzheimer's disease (ad), as well as in the aging brain (Harrison et al., Mol. Brain Res. 10: 71 (1991); Young et al. , Dev. Brain Res. 61: 243 (1991)), the obvious possibility of applying the specified path signal transmission in the treatment of BA based on the replacement of cholinergic. Thus provided by the present invention compounds may be useful in the treatment of Alzheimer's disease.

Some of these compounds are full agonists, while others are partial agonists when compared with the churches of Christ in terms of stimulation of PHOSPHOINOSITIDE hydrolysis in cells transfected gene m1AChR. Among partial agonists include the following compounds: AF160, AF160(Des), AF178, AF180, AF181, AF265 and AF267. Connection AF260, AF261, AF263 and AF266 showed in the specified test full agonistic activity. Curves "dose-effect", postreh 10 μm and 100 μm, respectively. Connection AF265 and AF267 behaved in this test as a partial agonist. Connection AF181 showed in this test, the weak activity of a partial agonist. So, at a concentration of 1 mm the specified connection activated PHOSPHOINOSITIDE hydrolysis, but weakened called the churches of Christ hydrolysis of PHOSPHATIDYLINOSITOL, which is expected in the case of a partial agonist. Judging by the fact that the compounds AF213 and AF184 blocked the churches of Christ mediated hydrolysis of PHOSPHATIDYLINOSITOL, these compounds exhibit antagonistic activity. Among enantiomers of compounds AF267 the greatest activity characteristic of the connection AF267B, representing the (-)-enantiomer; the specified connection activates the PHOSPHOINOSITIDE hydrolysis in cells transfected with the gene coding for the receptor m1, even at a concentration of 10 μm (40% of the churches of Christ) and 100 microns (80% of the churches of Christ).

We investigated the ability of new helical compounds to cause desensitization to the signal representing caused the churches of Christ hydrolysis of PHOSPHATIDYLINOSITOL. Connection AF261 caused less effect insensitivity compared with the churches of Christ: after 24-hour treatment with 100 ám or 1 mm connection AF261 hydrolysis of PHOSPHATIDYLINOSITOL in response to the addition of 1 mm of the churches of Christ were decreased by 45% compared to 60% reduction in pre-incubation with 1 mm of the churches of Christ. Connection AF267 manifesting is civilizatio. So, after an overnight incubation in the presence of 1 mm connection AF267 due to the churches of Christ response (PHOSPHOINOSITIDE hydrolysis) decreased by an average of 29%.

We also tested new spiraeoideae on their ability to cause PHOSPHOINOSITIDE hydrolysis in cells temporarily transformed genes m1AChR, m3AChR or m5AChR man in the parallel experiments described Pettalom and Vass (Pittel & Wess, Mol. Pharmacol. 45: 61- 64 (1994)). Connection AF267 showed the highest selectivity to the receptor subtype m1AChR, being with about 100 times more active in the case of cells that have been transformed genome m1AChR than m3AChR (the values of the ED50equal to 1.5 and 150 μm, respectively). This pattern is especially clearly visible when comparing the signal caused by 10 μm compound AF267; this signal reaches the maximum value in the case of cells that have been transformed genome m1AChR, but equal to only 15% of the maximum signal due to the churches of Christ in cells transformed genome m3AChR. Connection AF265 has been a bit more active and effective in the case of cells, the transformed genome m1AChR than m3AChR, and significantly less active when using gene m5AChR. Similar results were obtained for compounds AF267, especially for its most LASS="ptx2">

The release of arachidonic acid (AA) is another biochemical process activated receptor agonists m1AChR. As specified biochemical process can be associated with receptors m1AChR via G-proteins that differ from those involved in the hydrolysis of PHOSPHATIDYLINOSITOL, we explored spiraeoideae and in this test, using culture cells, stably transfected with a gene m1AChR rats. Connection AF263, AF265, AF266 and AF267 (at a concentration of 1 mm) showed in the above test, partial agonistic activity. Among enantiomers of compounds AF267 is active (-)-enantiomer, which when used in concentrations of 0.1 and 1 mm is a full agonist release of AK. Connection AF260 and AF261 had full agonistic activity when compared with 1 mm of the churches of Christ, which is consistent with the detection of PHOSPHOINOSITIDE hydrolysis in the same cell line. We found the tendency of these agonists to cause a more intense release of AK in comparison with the churches of Christ. This fact may indicate that during the test (20 min at a temperature of 37oC) called the churches of Christ, the signal undergoes desensitization. It seems plausible that the degree specified gum is laguepie compounds may be more effective as agonists of M1, than the churches of Christ, i.e. to be a "super-agonists". In this case, it is unclear why the considered property is not observed in the test for hydrolysis of PHOSPHATIDYLINOSITOL when using compounds AF260 and AF261. Conducting a 3-hour pre-incubation of cell cultures transfected gene m1AChR, in the presence of 1 mm of the churches of Christ, followed by extensive washing of the used ligand (6 x 1 ml), reach, greatly reducing the release of AK. Basal release rate AK is slightly altered, indicating the absence of traces of the churches of Christ after washing. For comparison, spending in an analogous experiment, pre-incubation with 1 mm connection AF265 or AF102B, received weakening induced by 1 mm release of the churches of Christ AK only to half of the initial response. In the case of longer periods of incubation, we preferred concentration of the agent 100 μm, since a concentration of 1 mm did not exert physiological effects in long-term impacts. After a 24 - hour pre-incubation in the presence of 100 µm appropriate the churches of Christ AK-response was lost almost completely. We compared the ability of new helical compounds to cause desensitization of the signal caused by the churches of Christ and resulting in the release of AK. 24-hour pre-incubation in the presence of 100 is logival special attention, suggesting that this compound is a partial agonist in this test. In contrast, stimulation with compounds AF260, AF261 and AF263 (when used within 24 h at a concentration of 100 μm) significantly attenuated the signal produced by the churches of Christ and resulting in the release of AK. Similar pre-incubation in the presence of compounds AF102B and AF267 caused relatively little effect on the subsequent signal, the signal produced by the churches of Christ and resulting in the release of AK, which again pointed to the similarity of these two compounds.

Test N. 4: Neurotrophin-like effects in cell cultures

Activation of M1 receptors agonists and the effect of Nerve Growth Factor (NGF) can lead to synergene effects in certain cell cultures, enriched by m1 receptors, including cell RC (cell pheochromocytoma rats), transfected gene m1AChR rats (cells RSM) (Pinkas-Kramarski et al., J. Neurochem. 59: 2158-2166 (1992)).

In accordance with another aspect of the present invention shows that the compounds corresponding to the formula AA and cause more than 25% PHOSPHOINOSITIDE metabolism that can lead to cinergena effect on due FNR axonal growth. Among the compounds provided by the present invention, m is ü axon elongation almost the same as the churches of Christ; and

2. connections, dramatically different from oxotremorine and can cause elongation of axons, or to induce even minimal morphological changes in the absence of NGF.

Both of the above class is suitable for treatment of ad. In the case of the second class may not be the uncontrolled growth of axons. Therefore, the preferred candidate for the role of the pharmaceutical agent in the treatment of BA should, for example, to cause the formation of axons only under strict control by locally synthesized and released growth factors such as NGF, nerve factor of cerebral origin (NFMP), NT-3, etc. are Examples of such unique activity can be detected in the case of compounds such as AF160, AF160(Des) and AF165 that are at least as effectively render synergenic effect on NEF mediated axonal growth, as the connection AF102B. Axons, the elongation of which was caused by the combined effects of NGF and new helical compounds, was found to be stable over long periods of cultivation. Based on this we can conclude that the transmission path of the signal used by the considered compounds in the induction of axon growth, not subject to rapid derivative effects of stimulation of axons in the cells RC, as well as in primary cultures of sympathetic neurons. However, it should be noted that the old is not processed culture of the investigated cells underwent degeneration and contained numerous dead cells detached from the surface of the Cup. Interestingly, this phenomenon of cell death was attenuated in cultures pre-treated with a combination of new helical compounds and NEF. It is well known that NGF rescues cells RX from programmed death (Rukenstein et al., J. Neuroscience, 11: 2552-2563 (1991)). Thus, the above observation indicates the potential ability of compounds provided by the present invention, to provide similar effects, favouring the survival of cells. All these features cause additional interest to the compounds in terms of their potential use for the treatment of patients suffering from Alzheimer's disease.

Neurotrophin-like activity, which is provided by the present invention agonists in relation to the nerve cells, and also depends on the presence of NEF, apparently suggests that the above-mentioned compounds exert their neurotrophic effects due to the mutual is m, that these effects are indirect through enhanced release protein precursor amyloid (BIA), is caused by the receptors m1AChR. It should be emphasized that the endogenous BIA necessary for normal growth fibroblastic cells, and exogenous BIA is able to stimulate proliferation of these cells (Saitoh et al., Cell, 58: 615 (1989); Mattson et al., TINS, 16: 409-414 (1993)). Interestingly, among other activities inherent in secretively forms BIA, well-known ability to regulate axonal growth and survival of nerve cells (Mattson et al., TINS, 16: 409-414 (1993)). Mortality of nerve cells in BA apparently leads to the reduction of education and/ or viability of neurotrophins, which in turn affects the viability of cholinergic neurons. In that case, if the above naratriptan events induced by agonists of M1 receptors, also present in the brain, this can be the most important clinical significance and, thus, to reflect the new method of treatment for ad.

Test n 5; the Secretion of the protein precursor of amyloid a (BPA) in brain slices and cell cultures.

There is growing evidence of the involvement of amyloid in PA is otherwise inappropriate metabolism protein precursor of amyloid a (BPA) (Mattson et al., TINS, 16: 409-414 (1993)). Activation of receptors m1AChR, especially m1-type, leads to increased secretion of BPA in vitro (Nitsch et al.. Science, 258: 304 (1992); Buxbaum et al., PNAS US, 89: 10075 (1992); Lahiri et al., Biochem. Int. 28: 853 (1992)). Thus, in accordance with another aspect of the present invention, the compounds corresponding to the formula AA, in particular those which exhibit selective agonistic activity towards the m1 receptors, and also enhance the release of the UAV can be suitable not only for the treatment of ad, but also to delay its progression or even to prevent BA.

To quantify the secretion of the unit use the method of Western blot turns on the Western (Nitsch et al., PNAS US, 90: 5191-5193 (1993)). We used monoclonal antibodies to BIA (S), which are widely used for the detection of BPA. These antibodies are obtained directly on the amino-terminal section of the unit, and identify all of the known isoforms of the unit. Considering the antibody does not react with amyloid or C-terminal parts of the unit formed after secretion of the unit. Exploring the influence of new helical compounds on the release of the unit, cell line, transfected with a gene of receptor m1, were cultured in 12-hole cups (in some experiments - 6-the data well) was added ligands of sterile 100-fold stock solution. Stopped the experiments, twice washing the above-mentioned holes serum-free medium. The cells were separated from the said cups in cold phosphate buffered saline solution (pH of 7.4; PBS). After centrifugation (10 min at 10000 d) was decanted supernatant, and the resulting precipitates suspended in 0.1 ml of cold lisanova buffer containing a mixture of protease inhibitors (50 mm TRIS-HCl (pH 7,4); 150 mm EDTA; 1% Triton X-100; 0.1 mm PMSF; 5 units/ml Aprotinin, and 5 μg/ml of pepstatin And; 5 µg/ml leupeptin). Then these precipitation within 5 was subjected to sonication at maximum intensity (Branson sonicator, model 130) and again centrifuged. The resulting supernatant (extracts of cell membranes) was transferred into a clean test tube. Samples of these supernatants were selected for detection of the protein using the Lowry method.

To test the unit equal amounts of protein (usually 100 μg per lane) were diluted in sample buffer containing 0,6% SDS and 1% 2-mercaptoethanol, and then put it on 10% acrylamide/SDS minigel (Hoeffer Scientific). Each gel was applied pre-stained molecular weight standards (Sigma). Electrophoresis was performed using alternating current of 25 mA/gel at a temperature of 4oC. the Hybrid stages were carried out at room temperature. Membrane for 1 h were fixed with the help of immersion in PBS containing 10% skim milk. After this phase, conducted 3x5-minute wash in PBS containing 0.05% Tween-20 and 0.1% BSA (PBST). Added monoclonal antibody (clone C; Boehringer Mannheim, #1285-262); dilution of 1:200 (0.25 microgram/ml IgG) in PBST buffer; incubation for 2 h at room temperature, or 18 hours at a temperature of 4oC. In some experiments, a similar hybridization was performed using a control mouse IgG at the same concentration. After this stage was carried out 3x5 minute wash in PBST. Used a secondary antibody to mouse IgG attached to horseradish peroxidase; dilution 1: 2000 in PBST; incubation time 1 h; then held above the washing. Staining of immunoblots carried out at room temperature for 15-30 min using a freshly prepared solution of 4 - chloronaphthalen (0.2 mg/ml in TBS containing 16% ethanol) and H2O2(0,01%). Photographed colored filters (negative film Kodak TM) and scanned by a laser scanner LKB Ultro-Scan KL when the size of the vertical interval of 40 μm and the width of the horizontal run 2.4 mm Each track scanned three times at slightly different positions; predstavitelskogo method, we measured the number of BIA remaining in the fraction of cell membranes after incubation (usually 24 hours) in the presence of investigated compounds. Small amounts of BPA, remaining on the membranes after the specified incubation in the presence of agonist, was considered as evidence that a large part of the unit was secreted by the relevant cells. This method of accounting is acceptable in that case, if the assumed ability of muscarinic agonists to attenuate the accumulation of - amyloid in Alzheimer's disease because, in contrast to the secreted form of the unit is only associated with the membrane of the unit can lead to the formation of - amyloid. To measure the amount of BPA that is secreted by cells into the culture medium ("conditioned medium"), the above medium was concentrated with the use of microfiltration membranes (Amicon, delay molecules weighing more than 30 kDa). Focusing the considered environment, the received samples using the method of Bradford (Bio-Rad kit #500-0006) determined the amount of protein using gamma globulin bull as standard. Equal amounts of protein were examined by electrophoresis in a poly-acrylamide gel (EPAG). The amount of BPA in the samples were detected using Western blot turns with montelino stained standards (Sigma), applied to each gel.

Preferred are compounds corresponding to the formula AA and characterized by the highest rate of secretion of the unit cell cultures (125% compared with 100% of basal level). The number of compounds exhibiting activity of this kind include the following compounds: AF160(Des), AF179, AF185, AF261, AF263, AF265 AND AF267.

In order to analyze large number of samples simultaneously, some studies were carried out using the method of the dot-blot (simultaneously analyzed 96 spots on the same filter). In this case, the environment, air-stimulated cells RSM, was applied in a vacuum directly on nitrocellulose filters (without concentration). Eventually, these filters were treated in accordance with the methods of Western blot turns. In addition, through the separation of secreted BIA using EPG, conducted a detailed analysis of dependence "dose - response and time dependence. The results obtained in the experiments described above and a dot-blot tests are in good agreement with each other. Thus, new spiraeoideae were investigated in terms of their influence on the secretion of the unit cells transfected with the s as AF261, AF263, AF265 and AF267 are full agonists at the specified test.

Desensitization secretion BPA-mediated receptor m1AChR, is stronger in the case of prolonged incubation in the presence of the churches of Christ, rather than partial agonists provided by the present invention. This conclusion is particularly attractive when considering the data obtained on the desensitization signals, resulting in the release of arachidonic acid (AA), in combination with recently nominated by the assumption that the release of AK is involved in the stimulation of the secretion of the unit (Emmerling et al., BBRC, 197: 292-297 (1993)). Our data seems to indicate that some of the new helical compounds preferable to high-agonists for clinical use for a long-term reduction in the number associated with the cells of the unit. Among the compounds characterized by the lowest level of desensitization in the test for release of AK include compounds AF265 and AF267. The measurement of the amount of the unit associated with the cells after 24-hour incubation in the presence of compounds AF265 or AF267 (each at a concentration of 100 μm), showed a decrease in the level of the unit; and the decrease was more sillo weaker effect.

Data on the secretion of the unit cell cultures stably transfected with a gene m1AChR, clearly indicate that some of the new helical compounds can stimulate the secretion of the unit, and therefore help to reduce the accumulation of amyloid in vivo. However, in order to extrapolate data obtained in cell culture, on the situation in vivo, it is necessary to demonstrate the ability of these compounds to stimulate the secretion of BPA in the brain. In this regard, investigated the secretion of BPA in cortical slices of rat brain. Typically the cortical slices of rat brain (300x300 μm) was obtained immediately from svezhepriobretennoy fabric, cut along two perpendicular directions using a McIlwain Tissue Chopper and rinsed three times with Krebs buffer, saturated with oxygen. The washed sections were saturated specified buffer to the equilibrium state for 50 min at a temperature of 37oC. After the specified saturation (which is necessary to remove residual cells, formed during cutting fabric) is used, the buffer was replaced with Krebs buffer containing 50 μg/ml BSA as a carrier protein) and a mixture of protease inhibitors (0.1 mm PMSF, and 5 μg/ml leupeptin, pepstatin and Aprotinin). Processed crazy for 1 h (in one experiment - 2H) at a temperature of 37oC. At the end of the incubation period were pooled sample buffer, centrifuged, and analyzed for protein content, were taken equal aliquots of protein and investigated their method of Western blot turns with monoclonal antibodies to BIA (S) when using EPG (see experiments with cultured cells). Secreted BIA were represented by two protein bands, characterized by a molecular weight of 117 kDa (major band) and 90 kDa (minor band). The bands BPA can meet secretively forms BPA and BPA (main Kunitz-containing Kunitz-nestorgames forms BIA). In the alternative case, the bands can meet Mature and immature (deglycosylation) secreted forms BPA. There is currently no sufficient data to preference for one of the above interpretations, since monoclonal antibodies S interact in this test, all forms of secreted BIA. The secretion of the unit slices of the cerebral cortex of rats was increased compared with the control level of secretion of the unit 2-3 times during the action as the churches of Christ, and, for example, the compounds of AF267 (both at a concentration of 0.1 mm). This pattern was carried out for both bellowing brain, are unique. Currently, the only evidence of secretion BIA brain tissue in vitro (Nitsch et al., PNAS US, 90: 5191-5193 (1993)), it is shown that electrical stimulation of hippocampal slices of rats leads to increased release of BPA. It should be emphasized that in this study did not use stimulation with agonists of the receptors. Thus, the data obtained by us are original and new, because they are the first to demonstrate the secretion regulation of BPA in the brain tissue by receptor ligand, in particular such agonist of the M1 receptor, such as the joint AF267. Preferred are compounds capable of stimulating the secretion of the unit by cortical slices of rat brain. As such compounds can be considered all provided by the present invention agonists, stimulating secondary messengers (type PHI) more than 25% compared with control. These unique observations seems to indicate that some of the above compounds can slow the allocation A4-amyloid peptides in the brain of patients with Alzheimer's disease.

Test N. 6: Phosphorylation of the Tau protein in cell cultures, transferiria specificeski, associated with microtubule protein, normal axons. Tau protein is expressed as multiple isoforms, each of which is derived from alternative splicing of the same gene (Tau-human protein has 6 isoforms, consisting of 352-441 amino acids). The function of the Tau protein is to stabilize microtubules axon of nerve cells; the specified binding protein with microtubules is regulated by phosphorylation at different sites (Mandelkow &Mandelkow, TIBS, 18: 480-483 (1993)). In turn, this phosphorylation regulates the growth and stability of the axon (Baas et al., J. Cell. Bi-ol. 115: 1333-1344 (1991). 55. Mattson, Brain Res. 582: 107-118 (1992)). Along with the release of amyloid in plaques, one of the distinctive features of Alzheimer's disease is the accumulation of the Tau protein or its derivatives in neurofibrillary knots (NFU). The presence of such sites probably reflects the end product resulting from the death of nerve cells in the affected nervous tissue. Several studies have shown changes in the phosphorylation of the Tau protein in brain samples taken after death in patients suffering from AD. This fact is best documented by increased immunoreactivity with monoclonal PNAS US, 87: 4840-4844 (1990); Vincent & Davies, Brain Res. 531: 127-135 (1990)). In this regard, it was suggested that changes delicate balance between Tau-specific kinases and phosphorylases can lead to the death of nerve cells in ad, and therefore the correction of such hypothetical imbalance may have significant therapeutic value.

In line with this, we analyzed the degree of phosphorylation of Tau protein in cultured cells, stably transformed genome m1AChR, after stimulation with the churches of Christ or one of the investigated compounds. The specified analysis was performed using monoclonal tel tau-1, which shows their ability to learn only dephosphorylation and non-phosphorylated isoforms of the Tau protein (Mandelkow &Mandelkow, TIBS, 18: 480-483 (1993)). After incubation of the cells or cortical slices of rat brain in the presence of appropriate agonists for different periods of time, these cells or three slices were washed in PBS, were collected in PBS containing 0.2 mm EDTA, and centrifuged for 5 min at 10000 g. The obtained precipitation membranes were analyzed for protein content, equal amounts of protein were used in EPG for Western blot turns with antibodies tau-1. Measurement immuno-reactives is to peroxidase secondary antibody was detected using the method of increasing Chemoluminiscence (UHL) (RPN-2109; Amersham, UK).

It should be emphasized that the churches of Christ, and studied agonists increased the immuno-reactivity against tau-1, the specified gain is completely blocked by atropine. A sharp increase in immuno-reactivity for antibodies tau-1, called the churches of Christ, was observed in cells cultured in the presence of NGF (50 ng/ml) for 3 days before stimulation of muscarinic ligands. As shown by similar experiments conducted using different concentrations of the churches of Christ or investigational agonist, to enhance the immuno-reactivity against tau-1 requires relatively high concentrations of ligand (10-100 μm).

In addition, we measured the degree of phosphorylation of the Tau protein in cortical slices of rat brain during stimulation of the churches of Christ in comparison with the activity of the compounds provided by the present invention. Investigated muscarinic agonists again was suddenly able to reduce the degree of phosphorylation of the Tau protein.

So, muscarinic agonists can reduce the degree of phosphorylation of the Tau protein through activation of receptor m1AChR in cultured cells, transfected with the corresponding gene, and, more importantly, in the slices of the brain. In addition, castiello in the presence of 100 μm of ligands for at least 3 days. These facts may indicate that selective agonists of M1 receptors are able to reduce the phosphorylation of the Tau protein.

These observations are new, as far as we know, so far not demonstrated any relationship between cholinergic deficiency in Alzheimer's disease and the accumulation and phosphorylation isoforms of the Tau protein in neurofibrillary knots, typical of brain suffering from the specified disease. These new facts can be useful in slowing the progress of Alzheimer's disease using agonists of M1 receptors.

Conclusions:

Agonists of m1 receptors can have different effects. Thus, the mechanism of action of agonists of m1 receptors unexpectedly turned out to be more complex than the traditional view. It is shown that agonists of m1 receptors characterized by the following activity in vitro (or in vivo):

1. The binding of m1 receptors and their activation;

2. Differential activation of some G-proteins (in particular, Gq, but not Gs;

3. Necrotroph-like and energenie effects in joint action with NEF;

4. Secretion of the protein-amyloid precursor (BIA), as well as oslablen CLASS="ptx2">

All of these effects can be explained by a model in which activation of the receptor m1AChR leads to a cascade of similar events. Such complex mechanisms based on the use of agonistic activity towards the m1 receptors may be associated with a common scenario involving abnormal processing of the unit, the shortage of NGF-like proteins, as well as the lack of holinolitikov when SSTA and BA. Accordingly, agonists of m1 receptors, similar to the compounds provided by the present invention, causing selective and positive activation of the secretory pathway of the processing unit, may be important in preventing the formation of amyloid; in addition, thanks to cinergena effect produced together with NGF, these compounds may enhance the action of neitrofilov at BA. Thus, agonists of m1 receptors may be applicable when using the strategy of replacement of holinolitikov, as well as in delaying the progress of ad. Provided by the present invention compounds that are agonists of m1 receptors, unlike peptides are relatively small molecules that can be neutrope-like effects, to cause visuo Tau proteins. Thus, treatment of cells with agonists of m1 receptors apparently can switch the processing of the unit with amyloidogenic lysosomal path on the normal secretory pathway, and the corresponding cholinergic therapy BA may have more prolonged effects on the formation of - amyloid (Lahiri et al., Biochem. Int. 28: 853-860 (1992)). Moreover, as some agonists provided by the present invention, showing NGF-like effects in a controlled way, especially in the presence of NGF, these compounds may have significant value in the treatment of BA/STA in the future. In this case, axonal growth can be controlled better. In addition, new spiraeoideae have a lasting beneficial effect on axonal growth in the presence of NGF. Thus, it can be assumed that the possible future treatment of patients with BA/STA, using these growth factors like NGF, may require less frequent application of a growth factor with its introduction together with the agonist of the m1 receptor. It should be noted that the introduction of NGF in the brain is the most complex problem because the specified connection is not able to cross the blood vessels. which of NGF-like compounds will be still necessary. Therefore, agonists of m1 receptors can reduce the number of required repeated applications NEF, because these agonists prolong the effects resulting from NEF (at least in vitro). Interestingly, clinical trials of NGF (icv), held on 2 patients showed the presence of serious side effects (severe pain, disorientation, disturbance of oral accounts) (3rd Springfield Symposium on Aizheimer's Desease, May 11 - 15, 1994, Springfield, Il., USA). The use of agonists of m1 receptors, similar to the compounds provided by the present invention, and is able to provide synergenic effect in a joint application with NGF, may be of great importance to reduce the side effects caused by the introduction of human NGF, with the ability to use smaller doses of NGF.

Test No. 7. Pharmacological and Toxicological properties.

For the analysis of observed animals that intravenous or oral (in the case of mice), or orally (intragastrically in the case of rats) was administered 3-6 dose levels of each compound. The results are shown in Table 5.

Through various time intervals after injection connection(10, 20, 30, 45, 60, 120, 240 min and 24 h) of obrabotannykh. After 24 h after injection of the tested compounds were considered the viability of experimental animals. The body temperature of the rats was measured using tele-thermometer (Model 46 TUC). The analyzed pharmacological and behavioural parameters were: salivation, redness around the nose and mouth, chromodacryorrhea, the degree of tranquility, ataxia, poisoning, tremor, convulsions, decreased temperature, opitonui, difficulty breathing, diarrhea, the constant movement of the jaws, the voltage of the intestine, mortality, changes in the pupil diameter, turning, Hypo - and hyperactivity, as well as izdavanje sounds. Some of the tested compounds were non-toxic at concentrations up to 500 mg/kg (in mice and rats; oral).

Discussion of results:

Connection AF160

Connection AF160 is a fairly effective muscarinic agonist, affecting to a greater extent in the Central than in the peripheral nervous system (for example, causes a lower temperature and not saliva), as well as at least 5 times less toxic than compound AF102B. A characteristic feature of the compounds is the absence of tremor in rats, itli at doses > 125 mg/kg orally. Similar features demonstrated for methylated analogue connection AF160 (AF178), the introduction of which, in doses up to 500 mg/kg (oral) not experience any tremors neither rats nor mice. The mouse is more sensitive to this compound than rats, as side effects on the peripheral and Central nervous system occur in mice at lower doses.

In rats connection AF160 manifests itself as an effective agonist. Clear evidence of this was found for doses of 25 mg/kg For the effect of low temperature and permanent Poglazov were calculated corresponding values ED50. The duration and severity of symptoms was dose dependent. The effect of temperature decrease was long, more than 4 hours

To connect AF160 characteristic 4 features:

1. The connection clearly shows greater activity against CNS than PNS (in particular, the temperature drop occurs at a lower dose than salivation).

2. There is a selectivity in relation to the Central nervous system, because there are not all associated with CNS effects (in particular, the temperature drops, but no tremors).

3. The duration of obnaruzhenny the

Connection AF160(Des)

One of the four experimental rats after 45 min after administration of 125 mg/kg detected salivation, lowering the temperature, as well as diarrhea. When increasing doses up to 500 mg/kg increased the number of animals in the group exhibiting these symptoms, in addition, there was a noticeable tension of the intestine and complacency. The effect of temperature decrease was not dose dependent. Consider the compound did not cause any other CNS-mediated or Autonomous effects. This connection is relatively inactive in terms cholinergic side effects. In the case of compound AF160(Des) side effects occur when using doses > 246 mg/kg (orally in rats).

When administered to mice 240 mg/kg connection AF160(Des) observed lacrimation, diarrhea and mydriasis. When using large doses up to 1000 mg/kg clearly manifested more symptoms: salivation, complacency, and lowering the temperature. The duration and severity of these symptoms (except for watery eyes) was dose dependent. Even at the highest dose levels (1000 mg/kg) did not observe mortality. Local injection of 1 mg (threshold level) connection AF160(Des) in the eye caused mydriasis after 15 minutes inogo mediastinoscope effect, induced mydriasis after 45 min with the introduction of 40 µg. The data suggests that the induced connection AF160(Des) mydriasis caused by the effects of the Central nervous system.

Connection AF163

The considered connection, administered at doses from 50 to 400 mg/kg in the rat caused only salivation, as well as redness around the nose and mouth. These effects quickly appeared (after 10 min) and were of short duration (20 min). At that time, as the number of animals exhibiting salivation, which was dose dependent, redness around the nose and mouth was only observed in 1 animal when using doses of 100 mg/kg Thus, with the introduction of the specified connection was observed only Autonomous effects.

Mice are considering the connection did not cause any side effects when used at the highest investigated dose (400 mg/kg), except for 1 animal, uttering sounds. When administered to mice connection AF163 up to the highest investigated dose (400 mg/kg orally) did not observe any side effects associated with Central or peripheral nervous system. Connection AF163 can be considered as a predecessor of the medicinal agent AF160, as in the case of dithio-analog R>
After 10 min after injection mice 31 mg/kg was observed the decrease of temperature, underactive, and tremors. When the dose is increased to 62 mg/kg increased and the number of symptoms. After 20 min for all 4 mice were marked complacency, ataxia, Straub-tail and downs from pivoting. After 30 min after administration of 1 out of 4 mice death. When the dose is increased to 125, 250 and 500 mg/kg was observed salivation, convulsions, and death all 4 mice, however, the time of onset of these symptoms progressively reduced, and the severity increased. The calculation of the values of the ED50showed that the connection AF177 mainly affects the Central nervous system. Connection AF177 can be considered as a predecessor of the medicinal agent, active against the Central nervous system and is effective muscarinic agonist.

Connection AF178

Within 1 h after injection mice 60 mg/kg of the compounds mentioned reduced motor activity, salivation, lacrimation, and diarrhea. When increasing doses up to 500 mg/kg was shown additional symptoms: shortness of breath, poor turning, tremors, and mydriasis. The duration and severity of most of these symptoms depended on to whom. what was prodolzhalis for about 4 hours it Should be emphasized that within 20 min after administration of 500 mg/kg of one of the animals showed a General tremor, which lasted for 10 minutes Despite the fact that the dose of 500 mg/kg caused a typical Toxicological symptoms, not marked by the death of animals. Thus, the estimated value of the ED50exceed the level of 500 mg/kg

Salivation was the only side effect identified in the rat at 62.5 mg/kg connection AF178. When increasing doses up to 125 mg/kg manifest and other effects: chromodacryorrhea, lowering the temperature, diarrhea and permanent pogryzi. The duration and severity of these symptoms increased with increasing dose. Shortness of breath was clearly only 1 animal in 5 min after injection of the dose of 500 mg/kg of the Considered connection is characterized by a rather wide limits of safe use.

Connection AF180

Connection AF180, administered at a dose of 125 mg/kg in the rat caused only salivation. This effect was evident after 20 min after application and continued for 40 minutes Further increase the dose to 500 mg/kg resulted in growth duration slushees from 15 to 240 min after injection. This dose did not cause any other mediated Central nervous system or autonomic effects.

After 20 min after injection mice 125 mg/kg was observed the decrease of temperature, and mydriasis. When increasing doses up to 500 mg/kg increased the number of animals showing these symptoms, in addition, clearly identified salivation, lacrimation and complacency. With the introduction of 500 mg/kg these symptoms lasted for more than 1.5 hours of the Considered compound did not cause any other mediated Central nervous system or autonomic effects.

Connection AF185

When administered to mice at 62.5 mg/kg connection AF185 watched mydriasis. At higher doses the specified connection up to 500 mg/kg clearly identified additional symptoms: salivation and a decrease in temperature. The highest dose (500 mg/ kg) did not cause death.

In rats, the highest dose of the compounds being considered (500 mg/ kg) did not cause any visible effects except for the redness around the nose and mouth, observed in 1 animal. It can be assumed that the connection AF185 is highly safe.

Connection AF261

The decrease in temperature (typical effect mediated CNS) was the unity of the rez 10 min after injection and lasted for 50 minutes When the dose is increased to 15.5 mg/kg in 10 min after injection were noted additional symptoms: salivation, chromodacryorrhea, underactive, ataxia and diarrhea. When using higher doses up to 250 mg/kg clearly manifested more symptoms: redness around the nose and mouth, tremors, convulsions, opistotonus, the constant movement of the jaws and the voltage of the intestine. These effects are observed in 5-10 min after injection. The severity and duration of most of these symptoms was dose dependent. After 45 minutes after administration of the highest dose (250 mg/kg) was marked by the death of all 4 rats.

Connection AF265

The decrease in temperature (typical effect mediated CNS) was the only symptom caused in rats by the introduction of 250 mg/ kg compound AF265. This effect became noticeable after 10 min after injection and lasted for 110 minutes When using the specified connection was not observed in any other mediated Central nervous system or autonomic effects. Consider the connection characterized by the action only on the Central nervous system.

Test N. 8; connection AF134 in untrained rats

Applying passive walking test avoidance, investigated the influence of described in (Fisher et al., Neurosci Lett. 102: 325 (1989)). 4 groups (20 rats in each group) untrained 3-4-month-old male rats Sprague-Dawley, weighing 200 - 300 g (Charles River Breeding, UK) was treated with one of the following doses connection AF134: 1, 5, 10 mg/kg intraperitoneal injection (C. B.), while one group received saline (1 ml/kg, V. 6.).

In rats treated with compound AF134, there were no significant deviations in terms of hidden reflex save with prior training shock if the specified connection was administered for 30 min or 60 min after the shock.

In another experimental model of action connection AF134 compared with the effects produced by scopolamine (anti-muscarinic connection), untrained rats in the 8-beam radial maze (Fisher et al., Neurosci Lett. 102: 325 (1989)). A group of 14 untrained rats were treated with scopolamine (0.2 mg/kg, W. B.), either saline (1 ml/kg, B. B.) for 20 minutes before beginning the test. All beams were bait. Each rat was exposed to both types of processing a three-day intervals between treatments. (After 10 days of training followed 10 days of testing.) On the same rats carried out a similar experiment using connection AF134 (5 mg/kg, century 6. the "anticosti" effect. However, the connection AF134 not led to any changes in the behavior of rats.

Connection AF134, showing antagonistic activity against muscarinic m1 receptors (judging by the analysis of binding), and m3 (judging by the preparations of ileum Guinea pigs), does not lead to impaired cognitive effects and, thus, may be suitable for the treatment of low mobility, Parkinson's disease, mixed Parkinson's disease and Alzheimer's disease, manic-depression, the effects of traumatic brain surgery person, as well as for the treatment of acute rhinitis, peptic ulcers and asthma in various disorders of the peripheral nervous system.

Test N. 9; Analysis of behavioral effects in animal models

Connection AF160 and AF102B - Test in a radial radial maze

Investigated the possible beneficial effects of doses of the compounds AF160 (3 and 5 mg/kg orally) and AF102B (3 mg/kg orally) on the treatment of memory disorders in rats injected connection AF64A (1.5 nm/2 µl per side). Described animal model to some extent resembles the cholinergic hypofunction observed in SST (Fisher et A1., J. Pharmacol. Expl. Therap. 257: 392-403 (1991)). In specified edenesque test was 2-3 months. For 1 week prior to the behavioral test, the rats were transferred to individual cages and were limited in the diet until they reach approximately 85% by weight, characteristic of free feeding. Then the rats received 5-6 pieces Altromin (15 g) per day to maintain weight at the same level. Rats had free access to water. The room was illuminated for 12 h per day (6:00-18: 00), behavioral tests were performed in the morning.

Behavioral test

2 groups of 40 rats were injected with respectively the connection AF64A and saline solution, after which each group was randomly subdivided into 4 subgroups, which were treated with compounds AF160 (3 mg/kg), AF160 (5 mg/kg), AF102B rats to the specified labyrinth, as well as to regularly enter the bait (45 mg). At this stage, rats were placed in the Central part of the maze and gave them free access to all 8 beams with bait. Each series was completed with the collection of all 8 lures, or after 15 minutes, depending on what event has happened before. During the 3rd and 4th days the bait was placed at the end of the rays. All other procedures were similar to that used in the initial training phase. During 5 days for 60 minutes before nyh

Noted all move through the maze, the time spent, as well as correct and incorrect responses. In order to evaluate the effect of a connection AF160 or AF102B during the test period, compared to the training period (which took over the baseline level of behavior), used a 3-dimensional factor analysis (HH) with the presence of repetitive variation (successive blocks of training days and days of testing), as well as non-recurring variation (injection connection AF64A or saline solution, or treatment with different doses of compounds AF160, AF102B or DDW). The comparison was performed according to the difference between the main effects.

Results

When analyzing the correct choice of 8 visits found a reliable relationship between the parameters of the group x treatment x week [F(2/48) = 3,95; P < 0,025] . Specifically, for the training and use of the medicinal agent, rats, injected connection AF64A, made significantly fewer correct election than rats injected with saline (P < 0,001). Both medicinal agent: connection AF102B (3 mg/kg), and the connection AF160 (3 mg/kg) improved the behavior of rats injected connection AF64A, during the course the group of rats treated with compounds AF160 (3 mg/kg) and AF102B (3 mg/kg), showed the same level of behavior, significantly exceeding those in the case of rats injected connection AF64A, but the treated water (P < 0,05). Connection AF160 at a concentration of 5 mg/kg had no reliable effect on this parameter. Compared with rats injected with saline, the behavior of rats, treated water, improved (5%) in the second week compared with the first week (P < 0,01). Similar improvement (7.5%) was also noted in the case of rats treated with compound AF160 (3 mg/kg; P < 0,001). Connection AF102B (3 mg/kg) had no effect on this parameter in rats injected with saline.

When analyzing full of errors detected significant Association between group x treatment x week [F(3/64) = 3,49; P < 0,025]. Within two weeks the number of total errors made by rats, injected connection AF64A, were significantly higher than the corresponding figure in the case of rats injected with saline (P < 0,001). The number of errors in all four groups of rats injected connection AF64A, was significantly decreased in the second week compared to the t; 0,001). Despite the improvement in the behavior of all four groups, only using the connection AF102B (3 mg/kg) behavioral indicators exceeded those in the case of the use of the compounds of AF64A in combination with water. Among rats injected with saline, using the connection AF160 (3 and 5 mg/kg) resulted in a significant improvement in the behaviour in the second week compared with the first week (P < 0,01; P < 0,001; respectively). Application connection AF102B caused behavioral disorders, this group of rats made more errors in the second week compared with the first week (P < 0,001).

In the analysis of time spent and found a significant relationship between the parameters of the group x treatment x week [F(2/48) = 3,29; P < 0,05]. The temporary improvement of performance was observed in the case of subgroups, the injected compound AF64A (except for option processing AF64A + AF160 [5 mg/kg]): AF64A + water - 31% (P < 0,01); AF64A + AF160 (3 mg/kg)- 47% (P < 0,001), as well as AF64A + AF102B (3 mg/kg) and 21% (P < 0,01). In the case of rats injected with saline, showed a significant improvement of temporal parameters in the following treatment options: physiologic solution + water - 54% (P < 0,001); physiologic solution + AF160 (3 mg/kg) - 37% (P < 0,001). Sedimentate "threshold effect". Despite a slight downward trend, the connection AF160 (5 mg/kg) did not cause significant changes in the behavior of rats injected with saline).

Conclusions

1. Rats injected connection AF64A (1.5 nm/2 µl per side) showed significant violations in terms of the proper selection, number of errors, and also spent time as compared with rats injected with saline.

2. Connection AF160 (3 mg/kg) resulted in a significant improvement in the behaviour of rats injected connection AF64A (compared with the control treatment) in terms of a correct choice, and it takes time. A higher dose (5 mg/kg) was effective only in relation to the number of errors compared with the basal level (but not control).

AF160(Des) Test in the maze Morris Water (LMW)

The purpose of this test was to use the test in LMU (Fisher et al., J. Pharmacol. Expl. Therap. 257: 392-403 (1991)) to study the ability of the compounds AF160 (Des) to pay violations cognitive activity in rats injected connection AF64A. Connection AF160(Des) was applied in two doses: 1 and 3 mg/kg orally. Two groups of rats (38 and 42 animals) were injected with, soo the emergency was subdivided into 4 subgroups, who have been treated with different doses of compounds AF160(Des) (1 or 3 mg/kg orally) or DDW (10 ml/kg orally). Medicinal agent once daily was applied for 60 min before testing within 5 days.

When considering behavior there were no significant side effects. Thus we obtained the following results:

1. According to the analysis of parameters such as latency avoidance and path length, injection connection AF64A (3 nm/2 µl per side) resulted in a significant breach of conduct.

2. Connection AF160(Des) (3 mg/kg) improved behavioral performance in the case of rats, injected as a connection AF64A and saline, in the third block of training.

2. Connection AF160(Des) (3 mg/kg) improved behavioral performance in the case of rats, injected as a connection AF64A and saline, in the third block of training.

A positive result suggests the possibility of testing in the future of different doses of the compounds to study their favorable effects in relation to impaired learning and memory.

Connection AF185 - Test passive avoidance s of passive avoidance response in rats processed by the connection AF64A.

Methods

Connection AF64A were injected with bilateral (3 nm/2 µl per side, icv) to create the original animal model (Fisher et al., J. Pharmacol. Expl. Therap. 257: 392-403 (1991)). Four weeks after this operation, two groups of rats, injected respectively connecting AF64A and saline was randomly subdivided into 4 subgroups according 10-11 rats; three subgroups were treated with various concentrations of compounds AF185 (1, 5 and 10 mg/kg orally), while one group received DDW (10 ml/kg). Connection AF185 was introduced directly after the shock, and after 72 h tested rats. The description of the details of this technique see (Fisher et al., J. Pharmacol. Expl. Therap. 257: 392-403 (1991)).

Results

The assessment of the initial and residual latency was performed using a 2-dimensional factor analysis (2x4): injection connection AF64A or saline solution in relation to the dose of a compound AF185 or DDW.

In terms of the source of the latency between any two groups was no significant difference.

In terms of residual latency revealed a relationship between groups and the nature of the processing (F(3/75) = 22,31; P < 0,001). On the basis of the simple differences between the eating AF64A, shorter than typical for rats injected with saline and processed DDW (P < 0,001). The residual period of latency in rats treated with compound AF185 (1,5, or 10 mg/kg) was significantly shorter compared with rats treated with compound AF64A + DDW (P < 0,001). It was not revealed any other significant differences.

Conclusions

1. Rats injected connection AF64A, find significant violations of the residual period of latency compared with rats injected with saline.

2. Rats injected compound and treated with AF64A connection AF185 (1.5 and 10 mg/kg) are characterized by the retention reactions, similar to those in the case of control animals.

3. Connection AF185 may be a compound effective in the treatment of memory disorders, such as those found in the case of Alzheimer's disease.

For specific compounds provided by the present invention, observed the following relationship between the used numbers of these compounds and their names in accordance with the systematic IUPAC nomenclature:

2,8-dimethyl-1-oxo-1-TSS-4, 8-diaza-Spiro[4.5]decane AF264;

3,8-dimethyl-1,4-dioxa-8-Aza-Spiro[4.5]Decan-3-one AF274;

2-ethyl-4,8-dimethyl-1-thia-4, 8-diaza-Spiro[4.5]Decan-3-one AF272;

3-methyl-1-oxo-4-thia-8-Aza-Spiro[4.5]decane-2-he AF269;

3-ethyl-8-methyl-1-oxo-4-thia-8-Aza-Spiro[4.5]decane-2-he AF262;

2-methyl-1-thia-4,8-diaza-Spiro[4.5]Decan-3-one AF263;

3,8-dimethyl-1-oxo-4-thia-8-Aza-Spiro[4.5]decane-2-he AF265;

2,8-dimethyl-1-oxo-4,8-diaza-Spiro[4.5]Decan-3-one AF260;

2,4,8-trimethyl-1-thia-4,8-diaza-Spiro[4.5]Decan-3-one AF266;

2-ethyl-8-dimethyl-1-thia-4,8-diaza-Spiro[4.5]Decan-3-one AF267;

2,8-dimethyl-1-thia-4, 8-diaza-Spiro[4.5]Decan-3-one AF261;

2,8-dimethyl-1-oxo-3,8-diaza-Spiro[4.5]Dec-2-EN-4-one AF238;

2,8-dimethyl-1,3,8-diaza-Spiro[4.5]Dec-1-EN-4-one AF230;

3-ethyl-8-methyl-1,3,8-diaza-Spiro[4.5]Dec-1-EN-4-one AF220;

1-ethyl-8-methyl-3-oxo-4,8-diaza-Spiro[4.5]decane-2-he AF174;

8-methyl-1-oxo-3,8-diaza-Spiro[4.5]decane-2-tion AF165;

3-ethyl-8-methyl-1-oxo-3,8-diaza-Spiro[4.5]decane-2-he AF172;

8-methyl-1-oxo-3,8-diaza-Spiro[4.5]decane-2,4-dione AF169;

3-ethyl-8-methyl-1-oxo-3,8-diaza-Spiro[4.5]decane-2,4-dione AF180;

4 butylamino-8-methyl-1,3,8-triaza-Spiro[4.5] DECA-1,3-diene-2,4-diamine AF194;

methyl-(8-methyl-2-methylsulfanyl-1,3,8-triaza-Spiro[4.5] DECA-1,3-Dien-4-yl)-amine AF193;

8-methyl-2-methylsulfanyl-1,3,8-triaza-Spiro[4.5] DECA-1,3 - Dien-4-ia-Spiro[4.5]Dec-1-ene AF190;

3-ethyl-2-ethylsulfanyl-8-methyl-1,3,8-triaza-Spiro[4.5] Dec-1-EN - 4-tion AF170;

3-ethyl-2-ethylsulfanyl-8-methyl-1,3,8-diaza-Spiro[4.5] Dec-1-EN - 4-one AF188;

8-methyl-2-methylsulfanyl-1,3,8-triaza-Spiro[4. 5]Dec-1-EN-4-one AF187;

8-methyl-2, 4-bis-methylsulfanyl-1,3,8-triaza-Spiro[4. 5] DECA-1,3-Dien AF177;

8-methyl-4-methylsulfanyl-1,3,8-triaza-Spiro[4. 5]Dec-3-EN-2-tion AF183;

4-ethylsulfanyl-8-methyl-1,3,8-triaza-Spiro[4.5]Dec - 3-EN-2-tion AF176;

3-ethyl-8-methyl-1,3,8-triaza-Spiro[4.5]decane-2,4-dition AF163;

8-methyl-1,3,8-triaza-Spiro[4.5]decane-2,4-dition AF173;

1-acetyl-8-methyl-1,3,8-triaza-Spiro[4.5]decane-2,4-dione AF164;

3-methyl-1,3,8-triaza-Spiro[4.5]decane-2, 4-dione AF179;

3,8-dimethyl-1,3,8-triaza-Spiro[4.5]decane-2, 4-dione AF178;

3-ethyl-1,3,8-diaza-Spiro[4.5]decane-2,4-dione AF160(Des);

3-ethyl-8-methyl-1,3,8-triaza-Spiro[4.5]decane-2,4-dione AF160;

3-ethyl-8-methyl-4-thioxo-1,3,8-triaza-Spiro[4.5]decane-2-he AF182;

8-methyl-3-(4-pyrrolidin-1 - yl-but-2-inyl)-1,3,8-triaza-Spiro[4.5] decane-2,4-dione AF197;

3-prop-2-inyl-1,3,8-diaza-Spiro[4.5]decane-2,4-dione AF186;

3-tert-butyl-8-methyl-1,3,8-diaza-Spiro[4.5]decane-2,4-dione AF213;

8-methyl-3-prop-2-inyl-1,3,8-triaza-Spiro[4.5]decane-2,4-dione AF185;

3-tert-butyl-8-methyl-2-thioxo-1,3,8-triaza-Spiro[4. 5]Decan-4-one AF184;

3-ethyl-8-methyl-2-thioxo-1,3,8-triaza-Spiro[4.5]decidsion-4-one AF195;

5-methyl-2-(1-methyl-piperidine-4-yl)-thiazolidin-4-tion AF275;

3-but-2-inyl-8-methyl-1,3,8-triaza-Spiro[4.5]decane-2,4-dione AF199.

Although the present invention is described in detail in this application using special links directly to these and other implicit methods of embodiments of the present invention, a specialist in this field must understand the numerous variations and modifications. Accordingly, the present invention is not restrictive and should not be limited to directly listed options embodiments, if only the ideology, the scope and essence of the present invention correspond to the following claims.

1. Spiraeoideae General formula (AA):

< / BR>
including their pharmaceutically acceptable salts, enantiomers, racemates and Quaternary compounds, representing derivatives of these compounds containing a tertiary nitrogen atom, which contain a five-membered group, in which spirostane is implemented by the carbon atom included as part of the five-membered groups, and the composition of the saturated ring system containing one nitrogen atom, and the specified five-membered on, 2,4-dirigeante, 2-thiohydantoin, oxazolidin-2-tion, 3-ethyloxazole-2-it, oxazolidin 2,4-dione, 3-ethyloxazole-2,4-dione, 2-methyl-1,4-oxazolidin-3-one, 2-methyl-1,4-thiazolidin-3-one, 2,4-dimethyl-1,4-thiazolidin-3-one, 2-ethyl-1,4-thiazolidin-3-one, 2-ethyl-4-methyl-1,4-thiazolidin-3-one, 3-methyl-1,4-oxathiolan-2-it, 2-ethyl-1,4-thiazolidin-3-one, 5-methyl-1,3-oxazolidine, 4-ethyl-1,3-oxazolidine, 3-ethyl-1,4-oxathiolan-2-it, 5-methyl-1,3-dioxolane-4-one, N-methylsuccinimide, N-ethylsuccinate, 3-tert-Butylimino, 3-(4-pyrrolidino-2-butynyl)-as 3-(2-butynyl-)-as 2,5-bis(methylthio)-4H-imidazole, 3-ethyl-4-thiohydantoin, 4-methylthiazole-2-tion, 3-ethyl-2,4-dirigeante, 4-ethylthio-3-imidazolin-2-tion, 1-ethyl-2-ethylthio-2-imidazolin-5-tion, 2-thio-4-hydroxyethylaminophenol, 2,5-bis(aminomethyl)-4H-imidazole, 2-methyl-2-thiazolin, 2-methyl-2-imidazoline, 2-methyl-2-oxazoline-4-one, 2-methyl-4H(5H)-imidazole-5(4) -, 2-methylthio-5-methoxy-4H-imidazole, 2-methylthio-5-amino-4H-imidazole, 2-methylthio-5-aminomethyl-4H-imidazole, 2-tion-3-utilizaton, 2 tion-3-tert-Butylimino, 2-methylthio-2-imidazolin-5(4) -, 1-ethyl-2-ethylthio-2-imidazolin-5-Oh and 1-ethyl-2-imidazolin-5-he indicated the saturated ring system containing one nitrogen atom, selected from a number of groups, including groups K, M, N

or 5; m is 2, n and p independently of one another are equal to 1, and n + p = 2; R1denotes a group selected from among the following: a hydrogen atom, a C1-6-alkyl, C2-6-quinil.

2. Spirochaetaceae under item 1, in which the specified saturated ring system containing one nitrogen atom, selected from the group comprising piperidine, 1-methylpiperidine, 1-propolypeptide, N-methylnitramine and Hinkley.

3. Spirochaetaceae under item 1 or 2, wherein it is chosen from the group consisting of:

1 methylpiperidin-4-Spiro-4' -(2',5' -bis(methylthio)-4H-imidazole)

1 methylpiperidin-4-Spiro-5'-(3'-ethyl-4'-thiohydantoin),

1 methylpiperidin-4-Spiro-5'-(4'-methylthioadenosine-2'-tion),

1 methylpiperidin-4-Spiro-5'-(3'-ethyl-2',4'-dirigeante),

1 methylpiperidin-4-Spiro-5'-(4'-ethylthio-3'-imidazolin-2-tion),

1 methylpiperidin-4-Spiro-4'-(1'-ethyl-2'-ethylthio-2'-imidazolin-5'-tion),

1 methylpiperidin-4-Spiro-5'-(2'-thio-4-hydroxy-etilaminopodandov),

1 methylpiperidin-4-Spiro-4'-(2',5'-bis(aminomethyl)-4H-imidazole)

1 methylpiperidin-4-Spiro-5'-(3'-ethyloxazole-2'-he),

1 methylpiperidin-4-Spiro-4'-(2'-methyl-2'-thiazolin),

1 methylpiperidin-4-Spiro-4'(5')-(2'-methyl-2'-imidazolin),

1 methylpiperidin-4-Spiro-5'-(2'-methyl-2'-oxa is'-methylthio-5'-methoxy-4 N-imidazole)

1 methylpiperidin-4-Spiro-4'-(2'-methylthio-5'-amino-4 N-imidazole)

1 methylpiperidin-4-Spiro-4'-(2'-methylthio-5'-aminomethyl-4 N-imidazole).

4. Spirochaetaceae under item 1 or 2, wherein it is chosen from the group consisting of,

1 methylpiperidin-4-Spiro-5'-(3'-utilization),

1 methylpiperidin-4-Spiro-5'-(1'-acetylimino),

piperidine-4-Spiro-5'-(3'-utilization),

1 methylpiperidin-4-Spiro-5'-(3'-methylhydantoin),

piperidine-4-Spiro-5'-(3'-methylhydantoin),

1 methylpiperidin-4-Spiro-5'-(3'-propargylglycine),

N-methylnitramine-3-Spiro-5'-as

N-methylnitramine-3-Spiro-5'-(3'-methylhydantoin),

N-methylnitramine-3-Spiro-5'-(3'-methylhydantoin),

1 methylpiperidin-4-Spiro-5'-(2', 4'-dirigeante),

1 methylpiperidin-4-Spiro-5'-(2'-thiohydantoin),

1 methylpiperidin-4-Spiro-5'-(oxazolidin-3'-tion),

1 methylpiperidin-4-Spiro-4'-(3'-ethyloxazole-2', 4'-he),

1 methylpiperidin-4-Spiro-5'-(oxazolidin-2',4'-dione),

1 methylpiperidin-4-Spiro-5'-(3'-ethyloxazole-2',4'-dione),

1 methylpiperidin-4-Spiro-5'-(2'-methyl-1',4'-oxazolidin-3-one),

1 methylpiperidin-4-Spiro-5'-(2'-methyl-1',4'-thiazolidine-3'-Oh),

1 methylpiperidin-4-Spiro-5'-(2',4'-dimethyl-1',4'-thiazolidine-3'-Oh),

1 methylpiperidin-4-Spiro-5'-(2 Spiro-5'-(3'-methyl-1',4'-oxathiolan-2'-he),

piperidine-4-Spiro-5'-(2'-methyl-1',4'-thiazolidine-3'-Oh),

1 methylpiperidin-4-Spiro-5'-(3'-methyl-1',4'-oxathiolan-2'-he),

1 methylpiperidin-4-Spiro-2'-(5'-ethyl-1',3'-oxazolidin),

1 methylpiperidin-4-Spiro-2'-(4'-ethyl-1',3'-oxazolidin),

1 methylpiperidin-4-Spiro-5'-(3'-ethyl-1',4'-oxathiolan-2'-he),

1 methylpiperidin-4-Spiro-5'-(2'-methyl-1',4'-thiazolidin -3'-he),

2 methylpiperidin-4-Spiro-5'-(2'-ethyl-1',4'-thiazolidin -3'-he),

1 methylpiperidin-4-Spiro-2'-(5'-methyl-1', 3'-dioxolane-4'-he),

1 methylpiperidin-4-Spiro-5'-(2'-methyl-1', 4'-thiazolidine-3'-tion),

2-N-methylspiro-(1,3-succinimide-4,3')Hinkley,

2-N-utilisera-(1,2-succinimide-4,3')Hinkley.

5. Spirochaetaceae under item 1 or 2, wherein it is chosen from the group consisting of:

1 methylpiperidin-4-Spiro-5'-(3'-(4-pyrrolidino-2-butynyl)-as),

1 methylpiperidin-4-Spiro-5'-(3'-tert-Butylimino),

1-propolypeptide-4-Spiro-5'-(3'-utilization),

1 methylpiperidin-4-Spiro-5'-(3'-(2-butynyl)-as), piperidine-4-Spiro-5'-(3'-propargylglycine),

2-methyl-1,4-thiazolidin-3-he-Spiro(5,3') Hinkley.

6. Spirochaetaceae under item 1 or 2, wherein it is chosen from the group consisting of:

1 methylpiperidin-4-Spiro-5'-(2'-tion-3'-utilization),

1-methylpiperidin the
1 methylpiperidin-4-Spiro-4'-(1'-ethyl-2'-ethylthio-2'-imidazolin-5'-he),

1 methylpiperidin-4-Spiro-4'-(1'-ethyl-2'-imidazolin-5'-he).

7. Spirochaetaceae under item 1 or 2, characterized in that it is selected from the group consisting of enantiomerically pure d - and l-1 methylpiperidin-4-Spiro-5'-(2'-methyl-1', 4', -thiazolidin-3'-Oh), and d and l-1 methylpiperidin-4-Spiro-5'-(2'-ethyl-1', 4'-thiazolidine-3'-one).

8. Pharmaceutical composition, characterized in that it contains at least one spirochaetaceae General formula (AA) PP.1-7 in an effective amount.

9. Spirochaetaceae General formula (AA) PP.1-7 having muscarinic activity.

10. Spirochaetaceae under item 9, which additionally has a biological activity selected from the following: naratriptan or synergistic activity when sovetskom action with NEF, the activity that causes the secretion of the protein precursor of amyloid a (BPA) and reduce the toxicity of amyloid, activity, increasing the share of dephosphorylating-proteins and NGF-like activity.

 

Same patents:

The invention relates to derivatives of 3-genocidally esters of the formula I or their pharmaceutically acceptable acid additive salts, where X is phenyl, Y is the group - (CH2)A- CR1R2- (CH2)BS(O)Z-R3where A and B are independently 0, 1 or 2, Z is 0, 1 or 2, R1and R2independently a hydrogen atom, (C1-C4)-alkyl, R3- (C1-C4)-alkyl or phenyl

The invention relates to new derivatives of hinoklidina, pharmaceutical compositions containing such compounds and to the use of such compounds for the treatment and prevention of inflammatory disorders and disorders of the Central nervous system, as well as some other disorders

The invention relates to new and useful derivatives of hinoklidina that are of interest in the field of medicinal chemistry

The invention relates to new derivatives of 1-phenyl-3-azabicycloalkanes-2-ones, to a method for producing them, to pharmaceutical compositions containing them and to their use as therapeutic agents

The invention relates to new compounds with pharmacological activity, in particular bicyclic 1-Aza-cycloalkanes General formula

(I) where R is lower alkyl, unsubstituted or substituted furan, thiophene or imidazole; alkenyl with 3-6 carbon atoms; quinil with 3-6 carbon atoms; phenyl, unsubstituted or substituted lower alkyl, alkoxygroup or by halogen; benzyl, unsubstituted or substituted by halogen; pyridyl; pyrimidinyl;

A, b and C independently of one another denote-CH2or a simple bond;

n is 0 or 1, mixtures of isomers, or individual isomers and their pharmacologically tolerable acid additive salts exhibiting holinoliticheskoy properties

,5,6]-6-amino-3-azabicyclo [3.1.0] gex-3-yl)- 6-fluoro-1-(2,4-differenl)-1,4 - dihydro-4-oxo-1,8 - naphthiridine-3-carboxylic and methanesulfonic acid and its preparation" target="_blank">

The invention relates to a new form of the anhydrous salt methanesulfonic acid and 7-([1,5,6]-6-amino-3-azabicyclo[3.1.0] Gex-3-yl)-6-fluoro-1-(2,4-differenl)- 1,4-dihydro - 4-oxo-1,8-naphthiridine-3-carboxylic acid, to a method of use of the compounds in the treatment of bacterial infections in mammals, especially humans, and to pharmaceutical compositions useful for him

The invention relates to quinolone and acridinium compounds of the formula I or their pharmaceutically acceptable salts, where R2- H, (C1-C6)alkyl (C1-C4)foralkyl; R3IS H, CN, (C1-C6)alkyl, (C1-C6)foralkyl, ethanol or R2and R3form 1,4-butandiol; R4means a group of formula II; R7- H, R8- H, (C1-C4)alkoxy, NO2CN, (C1-C4)foralkyl, halogen, (C1-C4)alkyl; R9- H, halogen, (C1-C4)alkyl or (C1-C4)foralkyl or R8and R9taken together form a (C1-C3)alkylenedioxy; R10and R11- H, (C1-C4)alkyl
The invention relates to neurology and can be used in the treatment of paralysis and paresis

Drug // 2121347

The invention relates to the field of medicine, to biologically active substances produced by chemical means, specifically derivatives acridone and monosaccharides, and is intended for use as anti-infective, anti-inflammatory and anti-cancer agents with a wide spectrum of biological action

The invention relates to derivatives of 3-genocidally esters of the formula I or their pharmaceutically acceptable acid additive salts, where X is phenyl, Y is the group - (CH2)A- CR1R2- (CH2)BS(O)Z-R3where A and B are independently 0, 1 or 2, Z is 0, 1 or 2, R1and R2independently a hydrogen atom, (C1-C4)-alkyl, R3- (C1-C4)-alkyl or phenyl

The invention relates to new pcoralcalciumonline derived nitrogen-containing heterocyclic compounds, in particular to compounds of the formula I mentioned in the description

FIELD: medicine, phthisiology.

SUBSTANCE: one should lymphotropically introduce the mixture of 5.0 ml 0.25%-novocaine solution and 2.0 ml 1%-dioxidine solution or the mixture of 5.0 ml 0.25%-novocaine solution and 0.5 g cefazoline subcutaneously into jugular cavity and deeply behind xiphoid process, successively 1 point once daily, 5-7 injections/course. After injection the site of injection should be treated either with heparin ointment or ultrasound (1-3 MHz, PPM 0.2 W/sq. cm, for 2 min, through Vaseline oil) followed by evaluating roentgenological dynamics of the process 10-14 d later.

EFFECT: higher efficiency of differential diagnostics.

3 ex

Up!