CN1069969A - Synthetic aryl polyamines as exicitatory/amino acid neurotransmitter antagonists - Google Patents

Abstract

Compounds of the formula and pharmaceutically acceptable acid addition salts thereof is a potent excitatory amino acid neurotransmitter antagonist,

R-(CH ₂ ) _m -CO-R′

The above-mentioned compounds can be used as drugs for treating psychosis in mammals, It can be used as an active ingredient in a pharmaceutical composition and used for the treatment of Excitatory Amino Acid Neurotransmitter Transmission in Mammalian Diseases, Control Invertebrates Vertebrate pests.

Description

The present invention relates to a class of arylpolyamines and pharmaceutically acceptable salts thereof, which are antagonists of excitatory amino acid neurotransmitters. The neurotransmitters affect neuronal cells of various organisms, including invertebrates and vertebrates. The polyamines of the present invention are synthetic analogs of certain polyamines present in the Agelenopsis aperta spider venom. The present invention also relates to the use of said polyamine and its salt in antagonizing excitatory amino acid neurotransmitters. The neurotransmitters affect cells such as those of the nervous system of an organism. The present invention also relates to the application of said polyamine and its salt in the treatment of diseases transmitted by excitatory amino acid neurotransmitters in mammals, as well as the application in controlling invertebrate pests. Salt composition. The invention also relates to a process for the preparation of said polyamines.

Jackson, H. et al. reported that Agelenopsis aperta spider venoms contain at least two toxins that affect calcium flux (Soc. Neu. Sci. Abstr. 12:1078 (1987)). Jackson, H. et al. disclose a toxin (termed AG2) having a molecular weight of less than 1,000 Daltons which appears to inhibit calcium flux in a wide range of tissues. In addition, Jackson, H. et al. also reported another toxin containing components with a molecular weight of about 6,000 Daltons obtained from Agelenopsis aperta (Soc. Neu. Sci. Abstr. 12: 730 (1986)). The toxin has been reported to block presynaptic transmission and is thought to block calcium channels associated with neurotransmitters.

Certain polyamines present in the Agelenopsis aperta spider venom are disclosed in U.S. Patent 5,037,846, filed April 28, 1989, and assigned to the assignee therein. Said polyamines and pharmaceutically acceptable salts thereof are disclosed as blockers of excitatory amino acid receptors in cells, and one of said polyamines (B ₁ ) is also disclosed as a blocker of calcium channels.

Excitatory amino acid neurotransmitter antagonists have a variety of uses. Excitatory amino acid neurotransmitter antagonists are useful in the treatment of conditions such as stroke, cerebral hemorrhage, neurodegenerative disorders such as Alzheimer's disease and epilepsy, and especially as psychotropic agents. See Excitatory Amino Acids In Health and Disease, D. Lodge, E., John Wiley and Sons Ltd., New York, NY 1988, the contents of which are incorporated herein by reference. In addition, the 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. Due to the development of glutamate receptor pharmacology in the past few decades, there has been remarkable and exciting progress in this area, and it is believed that it needs to be divided into several sub-disciplines. The subfamily of glutamate receptors belonging to the selective action of the exogenous agonist N-methyl-D-aspartate (NMDA) is the subject of intense research because of the role of these receptors in various neurological pathologies including stroke, epilepsy, and neurodegenerative diseases such as Alzheimer's disease). Currently, there are two main classes of NMDA receptor antagonists, which are actively promoting the research of clinically applied drugs. The first class includes competitive antagonists that interfere with the binding of glutamate to its receptor site. Such compounds are characterized by strong polar compounds, such as phosphonate compounds AP7 and AP5. The strongly charged structure of competitive antagonists renders them impermeable to the blood/brain barrier, thus limiting their therapeutic applications. The second class includes noncompetitive antagonists that block the action of NMDA receptors on ion channels associated with the NMDA receptor complex. The compounds include MK-801 and phencyclidine (PCP). The potent psychotomimetic action of this class of compounds is a distinct disadvantage of known drugs by the above-mentioned mechanism.

Recently, a third class of antagonists has been studied in detail based on the discovery of new glutamate antagonists from spider venoms. U.S. Patent Application Serial No. 554,311 (dated July 17, 1990) and U.S. Patent No. 5,037,846 (dated July 31, 1990) disclose mammalian NMDA isolated from the toxicant of Agelenopsis aperta The arylamine structure compound with effective and specific antagonistic effect on the receptor. The above-mentioned arylamine isolated from the poison of Agelenopsis aperta is a complex structure in which aromatic acid and polyamine fragments are linked together by amide bonds. In such structures, some of the amines in the polyamine moiety are functionalized as N-hydroxylamines or quaternary ammonium salts. The chemical structure of arylamine is different from the aforementioned general competitive drug AP5 or AP7 and the non-competitive compound MK-801. For example, the polyamine AGEL 416 disclosed in the aforementioned U.S. Patent 5,037,846 has the following structure:

The mechanism of the antagonism of the arylamines on NMDA is also different from the aforementioned competitive compound MK-801/PCP compounds. Arylamines in spider venoms thus provide a new class of compounds with antagonistic effects on NMDA receptors.

The benefit of revealing about naturally occurring compounds is that methods other than isolation/purification (from the total poison of Agelenopsis aperta) can now be applied to obtain said compounds, thus allowing the synthesis of non-naturally occurring analogous compounds of the same type.

The application relates to a class of aryl polyamines synthesized by the following formula:

Wherein R is a five- to seven-membered carbocyclic ring system or an eight- to eleven-membered carbocyclic ring system, or one or more members independently selected from F, Cl, Br, OH, C ₁ to C ₄ alkyl, C Any of the above carbocyclic ring systems substituted by ₁ -C ₄ alkoxy, CF ₃ , phenyl, amino, C ₁ -C ₄ alkylamino and di(C ₁ -C ₄ ) alkylamino substituents,

m is zero or 1

R' is -[NH(CH ₂ ) _n ] _X NH ₂ ; each n is independently 2-5; X is 1-6.

The present invention also relates to a compound of the following formula or a pharmaceutically applicable acid addition salt thereof,

R-(CH ₂ ) _m -CO-R′

Wherein R is a five- to seven-membered carbocyclic ring system or an eight- to eleven-membered carbocyclic ring system, or one or more members independently selected from F, Cl, Br, OH, C ₁ to C ₄ alkyl, C Any of the above carbocyclic ring systems substituted by ₁ -C ₄ alkoxy, CF ₃ , phenyl, amino, C ₁ -C ₄ alkylamino and di(C ₁ -C ₄ ) alkylamino substituents,

m is zero or 1,

Each n is independently 2-5; X is zero-4; Y and Z are each independently 1-5; and the larger sum of X and Y and Z is 1-5.

In addition, the present application also relates to a compound of the following formula or a pharmaceutically applicable acid addition salt thereof,

Wherein R is a five- to seven-membered carbocyclic ring system or an eight- to eleven-membered carbocyclic ring system, or one or more members independently selected from F, Cl, Br, OH, C ₁ to C ₄ alkyl, C Any of the above carbocyclic ring systems substituted by ₁ -C ₄ alkoxy, CF ₃ , phenyl, amino, C ₁ -C ₄ alkylamino and di(C ₁ -C ₄ ) alkylamino substituents,

m is zero or 1,

Here each a is the same and is 2 to 5; each b is the same and is 2 to 5; each n is independently 2 to 5; X is zero to 3; each Y is the same and is zero or 1; Z is zero to 3; and X+Y+Z is zero to 4.

The carbocyclic ring systems of the present invention may be saturated, unsaturated or aromatic, with aromatic systems being preferred. For monocyclic systems, phenyl is preferred. The bicyclic ring system can be a fused or bridged ring, and a 9- to 10-membered fused system is preferred, such as naphthyl or indene. Among the above bicyclic rings, naphthyl is particularly preferred.

Preferred R' groups are those wherein X is 4 or 5 and each n is independently 3 or 4. Particularly preferred R' groups are

The polyamines of the present invention and their pharmaceutically acceptable salts are antagonists of excitatory amino acid neurotransmitters. Thus, such polyamines themselves can be used to antagonize the excitatory amino acid neurotransmitters. The polyamines of the present invention are also useful in the control of invertebrate pests and in the treatment of mammalian diseases and conditions transmitted by excitatory amino acid neurotransmitters. The polyamines can also be used as drugs for treating psychosis in mammals.

The invention also relates to pharmaceutical compositions containing the above polyamines, methods of administering the above polyamines and methods of preparing such polyamines.

The synthesis method for preparing the polyamine of the following formula is shown in the following reaction formulas A to C.

Reaction A

According to the reaction formula A, the polyamine intermediate compound formula IV is prepared from diaminobutane through a series of steps. The suitable reaction conditions for preparing the intermediate compound VIII of Reaction Scheme A are shown in Example 5, Steps 1-7. Scheme B illustrates a method for preparing intermediate compounds of formula IX. See Example 5, Step 8 for suitable reaction conditions to prepare this intermediate. The method for preparing the polyamine compound formula XII of the present invention is shown in Reaction Scheme C. See Example 5, Steps 1-11 for suitable reaction conditions for coupling intermediate compounds of formulas VIII and IX and suitable reaction conditions for preparing compound of formula XII in the next step.

The polyamines of the present invention reversibly antagonize excitatory amino acid neurotransmitters that affect cells, such as the nervous system cells of a variety of organisms, including invertebrates and vertebrates. The term vertebrate as used herein is meant to include mammals. The term invertebrate as used herein is meant to include, for example, insects, ectoparasites and endoparasites.

The ability of the polyamine compounds of the present invention to antagonize excitatory amino acid neurotransmitters is represented by their ability to block the increase in cGMP caused by N-methyl-D-aspartic acid (NMDA) in the neonatal rat cerebellum, which The method is described below. The cerebellum of 10 Wistar rats aged 8-14 days was rapidly excised, placed in Krebs/sodium bicarbonate buffer (pH 7.4) at 4°C, and then sliced with a Mcllwain tissue slicer (The Nickle Laboratory Engineering CO. Gomshall, Surrey, England) and cut it into 0.5×0.5 mm slices. The resulting cerebellar slices were transferred to 100 ml of Krebs/sodium bicarbonate buffer at 37°C constantly equilibrated with 95:5 _O2 / _CO2 . Cerebellum slices were incubated in this manner for 90 minutes with 3 buffer changes. The buffer was then decanted, the tissue was centrifuged (3200 rpm, 1 min), and the tissue was resuspended in 20 ml of Krebs/sodium bicarbonate buffer. Take a 250 μl aliquot (approximately equivalent to 2 mg of tissue) from this and place in a centrifuge tube with a capacity of 1.5 ml. 10 [mu]l of the test compound was aspirated from the stock solution and added to these centrifuge tubes. Then 10 μl of 2.5 mM NMDA solution was added to start the reaction. The final concentration of NMDA was 100 μM. Control tubes had no NMDA added. The centrifuge tube was incubated at 37°C in a shaking water solution for 1 minute, and then 750 μl of 50 mM Tris-HCl, 5 mM EDTA solution was added to terminate the reaction. Immediately place the centrifuge tube in a boiling water bath for 5 minutes. The contents of each centrifuge tube were then sonicated for 15 seconds with a probe sonicator set at power level 3. 10 µl was taken out, and the protein concentration was determined by Lowry's method (Anal. Biochem 100: 201-220, 1979). Then each tube was centrifuged (10,000g, 5 minutes), and 100 μl of the supernatant was taken out, and the cGMP RIA detection kit produced by New England Nuclear Company (Boston, Massachusetts) was used to measure the level of cyclic GMP (cGMP) according to the manufacturer's method. . Results are expressed in pmol of cGMP produced per mg of protein.

In addition, the ability of the polyamine compounds of the present invention to antagonize excitatory amino acid neurotransmitters is represented by their blocking of the increase in the concentration of free [Ca ²⁺ ] in the cytosol caused by NMDA/glycine in dissociated cerebellar granule cells, and the method The description is as follows. Cerebellar granule cells were prepared from 8-day-old rat cerebellum (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 pipetted into 12-well plates containing 1 ml of Eagles Basal medium. Cells were dissociated and aliquots containing 6.25 x ¹⁰⁶ cells were added to wells containing Aclar squares. Twenty-four hours after coating, cytosine-β-D-arabinofuranoside was added (final concentration of 10 μM). Cells were analyzed for Fura 2 content at 6, 7 and 8 days after culture, respectively. Cells adhered to the square Aclar were transferred into HEPES buffer (containing 0.1% bovine serum albumin, 0.1% glucose, pH 7.4, without magnesium ions) in a 12-well dish. Cells were incubated at 37°C for 40 min; the buffer containing fura 2/AM was removed and replaced with 1 ml of the same buffer without fura 2/AM. Add 2 ml of pre-warmed 37°C buffer to the quartz cuvette. The cells on the square Aclar were transferred into the cuvette, and the cuvette was inserted into a constant temperature (37°C) cuvette holder equipped with a magnetic stirrer, and measured with a fluorescence classification photometer (Biomedical Instrument Group, University of Pennsylvania) The fluorescence intensity. The fluorescence signal stabilizes for about 2 minutes.

The increase in cytosolic free calcium concentration following the addition of 50 μM NMDA and 1 mM glycine was expressed as an increase in fluorescence. Then add 5-20 μl of a stock solution of the test compound at an appropriate concentration dissolved in phosphate-buffered saline (PBS, pH 7.4) into the cuvette. Correction of the fluorescent signal and correction of fura 2/AM overflow were performed using the method established by Grynkiewiez, G. et al. (J. Biol. Chem. 260:3440, 1985). At the end of each test, the maximum fluorescence value (Fmax) was measured after adding 35 μM ionomycin, followed by the addition of EGTA (12 mM) to chelate calcium, and the minimum fluorescence value (Fmin) was measured. Using the aforementioned operations, the ability of the target compound to antagonize the excitatory amino acid neurotransmitter can be demonstrated by taking the decrease in fluorescence after adding the target compound as an index.

The polyamines of the present invention are themselves useful for antagonizing excitatory amino acid neurotransmitters. Therefore, the polyamines of the present invention can also be used to control invertebrate pests, treat diseases and diseases transmitted by excitatory amino acid neurotransmitters in mammals, such as stroke, cerebral ischemia, neuron degenerative diseases (such as Alzheimer's disease) Murray's disease and epilepsy). The polyamines can also be used as psychotropic drugs in mammals. In addition, the polyamines of the present invention can also be used to study the physiology of cells, including but not limited to cells of the nervous system.

Pharmaceutically acceptable salts of the polyamines of the invention are also within the scope of the invention. Said salts can be obtained by methods well known to those skilled in the art. For example, the acid addition salts of the polyamines of the invention can be prepared according to the usual methods. The acid addition salts of the polyamines of this invention, such as the hydrochloride and trifluoroacetate salts, are preferred. The hydrochloride salts of the polyamines of the invention are particularly preferred.

When the pharmaceutically acceptable salt of the polyamine of the present invention is administered to mammals, it can be administered alone, or it can be mixed with a pharmaceutically acceptable carrier or diluent to form a pharmaceutical composition according to the general preparation method of pharmaceutical preparations. form to take. The polyamines of the present invention or pharmaceutically acceptable salts thereof can be taken orally or parenterally. Parenteral administration includes intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection and topical administration.

For oral administration of the polyamine of the present invention or a pharmaceutically acceptable salt thereof, the compound can be administered, for example, in the form of tablets or capsules, or in the form of an aqueous solution or suspension. For oral tablets, carriers which are commonly used include lactose and corn starch, and lubricating agents such as magnesium stearate are usually added. For oral capsules, lactose and dried cornstarch are commonly used diluents. For oral suspensions, the active ingredient is mixed with emulsifying and suspending agents. Certain sweetening and/or flavoring agents can be added, if desired.

For intramuscular, intraperitoneal, subcutaneous and intravenous injections, sterile solutions of the active ingredient are usually prepared and the pH of the solutions should be suitably adjusted and buffered. For intravenous injection, the total concentration of solutes should be controlled so that the preparation is isotonic.

When the polyamines of the present invention or salts thereof are used in humans, the daily dosage is usually determined by a prescribing physician. However, suitable dosages of the polyamines of the invention are about 1-30 mg/kg/day. Furthermore, the dosage of the polyamines of the present invention will vary with the age, weight of the patient, the response of the individual patient and the severity of the patient's condition and the effect of the particular compound being administered. Thus, dosages exceeding the dosage ranges given above are possible and are within the scope of the present invention.

When the polyamines of the present invention or salts thereof are used to control invertebrate pests, the compounds can be directly applied to the invertebrates, or provided to the environment of the invertebrates. For example, the compound of the invention is sprayed on the invertebrate as a solution. Dosages required to control invertebrate infestations will vary with invertebrates and environmental conditions and will be determined by the person applying the compound.

When the polyamine of the present invention or its salt is used in the study of cell physiology, the compound can be used in cells according to methods well known to those skilled in the art. For example, the compound can be administered to cells in an appropriate physiological buffer. A suitable concentration of the compound of the invention for this study is 100 [mu]M. However, the concentration of the compound in this study can be higher than 100 μM or lower than 100 μM. The dosage of the compound to be administered can be determined according to well-known methods by those skilled in the art.

example

Types of Experimental Methods

(A) Protection of secondary amines with di-tert-butylpyrocarbonate

(B) Cyanoethylation of primary amines

(C) Catalytic hydrogenation of nitriles to primary amines

(D) Formation of an amide bond reaction

(D ₁ ) Dimethylaminopropyl, Ethylcarbodiimide/Hydroxybenzotriazole

(D ₂ ) Dicyclohexylcarbodiimide/Hydroxybenzotriazole

(D ₃ ) Dimethylaminopropyl, Ethylcarbodiimide/Hydroxybenzotriazole/Triethylamine

(D ₄ ) Dicyclohexylcarbodiimide/Hydroxysuccinimide

(E) Deprotection of N-Boc substrates with HCl in dioxane

(F) Deprotection of N-Boc substrates with TFA (trifluoroacetic acid)

(G) Alkylation of amines with N-Boc-3-bromopropylamine

Example 1 Preparation of H ₂ N[(CH ₂ ) ₃ NH] _X polyamine side chain

Step 1 - Method B

A 103 g sample of 1,3-diaminopropane was combined with 45 mL of MeOH under stirring at 4°C. Acrylonitrile (100ml, 81g, 1.1eq) was added dropwise to the solution over 90 minutes via a pressure-balanced addition funnel. After 3 hours, 500 mg was removed and analyzed by ¹³ C NMR; no 1,3-diaminopropane was observed. The crude product, aminonitrile 8, was distilled under reduced pressure and 3 fractions were collected in the temperature range 100-125°C; all were pure enough to react with di-tert-butylcarbonate and then purified by silica gel chromatography.

¹ H NMR (250 MHz, CDCl ₃ ) δ2.67 (t, 2H, J=6.6Hz), 2.54-2.43 (m, 4H), 2.28 (t, 2H, J=6.6Hz), 1.37 (m, 2H , J=6.7Hz), 1.05 (s, 3H); ¹³ C NMR (63.1 MHz, CDCl ₃ ) δ118.8, 46.9, 44.9, 40.2, 33.4, 18.5.

Step 2 - Method A

To a solution of aminonitrile 8 (23 g, 0.18 mol) in 500 ml of dichloromethane was added di-tert-butyl dicarbonate (80 g, 0.36 mol, 2 eq) at 0°C. The reaction mixture was stirred at room temperature for 16 hours and treated with additional di-tert-butylpyrocarbonate (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 _K2CO3 , filtered and concentrated in vacuo _. The product was purified by flash chromatography ( _SiO₂ , 20→100% ethyl acetate in hexanes) to give the product N-Boc-nitrile 9 as a clear oil (14 g, 24% yield).

¹ H NMR (250 MHz, CDCl ₃ ) δ 3.40 (t, 2H, J=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 ) δ155.8, 155.1, 118.5, 80.7, 78.9, 45.7, 44.4 , 43.4, 32.4, 28.3, 28.2.

Step 3 - Method C

N-Boc nitrile 9 (49 g, 0.4 moles), 1000 mL acetic acid, and 20 g Pd(OH) ₂ /C (20% Pd(OH) ₂ by weight) were placed in a 2.6 L Parr shaker bottle. The shaker bottle was filled with hydrogen to 50 ^psig and shaken for 4 hours. The reaction solution was filtered through 0.47 μ filter paper and concentrated in vacuo. The residue was dissolved _in 1.5L _CH2Cl2 and washed with IN KOH (2 x 200ml). The base layer was extracted with _CH2Cl2 (400ml), the _{CH2Cl2 layers} were combined, dried over _K2CO3 , filtered and _concentrated in vacuo to afford N _- Boc amine 11 as a clear colorless oil (43 g, yielding rate of 86%).

¹ H NMR (250 MHz, CDCl ₃ ) δ3.28-3.12 (m, 2H), 3.11-3.00 (m, 4H), 2.64 (t, 2H, J=7Hz), 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, 31.6, 28.5, 28.2.

Step 4 - Method B

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 equiv) in 60 mL of methanol and stirred for 11 hours. Removal of the solvent afforded 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 (bs, 4H), 3.03 (m, 2H), 2.85 (t, 2H, J=6.6Hz), 2.57 (t, 2H, J=6.7Hz), 2.45 (t, 2H, J=6.7Hz), 1.72-1.53 (m, 4H), 1.41 (s, 9H), 1.38 (s, 9H); ¹³ C NMR (63.1 MHz, CDCl ₃ ) δ155.9, 118.6 , 79.6, 78.9, 46.3, 45.0, 43.9, 37.4, 28.8, 28.3, 18.6.

Step 5 - Method A

A 43 g sample of nitrile 12 (0.114 mol) prepared above was combined with di-tert-butylpyrocarbonate (25.6 g, 0.120 mol, 1.05 equiv) and ₃₅₀ ml _CH₂Cl₂ at 0°C and stirred for 9 hours. Thin layer chromatography (TLC) (EtOAc, KMnO ₄ ) showed that no starting material remained and the reaction was purified in the same manner as N-Boc-nitrile 9 to give N-Boc-nitrile 13 as a clear colorless oil ( 34 g, yield 63%).

¹ H NMR (250 MHz, CDCl ₃ ) δ3.45 (t, 2H, J=6.6Hz), 3.39-2.97 (m, 8H), 2.68-2.46 (2, 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 - Method C

N-Boc-amine 14 was prepared from N-Boc-nitrile 13 in 99% yield (30 g) following the procedure of N-Boc-nitrile 9 for N-Boc-amine 11.

¹ H NMR (300 MHz, CDCl ₃ ) δ3.32-2.94 (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 ) δ155.5, 79.5, 79.3, 45.5-43.7, 39.1, 37.3, 32.3, 28.3.

Step 7 - Method B

According to the method for preparing nitrile 12 from N-Boc-amine 11, nitrile 15 was prepared from N-Boc-amine 14 with a yield of 90%.

¹ H NMR (300 MHz, CDCl ₃ ) δ3.29-3.02 (m, 14H), 2.86 (t, 2H, J=6.7Hz), 2.57 (t, 2H, J=6.6Hz), 2.46 (t, 2H , J=6.6Hz), 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 ) δ155.5 , 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 spectrum (300 MHz, CDCl ₃ ) features of nitrile 15 and nitrile 12 are consistent.

Step 8 - Method A

Following the procedure of nitrile 12 to prepare N-Boc-nitrile 13, N-Boc-nitrile 16 was prepared from nitrile 15 as a clear colorless oil (30 g, 87% yield).

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 - Method C

N-Boc-amine 17 was prepared from N-Boc-nitrile 16 in 74% yield (2.61 g) following the procedure of N-Boc-nitrile 9 for N-Boc-amine 11.

¹ H NMR (250 MHz, CDCl ₃ ) δ3.39-2.97 (m, 14H), 2.63 (t, 2H, J=6.6Hz), 1.80-1.53 (m, 8H), 1.39 (s, 27H), 1.38 (s, 9H), 1.23 9 (s, 2H).

Step 10 - Method B

Nitrile 18 was prepared from N-Boc-amine 17 in 91% yield (19 g) following the procedure of N-Boc-amine 11 for nitrile 12 and was used without further purification.

¹ H NMR (300 MHz, CDCl ₃ ) δ3.24-2.94 (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 - Method A

Following the procedure of nitrile 12 for the preparation of N-Boc-nitrile 13, N-Boc-nitrile 19 was prepared from nitrile 18 (16 g, 74% yield).

¹ H NMR (250 MHz, CDCl ₃ ) δ3.43 (t, 2H, J=6.6Hz), 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 - Method C

N-Boc-amine 20 was prepared from N-Boc-nitrile 19 following the procedure of N-Boc-nitrile 9 for N-Boc-amine 11 (17 g, 99% yield).

¹ H NMR (300 MHz, CDCl ₃ ) δ3.24-2.95 (m, 18H), 2.6 (t, 2H, J≈6H), 1.72-1.52 (m, 10H), 1.42-1.32 (m, 45H).

Step 13 - Method B

According to the method for preparing nitrile 12 from N-Boc-amine 11, nitrile 21 was prepared from N-Boc-amine 20 with a yield of 99%.

¹ H NMR (250 MHz, CDCl ₃ ) δ3.32-3.03 (m, 16H), 2.91 (t, 2H, J=6.7Hz), 2.61 (t, 2H, J≈6Hz), 2.51 (t, 2H, J=6.6Hz), 1.82-1.57 (m, 10H), 1.44 (s, 36H), 1.43 (s, 9H).

Step 14 - Method A

According to the method for preparing N-Boc-nitrile 13 from nitrile 12, N-Boc-nitrile 22 was prepared from nitrile 21 with a yield of 99%.

3.45ppm (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 { ¹ H} (250 MHz, CDCl ₃ ) δ155.2, 79.3, 44.7, 28.3, 28.3, 28.2.

Step 15 - Method C

N-Boc-amine 23 was prepared from N-Boc-nitrile 22 following the procedure of N-Boc-nitrile 9 for N-Boc-amine 11 (8 g, 99% yield).

¹ H NMR (300 MHz, CDCl ₃ ) δ3.32-2.98 (m, 22H), 2.65 (t, 2H, J=6.6Hz), 1.78-1.53 (m, 12H), 1.41 (s, 54H), 1.40 (s, 9H).

Step 16 - Method B

According to the method for preparing nitrile 12 from N-Boc-amine 11, nitrile 24 was prepared from N-Boc-amine 23 with a yield of 99%.

¹ H NMR (250 MHz, CDCl ₃ ) δ3.32-3.04 (m, 22H), 2.90 (t, 2H, J=6.7Hz), 2.61 (t, 2H, J≈6Hz), 2.53 (t, 2H, J=6.7Hz), 1.82-1.57 (m, 12H), 1.44 (s, 45H), 1.43 (s, 9H).

Step 17 - Method A

N-Boc-nitrile 25 was prepared from nitrile 24 following the procedure of nitrile 12 for preparing N-Boc-nitrile 13 (5.5 g, 74% yield).

¹ H NMR (250 MHz, CDCl ₃ ) δ3.45 (t, 2H, J≈6.7Hz), 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 - Method C

N-Boc-amine 26 was prepared from N-Boc-nitrile 25 following the procedure of N-Boc-nitrile 9 for N-Boc-amine 11 (5.01 g, 91% yield).

¹ H NMR (250 MHz, CDCl ₃ ) δ3.30-2.97 (m, 26H), 2.60 (t, 2H, J=6Hz), 1.81 1.57 (m, 14H), 1.53-1.28 (m, 63H). ¹³ C NMR (250 MHz, CDCl ₃ ) δ155.2, 79.3, 44.7, 28.7, 28.4, 27.5.

Example 2

Step 1, Amide Bond Formation - Method D1

Under stirring and dry _N2 flow, 0.19 g of ferrocenecarboxylic acid (0.82 mmol, 1.1 eq), 0.12 g of hydroxybenzotriazole (0.89 mmol, 1.2 eq), 0.17 g of 1-(3- Dimethylaminopropyl)-3-ethylcarbodiimide (hydrochloride, 0.90 mmol, 1.2 equivalents) and 10 ml _CH2Cl2 were placed in 50 ml single-neck RBF for compounding _. After 30 minutes, 0.61 g of N-Boc-amine 27 (0.75 mmol, 1.0 eq, see Example 5a for preparation) was added to the solution. After 2 hours TLC (2xMeOH, _I2 ) showed that the N-Boc-amine was consumed. The reaction solution was diluted to 400ml with EtOAc and washed successively with pH 4 buffer (2 x 25ml), 25ml H ₂ O, IN KOH (2 x 25ml), 25ml H ₂ O and 50ml brine. The EtOAc layer was dried over _Na2SO4 , filtered, and _the solvent removed to give 712 mg (93%) of product as an orange oil.

¹ H NMR (250 MHz, CDCl ₃ ) δ4.78 (s, 2H), 4.32 (t, 2H, J=1.8Hz), 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, Polyamine Deprotection - Method F

Trifluoroacetic acid (30 ml) was degassed at 0 °C in a 100 ml single-necked RBF with a dry _N2 bubbling flow (through Teflon tubing). The ferroceneammonium tetraacylpolyamine from Step 1 above (712 mg, 0.69 mmol) was dissolved in 2 mL _CH2Cl2 and added to stirred _TFA , rinsed with 3 x ₂ mL _CH2Cl2 . After 30 minutes, the ice bath was removed; after an additional 30 minutes, the solvent was removed under reduced pressure, then concentrated by high vacuum. The residual reddish brown oil was triturated with _Et2O (3 x 30ml) to give a yellow solid and collected under nitrogen pressure on a porous "B" frit. The solid was rinsed with ether and the residual ether was purged with nitrogen pressure to give 690 mg of solid product (93% yield).

¹ H NMR (DMSO) δ4.71 (t, 2H, J = 1.73Hz), 4.38 (t, 2H, J = 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, _D2O ) 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 purity (not less than) 96.08%; Novapak C ¹⁸ column, within 60 minutes with 5-40% CH ₃ CN/2% T H ₂ O Elution, detection at 230nm, elution time: 23.2 minutes.

HRMS (FAB): (M+H), C ₂₇ H ₄₈ N ₆ O,

Calculated value: 529.3328845

Measured value: 529.33172

Example 3

Step 1, Amide Bond Formation - Method D3

2-Pyridylacetic acid hydrochloride (0.105 g, 0.60 mmol, 1.0 equiv) was combined with 0.16 mL of TEA (1.15 mmol, 2 equiv) and 4 mL _{of CH2Cl2} . After 10 minutes, DEC (0.12 g, 0.62 mmol, 1.0 eq) and 0.09 g of HOBt (0.66 mmol, 1.1 eq) were added, and the mixture was stirred for 2 h. N-Boc-amine 27 (0.44 g, 0.54 mmol, 0.9 equiv) was added and the reaction was stirred for an additional 10 hours. TLC (2xMeOH, _I2 ) showed that the N-Boc-amine was consumed. The reaction was diluted to 400ml with EtOAc, washed successively with IN KOH (40ml), brine (50ml), and dried over _MgSO4 . The EtOAc solution was filtered and the solvent was removed to give 0.4g of a clear green oil. The crude product was chromatographed on 10 g of silica gel (slurried with EtOAc) using EtOAc as eluent. The appropriate fractions were combined and the solvent removed to give 0.20 g (40% yield) of a clear pale green oil.

¹ H NMR (250 MHz, CDCl ₃ ) δ8.59-8.52 (m, ¹ H), 7.76-7.68) (m, 1H), 7.62-7.49 (m, ¹ H), 7.37 (d, ¹ H, J =8Hz), 3.78 (s, 2H), 3.31-3.02 (m, 20H), 1.80-1.56 (m, 12H), 1.56-1.34 (m, 45H).

Step 2, Polyamine Deprotection - Method F

Trifluoroacetic acid (30 ml) was degassed continuously at room temperature in a 100 ml single-necked RBF with a dry _N2 bubbling flow (through Teflon tubing). 2-Pyridylacetamide (180 mg) from Step 1 above was dissolved in ₂ mL _CH2Cl2 and added to stirred trifluoroacetic acid (TFA). After 1 hour, the solvent was removed under reduced pressure and the residue was concentrated under high vacuum. The residue was triturated with diethyl ether (3 x 30ml) to form a white solid and collected on a porous "B" sintered plate under nitrogen pressure to remove residual ether; 169mg (91% yield) was isolated product of.

¹ H NMR (250 MHz, D ₂ O) δ8.48 (d, ¹ H, J=15Hz), 7.98 (t, 1H, J=6Hz), 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 (m, 2H), 1.81-1.69 (m, 4H).

Example 4

Step 1, Amide Bond Formation - Method D4

Mix 4-biphenylacetic acid (53 mg, 0.25 mmol, 1.2 equiv.) with 5 ml CH ₂ Cl ₂ , 84 μl triethylamine (0.6 mmol, 3 equiv.), 70 mg dicyclohexylcarbodiimide (0.34 mmol mol, 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). After 16 hours TLC (2xMeOH, _KMnO₄ ) showed that the N-BOC-amine was consumed. The resulting solution was diluted to _100ml with _CH2Cl2 and washed with 20% aqueous _NH4OH (2 x 100ml). The base layer was extracted with _CH2Cl2 (3 x 50ml); all _CH2Cl2 extracts were _combined and washed with brine ( _50ml ), dried over _K2CO3 , filtered and _the solvent removed to give 281mg (prod. rate>100%) crude product. The pure _product was isolated as a white waxy _solid ( _{190 mg} , Yield 88%).

¹ H NMR (250 MHz, CDCl ₃ ) δ7.56-7.50 (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).

Step 2, Polyamine Deprotection - Method F

Trifluoroacetic acid (30 ml) was degassed at 0 °C with a continuous bubbling stream of _N2 (through Teflon tubing). Biphenylacetamide (150 mg, 0.15 mmol) as a dry powder from Step 1 above was added to stirring TFA. After 40 minutes the ice bath was removed; after another 20 minutes the solvent was removed under reduced pressure, then concentrated under high vacuum. After 2 hours, the resulting brown oil was triturated with _Et₂O (3 x 30 mL); a white solid formed which was collected on a porous "C" frit under nitrogen pressure. The solid was dissolved in water, washed through a sinter plate, and lyophilized to give 136 mg (99% yield) of product as a white solid.

¹ H NMR (300 MHz, D ₂ O) δ7.62-7.56 (m, 5H), 7.4 (t, 2H, J=7.5Hz), 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 1H-indole-3-acetamide-N-(16-amino-4,8,13-triazahexadecane-1-yl

step 1

According to the method published by Yamamoto, Hisashi (J.Am.Chem.Soc.103:6133-6136 (1981)), prepare formula 1 compound with diaminobutane and acrylonitrile

step 2

To a solution of N-cyanoethyl-1,4-diaminobutane (6.44 g, 0.0457 mol) in acetonitrile (200 mL) was added KF/celite (11 g) under nitrogen flow, followed by dropwise addition of N-(tert-butoxycarbonyl)-3-bromopropylamine (10.87 g, 0.0457 mol). The reaction was stirred at room temperature for 16 hours, then heated to 70°C for 24 hours. The reaction was cooled, filtered and concentrated in vacuo. The residue was dissolved in _CH2Cl2 ( _200ml ), washed with 1N NaOH (100ml), dried and concentrated in vacuo to give a crude product which _was chromatographed on silica gel (9:1 _CH2Cl2 /MeOH) to give 3.32 g of amine III.

¹ H NMR (CDCl ₃ ) δ1.19-1.59 (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 (br s, 1H); ¹³ C NMR (CDCl ₃ ) δ18.68, 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 real (M+H) m/z = 299.2434, C ₁₅ H ₃₁ N ₄ O ₂ (calculated 299.2447).

step 3

Under nitrogen flow, 4.7 g (15.8 mmol) of the compound of formula III prepared as described in Step 2 above was dissolved in 150 ml of dichloromethane. Then 7.56 g (34.7 mmol) of di-tert-butylpyrocarbonate were added and the reaction mixture was stirred overnight at room temperature. The mixture was then concentrated in vacuo and chromatographed on 400 g of silica gel eluting with 50:50 ethyl acetate/hexane solvent. Fractions were monitored by TLC (50:50 ethyl acetate/hexane). The reactions containing the product of formula IV were combined and concentrated in vacuo to give 7.9 g of product as an oil.

¹ H NMR (CDCl ₃ ) δ1.20-1.59 (m, 33H), 2.55 (m, 2H), 3.01-3.37 (m, 8H), 3.39 (t, J=6.6Hz, 2H), 5.25 (br s , 1H); ¹³ C NMRδ17.21, 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 FABMMS real values ( H) m/z = 499.3501 for C ₂₅ H ₄₇ N ₄ O ₆ (calculated 499.3496).

step 4

To 125 ml of acetic acid were added under nitrogen flow 7.85 g (15.8 mmol) of the compound of formula IV prepared as described in Step 3 above and 6.5 g of Pd(OH) ₂ /carbon. The mixture was hydrogenated at 50 psig for ² hours. The catalyst was filtered off, and the filter cake was washed with acetic acid. The filtrate was concentrated and dissolved in 250 ml of dichloromethane, washed _twice with 100 ml of 1N NaOH, and dried over _K2CO3 . The solution was filtered and the filtrate was concentrated in vacuo to yield 7.8 g of the compound of formula V.

¹ H NMR (CDCl ₃ ) δ1.24-1.59 (m, 35H), 2.14 (s, 2H), 2.61 (t, J=6.7Hz, 2H), 2.98-3.14 (m, 10H), 5.22 (br s , 1H); ¹³ C NMR (CDCl ₃ ) δ25.89, 28.42, 31.38, 32.36, 37.55, 38.95, 43.95, 46.65, 79.34, 79.48, 155.65, 156.03; HR FABMS real value (M+H) m/z = 503.3804, C ₂₅ H ₅₁ N ₄ O ₆ (calculated m/z = 503.3809).

step 5

Under nitrogen flow, 7.15 g (14.2 mmol) of the 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 solution was stirred at room temperature for 72 hours. The reaction mixture was concentrated, re-concentrated three times from dichloromethane and the solvent was removed in vacuo to give 7.65 g of the product of formula V as an oil.

¹ H NMR (CDCl ₃ ) δ1.26-1.73 (m, 36H), 2.44 (t, J=6.7Hz, 2H), 2.54 (t, J=6.7Hz, 2H), 2.83 (t, J=6.7Hz , 2H), 3.00-3.16 (m, 10H), 5.24 (br s, 1H); ¹³ C NMR (CDCl ₃ ) δ18.64, 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; HR FABMS real side value (M+H) m/z = 556.4064, C ₂₈ H ₅₄ N ₅ O ₆ (calculated value m/z = 556.4074 ).

step 6

6.45 g (11.6 mmol) of the compound of formula VI prepared as described in Step 5 above were dissolved in 125 ml of dichloromethane under nitrogen flow. To this solution was added 2.6 g (12 mmol) of di-t-butylpyrocarbonate, and the reaction mixture was stirred overnight at room temperature. The mixture was then concentrated in vacuo and chromatographed on 400 g of silica gel eluting with 50:50 ethyl acetate/hexane. Fractions containing product were combined and concentrated to give 6.6 g of product of formula VIII as an oil.

¹ H NMR (CDCl ₃ ) δ1.26-1.73 (m, 44H), 3.03-3.24 (m, 14H), 3.42 (t, J=6.6Hz, 2H), 5.25 (br s; 1H); ¹³ C NMR ( _CDCl3 ) δ17.20, 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, 115.571, 155.49, 15; HR FABMS real value (M+H) m/z = 656.4579, C ₃₃ H ₆₂ N ₅ O ₈ (calculated m/z = 656.4598).

step 7

To 150 ml of acetic acid were added 6.6 g (10.1 mmol) of the compound of formula VII prepared as described in Step 6 above and 6 g of Pd(OH) ₂ /carbon under a nitrogen stream. The mixture was hydrogenated at 50 psig for ² hours. The catalyst was filtered off, and the filter cake was fully washed with acetic acid. The filtrate was _concentrated and dissolved in 200 ml of dichloromethane, washed twice with 100 ml of 1N NaOH, and dried over _K2CO3 . The solution was filtered and the filtrate was concentrated in vacuo to yield 6.5 g of the compound of formula VIII.

¹ H NMR (CDCl ₃ ) 1.28-1.71 (m, 46H), 2.16 (br s, 2H), 2.65 (t, J=6.7Hz, 2H), 3.01-3.18 (m, 14H), 5.24 (br s, 1H); ¹³ C NMR (CDCl ₃ ) δ25.85, 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 real value (M+H) m/ z = 660.4914 for C ₃₃ N ₆₆ N ₅ O ₈ (calculated 660.4911).

Add 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 under a nitrogen stream . The reaction mixture was stirred at room temperature and after about 5 minutes a precipitate formed. After about 1.5 hours the precipitate was filtered, the filter cake was washed with 75 mL of tetrahydrofuran, and the filter cake was air dried to yield 1.84 g. The combined filtrates were concentrated and dissolved in ethyl acetate, filtered and washed with ethyl acetate. The filtrate was concentrated to give a foam. The foam was triturated with 75 mL of ether to give a hard gum. Then about 30 ml of ethyl acetate and diethyl ether were added successively. The solid was isolated by filtration, washed with ether and dried under nitrogen to yield 1.74 g of product of formula IX. It was also found that treatment of the mother liquor with petroleum ether afforded an additional 0.47 g of product.

step 9

Under nitrogen flow and stirring, 0.33 g (5 mmol) of the compound of formula VIII prepared as described in step 7 above was dissolved in 10 ml of dichloromethane, and then 0.136 g (5 mmol) of the compound of formula VIII prepared as described in step 8 above was added. millimole) compound of formula IX. The reaction was stirred overnight at room temperature. The reaction mixture was then diluted to 35ml with _{dichloromethane} , washed with 10ml 0.5N NaOH, dried over _K2CO3 and concentrated. The concentrate was chromatographed on silica gel, eluting with 4:1 ethyl acetate/hexane. Concentration of the product-containing fractions gave 0.37 g of a white foam containing the product of formula X with some ethyl acetate.

Step 10

Under nitrogen flow, 0.37 g (0.45 mmol) of the compound of formula X prepared as described in Step 9 above was dissolved in 10 ml of dichloromethane. Then 0.218 g (1 mmol) of di-tert-butylpyrocarbonate and 12 ml (0.1 mmol) of 4-(N,N-dimethylamino)pyridine were added successively. The reaction was stirred at room temperature for 1 hour, then allowed to stand overnight. The reaction mixture was chromatographed on silica gel, eluting with 4:1 ethyl acetate/hexane, and the product-containing fractions were concentrated to give 0.32 g of the product of formula XI as a white foam.

step 11

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

N-Boc-amine 27 having the following structure was prepared in a similar manner.

Following steps 1-7 above, N-Boc-amines of formula VIII are produced.

Step 8a

Nitrile XIII was prepared from N-Boc-amine VIII following the procedure for nitrile VI prepared from N-Boc-amine V (Example 5, Step 5) to give 1.00 g of product (93% yield).

¹ H NMR (CDCl ₃ ) δ1.26-1.66 (m, 47H), 2.45 (t, J=6.6Hz, 2H), 2.56 (t, J=6.7Hz, 2H), 2.85 (t, J=6.6Hz , 2H), 3.01-3.30 (m, 14H), 5.25 (br s, 1H); ¹³ C NMR (CDCl ₃ ) δ18.68, 25.92, 28.46, 28.48, 37.49, 44.19, 44.88, 45.16, 46.73, 78.93, 79.32, 79.44, 118.70, 155.46, 155.61, 156.04; HR FABMS real side value (M+H) m/z = 713.5191, C ₃₆ H ₆₉ N ₆ O ₈ (calculated m/z = 713.5177).

Step 9a

Using Method A, N-Boc-nitrile XIV is prepared from nitrile XIII by amine protection.

Step 10a

N-Boc-amine 27 was prepared by hydrogenation of N-Boc-nitrile XIV following the procedure for preparing N-Boc-amine VIII from N-Boc-nitrile VII (Step 7 of this Example).

Examples 6 and 7

Starting from polyamine 20 and the appropriate R-acetic acid (m = 1) or carboxylic acid (m = 0), a compound having R(CH ₂ ) _m CO[NH(CH ₂ ) ₃ ] ₅ NH ₂ ·5TFA compound.

Example m R

6 0 ferrocene

7 1 3-indole

Example 8-29

Starting from polyamine 27 and the appropriate R-acetic or carboxylic acid, the compounds having the following structures were prepared in the following manner.

R(CH ₂ ) _mCO [NH(CH ₂ ) ₃ ] ₃ NH(CH ₂ ) ₄ NH(CH ₂ ) ₃ NH ₂ 5HCl (Method E) or

R(CH ₂ ) _m CO[NH(CH ₂ ) ₃ ] ₃ NH(CH ₂ ) ₄ NH(CH ₂ ) ₃ NH ₂ 5TFA (Method F)

Example m R method

8 0 Use F after ferrocene D1,

9 0 2-pyridine D3 followed by F,

10 0 3-pyridine D3 followed by F,

11 0 4-pyridine D3 followed by F,

12 1 2-pyridine D3 followed by F,

13 1 3-pyridine D3 followed by F,

14 1 4-pyridine D3 followed by F,

15 0 2-quinoline D1 followed by F,

16 0 3-quinoline D2 followed by F,

17 1 3-indole D1 followed by E,

18 1 3-(5-oxindole) D2 followed by F,

19 1 3-(4-oxindole) D2 followed by E,

20 1 3-(5-bromoindole) D2 followed by F,

21 1 3-(4-fluoroindole) D2 followed by F,

22 0 2-(5-fluoroindole) D2 followed by F,

23 1 2-(5-fluoroindole) D2 followed by F,

24 1 3-(5-methoxyindole) D2 followed by F,

25 0 2-quinoxaline D2 followed by F,

26 0 Use F after hydroquinone D2,

27 0 4-resorcinol D2 followed by F,

28 1 p-biphenyl Use F after D4,

29 1 2-naphthalene D2 followed by F,

Examples 30 and 31

Starting from polyamine 17 and the appropriate R-acetic acid or carboxylic acid, compounds having the structure R(CH ₂ ) _m CO[NH(CH ₂ ) ₃ ] ₄ NH ₂ ·4TFA were prepared according to Methods D1 and F.

Example m R

30 0 ferrocene

31 1 3-indole

Examples 32 and 33

Starting from polyamine 14 and the appropriate R-acetic acid or carboxylic acid, a compound having the structure R(CH ₂ ) _m CO[NH(CH ₂ ) ₃ ] ₃ NH ₂ ·3TFA was prepared by Method D1 followed by Method F .

Example m R

32 0 Ferrocene

33 1 3-indole

Examples 34 and 35

Starting from polyamine 11 and the appropriate R-acetic acid or carboxylic acid, a compound having the structure R(CH ₂ ) _m CO[NH(CH ₂ ) ₃ ] ₂ NH ₂ ·2TFA was prepared by Method D1 followed by Method F . Instance m R

34 0 ferrocene

35 1 3-indole

Examples 36 and 37

Starting from polyamine 7 and the appropriate R-acetic acid or carboxylic acid, compounds having the structure R(CH ₂ ) _m CONH(CH ₂ ) ₃ NH ₂ ·TFA were prepared according to Methods D1 and F.

Example m R

36 0 Ferrocene

37 1 3-indole

Examples 38 and 39

Starting from polyamine 23 and the appropriate R-acetic acid or carboxylic acid, compounds having the structure R(CH ₂ ) _m CO[NH(CH ₂ ) ₃ ] ₆ NH ₂ ·6TFA were prepared according to Methods D1 and F.

Example m R

38 0 Ferrocene

39 1 3-indole

Examples 40 and 41

Starting from polyamine 26 and the appropriate R-acetic acid or carboxylic acid, compounds having the structure R(CH ₂ ) _m CO[NH(CH ₂ ) ₃ ] ₇ NH ₂ ·7TFA were prepared according to Methods D1 and F.

Example m R

40 0 ferrocene

41 1 3-indole

Preparation method A

A solution of 34.5 g (157.6 mmol) of 3-bromopropylamine·HBr in 600 ml of N,N-dimethylformamide was stirred under a stream of nitrogen. To this solution were successively added 34.4 g (157.6 mmol) of di-t-butylpyrocarbonate and 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 L with ethyl acetate, washed once with 500 1N HCl, three times with 500 ml _of water, once with brine, and dried over _Na2SO4 . After concentration, the product was chromatographed on 800 g of silica gel, eluting with 4:1 hexane/ethyl acetate, and the fractions were monitored by TLC (hexane/ethyl acetate, KMnO ₄ /I ₂ ). The product-containing fractions were combined, concentrated in vacuo, washed twice with 50 ml of dichloromethane, and purified by high vacuum concentration to yield 25.8 g of the product of this preparation.

Claims (6)

1, the method for preparing following formula: compound,

Wherein R be five to seven-element carbon ring system or eight to the undeca carbon ring system, perhaps for by being independently selected from F, Cl, Br, OH, C more than 1 or 1 ₁-C ₄Alkyl, C ₁-C ₄Alkoxyl group, CF ₃, phenyl, amino, C ₁-C ₄Alkylamino and two (C ₁-C ₄Alkyl) the arbitrary above-mentioned carbon-loop system of amino substituting group replacement,

M is zero or 1,

R ' is-[NH (CH ₂) _n] _xNH ₂,

Each n is 2-5 independently,

X is 1-6,

This method comprises

(a) based on the reagent of carbodiimide and form in the presence of the catalyzer of amido linkage,, make formula R compound and formula Boc-NH (CH in reaction is in the inert solvent ₂) _n[N (Boc) (CH ₂) _n] _X-1The reaction of N (Boc) H compound, wherein Boc represents tert-butoxycarbonyl, and the definition of R, n and x is the same,

(b) use the organic or inorganic acid treatment, slough tert-butoxycarbonyl.

2, in accordance with the method for claim 1, wherein the reagent system based on described carbodiimide is selected from dimethylamino-propyl, ethyl carbodiimide and dicyclohexylcarbodiimide, and the catalyzer system of described formation amido linkage is selected from hydroxybenzotriazole and N-Hydroxysuccinimide.

3, according to claim 1 or 2 described methods, wherein the solvent that reaction is inert is a methylene dichloride, and described organic or inorganic acid system is selected from trifluoroacetic acid and hydrochloric acid.

4, according to any one described method among the claim 1-3, wherein x is 5, and each n is 3 or 4 independently.

5, according to any one described method among the claim 1-4, wherein R is a phenyl or by the mono-substituted phenyl of F, Cl, Br, OH or MeOH, and m is 1.

6, according to any one described method among the claim 1-5, wherein R is-[NH(CH ₂) ₃] ₃-NH(CH ₂) ₄-NH(CH ₂) ₃-NH ₂