Radiolabelled inhibitors of glicine 1 transporters

FIELD: chemistry.

SUBSTANCE: invention relates to novel radiolabelled compounds of formula I in which R¹ represents isopropoxy or 2,2,2-trifluoro-1-methyl-ethoxy; and R² represents radiolabelled CH₃ group, where radionuclide represents ³H or ¹¹C. Invention also relates to pharmaceutical composition for diagnostic visualisation of GlyT1 transporter (glycine transporter type 1).

EFFECT: obtained are novel radiolabelled compounds, which can be applied in medicine as radioactive indicator in PET (positron emission tomography) for labeling and diagnostic molecular visualisation of functionality of glycine transporter type 1.

13 cl, 4 ex

The present invention relates to new radiolabelled inhibitors having the General formula I, the vector of glycine type 1 (GlyT1), useful for labeling and diagnostic imaging functionality vector of glycine type 1

where

R¹represents isopropoxy or 2,2,2-Cryptor-1-methyl-ethoxy; and

R²is a radiolabelled group CH3, where the radionuclide is a³N or¹¹C.

Found that the radiolabelled compounds of formula I can be used as a radioactive indicator in PET (positron emission tomography) for labeling and diagnostic molecular imaging functionality vector of glycine type 1. Molecular imaging is based on the selective and specific interaction of the molecular probe (for example, radioactive indicator) with the biological target (e.g., receptor, enzyme, ion channel, or any other cellular component, which is able to communicate with the molecular probe or keep a molecular probe), which is visualized by PET, nuclear magnetic resonance spectroscopy in the near infrared region or other methods. PET, renderer methods of nuclear medicine is s, ideal for obtaining three-dimensional images, which give important information about the distribution of the biological target in a given organ or metabolic activity of such a body or cells, or on the ability of drugs to penetrate into such a body, contact with the biological target and/or modify biological processes. Since PET is a noninvasive method of imaging, it can be used to study the pathophysiology of disease and drug action on a given molecular target or cellular processes in humans and animals. The availability of radioactive indicator for use in PET, specific to a given molecular targets that may contribute to drug development and understanding of the mechanism of action of drugs. In addition, a radioactive tracer for PET can contribute to the diagnosis of the disease by demonstrating the pathophysiological changes that occur as a result of disease.

Inhibitors vector glycine suitable for treatment of neurological and neuropsychiatric disorders. Most directly involved painful conditions are psychosis, schizophrenia (Armer RE and Miller DJ, Exp. Opin. Ther. Patents, 11 (4): 563-572, 2001), psychotic mood disorders, that is their as heavy main depressive disorder, mood disorders associated with psychotic disorders such as acute manic syndrome or depression associated with bipolar disorders, and mood disorders associated with schizophrenia (Pralong ET et al., Prog. Neurobioi, 67: 173-202, 2002), autistic disorder (Carlsson ML, J. Neural Trans. 105: 525-535, 1998), cognitive disorders such as dementia, including age-related dementia and senile dementia alzheimera type, memory disorders in a mammal, including man, syndromes attention deficit and pain (Armer RE and Miller DJ, Exp. Opin. Ther. Patents, 11 (4): 563-572, 2001).

The human brain is a complex organ consisting of millions of neurons engaged in internal communication. The understanding of disorders relating to diseases, is the key to the future development of effective new diagnostic and therapeutic tools. The study of biochemical disorders in humans is rapidly becoming an essential and inseparable component of the invention medicines and the process of their development. Traditionally, the invention and development of new medicines was carried out with special emphasis on in vitro techniques to best selection of promising candidates, which are then tested on live animals before introducing people. Due to the fact that in vitro otrajatelem part of the diversity of living systems, and animal models in vivo human disease often represent only some kind of approximation to human pathology, increasingly there is a growing realization that a clear understanding of the interaction of drugs with the receptor in a living person, at an early stage of this process will be a main driving force in giving additional impetus for the effective and timely invention and development of new therapeutic drugs. In recent years, an increasing number of cases of the use of diagnostic imaging in humans to assess pathologies, disease processes and the effects of medicines. These visualization tools include PET, MRI (magnetic resonance imaging), CT (computed tomography), ultrasound, EEG (electroencephalography), SPECT (single photon emission computed tomography) and others (British Medical Bulletin, 2003, 65, 169-177). Therefore, using non-invasive means of imaging, such as PET, is an invaluable tool for drug development in the future. Non-invasive nuclear imaging methods can be used to obtain basic and diagnostic information about the physiology and biochemistry of a wide variety of living entities. These methods rely on the application of sophisticated imaging equipment, coloradoson to detect radiation, emitted by radioactive indicators, introduced in such living entities. The information obtained can be converted by obtaining planar and tomographic images, which show the distribution of radioactive indicator as a function of time. The result of the use of radioactive tracers can be obtaining images that contain information about the structure, function, and, most importantly, physiology and biochemistry of the subject. Much of this information is impossible to obtain by other methods. Radioactive indicators used in these studies, developed with the ability to determine the behavior in vivo, which allows you to define specific information regarding the physiology or biochemistry of the subject. Currently radiotracers available for obtaining useful information related to cardiac activity, blood flow in the myocardium, pulmonary perfusion, liver function, blood flow in the brain regional glucose metabolism and oxygen in the brain (WO 2007/041025). In addition,

- PET imaging allows non-invasive and quantitative analysis of normal and abnormal neurochemistry laboratory in humans at an early stage of drug development in order to give additional impetus for productive and efficient invented the I of therapeutic agents;

- the use of indicator doses of labelled compounds allows you to perform an initial evaluation of new drugs: a study of the biological distribution; research degree employment receptors to optimize the dosage of medicines and the characteristic of the subsequent responses to the action of the medicinal product;

- understanding the mechanisms of disease in humans using non-invasive methods is directly related to future developments in the diagnosis and treatment of diseases and the development of new therapeutic agents.

Commonly used PET radionuclides include¹¹C,¹³N¹⁵O or¹⁸F. In principle, there is a possibility to enter a label to any drug with getting the equivalent of the initial connection, but found that only some of them are suitable as imaging agents in vivo in humans. The half-lives of radioactive elements¹¹C,¹³N¹⁵O and¹⁸F are 20, 10, 2, and 110 min, respectively. Such short half-lives provide a number of advantages in relation to their use as radiotracers for investigations of biological processes in vivo by PET. May be repeated studies of the same subject on the same day. PET is increasingly used is : as a tool to determine the relationship between the dose of the drug and the degree of employment of enzymes/receptors for particular compounds. Radiotracers for PET that are specifically associated with the target receptor or enzyme, can give information about

- the ability of drugs to penetrate into the brain and connect with customers-target

- the degree of employment of the target site, due to the effect of a given dose of the drug,

- based employment from time to time and

the relative kinetics in plasma and tissues are considered pharmaceuticals.

Research employment is performed with the use of radiotracers for PET, which is usually not identical to the investigational drug candidate (British Medical Bulletin, 2003, 65, 169-177).

The present invention was to find new radiotracers for in vivo PET imaging vector of glycine type 1. Found that the resulting radiolabelled compounds of formula I can be used as an imaging agent for visualizing vector of glycine type 1 in humans. The present invention is covered by the following radiolabelled compounds:

radiolabelled compound of the formula

radiolabelled compound of the formula

labeled radioactive the isotope compound of the formula

and

radiolabelled compound of the formula

Other embodiments of the present invention are the compounds of formula I for use as a ligand GlyT1, for use in the study of binding GlyT1 and for use as radiotracers for PET.

In addition, the compounds according to the present isoberlinia can be used for diagnostic imaging GlyT1 in the brain of a mammal.

The invention includes a method of diagnostic imaging vector GlyT1, including introduction to the mammal an effective amount of the compounds of formula I, and includes a method for determining the functionality of GlyT1 in the tissue of a mammal, comprising the administration to a mammal, for which it is desirable such a determination, the effective amount of the compounds of formula I.

The present invention is the use of the compounds of formula I for the manufacture of a medicine for diagnostic imaging GlyT1 in the brain of mammals and pharmaceutical compositions containing such a compound and pharmaceutically acceptable excipient.

Inhibitors vectors GlyT1 useful for treatment of diseases that are psychoses, pain, impaired memory and learning, schizophrenia, dementia and other diseases in which impaired cognitive processes, such as syndromes attention deficit or Alzheimer's disease. The preferred reading is the schizophrenia.

Schizophrenia is a progressive and causes clouding of the mind neurological disease characterized by episodic positive symptoms such as delusions, hallucinations, thought disorder and psychosis, and persistent negative symptoms such as emotional dullness, weak attention and social withdrawal, and cognitive impairment.

Its radioactive isotope compounds known in the prior art and described in WO 2006/082001 as inhibitors vectors GlyT1.

Scheme 1 represents the path of the synthesis of compounds of formula (I), where R²is a radiolabelled group

Scheme 1

The compound of formula II is put into communication with a base, such as cesium carbonate, and with a reagent of formula III, where R²represents a group containing a radionuclide selected from the³N or¹¹C and X represents a leaving group such as iodine. Reagents of formula III, such as [³N]methyliodide and [¹¹With]methyliodide known, and get them according to Larsen P., J. Ulin, K. Dhlstrom J. Label. Compds. Radiopharm. 37, 73-75, 1995.

Scheme 2 represents the path of the synthesis of the compounds of formula (II), where R¹represents (8)-2,2,2-Cryptor-1-methyl-ethoxy.

Scheme 2

The intermediate acid IV can be obtained by the interaction of commercially available ortho-fermenting acid with (S)-1,1,1-Cryptor-propane-2-I (CAS (Chemical abstract cervice): 3539-97-7) in the presence of a base such as sodium hydride, in a solvent such as dioxane. The combination of acid IV with known isoindoline V (WO 2006082001) to obtain the amide VI can be achieved in the presence of reagent combinations, such as TBTU, and bases, such as diisopropylethylamine, in a solvent such as DMF. Chlorosulfonylisocyanate compound VI with intermediate sulphonylchloride VII can be carried out in the presence of chlorosulfonic acid in a solvent such as dichloroethane. The reduction of compound VII to sulfinol acid II can be achieved by use of sodium sulfite as a reducing agent in a solvent type DMF and water.

Figure 3 presents the path of synthesis of the compounds of formula (II), where R1 represents isopropoxy.

Scheme 3

Intermediate compound IX may be obtained by interaction of the acid VIII (WO 2005014563) with p is adage alkylating agent: (iodomethyl)trimethylsilane, in the presence of a base, such as diisopropylamide lithium and excipients type TMEDA in a solvent such as THF. The combination of acid IX with known isoindoline V (WO 2006082001) to obtain the amide X can be achieved in the presence of reagent combinations, such as TBTU, and bases, such as aminobutiramida-ethylamine in a solvent such as DMF. The transformation of compound X in sulinowo acid II can be achieved in the presence of TBAF in a solvent type THF.

Reduction

As described above, found that the radiolabelled compounds of formula I can be used as a PET ligand for labelling and diagnosis is eskay molecular imaging functionality vector of glycine type 1.

Appropriate its compounds [5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon and (2 isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon active against carrier GlyT1 in vitro and have the value of IC50 (μm) 0,028 and of 0.014, respectively. The test method described in WO 2006/082001.

Autoradiographically studies on the rat brain

The distribution of binding sites [³N][5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone and [3H](2-isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone investigated in the rat brain.

For these experiments we used male Wistar rats. Rats were killed; the brain of rats were rapidly removed, frozen in powdered dry ice. Ten sagittal slices of a thickness of one micron cut on a cryostat and placed with the defrost on slides with adhesive coating. Brain slices were first incubated for 10 min in ringer's solution (NaCl 120 mm, KCl 5 mm, CaC₂2 mm MgCl₂1 mm, Tris-HCl 50 mm pH of 7.4) at 37°C and then for 60 min in ringer's solution at 37°C containing either [³N][5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tet is ahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon, or [³N](2-isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon at a concentration of 1 nm. To assess nonspecific binding (NSB) of radioactive indicator additional group of slices incubated with ringer's solution containing a radioactive tracer and reference GlyT1 inhibitor, Org 24598, at a concentration of 10 μm (60 min at 37°C). After incubation the sections were washed 2×5 min and 1×15 min in chilled on ice (4°C) ringer's solution and then three times quickly dipped in distilled water at 4ºC. Brain slices, placed on slides, dried in a stream of cold air and exhibited together with [³N]-standard ([³H]-microscale) on the imaging plate Fuji for 5 days. Then, the imaging plate was scanned on a scanner with high resolution for plates from FujiFilm. The total number of radioactive indicator, contacting of interest areas of the brain (TV (totally bound)), was measured using the MCID image analysis and expressed in fmol bound peroxidase radioactive indicator/mg protein. The number of [³N][5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone, specifically contacting GlyT1 carrier (SB (specifically bound)), was calculated according to the formula

The results showed that the distribution of the binding sites of [³N][5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone and [³N](2-isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone consistent with the known distribution for the vector GlyT1 (Cubelos C., Gimenez C, Zafra f, Cereb Cortex 15, 448-459, 2005; Zafra F., C. Aragon, L. Olivares, Danbolt NC, Gimenez C., Storm-Mathisen J., J Neuroscience. 15, 3952-69, 1995). High density of binding sites was observed in the thalamus, brain stem, Pons varolii, and the medulla and the cerebellum. Low density was observed in the striatum, cortical matter and the hippocampus. Joint incubation of radioactive indicators with specific GlyT1 inhibitor, Org 24598 or other specific inhibitors of GlyT1 in high concentrations has completely annihilated the binding of [³N][5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone and [³N](2-isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone with brain slices of the rat, confirming that these two radioactive indicator associated with the carrier of GlyT1.

RET-in vivo studies on baboons

1) PET imaging with [¹¹C](isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone and [ ¹¹With][5-methanesulfonyl-2-((S)-2.2.2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone

The experiments described below were carried out in male baboons (Papio anubis). Animals were not fed for 12 hours before the PET study. First of baboons was affected by sedative agent, using intramuscular injection of ketamine hydrochloride in the "deterrent" dosages (restraint dosages) 5-7 mg/kg to achieve a light stage of anesthesia and then maintained with continuous intravenous infusion of propofol in the mode of 0.3-0.4 mg/kg/h (emulsion for injection DIPRIVAN®). The volume of blood circulation was maintained by infusion of isotonic. For sampling of blood is injected with a catheter in the femoral artery. In all studies were performed continuous monitoring of key physiological parameters, including heart rate, ECG (electrocardiogram), blood pressure (Spacelabs Monitor, Issaquah, WA, USA) and oxygen saturation (pulse oximeter OxiMax® N-600™ from Nellcor, Pleasanton, CA, USA). The animal was placed in the PET scanner for studying brain HRRT® (from the English. high resolution research tomography) from ECAT (scanner with high resolution for scientific research, CPS Innovations, Inc., Knoxville, TN). To the head of the animal was affixed a mask made of a thermoplastic material, which was attached to the latch head for vosproizvodimost the commit. During the first 6 min spent a transmission scan using a point source of Cs-137 activity 1 MCI to correct shielding. [¹¹C](2-Isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon] and [¹¹With][5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon was injected intravenously as a bolus injection lasts 1 minute. PET scanning and sampling of arterial blood initiated after the start of injection of radioactive indicator and PET images recorded in the period from 0 to 90 minutes after injection of radioactive indicator.

The results of these studies using imaging showed that both radioactive indicator [¹¹C](2-isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon and [¹¹With][5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon quickly picked up by many sections of the brain in accordance with the curves of dependence of activity from time to time, which showed the maximum capture 20-30 min after injection and a slow decrease during the rest time of the study. The regional distribution of both radiotracers about what was Rajala known distribution of the vector of glycine type 1 (GlyT1) with higher accumulation in areas warriewood bridge, the brain stem, cerebellum and thalamus compared with areas of the cerebral cortex (Cubelos C., Gimenez C, Zafra f, Cereb. Cortex, 15, 448-459, 2005).

2) PET imaging in terms of pharmacological load in these experiments tested the ability of unlabeled GlyT1 inhibitors to block the capture of [¹¹C](2-isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone and [¹¹With][5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone in brain regions known to contain GlyTl. Since [5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon, and (2 isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon are selective GlyT1 inhibitors, for the experiments described below were selected one of these compounds [5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon.

Each animal received two consecutive injection of radioactive indicator in the same day. First, the introduction of radioactive indicator used to determine the basic capture of radioactive indicator. After scanning the baseline baboon received intravenous is elchinova [5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone (blocker). Infusion blocker was started 20 min before (second) injection of radioactive indicator. First, the blocker was administered by infusion at a dose of 0.2 mg/kg in 10 min flow rate was changed, so that during the remaining 100 minutes of research to deliver 0.5 mg/kg Preliminary pharmacokinetic experiments and modeling data indicated that such speed infusion give constant levels in plasma [5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone in the continuation of this time interval PET-scan.

Pre-processing of non-radioactive [5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-Metronom (selective inhibitor GlyTl) completely blocked the specific capture of both radiotracers [¹¹C-](2-isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone and [¹¹With][5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone and resulted in a homogeneous distribution of radioactive substances around the brain. These results confirmed the specificity of both radiotracers in relation to the carrier GlyT1 and clearly showed that associating them with the carrier of GlT-1 may be weakened by the presence of unlabeled drugs associated with the vector GlyT1.

Compounds of the present invention are diagnostic tools that can be useful in the diagnosis of disorders of the Central nervous system, for example, schizophrenia, cognitive impairment and Alzheimer's disease.

The compounds of formula I can be used together with pharmaceutically inert, inorganic or organic carriers in the process of making pharmaceutical compositions.

The dosage can vary within wide limits and, no doubt, in every case they will need to be adjusted in accordance with individual requirements.

Radiolabelled inhibitors are preferably administered intravenously.

The injection solution may have the following composition:

The following examples illustrate the invention but are not intended to limit its scope.

Example 1

[³N-Methyl]-[5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon

a) stage 1

2-((S)-2.2.2-Cryptor-1-methyl-ethoxy)-benzoic acid

36,0 g (316 mmol) of (3)-1,1,1-Cryptor-propan-2-ol (CAS: 3539-97-7) was added to a cold (0-5°C) suspension of 17.0 g (425 mmol) of NaH (practical use., 60%) in 200 ml of dioxane. The suspension was stirred at room temperature for 0.5 h, then was cooled (0-5°C) was added to a solution of 20.0 g (143 mmol) of 2-fluoro-benzoic acid in 100 ml of dioxane. The mixture was stirred for 0.5 h at room temperature and over 140 hours boiling under reflux. The mixture was poured into water (800 ml), washed with 300 ml of MTBE, then pakilala to pH 2 with hydrochloric acid and the product was extracted using MTBE. The solvent was concentrated in vacuo and the residue was led from a mixture of ethanol/water, getting 27,3 g (82%) indicated in the title compounds as white solids. MS (mass spectrometry) (m/e]: 234,1 [M]⁺.

b) stage 2

[5-(Tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl)-[2-((S)-2.2,2-Cryptor-1-methyl-ethoxy)-phenyl]-Manon

To a solution of 0.9 g (3.8 mmol) of 2-((8)-2,2,2-Cryptor-1-methyl-ethoxy)-benzoic acid in 9 ml of DMF in an argon atmosphere at room temperature was added 1.4 g (4.2 mmol) TBTU, and 3.3 ml (19.2 mmol) of N-ethyldiethanolamine and at the end of 0.8 g (3.8 mmol) of 5-(tetrahydro-Piran-4-yl)-2,3-dihydro-1H-isoindole (CAS: 905274-50-2). The mixture was stirred at room temperature overnight. The solvent was removed in vacuum. The residue was dissolved in ethyl acetate. The solution is washed twice with water and twice with saturated solution of NaHCO₃, dried over Na2SO₄, filtered and concentrated in vacuum. The crude oil was purified column flash chromatography on silica with elution with a gradient formed from heptane and ethyl acetate, receiving 1.5 g (93%) indicated in the title compound as a yellow oil. MS (m/e): 420,2 [M+H]⁺.

c) stage 3

3-[5-(Tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-carbonyl]-4-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-benzosulphochloride

A solution of 0.2 g (0.47 mmol) [5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-[2-((5)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-methanone in 2 ml of 1,2-dichloroethane was added dropwise to 0,32 ml (4.7 mmol) of chlorosulfonic acid while cooling in an ice bath. The mixture was stirred at room temperature for 30 minutes and then at 55°C for 30 minutes. The mixture ohla is given in an ice bath and extinguished, adding dropwise 2 ml of water. The mixture was diluted with dichloromethane. The organic layer was separated and the aqueous layer was twice extracted with dichloromethane. The combined dichloromethane extracts were dried over Na2SO₄) filtered and concentrated in vacuum. The resulting foam was stirred with ethyl acetate. The solid was filtered. The filtrate is washed twice with saturated solution of NaHCO₃, dried over Na2SO₄, filtered and concentrated in vacuum, obtaining 0.12 g (51%) indicated in the title compound as light yellow foam. MS (m/e): 517,1 [M]⁺.

d) stage 4

3-[5-(Tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-carbonyl]-4-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-benzosulfimide sodium salt

1,15 g (to 8.94 mmol) Na2SO₃and 1.70 g (a 9.60 mmol) of Na₂HPO₄hydrate was dissolved in 13 ml of water. Added an ethanolic solution of 2.40 g (4,63 mmol) 3-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-carbonyl]-4-((3)-2,2,2-Cryptor-1-methyl-ethoxy)-benzosulfimide. The reaction mixture was stirred at 35-40°C for 1 hour and then overnight at room temperature. Was added 1.3 g of spiders (Speedex), the reaction mixture was filtered and the filtrate was evaporated. The crude product was treated with an aqueous solution of citric acid/NaCl, then extracted with a mixture of 1:1 MTBE/THF. The organic solvent evaporated, estato is dissolved in a mixture of Meon/water (2:1) and treated, using 800 mg (9,52 mmol) and NaHCO₃. Added 1 g of spiders and the reaction mixture was filtered and concentrated in vacuum. The residue was purified chromatographically by using a reversed-phase column (RP-18, water/methanol)to give 1.12 g (48%) indicated in the title compound as a white foam. MS (m/e): 484,3 [M+H]⁺.

e) stage 5

[³N-Methyl]-[5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon

0.16 mg (1.2 mmol) Lil was added to a solution of 50 MCI (0.15 mg; 0.6 mmol) [³N]methylnitrate in 0.2 ml of DMF. After stirring the reaction mixture for 3 h at 20°C in a closed vial was added 0.6 mg (1.4 mmol) of 3-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-carbonyl]-4-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-benzosulfimide sodium salt and 1.0 mg (3.1 mmol) of cesium carbonate and stirring was continued for 2 h at 20°C. the Reaction mixture was treated with water and brine and then was extracted with ethyl acetate. After evaporation of the organic solvent the crude product was purified column chromatography (silica, ethyl acetate/heptane 4:1), receiving 23,9 MkI (48%), tritium-labeled specified in the connection header with specific activity 74 CI/mmol (according to MS analysis). HPLC analysis of radioactive compounds showed p is biohimicheskoe purity > 99%.

Example 2

[¹¹C-Methyl]-[5-methanesulfonyl-2-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon

3-[5-(Tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-carbonyl]-4-((S)-2,2,2-Cryptor-1-methyl-ethoxy)-benzosulfimide sodium salt (1 mg, 2 mmol) was dissolved in 100 μl of dimethylformamide. The vial was tightly closed; the solution was shaken for one minute was introduced into the system Bioscan AutoLoop and purged with argon (30 ml/min) for 5 seconds. [¹¹C]Methyliodide (obtained according to Larsen P., Ulin j, Dahlström, K. J. Label. Compds. Radiopharm. 37, 73-75, 1995) made in Bioscan Autoloop (Bioscan Inc., Washington, DC) gas flow (30 ml/min). The capture of [¹¹C]methyliodide in AutoLoop performed within 3.5 minutes, after which the flow was stopped. After 4.5 minutes, the reaction mixture is automatically sent to prepreparation HPLC and processed as described below. The product was collected in order to withstand the pressure tank, where it was diluted with 50 ml water, then sprayed on a C-18 SepPak Plus from Waters (see conditions below analytical and preparative HPLC). SepPak containing purified specified in the title compound, washed with 10 ml of isotonic solution, after which the product was suirable 1 ml of absolute ethanol, and then 10 ml of isotonic solution through a filter with pores of 0.22 micron for sterilization in sterile pyrogen-free vial, aderrasi 4 ml isotonic.

Conditions for HPLC: analytical: Onyx C18, a 4.6×100 mm; 35:65 MeCN:H₂O; TEA (triethylamine) pH of 7.2 at 3 ml/min; preparative: XTerra C₁₈, 5 μm, 19×100 mm; 40:60 MeCN:H₂O; 0,1M NH₄-formate at 18 ml/min

Example 3

[³H-Methyl]-(2-isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon

a) stage 1

2 Isopropoxy-5-(2-trimethylsilyl-econsultancy)benzoic acid

To stir (-70°C) suspension of 0.26 g (1 mmol) 2-isopropoxy-5-methanesulfonyl-benzoic acid (CAS: 845616-02-6) and 0.75 ml (5 mmol) of TMEDA in 2.6 ml of THF was added dropwise a solution of LDA (prepared from 1.3 ml (2.1 mmol) of a 1.6 M solution of n-utility in hexane and 0.3 ml (2.1 mmol) of Diisopropylamine in 2.5 ml THF at 0°C). The light yellow suspension was stirred at -70°C for 30 minutes. Dropwise over a period of time of 5 minutes was added to the solution to 0.19 ml (1.3 mmol) (iodomethyl)trimethyl-silane in 0.5 ml THF. The yellow suspension was stirred at -70°C for 15 minutes and then left to warm to room temperature. Light yellow solution was stirred at room temperature for 1 hour and then extinguished using 5 ml of brine. The mixture was diluted with 5 ml of water. The mixture was concentrated in vacuum. The aqueous layer was carefully acidified using 1 N. HCl, and was extracted 3 times with dichloromethane. Merge is installed extracts were dried over Na2SO ₄, filtered and concentrated in vacuum. The crude substance was purified column flash chromatography on silica with elution with a gradient formed from heptane and ethyl acetate, getting 0.21 g (63%) indicated in the title compound as a yellow oil. MS (m/e): 343,0 [M-H]⁺.

b) stage 2

[2 Isopropoxy-5-(2-trimethylsilyl-econsultancy)-phenyl-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon

By analogy with the procedure described for the synthesis of the compound from example 1, stage 2, is listed in the title compound was obtained from 5-(tetrahydro-Piran-4-yl)-2,3-dihydro-1H-isoindole (CAS: 905274-50-2) and 2-isopropoxy-5-(2-trimethylsilyl-econsultancy)-benzoic acid. MS (m/e): 529,3 [M]⁺.

c) stage 3

4 Isopropoxy-3-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-carbonyl]-benzosulfimide sodium salt

1.40 g (2,64 mmol) [2-isopropoxy-5-(2-trimethylsilyl-econsultancy)-phenyl]-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanone was dissolved in 14 ml of THF and was treated with 4.0 ml (4.0 mmol) of a 1 M solution of TBAF in THF at 60°C for 3.5 hours. The reaction mixture was poured into an aqueous solution of citric acid/NaCl, then extracted with a mixture of MTBE/THF 1:1. The organic solvent is evaporated, the residue was dissolved in a mixture of Meon/water (3:1) and was treated with 600 mg NaHCO 3. After evaporation the residue was purified by chromatographic reversed-phase column (RP-18, water/methanol)to give 0.66 g (55%) indicated in the title compound as a white foam. MS (m/e): 430,2 [M+H]⁺.

d) stage 4

[³H-Methyl]-(2-isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl]-methanon

0.16 mg (1.2 mmol) Lil was added to a solution of 50 MCI (0.15 mg; 0.6 mmol) [³H]methylnitrate in 0.2 ml of DMF. After stirring the reaction mixture for 3 h at 20°C in a closed vial was added 0.6 mg (1.3 mmol) of 4-isopropoxy-3-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-carbonyl]-benzosulfimide sodium salt and 1.0 mg (3.1 mmol) of cesium carbonate and stirring was continued for 2 h at 20°C. the Reaction mixture was treated with water and brine and then was extracted with ethyl acetate. After evaporation of the organic solvent the crude product was purified column chromatography (silica, ethyl acetate/heptane 4:1), receiving 29,2 MkI (59%), tritium-labeled specified in the connection header with specific activity 74 CI/mmol (according to MS analysis). HPLC analysis of radioactive compounds showed radiochemical purity >99%.

Example 4

[¹¹C-Methyl]-(2-isopropoxy-5-methanesulfonyl-phenyl)-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-yl-metano

By analogy with the procedure described for the synthesis of compounds of example 2, indicated in the title compound was obtained from 4-isopropoxy-3-[5-(tetrahydro-Piran-4-yl)-1,3-dihydro-isoindole-2-carbonyl]-benzosulfimide sodium salt and [¹¹C]methyliodide.

1. Radiolabelled compound of the formula

where
R¹represents isopropoxy or 2,2,2-Cryptor-1-methyl-ethoxy; and
R²is a radiolabelled group SN₃,
where the radionuclide is a³H or¹¹C.

2. Radiolabelled compound of formula I-A according to claim 1, which represents the

3. Radiolabelled compound of formula I-B according to claim 1, which represents the

4. Radiolabelled compound of formula I-C according to claim 1, which represents the

5. Radiolabelled compound of formula I-D according to claim 1, which represents the

6. The compound of formula I according to any one of claims 1 to 5 for use as a radiotracer for GlyT1 (vector glycine type 1).

7. The compound of formula I according to any one of claims 1 to 5 for use in the study of binding GlyT1.

8. The connection is of the formula I according to any one of claims 1 to 5 for use as a radiotracer in PET (positron emission tomography).

9. The compound of formula I according to any one of claims 1 to 5 for use in diagnostic imaging GlyT1 in the brain of a mammal.

10. The method of diagnostic imaging vector GlyT1, including introduction to the mammal an effective amount of a compound defined in any one of claims 1 to 5.

11. The method of determining the functionality of GlyT1 in the tissue of a mammal, comprising the administration to a mammal, for which it is desirable such a determination, the effective amount of the compound defined in any one of claims 1 to 5.

12. The use of compounds as defined in any one of claims 1 to 5, for the manufacture of a composition for diagnostic imaging GlyT1 in the brain of a mammal.

13. Pharmaceutical composition for diagnostic imaging vector GlyT1 containing compound defined in any one of claims 1 to 5, and pharmaceutically acceptable excipient.