CZ39994A3 - Synthetic aryl polyamines representing antagonists of excitation amino acid neurotransmitters, process of their preparation and use - Google Patents

Abstract

Compounds having the formula R-(CH2)m-CO-R' wherein R is a 5 to 7 member carbocyclic system or an 8 to 11 member carbobicyclic system or any such system substituted with one or more of F, Cl, Br, OH, C1 to C4 alkyl, C1 to C4 alkoxy, CF3, phenyl, amino, C1 to C4 alkylamino and di(C1 to C4 alkyl)amino; m is 0 or 1; R' is -[NH(CH2)n]xNH2; each n is independently 2 to 5; and x is 1 to 6, and their pharmaceutically acceptable acid addition salts are potent excitatory amino acid neurotransmitter antagonists. These compounds are useful as mammalian psychotherapeutants, as the active ingredient in pharmaceutical compositions for treating conditions in mammals which are mediated by excitatory amino acid neurotransmitters and in the control of invertebrate pests.

Description

Technical field

The present invention relates to a class of aryl polyamines and pharmaceutically acceptable salts thereof derived from excitatory amino acid neurotransmitter antagonists. These neurotransmitters act on neuronal cells of various organisms, including invertebrates and vertebrates. The polyamine compounds of the present invention are synthetic analogs of certain polyamines that have been found to be found in the venom of the Agelenopsis aperta spider. The present invention also includes the use of these polyamines and their salts in antagonizing excitatory amino acid neurotransmitters. These neurotransmitters have the ability to act on cells, such as cells of the nervous system of an organism. The invention also encompasses the use of these polyamines and their salts in the treatment of diseases and conditions that are mediated by excitatory amino acid neurotransmitters, and for use in controlling invertebrate insects. The present invention also includes compositions comprising said polyamines and salts derived therefrom, as well as methods for preparing said polyamines.

BACKGROUND OF THE INVENTION

It is known from the prior art that the spider venom of Agelenopsis aperta contains at least two toxins that act on the passage of calcium through the calcium channel, cf. Jackson H. et al., Soc. Neu. Sci. Abstr.

1078 (1987). In this publication the authors describe a toxin referred to herein as AG2 and having a molecular weight of less than 1000 daltons, which toxin is believed to suppress the passage of calcium through the calcium channel in a variety of tissues.

In addition, from another publication, cf. Jackson H. et al., Soc. Neu, Sci. Abstr. 12: 730 (1986), it is known that another toxin from the venom of the Agelenopsis aperta spider contains a molecular weight component of about 6000. This toxin is said to block presynaptic transmission, which toxin is believed to block calcium channels in association with releasing neurotransmitters.

Some polyamines found to be in the venom of the Agelenopsis aperta spider are disclosed in U.S. Patent No. 5,037,846, filed April 28, 1989 and filed by the same Applicant as the present invention. These polyamines and pharmaceutically acceptable salts derived from these compounds are said to be blockers of excitatory amino acid receptors in cells, and one of these polyamines, referred to as B _x , is also said to be a calcium channel blocker.

Compounds which are antagonists of excitatory amino acid neurotransmitters have a variety of uses. These excitatory amino acid neurotransmitter antagonists are useful for the treatment of a variety of conditions such as stroke, cerebral ischemia, neuronal degenerative disorders such as Alzheimer's disease and epilepsy, as well as psychotherapeutics and for other miscellaneous purposes. for example, Excitatory Amino Acids In Health and Disease, D. Lodge, E., John Wiley & Sons Ltd., New York, NY 1988, which publication is for reference only. Among other things, the above compounds are useful for studying physiological processes in cells, such as neuronal cells, and for use in controlling invertebrate insects.

Glutamate is the major excitatory neurotransmitter in the mammalian brain. Over the last decade, the development of the pharmacology of glutamate receptors has made considerable progress, with the great benefit of finding that these glutamate receptors can be distinguished into several subtypes. In particular, the glutamate receptor subtype, determined by the selective action of the exogenous N-methyl-D-aspartate agonist (NMDA), has been the subject of intensive scientific investigation since these receptors are thought to play a role in various neurological pathological processes such as stroke, epilepsy and neurodegenerative processes such as Alzheimer's disease. There are currently two classes of NMDA receptor antagonists that have recently been extensively investigated with clinically appropriate drugs. The first group consists of competing antagonists that prevent glutamate from binding to its receptor site. These compounds are characterized as highly polar compounds, examples of which are phosphonate compounds AP7 and AP5. This highly polarized structure of said competing agents prevents their penetration through the blood-brain barrier and thereby limits their therapeutic utility. A second class of NMDA receptor antagonists are non-competitive antagonists that block the function of NMDA receptors by acting on the ion channel that is part of the NMDA receptor complex. These compounds include MK-801 and phencyclidine (PCP).

The potential psychotomimetic effects of the class of these compounds present an obvious disadvantage for these known drugs which operate according to the above mechanism.

Recently, a third group of antagonists has been the subject of investigation, and the stimulus for these investigations has been the identification of novel glutamate antagonists in spider venom. United States Patent Application No. 554,311, filed July 17, 1990, and US Patent No. 5,037,846, filed July 31, 1990, disclose arylamine structure compounds that were isolated from poison. Agelenopsis aperta spider, which compounds are potent specific NMDA receptor antagonists in mammals. These arylamine compounds isolated from the venom of the Agelenopsis aperta spider are combined structures that consist of aromatic acid and polyamine segments linked to each other by amide bonds. Some of the amine moieties in these polyamine segments are functionally in the form of N-hydroxyamines or quaternary ammonium salts. The chemical structure of these arylamine compounds is different from the above standard competing reagents, i.e. from the above-mentioned AP5 or AP7, as well as from the above-mentioned non-competitive compound

MK-801 For example, the polyamine compound AGEL 416, which is described in the aforementioned U.S. Patent No. 5,037,846, has the following structure:

The mechanism of NMDA antagonistic action of these arylamine compounds is also different from the two groups mentioned, i.e. from the competing agents and the MK-801 / PCP compounds. Thus, it is apparent from the above that said arylamine compounds isolated from spider venom represent a new class of antagonist compounds at NMDA receptors.

It is therefore a benefit of the present invention that these naturally occurring compounds can now be prepared by methods other than isolating and purifying them from the venom of the Agelenopsis aperta spider, and analogous compounds belonging to the same class of compounds can also be synthetically prepared by similar methods. , which, however, do not occur in nature.

The essence of the invention

The invention relates to a group of synthetic aryl polyamine compounds of the general formula:

R / CH 2 -CO-R 'wherein:

R is a 5- to 7-membered carbocyclic system or an 8 to 11-membered carbocyclic system or any of these systems substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxy, C 1 -C 4 alkyl, C 1 -C 4 alkoxy carbon, CF ₃ , phenyl, amino, C 1 -C 4 alkylamino and C 1 -C 4 dialkylamino, m is 0 or 1,

R 'is - [NH (CH ₂ ) _n ] _x NH ₂ , wherein each n is independently 2 to 5, and x is 1 to 6.

The present invention also includes compounds of the formula:

R 6 CH 2 -CO-R ', or a pharmaceutically acceptable acid addition salt thereof, wherein:

R is a 5- to 7-membered carbocyclic system or an 8 to 11-membered carbocyclic system or any of these systems substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxy, C 1 -C 4 alkyl, C 1 -C 4 alkoxy carbon, CF ₃ , phenyl, amino, alkylamino of up to 4 carbon atoms and dialkylamino of 1 to 4 carbon atoms, m is 0 or 1,

R 'represents a group:

[(CH ₂ ) _n NH] _y H

- [NH (CH _{2) n] x} N \ _[(CH2) "NH] _z H wherein each n independently is 2-5, x is 0-4, y and z. Each independently are 1-5 and the sum of x? larger zyaz values are 1 to 5.

The present invention also includes compounds of the formula:

R / CH3 -CO-R ', or a pharmaceutically acceptable addition salt thereof, wherein:

R is a 5- to 7-membered carbocyclic system or an 8 to 11-membered carbocyclic system or any of these systems substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxy, C 1 -C 4 alkyl, C 1 -C 4 alkoxy carbon, CF ₃ , phenyl, amino, alkylamino of up to 4 carbon atoms and dialkylamino of 1 to 4 carbon atoms, m is 0 or 1,

R 'represents a group:

(CH ₂ ) _and [NH (CH ₂ ) ₅ ] _y +

- [NH (CH ₂ ) _n ] _x N \ N [(CH ₂ ) _n NH] _from H (CH ₂ ) _# [NH (CH ₂ ) n] / where all a are the same and may be in the range of 5, all b are the same, and may be in the range of 2 in ph and 5, each n is independently in the range of 2 to 5, x is in the range of 0 to 3, all y are the same and may be 0 or 1, is in the range of 0 to 3, and the sum of x + y + z is in the range of 0 to 4.

The carbocyclic systems of the present invention may be saturated, unsaturated or aromatic, with aromatic systems preferably being present. With regard to monocyclic systems, phenyl groups are preferred according to the invention. With respect to bicyclic systems, these may be fused systems or bridged systems, with 9- or 10-membered fused systems such as naphthyl or indene being preferred. Of the above bicyclic systems, naphthyl is particularly preferred.

Preferably, the radical R 'is such that x is 4 or 5 and each n is independently 3 or 4. Particularly preferred R' groups are - [NH (CH ₂ ) ₃ ] _S NH ₂ and - [NH (CH ₂ ) ₃ ] ₃ NH (CH ₂ ) ₄ NH (CH ₂ ) ₃ NH ₂ .

The polyamine compounds of the present invention and the pharmaceutically acceptable salts thereof are antagonists of excitatory amino acid neurotransmitters. Thus, it is preferred to use these compounds as such to antagonize excitatory amino acid neurotransmitters. The polyamine compounds of the present invention can also be used to control invertebrate insects and to treat diseases and conditions in mammals that are mediated by excitatory amino acid neurotransmitters. Further, the polyamine compounds of the present invention are useful as psychotherapeutic agents for mammals.

The present invention also includes pharmaceutical compositions comprising the above-mentioned polyamine compounds, methods of applying the polyamine compounds, and methods of preparing the polyamine compounds.

Scheme of synthetic preparation of polyamine compounds of general formula:

is shown in Reaction Schemes A through C which follow.

Reaction Scheme A

(AND)

BOC (II)

BOC — NH

CN (III) [BOC] ₂ O ▼

BOC

BOC

BOC

H ₂ / Pd (OH) ₂ , HOAc ▼

BOC

BOC (VI) [BOC] ₂ O ▼

BOC BOC

BOC

H ₂ / Pd (OH) ₂ , HOAc

AND

BOC BOC

(VIII)

Reaction scheme

(B)

(IX)

Reaction scheme c (IX) ₊ (VIII)

ZB (X) [BOC] ₂ O

IN

BOC (XI) ▼ n

(XII)

NH,

In carrying out the process of Reaction Scheme A above, the polyamine compound of formula IV, an intermediate of said process, is prepared by a series of sequential steps starting from diaminobutane. Reaction conditions suitable for the preparation of the above intermediate of Formula VIII according to Reaction Scheme A can be found in Examples 5, Parts 1 to 7. Reaction Scheme B illustrates the procedure for preparing the intermediate of Formula IX. The reaction conditions suitable for the preparation of this intermediate are shown in Example 5, Part 8. The preparation of the polyamine compounds of the present invention of formula XII is illustrated in Reaction Scheme C. Reaction conditions suitable for condensing the intermediates of formulas VIII and IX and sequential sequence of reaction steps to prepare the compound of Formula XII is illustrated in Example 5, Parts 1 to 11.

The polyamine compounds of the present invention reversibly antagonize excitatory amino acid neurotransmitters, which neurotransmitters affect the activity of cells, such as cells of the nervous system of various organisms, including invertebrates and vertebrates. Vertebrates also include mammals. The term invertebrates as used herein also includes insects, ectoparasites and endoparasites.

The ability of the polyamine compounds of the invention to antagonize excitatory amino acid neurotransmitters can be demonstrated by their ability to block N-methyl-D-aspartic acid (NMDA) induced elevation of cGMP in the cerebellum of neonatal rats (NMDA) as follows.

The brains of 10 Wistar rats, 8-14 days of age, are quickly removed and placed in Krebs / bicarbonate buffer at pH 7.4 and 4 < 0 > C, after which the brains are cut into particles of dimensions

0.5 millimeters x 0.5 millimeters using Mcllwain tissue scalpel (The Nickle Laboratory Engineering Co., Gomshall, Surrey, UK). The cerebellum particles thus prepared are transferred to 100 ml of Krebs / bicarbonate buffer at 37 ° C, which is continuously maintained in equilibrium with an O ₂ / CO ₂ 95: 5 mixture of oxygen and carbon dioxide. ninety minutes, changing buffer three times. Then the buffer is decanted, the tissue is centrifuged (centrifuged for 1 minute at 3200 rpm) and the separated tissue is then resuspended in 20 ml of said Krebs / bicarbonate buffer. Aliquots of 250 μΐ (approximately 2 milligrams) are then separated and placed in 1.5 ml microtubes. To these tubes, add 10 μΐ of test compound from the stock solution, followed by the addition of 10 μΐ of 2.5 mM NMDA solution to initiate the reaction. The final concentration of NMDA is 100 μΐ. Controls do not contain NMDA. The tubes are then incubated for one minute at 37 ° C and shaken on a water bath, followed by the addition of 750 μΐ of a 50 mM Tris-Cl solution followed by a 5 mM EDTA solution to stop the reaction. The tubes are immediately placed in a boiling water bath and left there for five minutes. The contents of each of these tubes are then sonicated for 15 * seconds using a probe ultrasonic device set to a setting intensity of three. Ten microliters are then removed and the protein content determined by the method of Lowry, Anal. Biochem. 100: 201-220 (1979). The tube is then centrifuged (centrifuged for 5 minutes at 10000 g), 100 μΐ of supernatant is collected and the cyclic GMP (cGMP) level is determined using the New England Nuclear (Boston, Massachusetts) cGMP RIA test. , following the procedure described by this supplier. Values are reported as pmoles of cMGP generated per milligram of protein.

In addition, the ability of the polyamine compounds of the invention to antagonize excitatory amino acid neurotransmitters can be demonstrated by their ability to block elevated levels of free calcium ions [Ca ²⁺ ] _even in cytosol scattered granular small brain cells induced by the NMDA / glycine combination. Granulated cerebellum cells are prepared from the cerebellum of 8 day old rats (see.

Wilkin, GP et al., Brain Res: 115: 181-199, 1976). Squares (1 cm ² ) of Aclar (Proplastics Inc, 5033 Industrial Ave., Wall, NJ, 07719) were coated with poly-L-lysine and placed in 12-well dishes containing 1 ml of Eagles Basal Medium. The above cells are dissociated and aliquots containing 6.25 are placed in each well containing Aclar squares. 10 ^s cells. Cytosine-beta-D-arabinofuranoside (final concentration 10 μΜ) is added 24 hours after inoculation. These cells are then used for fura2 analysis, which is performed after 6, 7 and 8 days of culture. Cells bound to Aclar squares are transferred to 12-well dishes containing 1 ml of 2 μΜ fura2 / AM (Molecular Probes Inc, Augene, OR) in HEPES buffer (containing 0.1% bovine serum albumin; 0.1% dextrosis; pH 7.4; buffer does not contain magnesium). The cells are incubated for 40 minutes at 37 ° C, then the fura2 / AM containing buffer is separated and replaced with 1 ml of the same fura2 / AM-free buffer. 2.0 ml of pre-warmed buffer (37 ° C) was then placed in a quartz cuvette. The cells are then placed on Aclar cells and placed in a thermostated holder (37 ° C) equipped with a magnetic stirrer, and fluorescence is measured using a fluoroscopic spectrophotometer (Biomedical Instrument Group, University of Pennsylvania). The fluorescence signal is allowed to stabilize for about 2 minutes.

The increase in free calcium in the cytosol, which is represented by an increase in fluorescence, is due to the addition of 50 μΜ NMDA and 1 μΜ glycine. Next, add a 5-20 μΐ stock solution of the test compound in saline with phosphate buffered saline (PBS; pH 7.4) to the cuvette at the appropriate concentration. Calibration of fluorescent signals and fura2 / AM leak correction are performed according to the standard method. Grynkiewicz, G. et al., J. Biol. Chem. 260: 3440 (1985). After completion of each test, the maximum fluorescence value (Fmax) is determined by the addition of ionomycin (35 μΜ) and the minimum fluorescence value (Fmin) is determined by the subsequent addition of EGTA (12 mM) to the calcium chelate. In carrying out the above procedure, the ability of the present compounds of the invention to antagonize excitatory amino acid neurotransmitters is determined by the decrease in fluorescence upon addition of the compounds.

The polyamine compounds of the present invention are useful for antagonizing excitatory amino acid receptors per se. As such, these polyamine compounds are also useful for controlling invertebrate insects, for treating diseases mediated by excitatory amino acid neurotransmitters and for treating conditions in mammals such as stroke, cerebral ischemia, neuronal degenerative disorders such as Alzheimer's disease and epilepsy. The polyamine compounds of the present invention are also useful as psychotherapeutics used in mammals, and are furthermore suitable for studying physiological processes in cells, including cells of the nervous system.

The present invention also includes pharmaceutically acceptable salts of the polyamine compounds of the invention. These salts are prepared by methods well known in the art for those skilled in the art. For example, acid addition salts of the polyamine compounds of the invention can be prepared by conventional techniques. Preferred acid addition salts of the polyamine compounds of the present invention are those with hydrochloric acid and trifluoroacetic acid. Hydrochloric acid addition salts of the compounds of the invention are particularly preferred.

When the polyamine compounds of the invention or pharmaceutically acceptable salts thereof are used for administration to mammals, they may be administered alone or in combination with a pharmaceutically acceptable carrier or diluent in the form of a pharmaceutical composition as is conventional in the art. field. The polyamine compounds or pharmaceutically acceptable salts thereof may be administered orally or parenterally. Methods of parenteral administration include intravenous administration, intramuscular administration, intraperitoneal administration, subcutaneous administration, and topical administration.

For oral administration of the polyamine compounds of the invention or pharmaceutically acceptable salts thereof, the compounds may be administered, for example, in the form of tablets or capsules, or in the form of an aqueous solution or suspension. In the case of tablets for oral administration, carrier materials which are commonly used in similar cases such as lactose and corn starch may also be used as carriers, and in these cases also the addition of lubricants such as magnesium stearate is usually used. In the case of oral administration of capsules, suitable diluents are lactose and dried corn starch. When aqueous suspensions are used for oral administration, the active ingredient is combined with emulsifying and suspending agents. If desired, a sweetener or fragrance may also be added.

In the case of intramuscular, intraperitoneal, subcutaneous and intravenous administration, sterile solutions containing the active ingredient are usually prepared, the pH of the solutions being adjusted to a suitable value and a buffer is added. In the case of intravenous administration, the total concentration of solutes is controlled in such a way that an isotonic solution is prepared.

When using the polyamine compounds of the present invention, or salts thereof, in humans, the daily dose should be determined by the attending physician. However, a suitable dosage of the polyamine compound is in the range of about 1 to 30 milligrams / kilogram / day. In addition, it should be noted that the dosage administered depends on the age of the patient, the weight of the patient and the response that occurs after administration of the individual patient, as well as the severity of the patient's symptoms and the efficacy of each particular compound administered. . It will be appreciated from the above that dosages outside the range may also be used, and such dosages are within the scope of the present invention.

When the polyamine compounds of the present invention or salts thereof are used to control invertebrate insects, the compounds are applied directly to said invertebrate insects or applied to the environment in which the insects occur. For example, the compounds of the invention may be used in the form of solutions and applied directly to said invertebrate insect. The amount of compound necessary to control the invertebrate insect will vary according to the particular species of insect and the conditions occurring in the environment in which it is to be administered, as determined by the person administering it.

When the polyamine compounds of the present invention or salts thereof are used for physiological studies of cells, the compounds are applied to the cells by conventional methods known in the art. For example, the compounds may be applied to the cells in a suitable physiological buffer. When performing these tests, a suitable concentration of the compounds of the present invention is about 100 μΜ. However, even here it should be noted that the concentration of said compounds in performing these tests may be considerably greater or much less than the reported 100 μΜ. The amount of compound administered will be determined by one of ordinary skill in the art according to conventional methods.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of the present invention and the processes for their preparation will now be illustrated with reference to the following non-limiting examples.

Types of procedures occurring in the experimental part:

(A) protecting the secondary amine with di-tert-butyldicarbonate;

(B) primary amine cyanoethylation;

(C) catalytic hydrogenation of the nitrile to the primary amine;

(D) reactions to form an amide bond;

(D1 dimethylaminopropyl, ethylcarbodiimide / hydroxybenztriazole;

(D2) dicyclohexylcarbodiimide / hydroxybenztriazole;

(D3) dimethylaminopropyl, ethylcarbodiimide / hydroxybenztriazole / triethylamine;

(D4) dicyclohexylcarbodiimide / hydroxysuccinimide;

(E) deprotection of N-BOC substrates with hydrochloric acid in dioxane;

(F) deprotection of N-BOC substrates with trifluoroacetic acid (TFA);

(G) alkylation of amines with N-BOC-3-bromopropylamine.

Example 1

Preparation of H ₂ N [(CH ₂ ) ₃ NH] _X polyamine side chain.

Stage 1 - Procedure Type B _► ^{H: N} \ z / \ / ^NH \ z ^// \ (8)

103 grams

1,3-diaminopropane was combined with 45 mL of MeOH at 4 ° C with stirring. Acrylonitrile (100 mL, 81 grams, or 1.1 equivalents) was added dropwise via addition funnel with pressure equalizer over 90 minutes.

After 3 hours a 500 mg portion was collected and analyzed by ¹³ C NMR, with no 1,3-diaminopropane detected. This crude material, which contained the aminonitrile compound (8) as a product, was subjected to distillation under reduced pressure, three fractions being collected over a temperature range of 100 to 125 ° C. All three fractions were sufficiently pure to be used for reactions with di-tert-butyl carbonate and subsequent chromatographic purification on silica gel.

@ ³ H NMR (250 MHz, CDCl3 .delta.):

2.67 (t, 2H, 6.6 Hz), 2.28 (t, 2H, J = 6.6Hz), 1.05 (with, 3H);

2.54 - 2.43 (m, 4H),

1.37 (m, 2H, J = 6.7 Hz), ¹³ C NMR (63.1 MHz, CDCl ₃ ) δ:

118.8, 46.9, 44.9, 40.2,

33.4, 18.5.

Stage 2 - Type A procedure (8)

Boc Boc

To a solution containing aminonitrile compound (8) (23 grams, 0.18 mole) in 500 mL methylene chloride at 0 ° C was added di-tert-butyl dicarbonate (80 grams, 0). , 36 mol, or equivalents). The reaction mixture was stirred at ambient temperature for 16 hours and then treated with an additional portion of di-tert-butyl dicarbonate (8 g, 0.036 mol). After stirring for an additional 4 hours, the reaction mixture was washed with 1 N KOH (2 x 60 mL), dried over potassium carbonate, filtered and concentrated in vacuo. The product was purified by flash chromatography (silica gel;

20 → 100% ethyl acetate in hexane as eluent) to give the final product N-BOC-nitrile compound (9) as a clear oil.

Yield: 14 g, 24%.

1 H NMR (250 MHz, CDCl 3 δ:

3.40 (t, 2H, 6.7Hz), 3.28 (t, 2H, J = 6.6Hz), 3.05 (bs, 2H) 2.63 - 2.46 (m, 2H) 1.70 - 1.56 (m, 2H), 1.42 (s, 9H); 1.38 (s, 9H). ¹³ C NMR (63.1 MHz, CDCl ₃ ) δ: 155.8 155.1, 118.5, 80.7, 78.9, 45.7, 44.4, 43.4, 32,4, 28.3, 28.2. Step 3 - Type C Procedure

Boc Boc

According to this procedure, the N-BOC-nitrile compound (9) (49 g, 0.4 mol), 1000 ml acetic acid and 20 g (20% palladium hydroxide Pd (OH) ₂ ) Pd (OH) ₂ / C was placed in a 2.6 liter Parr shaker vessel. The flask was filled with hydrogen and pressurized to 345 Pa and then shaken for 4 hours. The reaction mixture was then filtered on 0.47 µ filter paper and concentrated under vacuum. The residue was dissolved in 1.5 L of CH ₂ Cl ₂ and washed with 1 N KOH (2 x 200 mL). The basic layers were extracted with CH ₂ Cl ₂ (400 mL) and the combined dichloromethane layers were dried over K ₂ CO ₃ , filtered and concentrated in vacuo to give the desired N -BOC-amine (11) as a clear colorless oil.

Yield: 43 g, 86%.

¹ H NMR (250 MHz, CDCl ₃ ) δ:

3.28-3.12 (m, 2H), 3.11 - 3.00 (m, 4H); 2.64 (t, 2H, J = 7 Hz), 1.65-1.50 (m, 4H), 1.42 (s, 9H); 1.38 (s, 9H). ¹³ C NMR (63.1 MHz, CDCl ₃ ) δ: 155.8, 79.3, 78.5, 44.0, 43.4, 39.2, 32.3, 28.5, 28.2. Step 4 - Type B Procedure

Boc Boc

38 grams of N-BOC amine compound (11) (equivalent to 0.114 mol) was combined with 6.7 grams of acrylonitrile (0.126 mol, 1.1 equivalents) in 60 milliliters of methanol, and the mixture was stirred for 11 hours. The solvents were removed to give the desired nitrile compound (12) as a clear colorless oil, which was then used without further purification.

Yield eq: 43 grams, which Yippee ¹ H NMR (250 MHz, CDCl ₃ ) δ: 3.18 (bs, 4H) 2.85 (t, 2H, J = 6.6Hz), 2.45 (t, 2H, J = 6.7Hz) 1/41 (s, 9H) ¹³ C NM [R (63.1 MHz, CDCl3 ₎ ) δ 155.9 , 118.6, 79.6, 78, 28,8, 28.3, 18.6.

100%.

3.03 (m, 2H), 2.57 (t, 2H, J = 6.7Hz) 1.72 - 1 53 (m, 4 H), 1.38 (with, 9H). 46, 3, 45.0, 43.9, 37.4,

Step 5 Type A Procedure (12)

Boc Boc Boc

A 43 grams (0.114 mol) sample of nitrile compound (12) was combined with di-tert-butyldicarbonate (25.6 grams, 0.120 mmol, or 1.05 equivalents) and added 350 ml of CH ₂ Cl ₂ at 0 ° C and stirred for 9 hours. Thin Layer Chromatography (TLC) (EtOAc, KMnO 3) showed no residual starting material. The reaction mixture was purified as described above.

in the case of N-BOC-nitrile N-BOC-nitrile compound compound (9). The required ...................... (13) was obtained in the form. ^d '^; Clear, colorless oil. Yield: 34 grams which is 63%. _____. .------. . -Γ--,. ¹ H NMR (250 MHz, CDCl 3): 7 6 Λ N Z 3.45 (t, 2H, J = 6.6Hz), 3.39 - 2.97 (m, 8H) 2.68 - 2.46 (s, 2H), 1.82 - 1.56 (m, 4H) 1.44 (s, 18H); 1.87 (s, 9H); 1. '- ¹³ C NMR (75.7 MHz, CDCl ₃ ) δ J 155.9, 80.5, 79.7, 78.9 , 46.5, 44.5 , 43.9, 37.6,.

28.4, 28.3, 16.9.

Step 6 - Type C Procedure (13) -►

(14)

In this embodiment, the N-BOC-amine compound (14) was prepared from the N-BOC-nitrile compound (13) in the same manner as the N-BOC-amine compound (11) was prepared from the N-BOC-nitrile compound (9).

Yield: 99% (30 grams).

¹ H NMR (300 MHz, CDCl ₃ ) δ: 3.32 - 2.94 (m, 10H), 2.62 (t, 2H, J = 6.7Hz), 1.76 - 1.52 (m, 6H), 1.39 (s, 18H), 1.37 (s, 9H); 1.25 (s, 2 H). ¹³ C NMR (63.1 MHz, CDCl ₃ ) δ 155.5, 79.5, 79.3, 45.5 - 43.7, 39.1, 37.3, 32.3,

2ξ, 3.

(15)

Step 7 - Type B Procedure (14) -►

Boc Boc

The nitrile compound (15) was prepared from the N-BOC-amine compound (14) in the same manner as the nitrile compound (12) was prepared from the N-BOC-amine compound (11).

Yield: 90%. ¹ H NMR (300 MHz, CDCl ₃ ) δ: 3.29-3.02 (m, 14H), 2.86 (t, 2H, J = 6.7 Hz), 2.57 (t, 2H, J = 6.6Hz), 2.46 (t, 2H, J = 6.6 Hz), 1.72-1.57 (m, 6H), 1.41 (WITH, 9H), 1.40 (s, 9H); 1.39 (WITH, 9H). ¹³ C NMR (75.7 MHz, CDCl ₃₎ ) δ: 155.5, 155.0, 118.7, 79.6, 79 , 5, 46.7 - 46 , 0, 45.2 - 43.3, 38.0 - 36 , 9, 28.4, 18 , 7.

The ¹³ C NMR (300 MHz, CDCl ₃ ) values of the nitrile compound (15) were not distinguishable from the nitrile compound (12).

Step 8 - Type A procedure

Boc Boc

In this embodiment, the N-BOC-nitrile compound (16) was prepared from the nitrile compound (15) in a similar manner as the N-BOC-nitrile compound (13) was prepared from the nitrile compound (12) to give the product as a clear colorless oil.

Yield: 30 g, 87%.

¹ H NMR (300 MHz, CDCl ₃ ) δ:

3.36 (t, 2H, J = 6Hz), 3.18-2.90 (m, 14H),

2.57-2.42 (m, 2H), 1.72-1.48 (m, 6H),

1.40-1.28 (m, 27H).

Step 9 - Type C Procedure

Boc Boc

In this embodiment, the N-BOC-amine compound (17) was prepared from the N-BOC-nitrile compound (16) in a similar manner as the N-BOC-amine compound (11) was prepared from the N-BOC-nitrile compound (9).

Yield: 2.61 g 74% • ¹ H NMR (250 MHz, CDCl 3 δ: 3.39 - 2.97 (m, 14H), 2.63 (t, 2H, J = 6.6Hz), 1.80-1.53 (m, 8H), 1.39 (with, 27H), 1.38 (s, 9H); 1,23 (WITH, 2H).

Step 10 - Type B procedure (17)

Boc Boc

In this embodiment, the nitrile compound (18) was prepared from the N-BOC-amine compound (17) in the same manner as the nitrile compound (12) was prepared from the N-BOC-amine compound (11), and the compound thus prepared was used for the next procedure. without further cleaning.

Yield: 1 9 grams, which is 91%. ¹ H NMR (300 MHz, CDC1 ₃₎ δ: 3.24 - 2.94 (m, 14H) 2.87 (t, 2H, J ~ 6 Hz), 2.57 (t, 2 H) , J-6 Hz) 2.47 (t, 2H, J- 6 Hz), 1.74 - 1.54 (m, 8H) 1.45 - 1 36 m 36H). Stage 11 - Type A procedure

(18)

Boc Boc Boc Boc (19)

In the process of this embodiment, the N-Boc-nitrile compound (19) was prepared from nitrile compound (18) in the same manner as N-Boc-nitrile (13) is prepared Yield: 16 grams, ^X H NMR (250 MHz , CDCl ₃ ) δ:

3.43 (t, 2H, J = 6.6Hz), 2.62 - 2 50 (m, 2H); 1.43 (with, 9H), 1.41 (with, 18H),

from a nitrile compound (12).

%.

3.28-3.02 (m, 16H),

1.80 - 1.56 (m, 8H),

1.42 (s, 9H);

1.39 (s, 9H).

Stage 12 - Type C procedure (19)

Boc Boc

Boc

NH ₂ (21)

In this embodiment, the N-BOC-amine compound (20) was prepared from the N-BOC-nitrile compound (19) in the same manner as the N-BOC-amine compound (11) was prepared from the N-BOC-nitrile compound (9).

Yield; 17 grams, which is 99%.

Ή NMR (300 MHz, CDCl ₃ ) δ:

3.24-2.95 (m, 18H), 2.6 (t, 2H, J = 6H),

1.72-1.52 (m, 10H), 1.42-1.32 (m, 45H).

Step 13 - Type B procedure (20)

Boc Boc

CN (21)

In this embodiment, the nitrile compound (21) was prepared from the N-BOC-amine compound (20) in the same manner as the nitrile compound (12) was prepared from the N-BOC-amine compound (11).

Yield: 99%.

¹ H NMR (250 MHz, CDCl ₃ ) δ;

3.32-3.03 (m, 16H), 2.91 (t, 2H, J = 6.7 Hz),

2.61 (t, 2H, J = 6 Hz), 2.51 (t, 2H, J = 6.6 Hz),

1.82-1.57 (m, 10H);

1.43 (s, 9H).

Step 14 - Type A procedure

1.44 (s, 36H);

J 1 ,, J 'J (21) • f ·:

CN <22)

In this embodiment, the N-BOC-nitrile compound (22) was prepared from the nitrile compound (21) in the same manner as the N-BOC-nitrile compound (13) was prepared from the nitrile compound (12). Yield 99%.

¹ H NMR (CDCl 3) δ:

3.45 ppm (t, 2H, J = 6.6Hz), 3.26-3.02 (m, 18H), 2.65 - 2.53 (m, 2H), 1.79 - 1.55 (m, 10H), 1.43 (s, 9H); 1.42 (s, 9H); 1.41 (s, 18H); 1.40 (s, 9H). ¹³ C NMR { ^X H} (250 MHz, CDCl ₃ ) δ: 155.2, 79.3, 44.7, 28.3, 28.3, 28.2.

Stage 15 - Type C procedure (22)

Boc Boc Boc

In this embodiment, the N-BOC-amine compound (23) was prepared from the N- * BOC-nitrile compound (22) in the same manner as the N-BOC-amine compound (11) was prepared from the N-BOC-nitrile compound (9).

Yield: 8 g, 99%.

¹ H NMR (300 MHz, CDCl 3 δ:

3.32-2.98 (m, 22H), 2.65 (t, 2H, J = 6.6 Hz),

1.78 - 1.53 (m, 12H), 1.41 (s, 54H),

1.40 (s, 9H).

Stage 16 - Type B procedure (23)

Boc Boc Boc Boc Boc

In this embodiment, the nitrile compound (24) was prepared from the N-BOC-amine compound (23) in the same manner as the nitrile compound (12) was prepared from the N-BOC-amine compound (11).

Yield: 99%.

¹ H NMR (250 MHz, CDCl ₃ ) δ:

3.32 - 3.04 (m, 22H), 2.90 (t, 2H, J = 6.7 Hz), 2.61 (t, 2H, J = 6Hz), 2.53 (t, 2H, J = 6.7 Hz), 1.82-1.57 (m, 12H), 1.44 (with, 45H),

1.43 (s, 9H).

Step 17 - Type A procedure (24)

Italy (25)

In this embodiment, the N-BOC-nitrile compound (25) was prepared from the nitrile compound (24) in the same manner as the N-BOC-nitrile compound (13) was prepared from the nitrile compound (12).

Yield: 5.5 g, 74%.

¹ H NMR (250 MHz, CDCl ₃ ) δ:

3.45 (t, 2H, J = 6.7 Hz), 3.40-2.97 (m, 24H),

2.66 - 2.51 (m, 2H), 1.80 - 1.55 (m, 12H),

1.54-1.30 (s, 63H).

Step 18 - Type C Procedure (25)

Boc Boc Boc Boc Boc (26)

In this embodiment, the N-BOC-amine compound (26) was prepared from the N-BOC-nitrile compound (25) in the same manner as the N-BOC-amine compound (11) was prepared from the N-BOC-nitrile compound (9).

Yields] k: 5.01 grams, which is 91%. ¹ H NMR (250 MHz, CDCl3) δ: 3,30 - 2.97 (m, 26H); 2.60 (t, 2H, J = 6Hz); 1,81 - 1.57 (m, 14H); 1.53-1.28 (m, 63H). 1 H NMR (250 MHz, CDCl3) δ: 155.2, 79.3, 44.7, 28,7, 28.4, 27.5.

Example

O 'ΝΗ' ^{Χ Χ} ' ^/ Ζ ~ ^ν ' ΝΗ '

NH

NH, NH, NH _n

Fe

Zó \

Step 1: Amide Bonding - Type D1 Procedure.

According to this embodiment, 0.19 grams of ferrocenecarboxylic acid (representing 0.82 mmol or 1.1 equivalents) was placed in a 50 ml 1-neck RBF vessel, followed by 0.12 grams of hydroxybenzotriazole (0.89 mmol, 1.2 equivalents), 0.17 g of l- (3-dimethylaminopropyl) -3-ethylcarbodiimide (as the HCl salt, 0.90 mmol, 1.2 eq) and 10 ml of CH ₂ C1 _2, which was made with stirring and under a dry nitrogen atmosphere. After 30 minutes, 0.61 g of N-BOC-amine compound (27) (equivalent to 0.75 mmol, or 1.0 equivalent; see Example 5a below to prepare this compound) was added to this solution. Thin Layer Chromatography (TLC) (2 x MeOH,

₂ ) it was found that the N-BOC-amide compound was consumed within 2 hours. The reaction mixture was diluted to a total volume of 400 mL with ethyl acetate and then washed with pH 4 buffer (2 x 25 mL), followed by 25 mL of water, 1N KOH (2 x 25 mL), 25 mL of water. and 50 milliliters of brine. The ethyl acetate layer was dried over Na ₂ SO ₄ , filtered, and the solvents were removed to give the final product as an orange oil.

Yield: 712 mg, 93%.

¹ H NMR (250 MHz, CDCl ₃ ) δ:

4.78 (s, 2H), 4.32 (t, 2H, J = 1.8 Hz),

4.19 (s, 5H); 3.45-3.04 (m, 20H);

1.83-1.59 (m, 12H), 1.50 (s, 9H),

1.45 (s, 18H), 1.44 (s, 9H),

1.43 (s, 9H).

Step 2: Deprotection of the polyamine compound - Type F procedure

Trifluoroacetic acid (30 mL) was degassed by bubbling a dry nitrogen stream (using a Teflon tube) in a 100 mL single neck RBF at 0 ° C. Ferrocenkarboxamidpolyaminová compound obtained in the above Step 1 (in 712 mg, 0.69 mmol) was dissolved in 2 ml of CH ₂ C1 ₂ and then transferred to the stirring TFA with simultaneous flushing with dichloromethane CHjClj amount 3x2 milliliters. After 30 minutes, the ice bath was removed and after a further 30 minutes the solvents were removed under reduced pressure, after which Hi-Vac treatment was performed. The residual red-brown oil was stirred with Et ₂ O diethyl ether (3 x 30 mL) to isolate the yellow solid under nitrogen pressure on a porosity B frit. The solid was rinsed with ether and the residual ether was ejected under pressure. nitrogen to give the desired product as a solid.

Yield: 690 mg, 93%.

¹ H NMR (DMSO) δ: 4.71 (t, 2H, J = 1.73 Hz), 4.38 (t, 2H, J ~ 2Hz) 4.15 (s, 5H); 3.28-3.21 (m, 2H); 3.03 - 2.81 (m, 18H), 1.61 - 1.50 (m, 4H). ¹³ C NMR (250 MHz, D ₂ O): 2.04-1.72 (m, 8H),

177.3, 78.5, 76.1, 74.3, 49.7, 48.1, 47.3, 47.2,

47.1, 39.3, 38.8, 28.5, 26.5, 25.5, 25.4.

HPLC: (minimum) 96.08% purity; Novapak C18 column, 5-40% CH ₃ CN / 2% / TH ₂ O over 60 min, determined at 230 nm, elution time: 23.2 min.

HRMS (FAB)

(Μ + H) calcd for C ₂₇ H ₄₈ N _e O: 529.3328845, found: 529.33172.

Example 3 o

N

Step 1: Amide bond formation - Type D3 procedure.

The hydrochloride salt was used

Of 2-pyridylacetic acid (0.105 g, 0.60 mmol, or 1.0 equivalent) combined with 0.16 ml of TEA (1.15 mmol, or 2 equivalents) and 4 ml of CH ₂ Cl ₂ dichloromethane . After 10 minutes, DEC (0.12 g, 0.62 mmol, or 1.0 equivalent) and 0.09 g HOBt (0.66 mmol,

1.1 equivalents), and the mixture was stirred for 2 hours. To the resulting reaction mixture was added N-BOC-amine compound (27) (0.44 g, 0.54 mmol, or 0.9 equivalents) and stirred for an additional 10 hours . Thin Layer Chromatography (TLC) (2 x MeOH, I ₂ ) showed that the N-BOC-amine compound was consumed. The reaction mixture was then diluted to a volume of 400 mL with ethyl acetate EtOAc, then washed with 1 N KOH (40 mL), brine (50 mL), and then dried over MgSO ₄ . The ethyl acetate solution was filtered and the solvents were removed to give 0.4 g of pure green oil. The crude material was chromatographed on 10 g of silica gel, which was suspended in ethyl acetate with EtOAc, eluting with ethyl acetate. The appropriate fractions were combined and the solvent was removed to give the final product as a clear light green oil.

Yield: 0.20 g, 40%.

* H NM R (250MHz, CDC1 ₃ ) 8.59 - 8,52 (m, ^{Χ Η)} 7.62 - 7,49 (m, ^X H), 3.78 (s, 2H) 9 1.80 - 1,56 (m, 12),

7.76 - 7.68 (m, 1H), 7.37 (d, ^{Χ Η,} J = 8 Hz), 3.31 - 3.02 (m, 20H), 1.56 - 1.34 (m, 45H).

Step 2: Deprotection of the polyamine compound - Type F procedure

Trifluoroacetic acid (30 mL) was continuously degassed with a dry nitrogen stream bubbled through the acid (via a Teflon tube) in a 100 mL single neck RBF vessel at room temperature. The 2-pyridyl acetamide compound obtained in Step 1 above (180 g) was dissolved in 2 mL of CH ₂ Cl ₂ dichloromethane and then this solution was transferred to the above stirred trifluoroacetic acid TFA. After 1 hour, the solvent was removed under reduced pressure and the residue was placed on Hi-Vac. The residue was mixed with ether (3 x 30 mL) to give a white solid which was collected on a porosity B frit using pressurized nitrogen. The residual ether was then removed under nitrogen pressure. The desired product was obtained.

Yield: 169 g, 91%.

NMR (250 MHz, D ₂ O) δ:

8.48 (d, * H, J = 15Hz) 7.51 - 7,47 (m, 2H) 3.32 (t, 2H) . J ~ 9Hz) 2.17 - 1,98 (m, 2H) 1.81 - 1,69 (m, 4 H).

7.98 (t, 1H, J = 6Hz); 3.86 (s, 2H);

3.17 - 2.96 (m, 18H),

1.98-1.81 (m, 2H)

Example

O

NH

NH

NH

NH, NH, NH,

Step 1: Amide bond formation - Type D4 procedure.

In this embodiment, 4-biphenylacetic acid (53 grams = 0.25 mmol) was used

1.2 eq) was combined with 5 ml of CH ₂ C1 _2, 84 μΐ triethylamine (0.6 mmol, 3 eq) 70 mg dicyclohexylcarbodiimide (0.34 mmol, 1.6 eq), 11 milligrams of hydroxysuccinimide (0.09 mmol, 45 mol%) and 175 milligrams

N-BOC-amine compounds (27) (which represents 0.21 mmol, or 1.0 equivalent). TLC (two parts MeOH; potassium permanganate KMnO ₄ ) showed that after 16 hours

The N-BOC-amine compound consumed. The reaction mixture was diluted to 100 mL with CH ₂ Cl ₂ and washed with aqueous 20% NH ₄ OH (2 x 100 mL). The basic layers were extracted with CH ₂ Cl ₂ (3 x 50 mL) and the combined dichloromethane fractions were washed with brine (50 mL), dried with K ₂ CO ₃ , filtered, and the solvent was used. discarded to yield 281 mg (> 100% yield) of crude material. The pure product was isolated by flash chromatography on silica gel (12 grams of silica gel suspended in CH ₂ Cl ₂ dichloromethane eluting with a 0-10% MeOH / CH ₂ Cl ₂ gradient) to give the product a white, waxy solid.

Yield: 190 g which ^X j H NMR (250 MHz, CDC1 ₃₎ δ:

7.56 - 7.50 (m, 4H);

3.56 (s, 2H).

1.78 - 1.52 (m, 12H),

%.

7.43 - 7.28 (m, 5H), 3.26 - 2.98 (m, 20H), 1.48 - 1.36 (m, 45H).

Step 2: Deprotection of the polyamine compound - Type F procedure

Trifluoroacetic acid (30 mL) was continuously degassed using a nitrogen stream which was bubbled through the acid at 0 ° C (using a Teflon tube).

The biphenylacetamine compound obtained in Step 1 above (150 mg, 0.15 mmol) was added as dry powder to the stirred trifluoroacetic acid TFA. After 40 minutes the ice bath was removed and after another 20 minutes the solvents were removed under reduced pressure, the residue was placed on Hi-Vac. After 2 hours, the obtained reddish brown oil was mixed with Et 2 O (3 x 30 mL) and the resulting white solid was collected under nitrogen pressure on a porosity C frit. The solid thus obtained was dissolved in water, rinsed on a frit and dried freeze-drying to give the desired product as a white solid.

Yield: 136 mg ^X H NMR (300 MHz, D ₂ 0) δ:

7.62 - 7.56 (m, 5H);

7.38 - 7.31 (m, 2H),

3.13 (t, 2H, J = 6.7 Hz), 2.00-1.54 (m, 12H).

which is 99%.

7.4 (t, 2H, J = 7.5Hz); 3.45 (s, 2H);

3.02-2.80 (m, 18H),

Example 5

Preparation of 1H-indole-3-acetamide-N- (16-amino-4,8,13-triazahexadec-1-yl).

Stage 1:

NC ^{/ Z} ^

H ₂ N

CN (I)

Compound I was prepared from a diaminobutane compound and acrylonitrile according to the procedure of Yamamoto, Hisachi, J. Am. Chem. Soc. 103: 6133-61 (1981).

Stage 2:

(I) (Π) ▼

BOC

CN (III)

To a solution containing N-cyanoethyl-1,4-diaminobutane (6.44 g, 0.0447 mol) in acetonitrile (200 mL) under nitrogen was added KF / Celite (11 g), followed by the dropwise addition of N- (tert-butoxycarbonyl) -3-bromopropylamine (10.87 g, 0.0457 mol) over a period of 7 hours. The reaction mixture was allowed to stir for 16 hours at ambient temperature and then heated at 70 ° C for 24 hours. The next reaction mixture was allowed to cool, then filtered and concentrated under vacuum. The residue was dissolved in CH ₂ Cl ₂ (200 mL), washed with 1 N sodium hydroxide (100 mL), dried and concentrated in vacuo to give a crude product which was chromatographed on silica gel ( CH ₂ Cl ₂ / MeOH was used as eluent to give the desired amine compound III. Yield: 3.32 g.

1 H NMR (CDCl ₃ ) δ:

1.19 - 1.59 (m, 17H), 2.42 (t, J = 6.6Hz, 2H), 2.44 - 2.58 (m, 6H), 2.82 (t, J = 6.6Hz, 2H), 3.08 (m, 2H). 5.22 (brs, 1H); ¹³ C NMR (CDCl ₃ ) δ 18.68, 27.70, 27,74, 28.42, 29.94, 39.16, 45.03, 47.68, 48.99, 49,65, 78.78, 118.75, 156.11;

HR FABMS: observed (Μ + H) m / z = 299.2434, ^C 15 ^H 31 ^N 4 ° 2 (required 299.2447).

(III)

Stage 3:

BOC — bJ H t

[BOC] ₂ O

BOC — NH

BOC

BOC

The compound of formula III (4.7 g, 15.8 mmol), prepared as described in Step 2 above, was dissolved in 150 ml of dichloromethane under a nitrogen atmosphere. 7.56 g (34.7 mmol) of di-tert-butyldicarbonate were then added and the reaction mixture was stirred overnight at room temperature. The reaction mixture was concentrated in vacuo and chromatographed on 400 g of silica gel with ethyl acetate / hexane (50:50) as solvent. The individual fractions were monitored by thin layer chromatography (TLC). eluting with 50:50 ethyl acetate / hexane. The fractions containing the product of formula IV were combined and concentrated under vacuum to give the desired product as an oil.

Yield: 7.9 g.

¹ H NMR (CDCl ₃ ) δ 1.20 - 1.59 (m, 33H), 2.55 (m, 2H). 3.01 - 3.37 (m, 8H), 3.39 (t, J = 6.6 Hz, 2H), 5.25 (br s, 1H). • 1 ¹³ C NMR δ: 17.21, 25.73, 25,94, 28.22, 28.24, 28.27, 37,91, 43.78, 44.24, 46,60, 47.95, 78.96, 79.57, 80,44,

155.01, 155.75, 155.98;

HR FABMS: observed (Μ + H) m / z = 499.3501, C ₂₅ H ₄₈ N ₄ O ₆ (required 499.3496).

Stage 4:

BOC

BOC BOC (IV) ▼

BOC

BOC (A)

To 125 milliliters of acetic acid under a nitrogen atmosphere was added 7.85 g (15.8 mmol) of the compound of formula IV, prepared as described in Step 3 above, followed by 6.5 g of Pd (OH). ₂ / carbon. The reaction mixture was hydrogenated at 50 psi for 2 hours. The catalyst was removed by filtration and the filter cake was washed well with acetic acid.

The filtrate was concentrated, taken up in 250 mL of dichloromethane, washed twice with 100 mL of 1 N NaOH and dried over K ₂ CO ₃ . The solution was then filtered and the filtrate was concentrated in vacuo to give the title compound of formula V.

Yield: 7.8 g.

¹ H NMR (CDCl ₃ ) δ:

1.24-1.59 (m, 35H), 2.14 (s, 2H),

2.61 (t, J = 6.7 Hz, 2H), 2.98-3.14 (m, 10H),

5.22 (br s, 1H);

¹³ C NMR (CDCl ₃ ) δ:

25.89, 28.42, 31.38, 32.36, 37.55, 38.95, 43.95,

46.65, 79.34, 79.48, 155.65, 156.03;

HR FABMS: observed (Μ + H) m / z = 503.3 804,

C ₂₅ H ₅₁ N ₄ O _e (m / z 503.3809).

Stage 5:

(V) +

BOC

BOC (VI)

The compound of formula V (7.15 g, 14.2 mmol), prepared as described in step 4 above, was dissolved in 150 ml of methanol under a nitrogen atmosphere. 1.03 ml (15.6 mmol) of acrylonitrile was then added and the reaction mixture was stirred for 72 hours at room temperature. The reaction mixture was concentrated, further diluted and concentrated three times with dichloromethane, and then the solvent was stripped off under vacuum to give the title compound VI as an oil.

Yield: 7.65 g.

1 H NMR (CDCl ₃ ) δ:

1.26-1.73 (m, 36H), 2.44 (t, J = 6.7 Hz) 2H), 2.54 (t, J = 6.7Hz, 2H), 2.83 (t, J = 6.7 Hz) 2H), 3.00 - 3.16 (m, 10H); 5.24 (br s, 1 H). ¹³ C NMR (CDCl ₃ ) δ: 18.64, 25.84, 28.09, 28.43, 28 1.74, 37.84, 44,18, 44.68, 45.14, 46.29, 46.73, 46 i, 85, 49.70, 78,90,

79.29, 79.46, 118.52, 155.84, 155.98.

HR FABMS: observed (Μ + H) m / z = 556.4064, C ₂₈ H ₅₄ N ₅ O ₆ (desired m / z 556.4074).

Stage 6:

(VI) + [BOC] ₂ O -►

BOC BOC

BOC *

CN

BOC

The compound of formula VI (6.45 g, 11.6 mmol), prepared as described in Step 5 above, was dissolved under a nitrogen atmosphere in 125 mL of dichloromethane. 2.6 g (12 mmol) of di-t-butyldicarbonate were added to the obtained solution, and they were stirred overnight at room temperature. The reaction mixture was concentrated in vacuo and chromatographed on 400 g of silica gel using ethyl acetate / hexane (50:50) as eluent. The product containing fractions were combined and concentrated to give the desired product of formula VIII as an oil.

Yield: 6.6 g. ¹ H NMR (CDCl ₃ ) δ:

1,26 - 1.73 (m, 44H), 3, 03 - 3.24 (m, 14H) 3.42 (t , J = 6.6 Hz, 2H) 5, 25 (br, 1H). ¹³ C NMR (CDC1 ₃ ) δ: 17,20, 25.88, 27,83, 28, 12, 28,35, 28.45, 28.77, 37,87, 43,91, 44,20, 44, 77, 46,27, 46.88, 78.94, 79,42, 79,50, 80,54, 117 »91, , 154,96, 155.44, 155.74,

155.99.

HR FABMS: observed (Μ + H) m / z = 656.4579, ^C 33 ^H 62 ^N 5 ° 8 (desired m / z 656.4598).

Stage 7:

BOC BOC

I I

(VIII)

To 150 ml of acetic acid under a nitrogen atmosphere was added 6.6 g (10.1 mmol) of compound VII, prepared as described in step 6 above, followed by 6 g of Pd (OH) ₂ / carbon . The reaction mixture was hydrogenated at 50 psi for 2 hours. The catalyst was removed by filtration and the filter cake was washed thoroughly with acetic acid. The filtrate was concentrated, the residue was taken up in 200 ml of dichloromethane, washed twice with 100 ml of 1 N sodium hydroxide solution and dried over K ₂ CO ₃ . Next, the solution was filtered and the filtrate was concentrated in vacuo to give the desired compound of formula VIII. Yield: 6.5 g.

¹ H NMR (CDCl 3) δ:

1.28-1.71 (m, 46H), 2.16 (br s, 2H),

2.65 (t, J = 6.7 Hz, 2H), 3.01-3.18 (m, 14H),

5.24 (br s, 1H);

¹³ C NMR (CDCl 3) δ:

25.85, 27.66, 28.45, 28.76, 39.10, 44.21, 44.91,

46.80, 79.27, 79.46, 155.41, 155.67, 155.99.

HR FABMS: observed (Μ + H) m / z = 660.4914,

C ₃₃ H ₆₆ N ₅ ° 8 (M / z 660.4911).

Stage 8:

O

H

1.75 grams (10 mmol) of indole acetic acid, 1.15 grams (10 mmol) of N-hydroxysuccinimide and 2.06 grams (10 mmol) of dicyclohexylcarbodiimide were added to 75 milliliters under a nitrogen atmosphere. tetrahydrofuran. The reaction mixture was stirred at room temperature, whereupon a precipitate formed after about 5 minutes. After about 1.5 hours, the precipitate was removed by filtration, the filter cake was washed with 75 ml of tetrahydrofuran and dried to give 1.84 g of product. The combined filtrates were concentrated, taken up in ethyl acetate and filtered, and the filter was washed with ethyl acetate. The filtrate was concentrated to give a foam. The foam was triturated with 75 mL diethyl ether to give a hard gum. About 30 mL of ethyl acetate was added, followed by the addition of ethyl ether. The solid product was collected by filtration, washed with diethyl ether and dried under a nitrogen atmosphere to give 1.74 g of the product of formula IX. It was found that an additional 0.47 g of product could be obtained by treating the mother liquor with petroleum ether.

Stage 9:

(X)

N-BOC

H

The compound of formula VIII (0.33 g, 5 mmol), prepared as described in step 7 above, was dissolved under nitrogen atmosphere in 10 ml of dichloromethane with stirring, then 0.136 was added. g (5 mmol) of the compound of formula IX, prepared as described in Step 8 above. The reaction mixture was stirred overnight at room temperature. The reaction mixture was diluted to 35 mL with dichloromethane, washed with 10 mL of 0.5 N NaOH, dried with K ₂ CO ₃ and concentrated. The concentrate was chromatographed on silica gel using ethyl acetate / hexane (4: 1) as eluent. Product containing fractions were concentrated to give 0.37 g of the product as a white foam containing the compound of formula X with some ethyl acetate present. Stage 10:

(X) [BOC] ₂ O

H, N

BOC

BOC

N-BOC

H

BOC

The compound of formula X (0.37 g, 0.45 mmol), prepared as described in step 9 above, was dissolved under nitrogen atmosphere in 10 ml of dichloromethane. 0.218 g (1 mmol) of di-t-butyldicarbonate was then added, followed by 12 ml (0.1 mmol)

Of 4- (N, N-dimethylamino) pyridine. The reaction mixture was stirred at room temperature for 1 hour and then allowed to stand overnight. The reaction mixture was chromatographed on silica gel, eluting with ethyl acetate: hexane (4: 1), and the product containing fractions were concentrated to give the product of formula XI as a white foam.

Yield: 0.32 g.

Stage 11;

(XI)

(xii)

The compound of formula XI (0.32 g, 0.35 mmol), prepared according to Step 10 above, was added to 15 ml of trifluoroacetic acid under a nitrogen atmosphere and the mixture was stirred for 15 minutes. The reaction mixture was concentrated under vacuum and triturated with diethyl ether to give the desired product as a white powder.

Yield: 0.30 g.

In an analogous manner to the above, the N-BOC-amine compound (27) of the following formula was prepared:

Boc Boc

Following steps 1 to 7 above, the N-BOC-amine compound of formula VIII is obtained.

Stage 8a:

(VIII)

Boc Boc

The nitrile compound of formula XIII was prepared from the N-BOC-amine compound VIII in a similar manner as the nitrile compound VI was prepared from the N-BOC-amine compound V (see Example 5, step 5).

Yield: 1.00 g, 93%.

¹ H NMR (CDCl 3) δ:

1.26-1.66 (m, 47H),

2.56 (t, J = 6.7Hz, 2H),

2.45 (t, J = 6.6 Hz, 2H), 2.85 (t, J = 6.6 Hz, 2H),

3,01 - 3.30 (m, 14H), 5.25 (br s, - IH); ¹³ C NMR (CDClj) δ: 18,68, 25,92, 28,46, 28.48, 37.49, 44,19, 45,16, 46,73, 78,93, 79.32, 79.44, 118.70 155,61, 156.04 •

44.88,

155,46,

HR FABMS: observed (Μ + H) m / z = 713.5191,

C ₃₆ H ₆₉ N ₆ O ₈ (m / z 713.5177).

Stage 9a:

(XIII)

(XIV)

In this embodiment, the N-BOC-nitrile compound of formula XIV was prepared from the nitrile compound XIII using procedure A protected by an amine group.

Stage 10a:

In this embodiment, the N-BOC-amine compound (27) was prepared by hydrogenating the N-BOC-nitrile compound XIV in a similar manner to the preparation of N-BOC-amine compound VIII from N-BOC-nitrile compound VII (see step 7 in this example). ).

Examples 6 and 7

Starting from the examples, starting from a polyamine compound (20) and a suitable R-acetic acid (if m = 1) or a carboxylic acid (if m = 0) using a D1 type followed by followed by a type F procedure and compounds of formula:

R (CH ₂ ) _n CO [NH (CH ₂ ) ₃ ] ₅ NH ₂ . 5 TFA

Example m R 6 0 ferrocene 7 1 3-indole

Examples 8 to 29

Starting from the examples, starting from a polyamine compound (27) and a suitable R-acetic or carboxylic acid, the following formulas were prepared:

R (CH ₂ ) _n CO [NH (CH ₂ ) ₃ ] ₃ NH (CH ₂ ) ₄ NH (CH ₂ ) ₃ NH ₂ . 5 HCl (using procedure E) or

R (CH ₂ ) n CO [NH (CH ₂ ) ₃ ] ₃ NH (CH ₂ ) ₄ NH (CH ₂ ) ₃ NH ₂ . 5 TFA (using procedure F) using the procedures in the following table.

Example m R Progresses 8 0 ferrocene D1, then F 9 0 2-pyridine D3, then F 10 0 3-pyridine D3, then F 11 0 4-pyridine D3, then F 12 1 2-pyridine D3, then F 13 1 3-pyridine D3, then F 14 1 4-pyridine D3, then F 15 Dec 0 2-quinoline D1, then F 16 0 3-quinoline D2, then F 17 1 3-indole D1, then E 18 1 3- (5-hydroxyindole) D2, then F 19 Dec 1 3- (4-hydroxyindole) D2, then E 20 May 1 3- (5-bromoindole) D2, then F 21 1 3- (4-Fluoroindole) D2, then F 22nd 0 2- (5-Fluoroindole) D2, then F 23 1 2- (5-Fluoroindole) D2, then F 24 1 3- (5-methoxyindole) D2, then F 25 0 2-quinoxaline D2, then F 26 0 hydroxyquinone D2, then F 27 Mar: 0 4-resorcinol D2, then F 28 1 p-biphenyl D4, then F 29 1 2-Naphthalene D2, then F

Examples 30 and 31

Starting from the polyamine compound (17) and the appropriate R-acetic or carboxylic acid, the following procedures were prepared using process D1 followed by process F, compounds of the following formula: R (CH ₂ ) _and CO [NH (CH ₂ ) ₃ ] ₄ NH ₂ . 4 TFA

Example m R 30 0 ferrocene 31 1 3-indole

Examples 32 and 33

The procedures of these examples were based on a polyamine compound (14) and a suitable R-acetic or carboxylic acid, and were prepared using the procedure D1, followed by procedure F, of the following formula: R (CH ₂ ) n CO [ NH (CH ₂ ) ₃ ] ₃ NH ₂ . 3 TFA

Example m R 32 0 ferrocene 33 1 3-indole

Examples 34 and 35

The procedures of these examples were based on a polyamine compound (11) and a suitable R-acetic or carboxylic acid, prepared according to the procedure D1 followed by the procedure F, of the following formula: R (CH ₂ ) _b CO [ NH (CH ₂ ) ₃ ] ₂ NH ₂ . 2 TFA

Example m R 34 0 ferrocene 35 1 3-indole P I • Examples 36 and 37

In carrying out the procedures of these examples,

starting from a polyamine compound (7) and a suitable R-acetic or carboxylic acid, prepared using process D1 followed by process F, compounds of the following formula: R (CH ₂ ) _b CONH (CH ₂ ) ₃ NH ₂ . TFA Example m R 36 0 ferrocene 37 1 3-indole

Examples 38 and 39

The procedures of these examples were based on a polyamine compound (23) and a suitable R-acetic or carboxylic acid, and were prepared using the procedure D1, followed by the procedure F, of a compound of the following formula: R (CH ₂ ) _n CO [ NH (CH ₂ ) ₃ ] ₆ NH ₂ . 6 TFA

Example m R 38 0 ferrocene 39 1 3-indole

Examples 40 and 41

The procedures of these examples were based on a polyamine compound (26) and a suitable R-acetic or carboxylic acid, prepared using Procedure D1, followed by Procedure F, of a compound of the following formula:

R (CH ₂ ) _B CO (NH (CH ₂ ) ₃ ) ₇ NH ₂ 7 TFA

Example m R 40 0 ferrocene 41 1 3-indole

Example

Stage 1:

NC

nh ₂

NHBoc

NHBoc

To a solution containing N-cyanoethyl-1,4-diaminobutane (6.44 g, 0.0457 mol) in acetonitrile (200 mL) under a nitrogen atmosphere was added KF / Celite (11 g) followed by addition

N- (tert-butoxycarbonyl) -3-bromopropylamine (10.87 g, 0.0457 mol) was added dropwise over 7 hours. The reaction mixture was stirred for 16 hours at ambient temperature and then heated at 70 ° C for 24 hours and finally cooled, filtered and concentrated under vacuum. The residue was dissolved in CH ₂ C1 ₂ (200 ml), washed with 1 N NaOH (100 ml), dried and concentrated in vacuo to afford crude product which was chromatographed on silica gel (elution of CH ₂ Cl ₂ / MeOH 9: 1) to give the branched nitrile product.

Yield: 3.32 g.

¹ H NMR (CDCl ₃ ) δ:

1.31 - 1.65 (m, 27H), 2.36 - 2.44 (m, 6H), 2.49 (t, J = 6.6 Hz, 2H), 2.61 (m, 2H). 2.90 (t, J = 6.6 Hz, 2H), 3.09 - 3.16 (m, 4H); 5.28 (brs, 2H); ¹³ C NMR (CDCl ₃ ) δ: 18.74, 24.60, 27.05, 27.96, 28.50, 39.52, 45.14, 49.07, 52.21, 53.76, 78.90, 118.79, 156.09. HR FABMS: observed (Μ + H) m / z = 456.3549

C ₂₃ H ₄₆ N ₅ O ₄ (m / z 456.3550).

Stage 2:

NC

NHBoc

NHBoc

Di-tert-butyl dicarbonate (78 grams) was added to a solution containing the branched amine compound prepared according to Step 1 above (150 mg, 0.33 mmol) in dichloromethane (5 mL) under a nitrogen atmosphere. (0.36 mmol). The reaction mixture was stirred overnight and concentrated under reduced pressure to give a crude product which was chromatographed on silica gel (CH ₂ Cl ₂ / MeOH 9: 1) to give the title compound as a white solid. the desired N-BOC-nitrile compound was obtained.

Yield: 126 g, 69%.

Stage 3:

Boc

NHBoc

NHBoc

To the solution containing the N-BOC-nitrile compound prepared according to Step 2 above (126 mg, 0.277 mmol) was added Pd (OH) ₂ / carbon (200 mg), the reaction mixture thus prepared. The mixture was hydrogenated for 2 hours at a hydrogen pressure of 551.6 kPa, then filtered through celite and concentrated under vacuum. The crude product was taken up in dichloromethane (30 mL) and washed with 1 N NaOH (2 x 10 mL), dried over potassium carbonate, filtered, and concentrated under reduced pressure to give the desired N- BOC-amine compound

Yield: 129 mg, 100%.

The N-BOC-amine compound obtained in Step 3 above in dichloromethane (2 mL) was combined with indole acetic acid hydroxysuccinimide ester (14.1 g, 0.05 mmol) under nitrogen. The reaction mixture was stirred overnight and then diluted with CH ₂ Cl ₂ dichloromethane (25 mL). The organic extract was then dried over potassium carbonate, concentrated in vacuo and chromatographed over silica gel (CH ₂ Cl ₂ / MeOH 9: 1, then CH ₂ Cl ₂ / MeOH / PrNH _{2 as} eluent _). 9: 1: 0.1) to give the desired N-BOC-protected compound

Yield: 28 mg, 78%.

Stage 5:

Ν '

H

The indole compound from Step 4 above (28 mg, 0.039 mmol) was dissolved in trifluoroacetic acid (3 mL) and stirred for 1 hour.

The reaction mixture was concentrated under reduced pressure and triturated in diethyl ether to give the desired product.

Yield: 19 mg, 56%.

Preparation

It was mixed according to this procedure

3-bromopropylamine.HBr (34.5 g, 157.6 mmol) in 600 mL of Ν, Ν-dimethylformamide. 34.4 g (157.6 mmol) of di-tert-butyldicarbonate were added to the solution, followed by 32.3 ml (236 mmol) of triethylamine. In this way, a precipitate immediately formed. The reaction mixture was stirred overnight. The reaction mixture was diluted to 1.5 L with ethyl acetate, washed once with 500 mL of 1 N hydrochloric acid, three times with 500 mL of water, once with brine, and then dried over Na ₂ SO ₄ . After concentration, the product was chromatographed on silica gel (800 grams) using 4: 1 hexane / ethyl acetate as eluent, and the individual fractions were monitored by TLC (KMnO ₄ / I ₂ ).

The product containing fractions were combined, concentrated in vacuo, and washed twice with 50 mL dichloromethane and purified under high vacuum to afford the desired product.

Yield: 25.8 g.

Claims (15)

PATENTOVÉ Claims ro ro x) rj z =. o C / x co-i 1. A process for the preparation of compounds of the general formula: R-CHCH -CO-R 'in which it means; R is a 5- to 7-membered carbocyclic system or an 8 to 11-membered carbocyclic system or any of these systems substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxy, C 1 -C 4 alkyl, C 1 -C 4 alkoxy carbon, CF ₃ , phenyl, amino, C 1 -C 4 alkylamino and C 1 -C 4 dialkylamino, m is 0 or 1, R 1 represents a group - [NH (CH ₂ ) _n ] _x NH ₂ , wherein each n independently represents 2 to 5, and x is 1 to 6, characterized in that it comprises (a) reacting R with a compound of the formula: BOC-NH4CH3NNOCOCXCHy, ₄ N (BOC) H wherein: BOC t-butoxycarbonyl, a R, n and x have the same meaning as above, in the presence of a carbodiimide-based reagent ΊΟ coupling with an amide bond catalyst in an inert reaction solvent, and (b) removing the t-butoxycarbonyl group by treatment with an organic or inorganic acid. The method of claim 1, wherein said carbodiimide-based reagent is selected from dimethylaminopropyl, ethylcarbodiimide and dicyclohexylcarbodiimide and said amide bond catalyst is selected from hydroxybenzotriazole and hydroxysuccinimide. The process according to claim 1 or 2, wherein said inert reaction solvent is dichloromethane and said organic or inorganic acid is selected from trifluoroacetic acid and hydrochloric acid. A process according to claims 1 to 3, wherein in said compounds x is 5 and each n independently represents 3 or 4. Process according to any one of claims 1 to 4, characterized in that in said compounds R represents a phenyl group or a phenyl group monosubstituted by a fluorine, chlorine, bromine or hydroxyl group of OH or MeOH and m is 1. A process according to any one of claims 1 to 5, wherein in said compound R 'is - [NH (CH ₂ ) ₃ ] ₃ NH (CH ₂ ) ₄ NH (CH ₂ ) ₃ NH ₂ . 7. Compound of general formula: R / CH3 -CO-R ', or a pharmaceutically acceptable acid addition salt thereof, wherein: R is a 5- to 7-membered carbocyclic system or an 8 to 11-membered carbocyclic system or any of these systems substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxy, C 1 -C 4 alkyl, C 1 -C 4 alkoxy carbon, CF ₃ , phenyl, amino, C 1 -C 4 alkylamino and C 1 -C 4 dialkylamino, m is 0 or 1, R 'is - [NH (CH ₂ ) _n ] _x NH ₂ , wherein each n is independently 2 to 5, and x is 1 to 6. The compound of claim 7, wherein x is 5 and each n is independently 3 or 4. The compound of claim 8, wherein R is a phenyl group or a phenyl group monosubstituted with fluorine, chlorine, bromine, OH or MeOH, and m is 1. The compound of claim 8, wherein R 1 is - [NH (CH ₂ ) ₃ ] ₃ NH (CH ₂ ) ₄ NH (CH ₂ ) ₃ NH ₂ . 11. A compound of the formula: R-CH 2 CO-R 'or a pharmaceutically acceptable acid addition salt thereof, wherein: R is a 5- to 7-membered carbocyclic system or an 8 to 11-membered carbocyclic system or any of these systems substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxy, C 1 -C 4 alkyl, C 1 -C 4 alkoxy carbon, CF ₃ , phenyl, amino, C 1 -C 4 alkylamino and C 1 -C 4 dialkylamino, m is 0 or 1, R 'represents a group: [(CH ₂ ) _n NH] _y H - [NH (CH ₂ ) _n ] _x N - [(CH ₂ ) _n NH] ₂ H wherein each n independently represents 2 to 5, x is 0 to 4, y and z each independently represent 1 to 5, the sum of x and greater zyaz is 1 to 5. 12. Compound of general formula: R 1 -CH 2 -CO-R ', or a pharmaceutically acceptable addition salt thereof, wherein: R is a 5- to 7-membered carbobicyclic system or an 8 to 11-membered carbocyclic system or any of said systems substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxy, C 1 -C 4 alkyl, C 1 -C 4 alkoxy carbon, CF ₃ , phenyl, amino, C 1 -C 4 alkylamino and C 1 -C 4 dialkylamino, m is 0 or 1, R 'represents a group: / - [NH (CH ₂ ) _n 1 _x N \ (CH ₂ ) _and [NH (CH ₂ ) _b ] y \ N [(CH ₂ ) _n NH 3 _from H (CH ₂ ) _and [NH (CH ₂ ) _b ] / wherein all a are the same and may be in the range 2 to 5, all b are the same and may be in the range 2 in and 5, each n is independently from 2 to 5, x is from 0 to 3, all y are the same, and can be 0 or 1, z is from 0 to 3, and the sum of x + y + z ranges from 0 to 4. A pharmaceutical composition for antagonizing excitatory amino acid neurotransmitters in a mammal comprising the compound of claim 7, or a pharmaceutically acceptable addition salt thereof, in an amount for antagonizing the excitatory amino acid neurotransmitters in a mammal, and a pharmaceutically acceptable carrier or diluent. A pharmaceutical composition for antagonizing excitatory amino acid neurotransmitters in a mammal comprising the compound of claim 13, or a pharmaceutically acceptable addition salt thereof, in an amount for antagonizing the excitatory amino acid neurotransmitters in a mammal, and a pharmaceutically acceptable carrier or diluent. A compound according to claim 7 for the preparation of a medicament for antagonizing excitatory amino acid neurotransmitters in mammals.