PT100801A - PROCESS FOR THE PREPARATION OF UTILITY SYNTHETIC ARIL-POLYAMINS AS ANTAGONISTS OF NEUROTRANSMITTERS AMINOACIDES AND COMPOUNDS OBTAINED - Google Patents

Description

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Background of the Invention ;; , -

This invention relates to a class of aryl polyamines and to their pharmaceutically acceptable salts which are antagonists of excitatory amino acid neurotransmitters. These neurotransmitters affect the neuronal cells of a variety of organisms including invertebrates and vertebrates. The polyamines of the present invention are synthetic analogues of certain polyamines found in the venom of the spider Aqelenopsis aperta. This invention also relates to the use of such polyamines and their salts in the antagonism of excitatory amino acid neurotransmitters. These neurotransmitters affect cells such as the cells of an organism's nervous system. This invention further relates to the use of such polyamines and their salts in the treatment of diseases and conditions in a mammal mediated by excitatory amino acid neurotransmitters in the control of invertebrate pests and to compositions including said polyamines and their salts. The invention also relates to methods for the preparation of such polyamines.

It has been reported that the poison of the spider Aqelenopsis aperta contains at least two toxins that affect the calcium currents. Jackson, H., et al., Soc. Neu. Know. Abstr. 12: 1078 (1987). These authors disclose a toxin, referred to herein as AG2, which has a molecular weight of less than 1000 Daltons and appears to eliminate calcium currents in a wide range of tissues. In addition, Jackson, H. et al., Soc. Neu. Know. Abstr. 12: 730 (1986) discloses another toxin Aqelenopsis aperta which includes a component with a certain molecular weight of 6000. This toxin is referred to as blocking presynaptic transmissions and it has been suggested that the toxin blocks the associated calcium channels to the release of the neurotransmitter.

Certain polyamines found in the Aqelenopsis aperta vehfen are disclosed in U.S. Patent 5,037,846 filed April 28, 1989 and assigned to the assignees of the present patent. Such polyamines and their pharmaceutically acceptable salts are disclosed therein as excitatory amino acid receptor blocking agents in cells, and one such polyamine, B, is also shown as a calcium channel blocking agent.

Compounds that are antagonists of excitatory amino acid neurotransmitters have a variety of uses. The excitatory amino acid neurotransmitter antagonists are useful in the treatment of conditions such as seizures, cerebral ischemia, degenerative neuronal diseases such as Alzheimer's disease and epilepsy, and as therapeutic agents among other uses. See Excitator Amino Acids in Health and Disease. D. Lodge, E., John Wiley and Sons Ltd., New York, NY 1988, the teachings of which are incorporated herein by reference. In addition, such compounds are useful in the study of the physiology of cells such as neuronal cells and in the control of invertebrate pests. Glutamate is the major excitatory neurotransmitter in the mammalian brain. Great enthusiasm has been generated in the last decades, as the development of the pharmacology of glutamate receptors suggested their differentiation into several subtypes. The glutamate receptor subtype classified by the selective action of the exogenous agonist N-methyl-D-aspartate (NMDA) has been the subject of intense research since it was proposed that these receptors played a role in a variety of neurological pathologies including seizures , epilepsy, and neurodegenerative diseases such as Alzheimer's disease. There are currently two generic classes of. · ≫ ' NMDA receptor antagonists that are to be intensively studied for the discovery of clinically useful drugs. The first class consists of competitive antagonists that interfere with glutamate binding at its site at the receptor. These compounds are characterized as being highly polar compounds such as the phosphonate compounds AP7 and AP5. The highly charged structure of the competitive agents renders them unable to penetrate the blood / brain barrier, limiting their therapeutic utility. The second class consists of non-competitive antagonists that block NMDA receptor function by acting on the ion channel associated with the NMDA receptor complex. These compounds include MK-801 and phencyclidine (PCP). The potential psychotomimetic effects of these classes of compounds are obvious drawbacks of known medicaments that function by mechanisms of this type.

Recently, a third class of antagonists began to be studied, based on the identification of novel glutamate antagonists obtained from spider venoms. The arylamine structures isolated from the Aqelenopsis venom venom which exhibit potent and specific antagonism to mammalian NMDA receptors are disclosed in U.S. Patent Application Serial No. 554,311, filed July 17, 1990 and in U.S. Patent No. 5 037 846, filed July 31, 1990. The arylamines isolated from the venom of Aelenopsis aperta are composite structures constructed from an aromatic acid and amide bonded polyamine fragments. In these structures, some of the amines in the polyamine fragment are functionalized as N-hydroxyamines or quaternary ammonium salts. The chemical structures of the arylamines are distinct from those of the above-mentioned conventional competitive agents, AP7 and AP7r and that of the non-competitive compound MK-801. By f

For example, the AGEL 416 polyamine disclosed in the above-mentioned patent No. 5,037,846 is shown to have the following structure h /

The mechanism of NMDA antagonism by these arylamines is also distinct from both the competitive class and the MK-801 / PCP class. Accordingly, spider venom arylamines provide a novel class of antagonist compounds at the NMDA receptor.

Thanks to the disclosure herein of the compounds which exist in nature, it is now possible to obtain said compounds by methods other than isolation / purification from the integral venom of Aelenopsis aperta and, accordingly, it is also possible to synthesize compounds analogues of the same class which do not exist in nature.

Summary of the Invention

This patent application concerns a class of synthetic aryl polyamines of formula R- (CH) -CO-R ', wherein R represents a 5- to 7-membered carbocyclic system or an 8 to 11-membered carbobicyclic system, or any of these systems substituted with one or more substituents independently selected from F, Cl, Br, OH, Cl to C4 alkyl, C1 to C4 alkoxy, CF3, phenyl, amino, C1 to C4 alkylamino and di (Cl to C4) alkylamino; m is 0 or 1; R 'represents [i

- [NH (CHO)] NH-; each n is, independently of the others, equal to a number between 2 and 5; and x is equal to a number between 1 and 6.

This invention also relates to the compounds of formula R- (CH 2) m -CO-R ', or to the pharmaceutically acceptable acid addition salts thereof, wherein R represents a 5- to 7-membered carbocyclic system or an 8 to 11 membered carbobicyclic system, or any of these systems substituted with one or more substituents independently selected from F, Cl, Br, OH, Cl to C4 alkyl, C1 to C4 alkoxy, CF3, phenyl, amino, C1 to C4 alkylamino and di (C1 to C4) alkylamino; m is 0 or 1; R 'represents

t (CH 2) n NH] y H

- [NH (CH 2) n 3 x N

[(CH 2) n NH] z H; each n is, independently of the others, equal to a number between 2 and 5; x is equal to a number between 0 and 4; y and z are, independently of one another, equal to a number between 1 and 5; and the sum of x with the greater of y and z is equal to a number between 1 and 5.

This patent application further relates to compounds of the formula or a pharmaceutically acceptable acid addition salts thereof, wherein R represents a 5- to 7-membered carbocyclic system or a carbobicyclic system or any of these systems substituted with one or more substituents independently selected from F, Cl, Br, OH, Cl to C4 alkyl, C1 to C4 alkoxy, CF3, phenyl, amino, Cl to C4 alkylamino and di (C1 to C4) alkylamino; m is 0 or 1; R 'represents 7 r 7 r

(I.e.

(b) wherein each a is identical and equal to a number between 2 and 5, each b is identical and equal each n is independently of the others equal to 2 to 5, x is equal to a number between 0 and 3, each y is identical and equal to 0 or 1, z is a number between 0 and 3; ex + y + z equals a number between 0 and 4.

The carbocyclic systems of the present invention may be saturated, unsaturated or aromatic, with aromatic systems being preferred. With respect to monocyclic systems, phenyl is preferred. Bicyclic systems may be condensed or bridged, with 9- or 10-membered condensed systems, for example, naphthyl and indene. Of the aforementioned bicyclic systems, naphthyl is particularly preferred.

Preferred R 'groups are those wherein x is equal to 4 or 5 and each n is independently of the others equal to 3 or 4. Particularly preferred R' groups - [NH (CH 2)] 5NH 2 and - [NH CH 2) 3] 3 NH (CH 2) 4 NH (CH 2) 3 NH 2.

The polyamines of this invention and their pharmaceutically acceptable salts are antagonists of excitatory amino acid neurotransmitters. Accordingly, such polyamines are useful in antagonizing such excitatory amino acid neurotransmitters, per se. The polyamines of this invention are also useful in the control of invertebrate pests and in the treatment of diseases and conditions in a mammal mediated by excitatory amino acid neurotransmitters. Said polyamines are also useful as psychotherapeutic agents in mammals.

This invention also relates to pharmaceutical compositions including such polyamines, methods of administering the polyamines and methods of preparing said polyamines.

Detailed Description of the Invention

A synthetic scheme for the preparation of a polyamine of formula

is shown in Reaction Schemes A to C below. > i

REACTION SCHEME A

(III) IBOC] 20 * BOC

l (V) 10 - V f

BOC i

(VII) H 2 / Pd (OH) 2 / HOAc ▼

BOC -BOC

I

(IX) 11

V

REACTION SCHEME C

IX + VIII >

H BOC BOC I t

N-BOC H (X)

(XII)

According to Reaction Scheme A, the polyamine intermediate of formula IV is prepared by a sequence of steps starting with diaminobutane. Suitable reaction conditions for preparing the intermediate compound of formula VIII according to Reaction Scheme A are given in Example 5, parts 1 to 7. Reaction Scheme B illustrates a method for the preparation of the intermediate compound of formula IX. Suitable reaction conditions for the preparation of such intermediate are set forth in Example 5, part 8. The preparation of the polyamine compound of this invention of formula XII is shown in Reaction Scheme C. Suitable reaction conditions for the coupling of the intermediate compounds of formulas VIII and IX and subsequent preparation of the compound of formula XII are set forth in example 5, parts 1 to 11.

The polyamines of this invention reversibly antagonize neurotransmitter excitatory amino acids, which neurotransmitters affect cells such as cells of the nervous system of various organisms including invertebrates and vertebrates. The term vertebrates, as used herein, is to be understood as including mammals. The term invertebrates, as used herein, is to be understood as including, for example, insects, ectoparasites and endoparasites. The ability of the polyamines of the present invention to antagonize excitatory amino acid neurotransmitters is demonstrated by their ability to block N-methyl-D-aspartic acid (NMDA) -induced cGMP increases in newborn rat cerebellar cells according to the following procedure. Cerebellos of ten Wistar rats with 8-14 days are excised rapidly and placed in Krebs / bicarbonate buffer, pH 7.4 at 4 ° C and then cut into sections of 0.5 mm x 0.5 mm 13 13

using a Mcllwain tissue chopper (The Nickle Laboratory Engineering Co., Gomshall, Surrey, England). The resulting cerebellum pieces are transferred to 100 ml of Krebs / bicarbonate buffer solution at 37 ° C which is continuously equilibrated with 95: 5%. Cerebellum pieces are thereby incubated for ninety minutes with three changes of the buffer solution. The buffer solution is then decanted, the tissue is centrifuged (1 min, 3200 rpm) and the tissue is resuspended in 20 ml Krebs / bicarbonate buffer solution. Then, 250 μl aliquots (about 2 mg) are taken and placed in 1.5 ml microcentrifuge tubes. To these tubes

10 μΐ of the compound being studied is obtained from a stock solution followed by 10 μΐ of a 2.5 mM NMDA solution to elicit the reaction. The final NMDA concentration is 100μΜ. No NMDA is added to the controls. The tubes are incubated for one minute at 37 ° C in a shaking water bath, then 750 μΐ of a 50 mM Tris-Cl solution, 5 mM EDTA is added to stop the reaction. The tubes are immediately placed in a boiling water bath for five minutes. The contents of each tube are then sonicated for 15 seconds using a probe sonicator set to power level three. Ten microliters are withdrawn and the protein concentration is determined by the method of Lowry, Anal. Biochem. 100: 201-220 (1979). The tubes are then centrifuged (5 min, 10 000 xg), 100 μl of the supernatant is withdrawn, and the cyclic GMP level (cGMP) is determined using a New England Nuclear (Boston, Massachusetts) cGMP RIA device and following manufacturer's method. The data are recorded in terms of pMole of cGMP produced per mg of protein.

Additionally, the ability of the polyamines of the present invention to antagonize amino acid neurotransmitters 14

excitatory cells is demonstrated by their ability to block 2+ increases of [cytosolic Ca in NMDA / glycine-induced dissociated cerebellar granular cells according to the following procedure. Cerebellar granular cells are prepared from 8-day rat cerebellar cells (Wilkin, G.P. et al., Brain Res. 115: 181-199, 1976). (12 cm) of Aclar (Proplastics Inc., 5033 Industrial Ave., Wall, NJ, 07719) are coated with poly-L-lysine which are placed in 12-well dishes containing 1 ml Eagle Basal Medium. Cells are dissociated and added to each well containing the squares of aliquots containing 6.25 x 10 6 cells. Cytosine-beta-D-arabinofuranoside (10 μ concentração final concentration) is added 24 hours after this operation. Cells are used for fura2 analysis after 6, 7 and 8 days of culture. Cells (attached to the Aclar squares) are transferred to 12-well dishes containing 1 ml 2 μM fura2 / AM (Molecular Probes Inc., Eugene, OR) in HEPES buffer solution (containing 0.1% bovine serum albumin , 0.1% dextrose, pH 7.4, magnesium-free). The cells are incubated for 40 minutes at 37øC; the buffer solution containing fura2 / AM is removed and replaced with 1 ml of the same buffer containing no fura2 / AM. To a cuvette of quartz is added 2.0 ml of preheated buffer solution (37øC). The cells on the Aclar are placed in the cuvette and this is introduced into a device under thermostatic control (37øC) equipped with a magnetic stirrer, the fluorescence being measured with a fluorescence spectrophotometer (Biomedical Instrument Group, University of Pennsylvania). The signal resulting from the fluorescence measurement is allowed to stabilize for about 2 minutes.

An increase in free cytosolic calcium concentration, represented by an increase in fluorescence, is produced by the addition of 50 μM NMDA and 1 μg glycine. Then, 5-20 μΐ of a " stock " of the compound being studied in phosphate buffered saline (PBS, pH 7.4) in the appropriate concentration are added to the cuvette. Calibration of the fluorescence measurement signals and correction performed to take into account fura2 / AM leakage are performed using procedures recognized by Grynkiewicz, G. et al., J. Biol. Chem, 260: 3340 (1985). At the end of each test, the maximum fluorescence (Fmax) value is determined by addition of iono-mycin (35 μ) and the minimum fluorescence (Fmin) value is determined by the subsequent addition of EGTA (12 mM) in order to to chelate calcium. Using the above procedure, the ability of a compound under study to antagonize excitatory amino acid neurotransmitters is manifested by a decrease in fluorescence upon addition of the compound being studied.

The polyamines of this invention are useful in antagonizing excitatory amino acid neurotransmitters, per se. As such, polyamines are also useful in the control of invertebrate pests, in the treatment of diseases and conditions in a mammal mediated by excitatory amino acid neurotransmitters such as seizures, cerebral ischemia, neuronal degenerative diseases such as Alzheimer's disease and epilepsy. Said polyamines are also useful as psychotherapeutic agents in a mammal. In addition, polyamines are useful in the study of the physiology of cells including cells of the nervous system, but are not limited thereto.

Also encompassed by this invention are the pharmaceutically acceptable salts of the polyamines of this invention. Such salts are formed by methods well known to those skilled in the art. For example, acid addition salts of polyamines may be prepared according to conventional methods. Preferred acid addition salts of polyamines such as

addition salts of hydrochloric or trifluoroacetic acid. The hydrochloric acid addition salts of the polyamines are particularly preferred.

When it is desired to administer to a mammal a polyamine or a pharmaceutically acceptable salt thereof of this invention, those may be administered alone or in combination with pharmaceutically acceptable carriers or diluents in a pharmaceutical composition according to standard practice. The polyamines or their pharmaceutically acceptable salts may be administered orally or parenterally. Parenteral administration includes intravenous, intramuscular, intraperitoneal, subcutaneous and topical administration.

For purposes of oral administration of a polyamine or a pharmaceutically acceptable salt thereof of this invention, the compound may be administered, for example, in the form of tablets or capsules, or as an aqueous solution or suspension. In the case of tablets for oral use, carriers which are generally used include lactose and corn starch, and lubricating agents, such as magnesium stearate, are generally added. For oral administration in the form of capsules, lactose and dried corn starch are useful diluents. When aqueous suspensions are intended for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, sweetening or flavoring agents may be used.

For intramuscular, intraperitoneal, subcutaneous and intravenous use, sterile solutions of the active ingredient are generally prepared, and the pH of the solutions is suitably adjusted and buffered. For the purpose of

J I

J I

Intravenous use, the total concentration of solutes can be controlled in order to obtain an isotonic preparation.

When a polyamine or a pharmaceutically acceptable salt thereof of this invention is used in a human patient, the daily dose is usually determined by the attending physician. However, the suitable doses for the polyamines of this invention are between about 1 and 30 mg / kg / day. In addition, the dose will vary depending on the patient's age, weight and individual reaction, in addition to the severity of the patient's symptoms and the specific potency of the compound being administered. Accordingly, doses outside the indicated range of values, which are considered to be encompassed by the invention, are possible.

When a polyamine or a salt thereof of this invention is used in the control of invertebrate pests, such compounds are administered to such invertebrates directly or delivered to the environment of the invertebrates. For example, a compound of this invention may be sprayed as a solution under said invertebrates. The amount of compound required to control these invertebrates will vary depending on the invertebrate and environmental conditions and will be determined by the person applying the compound.

When a polyamine or a salt thereof of this invention is used in physiological studies of cells, such compounds are administered to the cells according to methods well known to those skilled in the art. For example, the compounds may be administered to the cells in a suitable physiological buffer. A concentration of 100 μΜ of the compounds of this invention is suitable for such studies. However, the concentration of said compounds in such studies may be greater than or much less than 100 μΜ. The amount of the compound to be administered will be determined

by the skilled person in the art according to well known methods. EXAMPLES Types of Procedures Found in Experiments (A)

Protection of secondary amine with di-tert-butyl dicarbonate (B) (C)

Primary amine cyanoethylation Catalytic hydrogenation of nitrile to give the primary amine (D)

(D2) Dicyclohexylcarbodiimide / (D3) Dimethylaminopropyl-ethylcarbodiimide / hydroxybenzotriazole / triethylamine (D4) Dicyclohexylcarbodiimide / (D3) Dimethylaminopropyl-ethylcarbodiimide / / hydroxysuccinimide (E) (F)

Deprotection of N-Boc substrates with HCl in dioxane Deprotection of N-Boc substrates with TFA (trifluoroacetic acid) (G)

Alkylation of amines with N-Boc-3-bromopropylamine

V

Example 1 - Preparation of Side Chains

Polyamine

Step 1 - Procedure Type B H2N (8)

A 103 g sample of 1,3-diaminopropane was combined with 45 ml of MeOH with stirring at 4 ° C. Acetonitrile (100 ml, 81 g, 1.1 g) was added dropwise to the solution via a pressure equilibrium addition funnel over 90 minutes. After 3 hours, a 500 mg portion was removed which was analyzed by means of C NMR; the presence of 1,3-diaminopropane was not detected. The crude material containing the aminonitrile product 8 was distilled under reduced pressure, collecting three fractions in the range of distillation temperature values between 100 and 125 ° C; all were sufficiently clean to be able to proceed with the reaction with di-tert-butyl carbonate and subsequent chromatographic purification on silica gel. 1 H NMR (250 MHz, CDCl 3) δ 2.67 (t, 2H, J = 6.6 Hz), 2.54-2.43 (m, 4H), 2.28 (t, 2H, J = 6.6 Hz), 1.37 (m, 2H, J = 6.7 Hz), 1.05 (s, 3H); 13 C NMR (63.1 MHz, CDCl 3) δ 118.8, 44.9, 40.2, 33.4, 18.5.

Step 2 - Procedure Type A

To a solution of aminonitrile 8 (23 g, 0.18 mol) in 500 ml of methylene chloride at 0øC was added di-tert-butyl carbonate (80 g, 0.36 mol, 2 eq) . The reaction mixture was stirred at room temperature for 16 hours and treated with an additional portion of di-tert-butyl dicarbonate (8 g, 0.036 mol). After stirring for an additional 4 hours, the reaction was washed with 1N KOH (2 x 60 mL), dried over K 2 CO 3, filtered and concentrated in vacuo. The product was purified by " flash chromatography " (SiO2, 20 -> 100% ethyl acetate in hexane) to afford the N-Boc-nitrile 9 product as a clear oil (14 g, 24% yield). 1 H NMR (250 MHz, CDCl 3) δ 3.40 (t, 2H, J = 6.7 Hz), 3.28 (t, 2H, J = 6.6 Hz) , 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

Parat of 2.6 L N-Boc-nitrile 9, (49 g, 0.4 mol), 1000 ml of acetic acid and 20 g (20% by weight Pd (OH) 2) Pd (OH) 2 / C. The vessel was filled with hydrogen gas until the value reached 3.45 x 105 Pa (50 psi) and stirred for 4 hours. The reaction was filtered through a 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 phases were extracted with CH 2 Cl 2 (400 mL); the CH2 Cl2 phases were combined, dried over K2 CO3, filtered and concentrated in vacuo to provide the N-Boc-amine 11 as a clear colorless oil (43 g, 86% yield). 1 H NMR (250MHz, CDCl 3) δ 3.28-3.12 (m, 2H), 3.11-3.00 (m, 4H), 2.64 (t, 2H, J = 7Hz), , 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, 31.6, 28.5, 28.2 .

Step 4 - Procedure Type B (")

02)

A 38 g sample of N-Boc-amine 11 (0.114 mol) was combined with 6.7 g of acrylonitrile (0.126 mol, 1.1 eq.) In 60 ml of methanol and stirred for 11 hours. The solvents were removed to give 43 g (100% yield) of nitrile 12 as a clear colorless oil which was used without further purification. 1 H NMR (250 MHz, CDCl 3) δ 3.18 (broad s, 4H), 3.03 (m, 2H), 2.85 (t, 2H, J = 6.6 Hz), 2.57 (t, 2H, J = 6.7 Hz), 2.45 (t, 1

J = 6.7 Hz), 1.72-1.53 (m, 4H), 1.41 (s, 9H), 1.38 (s, 9H); 13 C NMR (63.1 MHz, CDCl 3) δ 155.9, 118.6, 79.6, 78.9, 46.3, 45.0, 43.9, 37.4, 28.8, 28.3 , 18.6.

Step 5 - Procedure Type A 03) 02) -

Boc Boc Boc

A 43 g sample of the above nitrile 12 (0.114 mol) was combined with di-tert-butyl dicarbonate (25.6 g, 0.120 mmol, 1.05 eq.) And 350 ml of CH 2 Cl at 0 ° C and stirred for 9 hours. Thin layer chromatography (TLC) analysis (EtOAc, KMnO 4) did not reveal the existence of any remaining starting material; the reaction was purified in a manner analogous to that described for N-Boc-nitrile 9. N-Boc-nitrile 13 was recovered as a clear colorless oil (34 g, yield 63%). NMR (250 MHz, CDCl 3) δ 3.45 (t, 2H, J = 6.6 Hz), 3.39-2.97 (m, 8H), 2.68-2.46 (2H, 1.82-1.56 (m, 4H), 1.44 (s, 18H), 1.87 (S, 9H); 13 C NMR (75.7 MHz, CDCl 3) δ 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 2 5o procedure: (U)

Ecc 03) ϊ

N-Boc-amine 14 was prepared from N-Boc-nitrile 13 such as N-Boc-amine 11 was prepared from N-Boc-nitrile 9 in 99% yield (30 g). 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, 2H); 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, 28.3.

Step 7 - Type B Procedure

Boc E dc Boc 05)

Nitrile 15 was prepared from N-Boc-amine 14 such as nitrile 12 was prepared from N-Boc-amine 11 in 90% yield. 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 = J = 6.6 Hz-), 1.72-1.57 (m, 6H), 1.41 (s, 9H), 1.40 (s, 9H ), 1.39 (s, 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. 13 C NMR (300 MHz, CDCl 3) of nitrile 15 is not distinguishable from the nitrile spectrum 12.

Step 8 - Procedure Type A

N-Boc-nitrile 16 was prepared from nitrile 15 such as N-Boc-nitrile 13 was prepared from nitrile 12 as a clear colorless oil (30 g, 87% yield).

Hz), 1,40-

1 H-NMR (300MHz, CDCl 3) δ 3.36 (t, 2H, J = 6, 3.18-2.90 (m, 14H), 2.57-2.42 (m, 2H) 1.48 (m, 6H), -1.28 (m, 27H).

Step 9 - Type C Procedure

Boc-Boc-amine 17 was prepared from N-Boc-nitrile 16 such as N-Boc-amine 11 was prepared from N-Boc-nitrile 9 in a yield of 74% (2.61 g). NMR (250 MHz, CDCl 3) δ 3.39-2.97 (m, 14H), 2.63 (t, 2H, J = 6.6 Hz), 1.80-1.53 (m, 8H), 1.39 (s, 27H), 1.38 (s, 9H), 1.23 (s, 2H).

Step 10 - Procedure Type B (17) - (18)

Boc Eoc Boc Boc

Nitrile 18 was prepared from N-Boc-amine 17 such as nitrile 12 was prepared from N-Boc-amine 11 in 91% yield (19 g), and was used without further purification. Δ NMR (300 MHz, CDCl3) δ 3.24-2.94 (m, 14H), 2.87 (t, 2H, J = 6Hz), 2.57 (t, 2H, J = 6Hz), 2.47 (t, 2H, J = 6Hz), 1.74-1.54 (m, 8H), 1.45-1.36 (m, 36H).

Step 11 - Procedure Type A (18) (19)

Boc Boc

Boc-Boc-N-Boc-nitrile 19 was prepared from nitrile 18 such as N-Boc-nitrile 13, was prepared from nitrile 12 (16 g, 74% yield). 1 H NMR (250 MHz, CDCl 3) δ 3.43 (t, 2H, J = 6.6 Hz), 3.28-3.02 (m, 16H), 2.62-2.50 (m, 2H) , 1.80-1.56 (m, 8H), 1.43 (s, 9H), 1.42 (s, 9H), 1.41 (s, 18H), 1.39 (s, 9H).

Step 12 - Type C procedure

N-Boc-amine 20 was prepared from N-Boc-nitrile 19 such as N-Boc-amine 11 was prepared from N-Boc-nitrile 9 (17 g, yield 99%). 1 H NMR (300 MHz, CDCl 3) δ 3.24-2.95 (m, 18H), 2.6 (t, 2H, J = 6 Hz), 1.75-1.52 (m, 10H), 1. , 42-1.32 (m, 45H).

Step 13 - Type B Procedure (21). (20)

Boc Boc Boc Boc Boc

Nitrile 21 was prepared from N-Boc-amine 20 such as -nitrile 12 was prepared from N-Boc-amine 11 in 99% yield. (t, 6.6 1 H NMR (250 MHz, CDCl 3) δ 3.32-3.03 (m, 16H), 2.91 2H, J = 6.7 Hz), 2.61 (t, 2H, J (T, 2H, J = HZ), 1.82-1.57 (m, 10H), 1.44 (s, 36H), 1.43 (s, 9H).

Step 14 - Procedure Uncle A

N-Boc-nitrile 22 was prepared from nitrile 21 such as N-Boc-nitrile 13 was prepared from nitrile 12 in 99% yield. 3.45 ppm (t, 2H, J = 6.6 Hz), 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 (½) (250 MHz, CDCl 3) δ 155.2, 79.3, 44.7, 28.3, 28.3, 28.2.

Step 15 - Procedure Type c

N-Boc-amine 23 was prepared from N-Boc-nitrile 22 such as N-Boc-amine 1I was prepared from N-Boc-nitrile 9 (8 g, yield 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). 28 s · \

Step 16 - Type B Procedure

Nitrile 24 was prepared from N-Boc-amine 23 such as nitrile 12 was prepared from N-Boc-amine 11 in 99% yield. Δ NMR (250 MHz, CDCl3) δ 3.32-3.04 (m, 22H), 2.90 (t, 2H, J = 6.7Hz), 2.61 (t, 2H, J at 6 Hz ), 2.53 (t, 2H, J = 6.7Hz), 1.82-1.57 (m, 12H), 1.44 (s, 45H), 1.43 (s, 9H).

Step 17 - Procedure Type A

N-Bocnitrile 25 was prepared from nitrile 24 such as N-Boc-nitrile 13 was prepared from nitrile 12 (5.5 g, 74% yield). 1 H NMR (250 MHz, CDCl 3): δ 3.45 (t, 2H, J κ 6.7 Hz), 2H), 1.80-1.55 (m, 12H), 3.40-2.97 (m, 24H), 2.66-2.51 (m, 1.54-1.30 (s, 63H).

Step 18 - Type C Procedure

N-Boc-amine 26 was prepared from N-Boc-nitrile such as N-Boc-amine 11 was prepared from N-Boc-nitrile 9 (5.01 g, 91% yield). Δ KMN (250 MHz, CDCl 3) δ 3.30-2.97 (m, 26H), 2.60 (t, 2H, J = 6Hz), 1.81-1.57 (m, 14H) 53-1.28 (m, 63H). 13 C NMR (250MHz, CDCl 3) δ 155.2, 79.3, 44.7, 28.7, 28.4, 27.5.

Example 20

Step 1. Amide Binding Formation - Type Dl Procedure

To a 50 mL single neck round bottom flask (RBF) combined 0.19 g ferrocenecarboxylic acid (0.82 mmol, 1.1 eq.), 0.12 g of hydroxybenzotriazole (0.1 g, 89 mmol, 1.2 eq.), 0.17 g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (HCl salt, 0.90 mmol, 1.2 eq.) And 10 mL of CH Cl with 30 30

stirring under a dry N2 atmosphere. After 30 minutes, 0.61 g of N-Boc-amine 27 (0.75 mmol, 1.0 eq., See Example 5a below for the preparation) was added to the solution. TLC (2 x MeOH, I2) indicated that N-Boc-amine was consumed after 2 hours. The reaction was diluted to 400 mL with EtOAc and washed with a pH 4 buffer solution (2 x 25 mL), 25 mL H 2 O, 1 N KOH (2 x 25 mL), 25 H2 O and 50 mL water salted. The EtOAc layer was dried over Na 2 SO 4, filtered and the solvents were removed to give 712 mg (93%) of product as an orange oil. 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 (S, 9H), 1.45 (s, 18H), 1.44 (s, 9H), 1.40 (s, 43 (s, 9H).

Step 2 - Polyamine Deprotection - Procedure

Trifluoroacetic acid (30 ml) was degassed with a flow of bubbles of dry N2 (via a Teflon tube) in a 100 ml RBF with a single tubing at 0 ° C. The iron-xenocarboxamide polyamine from step 1 above (712 mg, 0.69 mmol) was dissolved in 2 mL of CH 2 Cl 2 and transferred to the TFA under stirring with 3 x washes of 2 mL of CH 2 Cl 2. After 30 minutes, the ice bath; after a further 30 minutes, the solvents were removed under reduced pressure, then by means of HiVac. The remaining reddish brown oil was macerated with Et 2 O (3 x 30 mL); a yellow solid formed which was collected under positive pressure of N in a porosity frit " B ". The solid was washed with ether and the residual ether was removed by positive N-pressure to give 690 mg (yield 93%) of the product as a solid. Δ NMR (DMSO) δ 4.71 (t, 2H, J = 1 / 73Hz), 4.38 (t, 2H, J2.2Hz), 4.15 (s, 5H), 3.28-3 , 21 (m, 2H), 3.03-2.81 (m, 18H), 2.04-1.72 (m, 8H), 1.61-1.50 (m, 4H); 13 C NMR (250 MHz, D2 O) 177.3, 78.5, 76.1, 74.3, 49.7, 48.1, 47.3, 47.2, 47.1, 39.3, 8, 28.5, 26.5, 25.5, 25.4. HPLC (not less than) 96.08% pure; Novapak C18 column, 5-40% CH3CN / 2% / T.H20 over 60 minutes, detection at 230 nm, elution time: 23.2 minutes. HRMS (FAB): (Μ + H) calc'd for CHHNNO, 529.3328845; found 529.33172.

Example 30

Step 1. Formation of Aroide Bindings - Type D3 Procedure The 2-pyridylacetic acid hydrochloride salt (0.105 g, 0.60 mmol, 1.0 eq.) Was combined with 0.16 ml of TEA (1.15 mmol, 2 eq.) and 4 ml of CH 2 Cl 2. After 10 minutes, DEC (0.12 g, 0.62 mmol, 1.0 eq.) And 0.09 g HOBt (0.66 mmol, 1.1 eq.) Were added and the mixture was stirred for 2 hours. hours. N-Boc-amine 27 (0.44 g, 0.54 mmol, 0.9 eq.) Was added and the reaction was stirred for another 10 hours. TLC analysis (2 x MeOH, 12) showed that N-Boc-amine had been consumed. The reaction was diluted to 400 mL with EtOAc, washed with 1 N KOH (40 mL), brine (50 mL) and dried over MgSO4. The EtOAc solution was filtered and the solvents were removed to give 0.4 g of a clear green oil. The crude material was chromatographed from 10 g of silica gel transformed into a slurry 32

with EtOAc and EtOAc as eluent. The appropriate fractions were combined and the solvents were removed to give 0.20 g (40% yield) of a clear green oil. 1 H NMR (250 MHz, CDCl 3) δ 8.59-8.52 (m, 1H), 7.76-7.68 (m, 1H), 7.62-7.49 (m, 1H), 7 (M, 2H), 3.78 (s, 2H), 3.31-3.02 (m, 20H), 1.80-1.56 (m, 12H) 56-1.34 (m, 45H).

Step 2, Polyamine Deprotection - Procedure

Type F Trifluoroacetic acid (30 ml) was degassed with a dry bubble stream (through a Teflon tube) in a 100 ml RBF and a single tubing at room temperature. The 2-pyridylacetamide from step 1 above (180 mg) was dissolved in 2 ml CHâ,,Clâ,, and transferred to the TFA under stirring. After one hour, the solvents were removed under reduced pressure and the residue was placed on-Hi-Vac. The residue was macerated with ether (3 x 30 mL); a white solid formed which was collected under N 2 positive pressure on a porosity frit " B ". The remaining ether was removed by positive pressure of N 2; 169 mg (91% yield) of the product was isolated. 1 H NMR (250 MHz, D 2 O) δ 8.48 (d, J = 15 Hz), 7.98 (t, 1H, J = 6 Hz), 7.51-7.47 (m, 2H) , 3.86 (s, 2H), 3.32 (t, 2H, J = 9Hz), 3.17-2.96 (m, 18H), 2.17-1.98 (m, 6H), 1.98-1.81 (br, 2H), 1.81-1.69 (m, 4H).

N

Exemolo 4

Step 1, Amide Binding Formation - Procedure

Type D4 4-biphenylacetic acid (53 mg, 0.25 mmol, 1.2 eq.) Was combined with 5 ml of 84 μΐ of triethylamine (0.6 mmol, 3 eq.) / 70 mg dicyclohexylcarbodiimide (0.34 mmol, 1.6 eq.), 11 mg of hydroxysuccinimide (0.09 mmol, 45 mol%) and 175 mg of N-Boc-amine 27 (0.21 mmol, 1.0 eq.). The TCL analysis (2 x

MeOH, KMnOâ, ") revealed that the N-Boc-amine had been consumed after 16 hours. The reaction was diluted to 100 ml with CH2 Cl2 and washed with 20% aqueous NH4 OH solution (2 x 100 ml). The basic phases were extracted with CH 2 Cl 2 (3 x 50 mL); all fractions of CH 2 Cl 2 were combined and then washed with brine (50 ml), dried over K 2 CO 4, filtered, and the solvents were removed to give 281 mg (yield > 100%) of gross. The pure product was isolated by " flash chromatography " on silica gel (12 g transformed into a slurry in CH 2 Cl 2 and eluted with a gradient 0.10% MeOH / CH 2 Cl 2) as a waxy white solid (190 mg, 88% yield). Δ NMR (250 MHz, CDCl 3) δ 7.56-7.50 (m, 4H), 7.43-7.28 (m, 5H), 3.56 (s, 2H), 3.26-2, 98 (m, 20H), 1.78-1.52 (m, 12H), 1.48-1.36 (m, 45H). - .34 - .34

Step 2. Polyamine Deprotection - Procedure

Type F Trifluoroacetic acid (30 ml) was degassed with a continuous stream of N2 bubbles (via a Teflon tube) at 0 ° C. The biphenylacetamine from step 1 above (150 mg, 0.15 mmol) was added as a dry powder to the TFA under stirring. After 40 minutes, the ice bath was removed; after a further 20 minutes, the solvents were removed under reduced pressure, then Hi-Vac. After 2 hours, the resulting brownish oil was macerated with Et 3 O (3 x 30 mL); a white solid formed which was collected under N 2 positive pressure on a porosity frit " C ". The solid was dissolved in water, rinsed through the above-mentioned device and freeze-dried to afford 136 mg (yield 99%) of product as a white solid. 1 H NMR (300 MHz, D 2 O) δ 7.62-7.56 (m, 5H), 7.4 (t, 2H, J = 7.5 Hz), 7.38-7.31 (m, 2H) , 3.45 (s, 2H), 3.13 (t, 2H, J = 6.7Hz), 3.02-2.80 (m, 18H), 2.00-1.54 (m, 12H ).

Example 5 H-indole-3-acetamide-N- (16-amino-4,8,13-triazahexadec-1-yl) Step 1

(1) The compound of formula I was prepared from diaminobutane and acrylonitrile according to the procedure published by Yamamoto, Hisashi, J. Am. Chem. Soc. 103: 6133-6136 (1981).

Step 2

H

(III) To a solution of 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 dropwise addition 36 -

over a 7 hour period of N- (tert-butoxycarbonyl) -3-bromopropylamine (10.87 g, 0.0457 mol). The reaction was allowed to stand for 16 hours at room temperature and then was heated at 70 ° C for 24 hours. The reaction was allowed to cool and was filtered and concentrated in vacuo. The residue was dissolved in CH 2 Cl 2 (200 mL), washed with 1 N NaOH (100 mL), dried and concentrated in vacuo to provide the crude product which was chromatographed on silica gel (using CH 2 Cl 2 / MeOH 9: 1) to provide 3.32 g of amine III. 1 H NMR (CDCl 3) δ 1.19-1.59 (m, 17H), 2.42 (t, J = 6.6 Hz, 2H), 2.44-2.58 (m, 6H) 82 (t, J = 6.6 Hz, 2H), 3.08 (m, 2H), 5.22 (broad s, 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; HRMS -bar observed (Μ + H) m / z = 299.2434, C 15 H 31 N 4 C> 2 (req 299.2447).

Step 3

BOC * f

Under a nitrogen atmosphere, 4.7 g (15.8 mmol) of compound of formula III, prepared as described above in step 2, were dissolved in 150 ml of dichloromethane. Thereafter, 7.56 g (34.7 mmol) of

di-tert-butyl ester and the reaction mixture was stirred overnight at room temperature. The mixture was then concentrated under vacuum and chromatographed over 400 g of silica gel using 50:50 ethyl acetate / hexane as the solvent. The fractions were analyzed by TLC (50:50 ethyl acetate / hexane). The fractions containing the product of formula IV were combined and concentrated in vacuo to afford 7.9 g of product as an oil. 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 (broad s, 1H); 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; Observed FN-Bar-HRC (M + H) m / z = 499.3501, C25H47N4C> 6 (Req 499.3496).

Step 4

BOC-N

BOC BOC-N I

CN BOC BOC (IV) ▼

BOC

BOC

N

NH 2

(V) To 125 ml of acetic acid under a nitrogen atmosphere was added 7.85 g (15.8 mmol) of compound of formula IV, prepared as described in step 3 above, and 6.5 g of 38 - - * f * ι

X *

Pd (OH) 2 / carbon. The 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 1N NaOH and dried over K 2 CO 4 > 3. The solution was filtered and the filtrate was concentrated under vacuum to provide 7.8 g of compound of formula V. 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 (broad 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; Observed FN-Bar-HRC (Μ + H) m / z = 503.3804, C25H51N406 (req m / z = 503.3809).

Step 5 > V +

Under a nitrogen atmosphere, 7.15 g (14.2 mmol) of compound of formula V, prepared as described in step 4 above, were dissolved in 150 ml of methanol. Then, 1.03 ml (15.6 mmol) of acrylonitrile was added and the reaction was stirred for 72 hours at room temperature. The reaction mixture was then concentrated, reconcentrated three times from dichloromethane and the solvent was extracted under vacuum to provide 7.65 g of product of formula VI as an oil. Δ NMR (CDCl3) δ 1.26-1.73 (m, 36H), 2.44 (t, J = 6.7 Hz, 2H), 2.54 (t, J = 6.7 Hz, 2H) , 2.83 (t, J = 6.7 Hz, 2H), 3.00-3.16 (m, 10H), 5.24 (broad s, 1H); 13 C NMR (CDCl 3) δ 18.64, 25.84, 28.09, 28.43, 28.74, 37.84, 44.18, 44.68, 45.14, 46.29, 46.73, 46.85, 49.70, 78.90, 79.29, 79.46, 118.52, 155.84, 155.98; HRMS -bar observed (Μ + H) m / z = 556.4064, C26H54N506 (req m / z = 556.4074).

Step 6 * [BOC] 20

BOC (VII) VI + boc-n

H

Under a nitrogen atmosphere, 6.45 g (11.6 mmol) of compound of formula VI, prepared as described in step 5 above, were dissolved in 125 ml of dichloromethane. To this solution was added 2.6 g (12 mmol) of di-t-butyl dicarbonate and the reaction mixture was stirred overnight at room temperature. The mixture was then concentrated in vacuo and chromatographed over 400 g of silica gel using 50:50 ethyl acetate / hexane as eluent. The product fractions were combined and concentrated to afford 6.6 g of the product of formula VII as an oil. - 40 - r tr t

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 (broad s, 1H); 13 C NMR (CDCl 3) δ 17.20, 25.88, 27.83, 28.12, 28.35, 28.45, 28.77, 37.87, 43.91, 44.20, 44.77, 27, 46.88, 78.94, 79.42, 79.50, 80.54, 117.91, 154.96, 155.44, 155.74, 155.99; Observed FN-Bar-HRC (M + H) m / z = 656.4579, (req m / z = 656.4598).

Step 7

BOC BOC

NH 2

VII * BOC-N '

H

BOC (VIII) To 150 ml of acetic acid under a nitrogen atmosphere was added 6.6 g (10.1 mmol) of the compound of formula VII, prepared as described in step 6 above, and 6 g of ethyl acetate. 4

Pd (OH) / carbon. The mixture was hydrogenated at 3.45 x 10 (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 200 ml of dichloromethane, washed twice with 100 ml of 1N NaOH and dried over K 2 CO 3.

The solution was filtered and the filtrate was concentrated under vacuum to provide 6.5 g of product of formula VIII. 1 H NMR (CDCl 3) δ 1.28-1.71 (m, 46H), 2.16 (broad 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; Observed FN-Bar-HRC (Μ + H) m / z = 660.4914, C33N66N5Og (req 660.4911).

J t · * I

J t · * I

Under a nitrogen atmosphere, 1.75 g (10 mmol) of indoleacetic acid, 1.15 g (10 mmol) of N-hydroxysuccinimide and 2.06 g (10 mmol) of dicyclohexylcarbodiimide to 75 ml of tetrahydrofuran. The reaction mixture was stirred at room temperature, a precipitate forming 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 the cake was air-dried to give 1.84 g. The combined filtrate was concentrated, collected in ethyl acetate and filtered, the filter washed with ethyl acetate. The filtrate was concentrated to provide a foam. The foam was triturated with 75 mL of diethyl ether to provide a hard gum. Then, about 30 ml of ethyl acetate were added, followed by ethyl ether. The solids were isolated by filtration, washed with diethyl ether and dried under nitrogen to give 1.74 g of 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. - > . ≫

Step 9

IX BOC BOC

N'-30C H

VIII

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

Step 10

Under a nitrogen atmosphere, 0.37 g (0.45 mmol) of compound of formula X, prepared as described in step 9 above, were dissolved in 10 ml of dichloromethane. Then 0.218 g (1 mmol) of di-t-butyl dicarbonate was added, followed by 12 ml (0.1 mmol) of 4- (Ν, Ν-dimethylamino) pyridine. The reaction was stirred at room temperature for 1 hour and then allowed to stand overnight. The reaction mixture was chromatographed on silica gel using 4: 1 ethyl acetate / hexane and the product fractions were concentrated to provide 0.32 g of product of formula XI as a white foam.

Step ll >

XI

(XII) - 44 - 1

Under a nitrogen atmosphere, 0.32 g (0.35 mmol) of compound of formula XI, prepared as described in part 10 above, was added to 15 ml of trifluoroacetic acid, stirring for 15 minutes. The reaction mixture was then concentrated in vacuo and triturated with diethyl ether to provide 0.30 g of product as a white powder.

By an analogous procedure, N-Boc-amine 27 was prepared, having the following structure.

Steps 1 to 7, above, were followed which resulted in the N-Boc-amine of formula VIII.

Step 8a (m)

(XIII)

Scc Soc

Nitrile XIII prepared from N-Boc-amine VIII, such as nitrile VI was prepared from N-Boc-amine V (Example 5, Step 5) to give 1.00 g of product (yield 93% ). 1 H NMR (CDCl 3) δ 1.26-1.66 (m, 47H), 2.45 (t, J = 6.6

2H), 2.56 (t, J = 6.7 Hz, 2H), 2.85 (t, J = 6.6 Hz, 2H), 3.01-3.30 (m, 14H), 5.25 (broad s, 1H); 13 C NMR (CDCl 3) δ 18.68, Ϊ ι '>

25.92, 28.46, 28.48, 37.49, 44.19, 44.88, 45.16, 46.73, 73.93, 79.32, 79.44, 118.70, 155, 46, 155.61, 156.04; Observed HRMS -bar (Μ + Η) m / z = 713.5191, C HggH₂NNOOg (req ια / ζ = 713.5177).

Step 9a (χιιι)

N-Boc-nitrile XIV was prepared from nitrile XIII by amine protection using procedure A.

Step 10a N-Boc-amine 27 was prepared by hydrogenation of N-Boc-nitrile XIV, such as N-Boc-amine VIII was prepared from N-Boc-nitrile VII (step 7 of this example).

Examples 6 and 7

Starting from the appropriate polyamine 20 and R-acetic acid (for m = 1) or carboxylic acid (for m = 0), compounds of structure R (CH 2) m CO [NH (CH 2) 3] 5NH 2 .5 TFA were prepared by means of procedure D followed by procedure F.

Example m R 60 Ferrocene 7-1-indole (C '); t C '>

Examples 8 to 29

Starting from polyamine 27 and the appropriate R-acetic or carboxylic acid, compounds of structure R (CH 2) m CO [NH (CH 2) 3] NHN (CH 2) 4 NH (CH 2) or R (CH 2) m CO [NH (CH 2) 3] 3 NH (CH 2) 4 NH (CH 2) 3 NH 2 TFA (from procedure F) by the following procedures. .

χ

The title compound was prepared according to the procedure described in Example 13, except that the compound was prepared according to the procedure described in Example 13, above. -pyridine D3, then F1414-pyridine D3, then F15202-quinoline D1, then F163-3-quinoline D2, then F173-indole D1, then F181 3- (5-hydroxy -indole) D1, then F 191 3- (4-hydroxyindole) D2, then F201 3- (5-bromoindole) D2, then F 211 3- (4-fluoroindole) D2, then F 22 0 2- (5-fluoroindole) D2, then F- r ι

23 1 2- (5-fluoroindole) D 2, then F 24 1 3- (5-methoxyindole) D 2, then F 25 O 2 -quinoxaline D 2, then F 260 hydroquinone D 2, then F 27 0 4 -resorcinol D 2 , then F 28 1 p-biphenyl D 4, then F 29 1 2-naphthalene D 2, then F Exemples 30 and 31 Starting from polyamine 17 and the appropriate R-acetic carboxylic acid, compounds of structure R (CH 2) m CO [ NH (CH 2) 3] 4 NH 2 .4 TFA via procedure D1 followed by procedure F.

Example 30 R 30 Ferrocene 31 1 3 -indole Exempoles 32 and 33 Starting from the appropriate carboxylic polyamine, R-acetic compounds of structure

R (CH 2) m CO [NH (CH 2) 3] 3 · 2 · 3TFA Example 1 of procedure D1 followed by procedure F.

Example m R 32 Ferrocene 33 13-indole

Exemples 34 and 35

Starting from polyamine 11 and the appropriate R-acetic or carboxylic acid, compounds of structure R (CH 2) mC [NH (CH 2) 3] 2 NH 2 · 2 TFA were prepared by procedure D 1 followed by procedure F.

Example m R 34 Ferrocene 35 13-indole

Examples 36 and 37

Starting from polyamine 7 and the appropriate R-acetic or carboxylic acid, compounds of structure R (CH 2) 3 were prepared by procedure D 1 followed by procedure F 2. .

Example m 36 Ferrocene 373-indole

Examples 38 and 39

Starting from polyamine 23 and the appropriate R-acetic or carboxylic acid, compounds of structure R (CH 3) CO [NH (CH 2) 1.6 TFA were prepared by procedure D 1 followed by 2 ΤΓΙ 2 * -3 o 2 of the procedure F.

Example m R 38 Ferrocene 391 3-indole

Examples 40 and 41

Starting from polyamine 26 and the appropriate R-acetic or carboxylic acid, compounds of structure R (CH 2) m CO [NH (CH 2) 3] 7 NH 2 VTFA were prepared by procedure D 1 followed by procedure F.

Example m

R 40 0 ferrocene 41 1 3 -indole

Preparation A

Under a nitrogen atmosphere, 34.5 g (157.6 mmol) of 3-bromopropylamine.HBr were stirred in 600 ml of N, N-dimethylformamide. To this solution was added 34.4 g (157.6 mmol) of di-tert-butyl dicarbonate followed by 32.3 ml (236 mmol) of triethylamine. A precipitate formed immediately. The reaction was stirred overnight. The reaction mixture was then diluted to 1.5 liters with ethyl acetate, washed once with 500 ml of 1N HCl, three times with 500 ml of water, once with brine and dried over Na2 / SO4. After concentration , the product was chromatographed on 800 g of silica gel using 4: 1 hexane / ethyl acetate, and the fractions were followed by TLC (KMNO 3/4). The fractions containing the product were combined, concentrated in vacuo, twice with dichloromethane (50 mL) and purged under high vacuum to provide 25.8 g of the product of this preparation.

Lisbon, August 21, 1992

J. PEREIRA DA CRUZ Official Agent of Industrial Property RUA VICTOR CORDON, 10-A 3. »1200 USBOA

Claims (1)

% A process for the preparation of a compound of the formula wherein: R represents a 5 to 7 membered carbocyclic system or an 8 to 11 membered carbobicyclic system, or any of these systems substituted with one or more substituents independently selected from F, Cl, Br, OH, Cl to C4 alkyl, C1 to C4 alkoxy, CF3, phenyl, amino, C1 to C4 alkylamino and di (C1 to C4 alkyl) ) amino, m is equal to 0 or 1, R 'represents - [NH (CH)] NH, each n is independently of the others equal to a number between 2 and 5, ex is equal to a number between 1 and 6 , which comprises: (a) reacting a compound of formula R with a compound of formula Boc-NH (CH) [N (Boc) (CH3)] N (Boc) H, wherein Boc represents t-butyloxycarbonyl and R 2, n and n are as defined above in the presence of a carbodiimide base reagent together with an amide bond forming catalyst in an inert reactant and (b) removal of the t-butyloxycarbonyl groups by treatment with an organic or inorganic acid. 2. A process according to claim 1, wherein the said carbodiimide-based reactant is selected from dimethylaminopropyl-ethylcarbodiimide and dicyclohexylcarbodiimide and the said amide bond formation catalyst is selected from hydroxybenzotriazole and hydroxysuccinimide. 3. A process according to Claim 1 or 2, wherein said solvent inert to the reaction is dichloromethane and said organic or inorganic acid is selected from trifluoroacetic acid and hydrochloric acid. 4. A process according to one of Claims 1 to 3, characterized in that x is equal to 5; and each n is independently of the others, equal to 3 or 4. The process according to any one of Claims 1 to 4, wherein: R is phenyl or phenyl monosubstituted with F, Cl, Br, OH or MeOH; and m is equal to 1. > A process according to one of Claims 1 to 5, wherein R 'is - [NH (CH 2) 3] 3 -NH (CH 2) 4 -NH (CH 2) 3 -NH 2. A compound having the formula R- (CH 2) 2 -CO-R 2, or a pharmaceutically acceptable acid addition salt thereof, wherein: R represents a 5- to 7-membered carbocyclic system or a or any of these systems substituted with one or more substituents independently selected from F, Cl, Br, OH, Cl 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 (CHO)] NH2; Z Π X c * each n is, independently of the others, equal to a number between 2 and 5; and x is a number from 1 to 6. A compound according to Claim 7, characterized in that x is equal to 5; and each n is, independently of the others, equal to 3 or 4. X '. X '. A compound according to Claim 8 wherein: R 2 represents phenyl or phenyl monosubstituted with F, Cl, Br, OH or MeOH; and m is equal to 1. The compound according to Claim 8, wherein R 'is - [NH (CH 2) 3] 3 -NH (CH 2) 4 -NH (CH 2) 3 -NH 2. (CH2) m-CO-R ', or a pharmaceutically acceptable acid addition salt thereof, wherein: R represents a 5- to 7-membered carbocyclic system or a system or any of these systems substituted with one or more substituents independently selected from F, Cl, Br, OH, Cl 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 'represents [(CH 2) n NH] and H - [NH (CH 2) η] χΝ (CH 2) n NH] z H; To the Each η is, independently of the others, equal to a number between 2 and 5; x is equal to a number between 0 and 4; y and z are, independently of one another, equal to a number between 1 and 5; and the sum of x with the greater of e and z is equal to a number between 1 and 5. A compound of the formula: in which: R represents a 5- to 7-membered carbocyclic system or an 8 to 11-membered carbobicyclic system, or any such system substituted with one or more substituents independently selected from F , Cl, Br, OH, Cl to C4 alkyl, C1 to C4 alkoxy, CF3, amino, C1 to C4 alkylamino and di (C1 to C4 alkyl) amino; m is 0 or 1; R 'represents - [NH (CH ") 1 2' nJxN (CH 2) a [NH (CH 2) b] (CH2) a [NH (CH2) b] y wherein: each a is identical and is equal to a number between 2 and 5; 6 each b is identical and is equal to a number between 2 and 5; each n is, independently of the others, equal to a number between 2 and 5; x is equal to a number between 0 and 3; each y is identical and is equal to 0 or 1; z is an integer from 0 to 3; ex + y + z is equal to a number between 0 and 4. A pharmaceutical composition for antagonizing mammalian excitatory amino acid neurotransmitters, characterized in that it comprises an antagonizing amount of neurotransmitter-retaining amino acids in mammals of a compound according to or a pharmaceutically acceptable acid addition salt thereof and a pharmaceutically acceptable diluent or carrier. A pharmaceutical composition for antagonizing mammalian excitatory amino acid neurotransmitters comprising an antagonizing amount of mammalian neurotransmitter amino acids of a compound of Claim 9 or a pharmaceutically acceptable acid addition salt thereof and a pharmaceutically acceptable acid addition salt thereof. diluent or carrier. Lisbon, August 21, 1992 J. PEREIRA DA CRUZ Official Agent of Industrial Property RUA VICTOR CORDON, 10-A 3 * 1200 LISBOA