HK40060177B - Derivative compound introducing biphenyl group into novel aminoalkanoic acid and antifungal pharmaceutical composition comprising same - Google Patents

Description

Biphenyl-introduced derivative compounds of novel aminoalkyl acids and antifungal pharmaceutical compositions containing them

Technical Field

This invention relates to derivative compounds of aminoalkyl acids containing biphenyl groups, their stereoisomers, or pharmaceutically acceptable salts thereof; and pharmaceutical compositions comprising them as active ingredients for the prevention or/and treatment of fungal infections.

Background Technology

With increased life expectancy in modern people, opportunistic infections caused by fungi are increasing in the elderly due to decreased immune function. Furthermore, infections caused by opportunistic fungi are increasing worldwide in patients with weakened immune systems due to immunosuppressants reducing transplant rejection, organ transplant recipients with impaired immune function, or those with weakened immunity due to chemotherapy and acquired immunodeficiency syndrome (AIDS). In the past, fungal infections were mainly caused by localized infections such as athlete's foot, tinea cruris, and thrush; however, in recent years, systemic fungal infections have become so prevalent that they rank fourth in frequency among all hospital-acquired infections. Representative opportunistic fungi reported include *Candida albicans*, *Candida glabrata*, *Candida krusei*, and *Cryptococcus neoformans*. *Cryptococcus neoformans*, a pathogenic fungus causing systemic infections, is commonly found in soils worldwide; its basidiospores are inhaled from the surrounding environment into the lungs via the respiratory tract. In immunocompromised patients, such as organ transplant recipients or AIDS patients, latent fungi in the lungs can cause lung infections and cross the blood-brain barrier (BBB) into the central nervous system, leading to life-threatening meningitis. Cryptococcal meningitis, in particular, has the highest mortality rate among meningitis types, reportedly causing over 600,000 deaths worldwide each year. However, because fungi, like mammals, are composed of eukaryotic cells, their biochemical and metabolic pathways are very similar, making it difficult to identify fungal-specific targets. Therefore, drugs used to treat cryptococcosis to date have several limitations, resulting in only limited treatment options. To date, developed antifungal agents for inhibiting Cryptococcus fungi can be mainly classified into polyenes, including amphotericin B; azoles, including ketoconazole, fluconazole, itraconazole, and voriconazole; and echinocandins, including non-azoles such as terbinafine, flucytosine, and caspofungin. Amphotericin B, belonging to the polyene class, binds to ergosterol in the cell membrane of Cryptococcus, inducing apoptosis through oxidative damage, thereby inhibiting the bacteria. However, it exhibits high toxicity to the body, causing side effects. Aazole drugs inhibit the 14α-demethylase enzyme that converts lanosterol to ergosterol, a necessary component of fungal cell membranes, thus inhibiting ergosterol production, weakening the cell membrane, and inducing apoptosis, thereby exerting an antifungal effect. Reports indicate that azole drugs are prone to drug resistance development. Terbinafine inhibits ergosterol synthesis by preventing the conversion of squalene to squalene epoxy. Flucytosine, a metabolic antagonist that inhibits nucleic acid synthesis, exerts its antifungal effect by inducing miscoding of fungal RNA and antagonizing DNA synthesis. Echinocandins inhibit fungal cell wall formation; unlike the previously mentioned antifungal agents that act on the cell membrane, these drugs target the cell wall. Because previously developed antifungal agents exhibit various side effects, such as toxicity and resistance, depending on the drug type, it is necessary to develop novel antifungal agents that can minimize these side effects while improving antifungal efficacy.

Summary of the Invention

Technical issues

The object of this invention is to provide derivatives, salts and/or solvates of novel aminoalkyl acids incorporating biphenyl groups.

Another object of the present invention is to provide an antifungal pharmaceutical composition comprising, as an active ingredient, a derivative of the present invention containing biphenyl-introduced aminoalkyl acid, its salt and/or solvate.

Another object of the present invention is to provide an antifungal pesticide formulation comprising, as an active ingredient, the derivative of the present invention containing biphenyl-introduced aminoalkyl acid, its salt and/or solvate.

Another object of the present invention is to provide an antifungal preparation for animals comprising, as an active ingredient, a derivative of the present invention containing biphenyl-introduced aminoalkyl acid, its salt and/or solvate.

Another object of the present invention is to provide an antifungal composition comprising the derivatives of the present invention that introduce biphenyl into aminoalkyl acids, their salts and/or solvates.

Another object of the present invention is to provide a personal cleansing composition, cosmetic composition, or shampoo composition comprising the derivative of the present invention that introduces biphenyl into an aminoalkyl acid, its salt, and/or solvate.

Furthermore, the purpose of this invention is to provide a method for preparing the benzyloxybenzylamine amino acid derivative of this invention.

Solution to the problem

As one aspect for achieving the above objectives, the present invention provides compounds represented by Chemical Formula 1, their stereoisomers, or pharmaceutically acceptable salts thereof:

Chemical Formula 1:

In the above chemical formula 1,

n is 0, 1, 2, 3, 4, or 5.

_R1 , _R2 , and _R3 are each independently the same or different, and are each independently selected from hydrogen, _C1-7 alkyl, hydroxyl, halogen, halogenated _C1-7 alkyl, _C1-7 alkoxy, and halogenated _C1-7 alkoxy.

X is selected from m identical or different substituents (m is an integer from 1 to 5) selected from the group consisting of halogen groups, halogenated _C1-7 alkyl groups and halogenated _C1-7 alkoxy groups.

Furthermore, the compounds represented by Formula 1 are compounds with no limitation on the three-dimensional arrangement of substituents bonded to chiral carbons, and may contain all structurally possible optical isomers. Specifically, the compounds represented by Formula 1 can be provided in the form of their (R) or (S) isomers alone or mixtures thereof (e.g., racemates).

In this invention, the halogen can be selected from fluorine, chlorine, bromine and iodine.

The _C1-7 alkyl groups mentioned above can be straight-chain, branched, or cyclic alkyl groups, and can be selected from methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, and octyl.

The _C1-7 alkoxy groups mentioned above can be selected from methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy, heptoxy, and octoxy.

The aforementioned halogenated _C1-7 alkyl groups may be selected from difluoromethyl, trifluoromethyl, difluoroethyl, trifluoroethyl, trifluoropropyl, trifluoropentyl, trifluorohexyl, and trifluoroheptyl.

The aforementioned halogenated _C1-7 alkoxy groups may be selected from difluoromethoxy, trifluoromethoxy, difluoroethoxy, trifluoroethoxy, trifluoropropoxy, trifluoropentoxy, trifluorohexyloxy, and trifluoroheptoxy.

This invention includes not only the above-mentioned compound 1 or its pharmaceutically acceptable salt, but also solvates or hydrates prepared therefrom having the same effects, all of which fall within the scope of this invention.

The compounds of the present invention are based on aminoalkyl acids, and may be introduced into biphenyl derivative compounds.

For example, the aforementioned aminoalkyl acid can be an α-amino acid derivative containing a straight-chain hydrocarbon with _C2-4 in the side chain, such as α-aminobutyric acid, valine, or leucine.

The term "α-Aminobutyric acid (AABA)" in this invention refers to the compound shown in Chemical Formula 6 below, which has the IUPAC name 2-Aminobutanoic acid. AABA is a non-protein α-amino acid with the chemical formula _C4H9NO2 , known in biochemistry as a homoalanine. It has a longer _C1 -chain than alanine and contains a _C2 straight-chain hydrocarbon in its side chain.

Chemical Formula 6:

The term "Norvaline (Nva)" in this invention refers to the compound shown in Chemical Formula 7 below, which has the IUPAC name of 2-Aminopentanoic acid. It is a branched chain amino acid (BCAA) and is a water-soluble amino acid as an isomer of valine, with the chemical formula _CH3 ( _CH2 ) _2CH ( _NH2 ) _CO2H .

Chemical Formula 7:

The term "Norleucine (Nle)" in this invention refers to the compound shown in Chemical Formula 8 below, which has the IUPAC name 2-aminohexanoic acid, and is an amino acid with the chemical formula _CH3 ( _CH2 ) _3CH ( _NH2 ) _CO2H .

Chemical formula 8:

For example, the compounds of the present invention can be compounds in which _R1 and _R2 are each independently H or methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, cyclobutyl, n-pentyl, cyclopentyl, n-hexyl or cyclohexyl and _R3 is methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

Furthermore, the compounds of the present invention can be compounds in which _R1 and _R2 are each independently H or methyl and _R3 is methyl, ethyl or n-propyl, but are not limited thereto.

For example, in the compounds of the present invention, X can be one or two identical or different substituents from the group consisting of fluorine, chlorine, trifluoromethyl, and trifluoromethoxy. For example, the above substituents can be one or more identical or different.

For example, in the compounds of the present invention, X can be fluorine, chlorine, trifluoromethyl or trifluoromethoxy. Specifically, X can be p-fluorine, m-fluorine, p-, m-difluoro, p-chlorine, m-chlorine, p-, m-dichloro, p-trifluoromethyl or p-trifluoromethoxy, but is not limited thereto.

Specifically, the compounds described above can be as shown below, but are not limited to these.

1) 2-Amino-N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)butyramide;

2) 2-Amino-N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)pentanamide;

3) 2-Amino-N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)hexamethylenediamide;

4) 2-Amino-N-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)pentanamide;

5) N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)-2-(methylamino)butyramide;

6) N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)-2-(methylamino)pentanamide;

7) N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)-2-(methylamino)hexamamide;

8) N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)-2-(dimethylamino)pentanamide;

9) 2-Amino-N-((3',4'-dichloro-[1,1'-biphenyl]-4-yl)methyl)pentanamide;

10) 2-Amino-N-((3',4'-dichloro-[1,1'-biphenyl]-4-yl)methyl)hexamamide;

11) 2-Amino-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)pentanamide;

12) 2-Amino-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)hexamamide;

13) 2-Amino-N-((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methyl)pentanamide;

14) 2-Amino-N-((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methyl)hexamamide;

15)N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)-2-(methylamino)pentanamide;

16) 2-(methylamino)-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)pentanamide;

17) N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)-2-(dimethylamino)pentanamide;

18) 2-Amino-N-(2-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)ethyl)butyramide;

19) 2-Amino-N-(2-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)ethyl)pentanamide;

20) 2-Amino-N-(2-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)ethyl)hexamamide;

21) 2-Amino-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)butyramide;

22) 2-Amino-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)pentanamide;

23) 2-Amino-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)hexamamide;

24) 2-Amino-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)butyramide;

25) 2-Amino-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)pentanamide;

26) 2-Amino-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)hexamamide;

27) 2-Amino-N-(2-(3',4'-difluoro-[1,1'-biphenyl]-4-yl)ethyl)pentanamide;

28)N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)butyramide;

29)N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)pentanamide;

30)N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)hexamamide;

31) 2-(methylamino)-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)butyramide;

32)2-(methylamino)-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)pentanamide;

33) 2-(methylamino)-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)hexamamide;

34) 2-(methylamino)-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)butyramide;

35) 2-(methylamino)-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)pentanamide;

36) 2-(methylamino)-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)hexamamide; or

37)N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(dimethylamino)pentanamide;

The compounds of the present invention can exist in the form of pharmaceutically acceptable salts. As salts, acid salts formed from pharmaceutically acceptable free acids are useful. The term "pharmaceutically acceptable salt" in this invention refers to any organic or inorganic addition salt of the above-mentioned compounds in which the side effects caused by these salts do not diminish the beneficial efficacy of the compounds represented by Formula 1 at concentrations that are relatively non-toxic and harmless to patients.

Acid addition salts are prepared by conventional methods, such as dissolving the compound in an excess of an aqueous acid solution and precipitating the salt with a water-miscible organic solvent such as methanol, ethanol, acetone, or acetonitrile. An equimolar amount of the compound and the acid or alcohol (e.g., ethylene glycol monomethyl ether) in water can be heated, followed by evaporation and drying or filtration of the precipitated salt.

At this point, both organic and inorganic acids can be used as free acids. For example, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, and tartaric acid can be used. Other organic acids that can be used include methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, and hydroiodic acid, but are not limited to these.

Furthermore, pharmaceutically acceptable metal salts can be prepared using alkalis. For example, alkali metal salts or alkaline earth metal salts are obtained by dissolving the compound in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering out the undissolved salt, and then evaporating and drying the filtrate. In this case, the preparation of sodium, potassium, or calcium salts as metal salts is particularly suitable for pharmaceutical use, but not limited to these. Corresponding silver salts can also be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

Unless otherwise stated, pharmaceutically acceptable salts of the compounds of the present invention include salts of acidic or basic groups that may be present in the compounds of Formula 1 above. For example, pharmaceutically acceptable salts may include sodium, calcium, and potassium salts of hydroxyl groups, and other pharmaceutically acceptable salts of amino groups include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate, acetate, dihydrogen phosphate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate, and tosylate, etc., which can be prepared by methods for preparing salts according to formulas in the art.

Salts of compounds of Formula 1 of the present invention are pharmaceutically acceptable salts that exhibit pharmacological activities equivalent to those of compounds of Formula 1. For example, salts of compounds of Formula 1 that exhibit antifungal activity can be used without restriction.

In another aspect, the present invention provides a method for preparing the above-mentioned derivative compounds of aminoalkyl acids containing biphenyl groups, their stereoisomers, or pharmaceutically acceptable salts thereof, comprising a first step of reacting an aminoalkyl acid derivative compound of formula 2 protected by a tert-butyloxycarbonyl group with a biphenyl derivative compound of formula 3 containing a primary amino group to form a peptide bond; and a second step of reacting the compound obtained in the first step with an acid to remove the tert-butyloxycarbonyl group.

Chemical Formula 1:

Chemical formula 2:

Chemical formula 3:

In the above chemical formula 1,

n is 0, 1, 2, 3, 4, or 5.

_R1 , _R2 , and _R3 are each independently the same or different, and are each independently selected from hydrogen, _C1-7 alkyl, hydroxyl, halogen, halogenated _C1-7 alkyl, _C1-7 alkoxy, and halogenated _C1-7 alkoxy.

X is selected from m identical or different substituents (m is an integer from 1 to 5) selected from the group consisting of halogen groups, halogenated _C1-7 alkyl groups and halogenated _C1-7 alkoxy groups.

In the preparation method of the present invention, the amino acid derivative shown in Formula 4 can be reacted with di-tert-butyl dicarbonate (also known as tert-butyl carbonyl anhydride) to prepare the aminoalkyl acid derivative compound protected by the tert-butyl carbonyl group shown in Formula 2:

Chemical formula 4:

In the above chemical formula,

_R1 ', _R2 and _R3 are each independently the same or different, and are each independently selected from hydrogen, _C1-7 alkyl, hydroxyl, halogen, halogenated _C1-7 alkyl, _C1-7 alkoxy and halogenated _C1-7 alkoxy.

At this point, if _R2 of the final compound is an alkyl group, an alkylation step involving reaction with a haloalkane in the presence of a base can be carried out after the above reaction. For example, the above alkylation can be carried out, but is not limited to, by dissolving the compound shown in Formula 4 and a haloalkane compound such as iodinated alkanes in an organic solvent such as tetrahydrofuran, adding sodium hydride as a base dropwise at a low temperature such as 0°C, and allowing the reactants to react at a temperature of 15°C to 30°C for 12 to 48 hours, and can be carried out without limitation by utilizing or modifying alkylation reactions of amines known in the art.

On the other hand, in the preparation method of the present invention, a _C0-2 alkylamine derivative with a terminal substituted halophenyl group (as shown in Formula 5) can be reacted with di-tert-butyl dicarbonate to introduce a tert-butyloxycarbonyl protecting group into the amine group, followed by reaction with a phenylboronic acid derivative (as shown in Formula 6), and then reaction with an acid to remove the tert-butyloxycarbonyl protecting group, thereby preparing a biphenyl derivative compound containing a primary amine group.

Chemical formula 5:

Chemical Formula 6:

In the above chemical formula,

X’ is a halogen.

X is selected from m identical or different substituents (m is an integer from 1 to 5) selected from the group consisting of halogen groups, halogenated _C1-7 alkyl groups and halogenated _C1-7 alkoxy groups.

In this case, the reaction with the aforementioned phenylboronic acid derivative can be achieved through a cross-linking reaction based on a metal catalyst in the presence of a base. For example, the reaction can be carried out in the presence of a base using a metal catalyst such as palladium or nickel. The aforementioned metal catalyst can be a catalyst formed by phosphine ligands bound to a _metal . For example, the reaction can be a Suzuki-Miyaura cross-coupling reaction carried out by Pd( _PPh3 ) ₄ in the presence of _Na2CO3 , but is not limited to this.

For example, in the preparation method of the present invention, the first step can be achieved by an anhydrous coupling reaction in an organic solvent in the presence of N-methylmorpholine (NMM) and isobutyl chloroformate (IBCF). Tetrahydrofuran can be used as the organic solvent, but is not limited thereto.

For example, in the preparation method of the present invention, the second step for removing the tert-butyloxycarbonyl protecting group can be carried out by reacting with hydrochloric acid, but is not limited thereto.

Furthermore, in the preparation method of the present invention, when both _R1 and _R2 of the final prepared compound are alkyl, a third step of alkylating the amine to form a secondary amine may be included after the second step. The amination can be carried out by reacting formaldehyde with hydrogen in the presence of Pd/C as a reducing agent while supplying hydrogen. For example, the reaction can be carried out at a temperature of 15°C to 30°C for 6 to 24 hours, but is not limited thereto, and can be carried out without limitation by utilizing or modifying alkylation reactions of amines known in the art.

In another aspect, the present invention provides antifungal compositions comprising, as an active ingredient, a derivative compound of an aminoalkyl acid incorporating a biphenyl group, its stereoisomer, or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention relates to pharmaceutical compositions for treating or preventing fungal infections, comprising, as an active ingredient, a derivative compound of an aminoalkyl acid incorporating a biphenyl group, its stereoisomer, or a pharmaceutically acceptable salt thereof.

For example, the derivative compounds of novel aminoalkyl acids, stereoisomers thereof, or pharmaceutically acceptable salts thereof, which incorporate biphenyls according to the present invention, can exert antifungal activity against opportunistic fungi and are therefore used as antifungal compositions for the prevention or treatment of fungal infections.

In this invention, the term "prevention" refers to all actions that inhibit or delay the occurrence, spread, and recurrence of the target disease by administering the above-described pharmaceutical composition, and "treatment" refers to actions that improve or beneficially alter the symptoms of the target disease by administering the above-described pharmaceutical composition.

For example, non-limiting examples of fungal infections that can be prevented or treated with the pharmaceutical compositions of the present invention may include infections caused by Cryptococcus neoformans, Candida albicans, Candida auris, Candida glabrata, and Aspergillus fumigatus. Specifically, the aforementioned fungal infections may include meningitis caused by Cryptococcus, but are not limited thereto.

Examples of suitable carriers, excipients, or diluents that may be included in such compositions include lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil, etc. Furthermore, the compositions of the present invention may also contain fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc.

Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc. These solid dosage forms are formulated by mixing at least one excipient with the aforementioned composition, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. Mixing and formulation are also involved. Furthermore, in addition to simple excipients, lubricants such as magnesium stearate and talc can also be used.

Examples of liquid formulations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, they may also include a variety of excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives.

Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, can include Wiptsol, polyethylene glycol, Tween 61, cocoa butter, laurinum, and glycerin gelatin. On the other hand, injectable preparations may include existing additives such as solvents, isotonic agents, suspending agents, emulsifiers, stabilizers, and preservatives.

The compositions of the present invention are administered in pharmaceutically effective amounts. The term "pharmaceutically effective amount" in this invention refers to an amount sufficient to treat a disease with a reasonable benefit/risk ratio suitable for medical treatment without causing side effects. Effective dose levels depend on factors such as the patient's health condition, disease type, severity, drug activity, drug sensitivity, method of administration, time of administration, route of administration and excretion rate, treatment duration, concurrent or combined medications, and other factors well-known in the medical field. The compositions of the present invention can be administered as a single therapeutic agent or in combination with other therapeutic agents, can be administered sequentially or simultaneously with existing therapeutic agents, and can be administered once or multiple times. Considering all the above factors, it is important to administer the dosage with minimal side effects to achieve maximum effect, which is readily determined by those skilled in the art.

For example, the dosage described above may be increased or decreased depending on the route of administration, disease severity, gender, weight, age, etc. Therefore, the dosage described above does not limit the scope of the present invention in any way.

Furthermore, the present invention provides a treatment method for fungal infections that includes the step of administering the above-described pharmaceutical composition to an individual who requires it.

In this invention, the term "individual" refers to all animals, including humans, monkeys, cattle, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits, or guinea pigs, that have or may be susceptible to fungal infections. These diseases can be effectively prevented or treated by administering the pharmaceutical composition of this invention to the individual. Furthermore, the pharmaceutical composition of this invention exerts a therapeutic effect on diseases induced by fungal infections through its antifungal activity; therefore, a synergistic effect is produced when it is administered in combination with existing therapeutic agents.

The term "administration" in this invention refers to the delivery of a predetermined substance to a patient by any suitable method. The routes of administration for the compositions of this invention can be any general route capable of reaching the target tissue. Intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, local, intranasal, intrapulmonary, and rectal administration are possible, but not limited to these. Furthermore, the pharmaceutical compositions of this invention can be administered via any device capable of moving the active substance to target cells. Preferred administration methods and formulations include intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, and intravenous infusions. Injectable formulations can be prepared using aqueous solvents such as physiological saline and Ringer's solution, and non-aqueous solvents such as vegetable oils, higher fatty acid esters (e.g., ethyl oleate), and alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, glycerol, etc.). They may also include pharmaceutical carriers such as stabilizers to prevent deterioration (e.g., ascorbic acid, sodium bisulfite, sodium metabisulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers to adjust pH, and preservatives to inhibit microbial growth (e.g., phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

In this invention, the term "therapeutic effective amount" used in conjunction with the active ingredient refers to the amount of a biphenyl-introduced aminoalkyl acid derivative compound, its stereoisomer, or a pharmaceutically acceptable salt thereof that is effective in treating or preventing the target disease.

The effects of the invention

According to various embodiments of the present invention, therapeutic and preventive agents for a variety of fungal infections are provided. These agents overcome the shortcomings of existing antifungal drugs by using compounds with a basic skeleton containing biphenyl aminoalkyl acids such as α-aminobutyric acid, valine, or leucine as a basic skeleton. Specifically, the side effects of existing drugs can be alleviated or eliminated by improving the stability and efficacy of the drug, thereby improving the therapeutic effect. Furthermore, the compounds of the present invention can be used in the preparation of antimicrobial compositions against Gram-positive, Gram-negative, and MRSA-resistant bacteria. Additionally, the compounds of the present invention can be used in the development of anti-inflammatory therapeutic agents.

Attached Figure Description

Figure 1 shows a comparison of the antifungal activity of compound 74 of the present invention with that of a commercially available comparative drug.

Figure 2 shows a comparison of the antifungal activity of compound 74 of the present invention with that of a commercially available comparative drug.

Figure 3 shows a comparison of the biofilm removal effects of compound 74 of the present invention and a commercially available comparative drug.

Detailed Implementation

The present invention will now be described in more detail through the following preparation examples and embodiments. However, the following preparation examples and embodiments are merely illustrative of the present invention, and the scope of the present invention is not limited thereto.

First, the reactions used in the synthesis of the compounds of this invention are summarized below.

Reaction a: Introduction of the tert-butyloxycarbonyl protecting group (Boc protecting group)

Leucine (1.0 equivalent), Boc anhydride (1.5 equivalent), and sodium bicarbonate (1.5 equivalent) were dissolved in a 1:1 mixture of distilled water and methanol and reacted at room temperature for 36–48 hours. After concentrating the mixture under vacuum, the pH of the aqueous layer was adjusted to 2 with 1.0 M hydrochloric acid. Then, the water in the organic layer obtained from ethyl acetate extraction was removed using sodium sulfate, and the solvent was evaporated under vacuum to give the title compound.

Reaction b: Methylation of amino groups

The compound obtained from reaction a above (1.0 equivalent) and iodomethane (10 equivalent) were dissolved in tetrahydrofuran solvent, and sodium hydride (10 equivalent) was slowly added dropwise at 0°C. The reaction mixture was allowed to react at room temperature for 24 hours. After the reaction was complete, the mixture was diluted with ether solvent and distilled water was added. The pH of the aqueous layer was adjusted to 2 with 20% citric acid solution. Then, the water in the organic layer obtained by extraction with ethyl acetate was removed using sodium sulfate, and the solvent was evaporated under vacuum. The resulting residue was separated and purified by silica gel chromatography to obtain the title compound.

Reaction c: Introducing a tert-butyloxycarbonyl protecting group into the primary amine group.

4-Bromophenethylamine (1.0 equivalent) was dissolved in dimethyl chloride solvent, followed by the addition of potassium carbonate (1.5 equivalent) and Boc anhydride (1.05 equivalent), and the reaction was carried out at room temperature for approximately 12–18 hours. The reaction mixture was diluted with dimethyl chloride and washed twice. The organic layer was dried with sodium sulfate and concentrated under vacuum. The residue was washed with hexane and then evaporated under vacuum to give the title compound.

Reaction d: Synthesis of benzidine hydrochloride derivatives

The compound obtained from reaction c above, tert-butyl (4-bromobenzyl)carbamate or tert-butyl (4-bromophenyl)carbamate (1.0 equivalent), phenylboronic acid (1.5 equivalent), sodium carbonate (5.0 equivalent), and tetrakis(triphenylphosphine)palladium (0.04 equivalent) were dissolved in a mixture of degassing toluene and distilled water in a ratio of 2:1 to 2.5:1, and refluxed at 140°C for approximately 12–18 hours. After the reaction, the catalyst was removed by filtration through diatomaceous earth, and the solvent in the filtered organic layer was evaporated under vacuum. The resulting residue was separated and purified by silica gel chromatography. The purified product was dissolved in ethyl acetate, and 4.0 M hydrochloric acid (6.0–10.0 equivalent) was added while stirring at room temperature. The resulting white solid in salt form was washed with ethyl acetate and then completely dried under vacuum to give the title compound.

Reaction e: Mixed anhydride coupling (MAC) reaction

Add the compound synthesized according to reaction a or reaction b (1.0 equivalent) and N-methylmorpholine (NMM, 2.5-2.8 equivalents) to distilled tetrahydrofuran solvent. After stirring for 15 minutes, add isobutyl chloroformate (IBCF, 1.3 equivalents), stir for another 15 minutes, and then add the compound obtained from reaction d (1.05 equivalents). Allow the reaction mixture to react at room temperature for about 3-5 hours. Filter the mixture and evaporate the solvent under vacuum. Separate and purify the resulting residue by silica gel chromatography to obtain the title compound.

Reaction f: Removal of the tert-butyloxycarbonyl protecting group

The compound derivative (1.0 equivalent) obtained from reaction e above was dissolved in ethyl acetate, and then stirred at room temperature with 4.0 M hydrochloric acid (6.0-10.0 equivalent). After washing the resulting white solid in salt form with ethyl acetate, it was completely dried under vacuum to give the title compound.

Reaction formula g: Dimethylation of amino groups

The compound obtained from reaction f above (1.0 equivalent) was dissolved in methanol, followed by the addition of triethylamine (6.0 equivalent), then formaldehyde (37% by weight solution, 1.0–2.5 equivalent) and 10% palladium catalyst (0.1–0.5 equivalent). The reaction mixture was allowed to react at room temperature for 18 hours. After the reaction, the catalyst was removed by filtration with diatomaceous earth, and the filtered organic layer was evaporated under vacuum to obtain a white solid. The resulting product was recrystallized from methanol and diethyl ether to give the title compound.

Examples of preparation for synthesizing the compounds of the present invention are shown below.

Preparation Example

Preparation Example 1: Preparation of (R)/(S)-2-((tert-butoxycarbonyl)amino)butyric acid (4)

Using reaction formula a, compound 1 (2-aminobutyric acid, 5.00 g, 48.5 mmol), Boc anhydride (19.9 ml, 72.7 mmol) and _NaHCO3 (6.11 g, 72.7 mmol) were reacted to synthesize compound 4, (R)/(S)-2-((tert-butoxycarbonyl)amino)butyric acid (8.25 g, 83%), which is a white powder.

_Rf = 0.00 (DCM 9.5: 0.5g methanol and a few drops of acetic acid);

¹ H NMR (DMSO-d ₆ , 300MHz) 12.40 (C(O)OH), 7.02 (d, J=7.9Hz, Boc-NH), 3.69-3.82 (m, Chiral-H), 1.48-1.72 (m, CH ₂ CH ₃ ), 1.38 (s, Boc), 0.87 (t, J=7.3Hz, CH ₂ CH ₃ ).

Preparation Example 2: Preparation of (R)/(S)-2-((tert-butoxycarbonyl)amino)valeric acid (5)

Using reaction formula a, compound 2 (2-aminovaleric acid, 10.00 g, 25.6 mmol), Boc anhydride (35.1 ml, 128.0 mmol), and _NaHCO3 (10.8 g, 128.0 mmol) were reacted to synthesize a white powder compound 5, (R)/(S)-2-((tert-butoxycarbonyl)amino)valeric acid (13.40 g, 83%).

_Rf = 0.85 (DCM 3: Methanol 17);

¹ H NMR (DMSO-d ₆ , 400MHz) 12.40 (C(O)OH), 7.03 (d, J=8.0Hz, Boc-NH), 3.75-3.89 (m, Chiral-H), 1.50-1.65 (m, CH ₂ CH ₂ CH ₃ ), 1.20-1.38 (m, CH ₂ CH ₂ CH ₃ , Boc), 0.85 (t, J=7.4Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 3: Preparation of (R)/(S)-2-((tert-butoxycarbonyl)amino)hexanoic acid (6)

Using reaction formula a, compound 3 (2-aminohexanoic acid, 5.00 g, 38.1 mmol), Boc anhydride (15.7 ml, 57.2 mmol) and _NaHCO3 (4.80 g, 57.2 mmol) were reacted to synthesize a white powder compound 6, (R)/(S)-2-((tert-butoxycarbonyl)amino)hexanoic acid (7.14 g, 81%).

_Rf = 0.40 (DCM 9: Methanol 1);

¹ H NMR (CDCl ₃ , 400MHz) 10.26 (C(O)OH), 5.00 (d, J=7.6Hz, Boc-NH), 4.32-4.33 (m, Chiral-H), 1.63-1.87 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.47 (s, Boc), 1.31-1.38 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.93 (t, J=7.0Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 4: Preparation of (R)/(S)-2-((tert-butoxycarbonyl)(methyl)amino)butyric acid (7)

Using reaction b, compound 4 (3.00 g, 14.8 mmol), _CH3I (9.2 ml, 147.6 mmol), and NaH (3.54 g, 147.6 mmol) were reacted to synthesize a yellow oily compound 7, (R)/(S)-2-((tert-butoxycarbonyl)(methyl)amino)butyric acid (2.84 g, 88%).

_Rf = 0.45 (DCM 9: Methanol 1 and a few drops of acetic acid);

¹ H NMR (DMSO-d ₆ , 300 MHz) 12.7 (C(O)OH), 4.14-4.43 (m, Chiral-H), 2.71 (s, NCH ₃ ), 1.50-1.73 (m, CH ₂ CH ₃ , Boc), 0.79-0.87 (m, CH ₂ CH ₃ ).

Preparation Example 5: Preparation of (R)/(S)-2-((tert-butoxycarbonyl)(methyl)amino)valeric acid (8)

Using reaction b, compound 5 (1.50 g, 6.90 mmol), _CH3I (4.3 ml, 69.0 mmol), and NaH (1.66 g, 69.0 mmol) were reacted to synthesize a yellow oily compound 8, (R)/(S)-2-((tert-butoxycarbonyl)(methyl)amino)valeric acid (1.34 g, 83%).

_Rf = 0.45 (DCM 9: Methanol 1 and a few drops of acetic acid);

¹ H NMR (DMSO-d ₆ , 300MHz) 12.7 (C(O)OH), 4.54-4.28 (m, Chiral-H), 2.70 (s, NCH ₃ ), 1.79-1.64 (m, CH ₂ CH ₂ CH ₃ ), 1.41-1.37 (m, CH ₂ CH ₂ CH ₃ , Boc), 1.37-1.29 (m, CH ₂ CH ₂ CH ₃ ).

Preparation Example 6: Preparation of (R)/(S)-2-((tert-butoxycarbonyl)(methyl)amino)hexanoic acid (9)

Using reaction b, compound 6 (3.00 g, 13.0 mmol), _CH3I (8.1 ml, 129.7 mmol), and NaH (5.19 g, 129.7 mmol) were reacted to synthesize a yellow oily compound 9, (R)/(S)-2-((tert-butoxycarbonyl)(methyl)amino)hexanoic acid (3.18 g, 100%).

_Rf = 0.38 (DCM 9: Methanol 1);

¹ H NMR (CDCl ₃ , 400MHz) 12.6 (C(O)OH), 4.25-4.52 (m, Chiral-H), 2.70 (s, NCH ₃ ), 1.66-1.79 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.18-1.40 (m, CH ₂ CH ₂ CH ₂ CH ₃ , Boc), 0.86-0.89 (m, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 7: Preparation of (4-bromophenylethyl) tert-butyl carbamate (12)

Using reaction c, 4-bromophenylethylamine (3.9 ml, 25.1 mmol), _K₂CO₃ (5.21 g, 37.7 mmol) _, and Boc anhydride (7.2 ml, 26.4 mmol) were reacted to synthesize a white powder compound 12, (4-bromophenylethyl)carbamate tert-butyl ester (6.23 g, 83%).

_Rf = 0.36 (EtOAc 1: n-hexane)5;

¹ H NMR (DMSO-d ₆ , 400MHz) 7.46 (d, J=8.6Hz, ArH), 7.15 (d, J=8.2Hz, ArH), 6.87 (s, NH), 3.09-3.14 (m, NHCH ₂ CH ₂ ), 2.64-2.67 (m, NHCH ₂ CH ₂ ), 1.35 (s, Boc).

Preparation Example 8: Preparation of 3',4'-dichloro-[1,1'-biphenyl]-4-amine hydrochloride (13)

Using reaction formula d, compound 10 (tert-butyl 4-bromophenylcarbamate, 4.00 g, 14.7 mmol), 3,4-dichlorophenylboronic acid (3.37 g, 17.6 mmol), tetrakis(triphenylphosphine)palladium (0.68 g, 0.59 mmol), and _Na₂CO₃ (7.80 _g , 73.5 mmol) were reacted to obtain the compound. Then, the Boc group was removed using 4.0 M HCl (7.9 ml, 31.5 mmol in dioxane) to synthesize compound 13,3',4'-dichloro-[1,1'-biphenyl]-4-amine hydrochloride (1.27 g, 34%), which was a white powder.

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400 MHz) 9.94 (s, NH ₃ ), 7.95 (d, J = 2.0 Hz, ArH), 7.40-7.80 (m, ArH), 7.39 (d, J = 8.5 Hz, ArH).

Preparation Example 9: Preparation of 4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-amine hydrochloride (14)

Using reaction formula d, compound 10 (3.99 g, 14.7 mmol), 4-(trifluoromethoxy)phenylboronic acid (7.77 _g , 22.0 mmol), tetra(triphenylphosphine)palladium (0.68 g, 0.59 mmol) and _Na₂CO₃ (7.77 g, 73.3 mmol) were reacted to obtain the compound. Then, the Boc group was removed by 4.0 M HCl (12.8 ml, 51.1 mmol in dioxane) to synthesize compound 14, 4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-amine hydrochloride (1.99 g, 48%), which was a white powder.

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 9.45 (br s, NH ₃ ), 7.77 (d, J=8.7Hz, ArH), 7.71 (d, J=8.4Hz, ArH), 7.45 (d, J=8.4Hz, ArH), 7.28 (d, J=8.2Hz, ArH).

Preparation Example 10: Preparation of 2-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)methyl-1-amine hydrochloride (15)

Using reaction formula d, compound 11 (tert-butyl 4-bromobenzylcarbamate, 6.00 g, 21.0 mmol), 3,4-dichlorophenylboronic acid (4.80 g, 25.2 mmol), tetrakis(triphenylphosphine)palladium (0.97 g, 0.84 mmol), and _Na₂CO₃ (111.1 _g , 104.8 mmol) were reacted to obtain the compound. Then, the Boc group was removed using 4.0 M HCl (3.1 ml, 12.3 mmol in indioxane) to synthesize compound 15,2-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)methyl-1-amine hydrochloride (1.08 g, 17%), which was a white powder.

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.71 (s, NH ₃ ), 7.97 (s, ArH), 7.63-7.83 (m, ArH), 4.07 (s, NH ₃ CH ₂ ).

Preparation Example 11: Preparation of (4'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)methylamine hydrochloride (16) Using reaction formula d, compound 11 (6.00 g, 21.0 mmol), 4-(trifluoromethyl)phenylboronic acid (5.97 g, 31.5 mmol), tetrakis(triphenylphosphine)palladium (0.97 g, 0.84 mmol) and _Na₂CO₃ (11.1 _g , 104.8 mmol) were reacted to obtain the compound. Then, the Boc group was removed by 4.0 M HCl (17.9 ml, 71.7 mmol in dioxane) to synthesize compound 16, (4'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)methylamine hydrochloride (1.08 g, 66%), which was a white powder.

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400 MHz) 8.49 (s, NH ₃ ), 7.93 (d, J = 8.2 Hz, ArH), 7.83 (t, J = 9.0 Hz, ArH), 7.64 (d, J = 8.2 Hz, ArH), 4.09 (s, NH ₃ CH ₂ ).

Preparation Example 12: Preparation of (4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methylamine hydrochloride (17)

Using reaction formula d, compound 11 (4.00 g, 14.0 mmol), 4-(trifluoromethoxy)phenylboronic acid (4.32 _g , 21.0 mmol), tetra(triphenylphosphine)palladium (0.65 g, 0.56 mmol) and _Na₂CO₃ (7.41 g, 69.9 mmol) were reacted to obtain the compound. Then, the Boc group was removed by 4.0 M HCl (13.9 ml, 55.6 mmol in dioxane) to synthesize compound 17, (4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methylamine hydrochloride (2.73 g, 65%), which was a white powder.

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.33 (s, NH ₃ ), 7.81-7.83 (m, ArH), 7.75 (d, J=8.2Hz, ArH), 7.59 (d, J=8.2Hz, ArH), 7.48 (d, J=8.3Hz, ArH), 4.08 (s, NH ₃ CH ₂ ).

Preparation Example 13: Preparation of 2-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)ethyl-1-amine hydrochloride (18)

Using reaction formula d, compound 12 ((4-bromophenylethyl)carbamate tert-butyl ester, 1.00 g, 3.33 mmol), 3,4-dichlorophenylboronic acid (0.76 g, 4.00 mmol), tetrakis(triphenylphosphine)palladium (0.15 g, 0.15 mmol), and _Na₂CO₃ ( _1.77 g, 16.7 mmol) were reacted to obtain the compound. Then, the Boc group was removed using 4.0 M HCl (2.50 ml, 10.0 mmol in indioxane) to synthesize compound 18,2-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)ethyl-1-amine hydrochloride (2.73 g, 65%).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.33 (s, NH ₃ ), 7.93 (d, J = 1.9 Hz, ArH), 7.66-7.72 (m, ArH), 7.39 (d, J = 8.2 Hz, ArH), 2.98-3.07 (m, NH ₃ CH ₂ CH ₂ ).

Preparation Example 14: Preparation of 2-(4'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)ethyl- ₁ -amine hydrochloride (19) Using reaction formula d, compound 12 (0.50 g, 1.67 mmol), 4-(trifluoromethyl)phenylboronic acid (0.38 g, 2.00 mmol), tetra(triphenylphosphine)palladium (0.08 g, 0.07 mmol) and _Na₂CO₃ (0.88 g, 8.33 mmol) were reacted to obtain the compound. Then, the Boc group was removed by 4.0 M HCl (1.25 ml, 5.00 mmol in dioxane) to synthesize compound 19, 2-(4'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)ethyl-1-amine hydrochloride (0.28 g, 56%).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400 MHz) 8.37 (s, NH ₃ ), 7.71-7.91 (m, ArH), 7.44 (d, J = 8.1 Hz, ArH), 3.01-3.11 (m, NH ₃ CH ₂ CH ₂ ).

Preparation Example 15: Preparation of 2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl-1-amine hydrochloride (20) Using reaction formula d, compound 12 (1.50 g, 5.00 mmol), 4-(trifluoromethoxy)phenylboronic acid (1.23 g, 6.00 mmol), tetrakis(triphenylphosphine)palladium (0.23 g, 0.20 mmol) and _Na₂CO₃ ( _2.65 g, 25.0 mmol) were reacted to obtain the compound. Then, the Boc group was removed by 4.0 M HCl (3.75 ml, 15.0 mmol in dioxane) to synthesize compound 20, 2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl-1-amine hydrochloride (0.88 g, 55%).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.31 (s, NH ₃ ), 7.79 (d, J=8.7Hz, ArH), 7.66 (d, J=8.1Hz, ArH), 7.45 (d, J=8.2Hz, ArH), 7.40 (d, J=8.1Hz, ArH), 2.97-3.10 (m, NH ₃ CH ₂ CH ₂ ).

Preparation Example 16: Preparation of 2-(3',4'-difluoro-[1,1'-biphenyl]-4-yl)ethyl-1-amine hydrochloride (21)

Using reaction formula _d , compound 12 (1.00 g, 3.33 mmol), 3,4-dichlorophenylboronic acid (0.76 g, 4.00 mmol), tetrakis(triphenylphosphine)palladium (0.15 g, 0.15 mmol) and _Na₂CO₃ (1.77 g, 16.7 mmol) were reacted to obtain the compound. Then, the Boc group was removed by 4.0 M HCl (2.50 ml, 10.0 mmol in dioxane) to synthesize compound 21,2-(3',4'-difluoro-[1,1'-biphenyl]-4-yl)ethyl-1-amine hydrochloride (2.73 g, 65%).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 7.95 (s, NH ₃ ), 7.74-7.79 (m, ArH), 7.67 (d, J=8.1Hz, ArH), 7.48-7.54 (m, ArH), 7.37 (d, J=8.1Hz, ArH), 2.90-3.09 (m, NH ₃ CH ₂ CH ₂ ).

Preparation Example 17: Preparation of (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxobut-2-yl)tert-butyl carbamate (22)

Using reaction e, compound 4 (0.63 g, 3.12 mmol), NMM (0.96 ml, 8.74 mmol), IBCF (0.53 ml, 4.06 mmol) and compound 13 (0.90 g, 3.28 mmol) were reacted to synthesize a pale yellow powder, compound 22, (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxobut-2-yl)tert-butyl carbamate (1.09 g, 82%).

_Rf = 0.33 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 8.55 (s, C(O)NH), 7.58 (d, J=7.3Hz, ArH), 7.44-7.47 (m, ArH), 7.34 (dd, J=1.8Hz, 8.3Hz, ArH), 5.12 (s, Boc-NH), 4.18 (s, Chiral-H), 1.67-2.05 (m, CH ₂ CH ₃ ), 1.47 (s, Boc), 1.03 (t, J=7.4Hz, CH ₂ CH ₃ ).

Preparation Example 18: Preparation of (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxopent-2-yl)carbamate tert-butyl ester (23)

Using reaction e, compound 5 (0.30 g, 1.52 mmol), NMM (0.42 ml, 3.80 mmol), IBCF (0.26 ml, 1.98 mmol) and compound 13 (0.44 g, 1.60 mmol) were reacted to synthesize a white powder compound 23, (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxopent-2-yl)carbamate tert-butyl ester (0.61 g, 92%).

_Rf = 0.37 (EtOAc 1: n-hexane)3;

* ¹ H NMR (CDCl ₃ , 400MHz) 8.53 (s, C(O)NH), 7.62 (d, J=8.6Hz, ArH), 7.48-7.50 (m, ArH), 7.38 (dd, J=2.0Hz, 8.3Hz, ArH), 5.08 (s, Boc-NH), 4.24 (s, Chiral-H), 1.63-1.99 (m, CH ₂ CH ₂ CH ₃ ), 1.47-1.50 (m, Boc, CH ₂ CH ₂ CH ₃ ), 0.99 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 19: Preparation of (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxohex-2-yl)tert-butyl carbamate (24)

Using reaction e, compound 6 (0.80 g, 3.46 mmol), NMM (0.95 ml, 8.69 mmol), IBCF (0.58 ml, 4.50 mmol) and compound 13 (1.00 g, 3.64 mmol) were reacted to synthesize a white powder compound 24, (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxohex-2-yl)carbamate tert-butyl ester (1.11 g, 71%).

_Rf = 0.50 (EtOAc 1: n-hexane)3;

¹ H NMR (DMSO-d ₆ , 400MHz) 10.10 (s, NH), 7.91 (d, J=1.9Hz, ArH), 7.64-7.74 (m, ArH), 7.04 (d, J=7.8Hz, NH), 4.02-4.07 (m, NHCHCH ₂ ), 1.57-1.64 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.39 (s, Boc), 1.26-1.32 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.86 (t, J=6.8Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 20: Preparation of (R)/(S)-(1-oxo-1-((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)amino)pent-2-yl)carbamate tert-butyl ester (25)

Using reaction e, compound 5 (0.43 g, 1.97 mmol), NMM (0.61 ml, 5.52 mmol), IBCF (0.33 ml, 2.56 mmol) and compound 14 (0.60 g, 2.07 mmol) were reacted to synthesize a white powder, compound 25, (R)/(S)-(1-oxo-1-((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)amino)pent-2-yl)carbamate tert-butyl ester (0.66 g, 73%).

_Rf = 0.30 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 8.62 (s, C(O)NH), 7.61 (d, J=7.3Hz, ArH), 7.54 (d, J=7.6Hz, ArH), 7.49 (d, J=7.9Hz , ArH), 7.26-7.29 (m, ArH), 5.19 (d, J=7.4Hz, Boc-NH), 4.29 (s, Chiral-H), 1.65-1.98 (m, CH ₂ CH ₂ CH ₃ ), 1.43-1.56 (m, Boc, CH ₂ CH ₂ CH ₃ ), 0.99 (t, J = 7.1 Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 21: Preparation of (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxobut-2-yl)(methyl)carbamate tert-butyl ester (26)

Using reaction e, compound 7 (0.68 g, 3.12 mmol), NMM (0.96 ml, 8.74 mmol), IBCF (0.53 ml, 4.06 mmol) and compound 13 (0.90 g, 3.28 mmol) were reacted to synthesize an oily compound 26, (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxobut-2-yl)(methyl)carbamate tert-butyl ester (0.74 g, 54%).

_Rf = 0.50 (EtOAc 1: n-hexane)3;

* ¹ H NMR (CDCl ₃ , 400MHz) 8.50 (s, C(O)NH), 7.58-7.63 (m, ArH), 7.46-7.50 (m, ArH), 7.38 (dd, J=1.8Hz, 8.2Hz, ArH), 4.57 (s, Chiral-H), 2.83 (s, NCH ₃ ), 1.71-2.04 (m, CH ₂ CH ₃ ), 1.51 (d, J=6.8Hz, Boc), 0.97 (t, J=7.3Hz, CH ₂ CH ₃ ).

Preparation Example 22: Preparation of (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxopent-2-yl)(methyl)carbamate tert-butyl ester (27)

Using reaction e, compound 8 (0.86 g, 3.72 mmol), NMM (1.14 ml, 10.4 mmol), IBCF (0.63 ml, 4.83 mmol) and compound 13 (1.07 g, 3.90 mmol) were reacted to synthesize a yellow powder, compound 27, (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxopent-2-yl)(methyl)carbamate tert-butyl ester (0.79 g, 47%).

_Rf = 0.48 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 8.49 (s, C(O)NH), 7.58-7.64 (m, ArH), 7.47-7.51 (m, ArH), 7.38 (d, J=8.3Hz, ArH), 4.66 (s, Chiral-H), 2.82 (s, NCH ₃ ), 1.67-2.04 (m, CH ₂ CH ₂ CH ₃ ), 1.51 (s, Boc), 1.33-1.39 (m, CH ₂ CH ₂ CH ₃ ), 0.99 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 23: Preparation of (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxohex-2-yl)(methyl)carbamate tert-butyl ester (28)

Using reaction e, compound 9(R)/(S)-2-((tert-butoxycarbonyl)(methyl)amino)hexanoic acid (0.84 g, 3.47 mmol), NMM (1.10 ml, 9.71 mmol), IBCF (0.58 ml, 4.51 mmol) and compound 13 (1.00 g, 3.64 mmol) were reacted to synthesize an oily compound 28, (R)/(S)-(1-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)amino)-1-oxohex-2-yl)(methyl)carbamate tert-butyl ester (0.86 g, 53%).

_Rf = 0.55 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 8.49 (s, C(O)NH), 7.58-7.64 (m, ArH), 7.47-7.51 (m, ArH), 7.38 (dd, J=2.1Hz, 8.4Hz, ArH), 4.64 (s, Chiral-H), 2.82 (s, NCH ₃ ), 1.67-2.01 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.52 (s, Boc), 1.24-1.44 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.93 (t, J=7.1Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 24: Preparation of (R)/(S)-2-amino-N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)butyramide hydrochloride (29)

Compound 29, (R)/(S)-2-amino-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)butyramide hydrochloride (0.87 g, 97%), was synthesized by reacting compound 22 (1.06 g, 2.50 mmol) with 4.0 M HCl (3.80 ml, 15.0 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (CDCl ₃ , 400MHz) 11.05 (s, C(O)NH), 8.38 (s, NH ₃ ), 7.93 (d, J=1.9Hz, ArH), 7.66-7.79 (m, ArH), 4.01-4.04 (m, Chiral-H), 1.86-1.91 (m, CH ₂ CH ₃ ), 0.96 (t, J=7.5Hz, CH ₂ CH ₃ ).

Preparation Example 25: Preparation of (R)/(S)-2-amino-N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)pentanamide hydrochloride (30)

Compound 30, (R)/(S)-2-amino-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)pentanamide hydrochloride (0.40 g, 81%), was synthesized by reacting compound 23 (0.58 g, 1.33 mmol) with 4.0 M HCl (2.00 ml, 7.95 mmol in dioxane) using reaction f.

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 11.05 (s, C(O)NH), 8.40 (s, NH ₃ ), 7.93 (d, J=1.5Hz, ArH), 7.66-7.79 (m, ArH), 4.06 (s, Chiral-H), 1.79-1.85 (m, CH ₂ CH ₂ CH ₃ ), 1.36-1.43 (m, CH ₂ CH ₂ CH ₃ ), 0.91 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 26: Preparation of (R)/(S)-2-amino-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)hexamethylene hydrochloride (31)

Compound 31, (R)/(S)-2-amino-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)hexamethylene hydrochloride (0.81 g, 86%), was synthesized by reacting compound 24 (1.10 g, 2.44 mmol) with 4.0 M HCl (3.66 ml, 14.6 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 10.97 (s, C(O)NH), 8.38 (s, NH ₃ ), 7.93 (d, J=2.0Hz, ArH), 7.66-7.78 (m, ArH), 4.03 (s, Chiral-H), 1.81-1.87 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.33-1.39 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.87 (t, J=6.9Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 27: Preparation of (R)/(S)-2-amino-N-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)pentanamide hydrochloride (32)

Compound 32, (R)/(S)-2-amino-N-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)pentanamide hydrochloride (0.51 g, 93%), was synthesized by reacting compound 25 (0.64 g, 1.41 mmol) with 4.0 M HCl (2.12 ml, 8.46 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 10.90 (s, C(O)NH), 8.35 (s, NH ₃ ), 7.76-7.79 (m, ArH), 7.70 (d, J=8.8Hz, ArH), 7.45 (d, J=8.2Hz, ArH), 4.03 (s, Chiral-H), 1.82 (q, J=6.9Hz, 7.9Hz, CH ₂ CH ₂ CH ₃ ), 1.34-1.47 (m, CH ₂ CH ₂ CH ₃ ), 0.92 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 28: Preparation of (R)/(S)-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)-2-(methylamino)butyramide hydrochloride (33)

Compound 33, (R)/(S)-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)-2-(methylamino)butyramide hydrochloride (0.55 g, 93%), was synthesized by reacting compound 26 (0.69 g, 1.58 mmol) with 4.0 M HCl (2.40 ml, 9.50 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 11.00 (s, C(O)NH), 9.13 (s, NH ₂ ), 7.94 (d, J=1.7Hz, ArH), 7.66-7.78 (m, ArH), 3.96 (t, J=5.5Hz, Chiral-H), 2.57 (s, NCH ₃ ), 1.87-2.05 (m, CH ₂ CH ₃ ), 0.94 (t, J=7.5Hz, CH ₂ CH ₃ ).

Preparation Example 29: Preparation of (R)/(S)-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)-2-(methylamino)pentanamide hydrochloride (34)

Compound 34, (R)/(S)-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)-2-(methylamino)pentanamide hydrochloride (0.23 g, 71%), was synthesized by reacting compound 27 (0.38 g, 0.83 mmol) with 4.0 M HCl (1.25 ml, 4.98 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 10.98 (s, C(O)NH), 9.09 (s, NH ₂ ), 7.94 (s, ArH), 7.66-7.76 (m, ArH), 3.96 (t, J=6.1Hz, Chiral-H), 2.57 (s, NCH ₃ ), 1.80-1.93 (m, CH ₂ CH ₂ CH ₃ ), 1.31-1.39 (m, CH ₂ CH ₂ CH ₃ ), 0.91 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 30: Preparation of (R)/(S)-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)-2-(methylamino)hexamethylene amide hydrochloride (35)

Compound 35, (R)/(S)-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)-2-(methylamino)hexamethylene diamide hydrochloride (0.60 g, 84%), was synthesized by reacting compound 28 (0.82 g, 1.77 mmol) with 4.0 M HCl (2.65 ml, 10.6 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 10.81 (s, C(O)NH), 9.05 (s, NH ₂ ), 7.94 (d, J=2.0Hz, ArH), 7.66-7.77 (m, ArH), 3.92 (s, Chiral-H), 2.57 (s, NCH ₃ ), 1.87-1.99 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.29-1.33 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.86 (t, J=6.8Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 31: Preparation of (R)/(S)-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)-2-(dimethylamino)pentanamide (36)

Compound 36, (R)/(S)-N-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)-2-(dimethylamino)pentanamide, was synthesized by reacting compound 30 (1.0 equivalent), triethylamine (6.0 equivalent), formaldehyde (2.0 equivalent), and palladium catalyst (0.4 equivalent) using reaction formula g.

¹ H NMR (DMSO-d ₆ , 400MHz) 10.98 (s, C(O)NH), 7.94 (s, ArH), 7.66-7.76 (m, ArH), 3.96 (t, J=6.1Hz, Chiral-H), 2.57 (s, N(CH ₃ )2), 1.80-1.93 (m, CH ₂ CH ₂ CH ₃ ), 1.31-1.39 (m, CH ₂ CH ₂ CH ₃ ), 0.91 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 32: Preparation of (R)/(S)-(1-(((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)amino)-1-oxopent-2-yl)carbamate tert-butyl ester (37)

Using reaction e, compound 5 (0.57 g, 2.64 mmol), NMM (0.73 ml, 6.60 mmol), IBCF (0.45 ml, 3.43 mmol) and compound 15 (0.80 g, 2.77 mmol) were reacted to synthesize a white powder compound 37, (R)/(S)-(1-(((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)amino)-1-oxopent-2-yl)tert-butyl carbamate (1.20 g, 100%).

_Rf = 0.04 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.64 (d, J=2.0Hz, ArH), 7.49 (dd, J=2.6Hz, 8.5Hz, ArH), 7.33-7.40 (m, ArH), 6.48 (s, C(O)NH), 4.93 (s, Boc-NH), 4.49 (d, J=4.2Hz, NHCH) ₂ ), 4.07-4.09 (m, Chiral-H), 1.36-1.43 (m, CH ₂ CH ₂ CH ₃ , Boc), 0.95 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 33: Preparation of (R)/(S)-(1-(((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)amino)-1-oxohex-2-yl)carbamate tert-butyl ester (38)

Using reaction e, compound 6 (0.38 g, 1.65 mmol), NMM (0.45 ml, 4.13 mmol), IBCF (0.28 ml, 2.15 mmol) and compound 15 (0.50 g, 1.74 mmol) were reacted to synthesize a white powder compound 38, (R)/(S)-(1-(((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)amino)-1-oxohex-2-yl)tert-butyl carbamate (0.46 g, 60%).

_Rf = 0.19 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.63 (d, J=2.0Hz, ArH), 7.49 (dd, J=4.3Hz, 8.2Hz, ArH), 7.33-7.39 (m, ArH), 6.55 (s, C(O)NH), 4.98 (d, J=3.8Hz, Boc-NH), 4.49 (d, J=5.5Hz, NHCH ₂ ), 4.07-4.11 (m, Chiral-H), 1.58-1.90 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.42 (s, Boc), 1.34 (d, J=2.2Hz, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.88-0.94 (m, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 34: Preparation of (R)/(S)-(1-oxo-1(((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)amino)pent-2-yl)carbamate tert-butyl ester (39)

Using reaction e, compound 5 (0.36 g, 1.66 mmol), NMM (0.46 ml, 4.14 mmol), IBCF (0.28 ml, 2.15 mmol), and compound 16 (0.50 g, 1.74 mmol) were reacted to synthesize a white powder compound 39, (R)/(S)-(1-oxo-1(((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)amino)pent-2-yl)carbamate tert-butyl ester (0.72 g, 96%).

_Rf = 0.17 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 300MHz) 7.62-7.70 (m, ArH), 7.44 (dd, J=8.1Hz, 44.7Hz, ArH), 6.85 (s, C(O)NH), 5.16-5.18 (m, Boc-NH), 4.47-4.49 (m, ArCH ₂ ), 4.11-4.15 (m, Chiral-H), 1.54-1.89 (m, CH ₂ CH ₂ CH ₃ ), 1.41 (s, Boc, CH ₂ CH ₂ CH ₃ ), 0.93 (t, J=7.2Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 35: Preparation of (R)/(S)-(1-oxo-1(((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)amino)hex-2-yl)tert-butyl carbamate (40)

Using reaction e, compound 6 (0.54 g, 2.32 mmol), NMM (0.71 g, 6.49 mmol), IBCF (0.39 ml, 3.01 mmol) and compound 16 (0.70 g, 2.43 mmol) were reacted to synthesize a white powder compound 40, (R)/(S)-(1-oxo-1(((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)amino)hex-2-yl)tert-butyl carbamate (0.87 g, 81%).

_Rf = 0.16 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.70 (q, J=4.6Hz, 8.6Hz, ArH), 7.57 (d, J=8.2Hz, ArH), 7.39 (d, J=8.2Hz, ArH), 6.50 (s, C(O)NH), 4.96 (s, Boc-NH), 4.53 (d, J=4.7Hz, NHCH ₂ ), 4.09-4.10 (m, Chiral-H), 1.59-1.94 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.45 (s, Boc), 1.37-1.38 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.93 (t, J=7.0Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 36: Preparation of (R)/(S)-(1-oxo-1(((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methyl)amino)pent-2-yl)carbamate tert-butyl ester (41)

Using reaction e, compound 5 (0.55 g, 2.51 mmol), NMM (0.77 ml, 7.02 mmol), IBCF (0.42 ml, 3.26 mmol) and compound 17 (0.80 g, 2.63 mmol) were reacted to synthesize a white powder compound 41, (R)/(S)-(1-oxo-1(((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methyl)amino)pent-2-yl)carbamate tert-butyl ester (1.05 g, 90%).

_Rf = 0.09 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.57-7.61 (m, ArH), 7.53 (d, J=8.2Hz, ArH), 7.37 (d, J=8.2Hz, ArH), 7.30 (d, J=8.2Hz, ArH), 6.47 (s, C(O)NH), 4.96 (s, Boc-NH), 4.52 (s, NHCH ₂ ), 4.10-4.11 (m, Chiral-H), 1.59-1.94 (m, CH ₂ CH ₂ CH ₃ ), 1.37-1.45 (m, CH ₂ CH ₂ CH ₃ , Boc), 0.97 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 37: Preparation of (R)/(S)-(1-oxo-1(((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methyl)amino)hex-2-yl)tert-butyl carbamate (42)

Using reaction e, compound 6 (0.58 g, 2.51 mmol), NMM (0.77 ml, 7.02 mmol), IBCF (0.42 ml, 3.26 mmol) and compound 17 (0.80 g, 2.63 mmol) were reacted to synthesize a white powder compound 42, (R)/(S)-(1-oxo-1(((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methyl)amino)hex-2-yl)tert-butyl carbamate (1.06 g, 88%).

_Rf = 0.18 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.58-7.61 (m, ArH), 7.53 (d, J=8.1Hz, ArH), 7.37 (d, J=8.1Hz, ArH), 7. 30 (d, J=8.5Hz, ArH), 6.52 (s, C(O)NH), 4.99 (s, Boc-NH), 4.53 (d, J=5.1Hz, NHCH ₂ ), 4.09-4.11 (m, Chiral-H), 1.62-1.94 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.45 (s, Boc), 1.36-1.37 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.92 (t, J=6.9Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 38: Preparation of (R)/(S)-(1-(((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)amino)-1-oxopent-2-yl)(methyl)carbamate tert-butyl ester (43)

Using reaction e, compound 8 (0.38 g, 1.65 mmol), NMM (0.46 ml, 4.13 mmol), IBCF (0.28 ml, 2.15 mmol) and compound 15 (0.50 g, 1.73 mmol) were reacted to synthesize an oily compound 43, (R)/(S)-(1-(((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)amino)-1-oxopent-2-yl)(methyl)carbamate tert-butyl ester (0.57 g, 73%).

_Rf = 0.18 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 300MHz) 7.30-7.68 (m, ArH), 6.27-6.64 (m, C(O)NH), 4.41-4.59 (m, NHCH ₂ , Chrial-H), 2.78 (s, NCH ₃ ), 2.04-1.63 (m, CH ₂ CH ₂ CH ₃ ), 1.44 (s, Boc), 1.32-1.26 (m, CH ₂ CH ₂ CH ₃ ), 0.96 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 39: Preparation of (R)/(S)-methyl(1-oxo-1(((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)amino)pent-2-yl)carbamate tert-butyl ester (44)

Using reaction e, compound 8 (0.38 g, 1.66 mmol), NMM (0.46 ml, 4.14 mmol), IBCF (0.28 ml, 2.15 mmol) and compound 16 (0.50 g, 1.74 mmol) were reacted to synthesize an oily compound 44, (R)/(S)-methyl(1-oxo-1(((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)amino)pent-2-yl)carbamate tert-butyl ester (0.49 g, 64%).

_Rf = 0.22 (EtOAc 1: n-hexane)3);

¹ H NMR (CDCl ₃ , 300MHz) 7.64-7.71 (m, ArH), 7.44 (dd, J=7.9Hz, 53.4Hz, ArH), 6.28-6.64 (m, C(O)NH), 4.43-4.61 (m, NHCH ₂ , Chiral-H), 2.78 (s, NCH ₃ ), 1.63-2.04 (m, CH ₂ CH ₂ CH ₃ ), 1.44 (s, Boc), 1.26-1.35 (m, CH ₂ CH ₂ CH ₃ ), 0.96 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 40: Preparation of (R)/(S)-2-amino-N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)pentanamide hydrochloride (45)

Compound 45, (R)/(S)-2-amino-N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)pentanamide hydrochloride (0.73 g, 71%), was synthesized by reacting compound 37 (1.19 g, 2.64 mmol) with 4.0 M HCl (3.95 ml, 15.8 mmol in dioxane) using reaction f.

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 300MHz) 9.04 (s, C(O)NH), 8.21 (s, NH ₃ ), 7.94 (s, ArH), 7.66-7.73 (m, ArH), 7.40 (d, J=8.1Hz, ArH), 4.39-4.41 (m, NHCH ₂ ), 3.78-3.82 (t, J=6.3Hz, Chiral-H), 1.68-1.76 (m, CH ₂ CH ₂ CH ₃ ), 1.29-1.39 (m, CH ₂ CH ₂ CH ₃ ), 0.89 (t, J=7.2Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 41: Preparation of (R)/(S)-2-amino-N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)hexamethylene hydrochloride (46)

Compound 46, (R)/(S)-2-amino-N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)hexamethylene hydrochloride (0.27 g, 85%), was synthesized by reacting compound 38 (0.46 g, 0.99 mmol) with 4.0 M HCl (1.48 ml, 1.48 mmol in dioxane) using reaction f.

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 9.17 (m, C(O)NH), 8.32 (s, NH ₃ ), 7.94 (d, J=2.0Hz, ArH), 7.66-7.73 (m, ArH), 7.41 (d, J=8.2Hz, ArH), 4.34-4.45 (m, NHCH ₂ ), 3.81 (t, J=6.1Hz, Chiral-H), 1.75-1.76 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.27-1.28 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.85 (t, J=6.6Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 42: Preparation of (R)/(S)-2-amino-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)pentanamide hydrochloride (47)

Compound 47, (R)/(S)-2-amino-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)pentanamide hydrochloride (0.60 g, 99%), was synthesized by reacting compound 39 (0.70 g, 1.55 mmol) with 4.0 M HCl (2.33 ml, 9.32 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 300MHz) 9.37 (t, J=5.7Hz, C(O)NH), 8.46 (s, NH ₃ ), 7.71-7.92 (m, ArH), 7.45-7.51 (m, ArH), 4.40-4.43 (m, NHCH ₂ ), 3.89 (s, Chiral-H), 1.74-1.82 (m, CH ₂ CH ₂ CH ₃ ), 1.16-1.42 (m, CH ₂ CH ₂ CH ₃ ), 0.89 (t, J=7.2Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 43: Preparation of (R)/(S)-2-amino-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)hexamethylene hydrochloride (48)

Compound 48, (R)/(S)-2-amino-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)hexamethylene hydrochloride (0.72 g, 98%), was synthesized by reacting compound 40 (0.85 g, 1.83 mmol) with 4.0 M HCl (2.75 ml, 11.0 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 9.09 (s, C(O)NH), 8.23 (s, NH ₃ ), 7.89 (d, J=8.2Hz, ArH), 7.82 (d, J=8.4Hz, ArH), 7.73 (d, J=8.1Hz, ArH), 7.44 (d, J=8.1Hz, ArH), 4.36-4.46 (m, NHCH ₂ ), 3.80 (t, J=6.4Hz, Chiral-H), 1.74-1.76 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.27-1.29 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.85 (t, J=6.5Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 44: Preparation of (R)/(S)-2-amino-N-((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methyl)pentanamide hydrochloride (49)

Compound 49, (R)/(S)-2-amino-N-((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methyl)pentanamide hydrochloride (0.82 g, 92%), was synthesized by reacting compound 41 (1.04 g, 2.22 mmol) with 4.0 M HCl (3.34 ml, 13.3 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 9.14 (t, J=5.7Hz, C(O)NH), 8.30 (s, NH ₃ ), 7.77-7.81 (m, ArH), 7.67 (d, J=8.2Hz, ArH), 7.46 (d, J=8.1Hz, ArH), 7.41 (d, J=8.2Hz, ArH), 4.40 (d, J=5.8Hz, NHCH ₂ ), 3.82 (t, J=6.5Hz, Chiral-H), 1.71-1.76 (m, CH ₂ CH ₂ CH ₃ ), 1.28-1.38 (m, CH ₂ CH ₂ CH ₃ ), 0.89 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 45: Preparation of (R)/(S)-2-amino-N-((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methyl)hexamethylene hydrochloride (50)

Compound 50, a white powder, was synthesized by reacting compound 42 (1.04 g, 2.17 mmol) with 4.0 M HCl (3.26 ml, 13.0 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 9.09 (t, J=5.8Hz, C(O)NH), 8.25 (s, NH ₃ ), 7.78 (d, J=8.7Hz, ArH), 7.66 (d, J=8.2Hz, ArH), 7.45 (d, J=8.3Hz, ArH), 7.41 (d, J=8.2Hz, ArH), 4.35-4.44 (m, NHCH ₂ ), 3.80 (t, J=6.4Hz, Chiral-H), 1.72-1.75 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.27-1.28 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.85 (t, J=6.5Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 46: Preparation of (R)/(S)-N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)-2-(methylamino)pentanamide hydrochloride (51)

Compound 51, (R)/(S)-N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)-2-(methylamino)pentanamide hydrochloride (0.64 g, 88%), was synthesized by reacting compound 43 (0.84 g, 1.81 mmol) with 4.0 M HCl (2.80 ml, 10.9 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 300MHz) 9.10-9.78 (m, NH ₂ ), 9.57 (t, J=5.5Hz, C(O)NH), 7.96 (s, ArH), 7.69-7.72 (m, ArH), 7.43 (d, J=10.7Hz, ArH), 4.42-4.44 (m, NHCH ₂ ), 3.85-3.90 (m, Chiral-H), 2.48 (s, NCH ₃ ), 1.76-1.90 (m, CH ₂ CH ₂ CH ₃ ), 1.26-1.38 (m, CH ₂ CH ₂ CH ₃ ), 0.90 (t, J=7.1Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 47: Preparation of (R)/(S)-2-(methylamino)-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)pentanamide hydrochloride (52)

Compound 52, (R)/(S)-2-(methylamino)-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)pentanamide hydrochloride (0.32 g, 82%), was synthesized by reacting compound 44 (0.45 g, 0.97 mmol) with 4.0 M HCl (1.45 ml, 5.81 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 300MHz) 9.03-9.81 (m, NH ₂ ), 9.52 (t, J=5.5Hz, C(O)NH), 7.72-7.92 (m, ArH), 7.46 (d, J=8.2Hz, ArH), 4.43-4.45 (m, NHCH ₂ ), 3.86 (s, Chiral-H), 2.48 (s, NCH ₃ ), 1.77-1.89 (m, CH ₂ CH ₂ CH ₃ ), 1.25-1.38 (m, CH ₂ CH ₂ CH ₃ ), 0.90 (t, J=7.2Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 48: Preparation of (R)/(S)-N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)-2-(dimethylamino)pentanamide (53)

Compound 53, (R)/(S)-N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)-2-(dimethylamino)pentanamide, was synthesized by reacting compound 45 (1.0 equivalent), triethylamine (6.0 equivalent), formaldehyde (1.05 equivalent) and palladium catalyst (0.2 equivalent) using reaction formula g.

¹ H NMR (DMSO-d ₆ , 300MHz) 9.57 (t, J=5.5Hz, C(O)NH), 7.96 (s, ArH), 7.69-7.72 (m, ArH), 7.43 (d, J=10.7Hz, ArH), 4.42-4.44 (m, NHCH ₂ ), 3.85-3.90 (m, Chiral-H), 2.48 (s, N(CH ₃ )2), 1.76-1.90 (m, CH ₂ CH ₂ CH ₃ ), 1.26-1.38 (m, CH ₂ CH ₂ CH ₃ ), 0.90 (t, J=7.1Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 49: Preparation of (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxobut-2-yl)carbamate tert-butyl ester (54)

Using reaction e, compound 4 (0.32 g, 1.57 mmol), NMM (0.43 ml, 3.93 mmol), IBCF (0.27 ml, 2.05 mmol) and compound 18 (0.50 g, 1.65 mmol) were reacted to synthesize a white powder compound 54, (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxobut-2-yl)tert-butyl carbamate (0.66 g, 93%).

_Rf = 0.05 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.65 (s, ArH), 7.47-7.50 (m, ArH), 7.39 (d, J=8.2Hz, ArH), 7.26-7.28 (m, ArH ), 6.07 (s, C(O)NH), 4.94 (s, Boc-NH), 3.93 (d, J=6.6Hz, Chiral-H), 3.50-3.94 (m, NHCH ₂ CH ₂ ), 2.86 (t, J=6.9Hz, NHCH ₂ CH ₂ ), 1.55-2.88 (m, CH ₂ CH ₃ ), 1.43 (s, Boc), 0.91 (t, J=7.4Hz, CH ₂ CH ₃ ).

Preparation Example 50: Preparation of (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxopent-2-yl)carbamate tert-butyl ester (55)

Using reaction e, compound 5 (0.50 g, 2.30 mmol), NMM (0.51 ml, 4.60 mmol), IBCF (0.39 ml, 2.99 mmol) and compound 18 (0.73 g, 2.42 mmol) were reacted to synthesize a white powder compound 55, (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxopent-2-yl)tert-butyl carbamate (0.96 g, 89%).

_Rf = 0.26 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.46-7.64 (m, ArH), 7.26-7.40 (m, ArH), 6.36 (s, C(O)NH), 5.07 (s, Boc-NH), 4.01-4.03 (m, Chiral-H), 3.47-3.61 (m, NHCH ₂ CH ₂ ), 2.85 (t, J=7.0Hz, NHCH ₂ CH ₂ ), 2.67 (s, NCH ₃ ), 1.48-1.80 (m, CH ₂ CH ₂ CH ₃ ), 1.42 (s, Boc), 1.26-1.36 (m, CH ₂ CH ₂ CH ₃ ), 0.90 (t, J=7.2Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 51: Preparation of (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxohex-2-yl)carbamate tert-butyl ester (56)

Using reaction e, compound 6 (0.36 g, 1.57 mmol), NMM (0.43 ml, 3.93 mmol), IBCF (0.27 g, 2.05 mmol) and compound 18 (0.50 g, 16.5 mmol) were reacted to synthesize a white powder compound 56, (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxohex-2-yl)tert-butyl carbamate (0.76 g, 100%).

_Rf = 0.07 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.65 (s, ArH), 7.48 (d, J=6.3Hz, ArH), 7.39 (d, J=8.0Hz, ArH), 7.26-7.28 (m, Ar H), 6.07 (s, C(O)NH), 4.91 (s, Boc-NH), 3.97 (d, J=5.6Hz, Chiral-H), 3.51-3.62 (m, NHCH ₂ CH ₂ ), 2.86 (t, J=6.1Hz, NHCH ₂ CH ₂ ), 1.51-1.81 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.42 (s, Boc), 1.27 (d, J=6.7Hz, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.87 (d, J=5.5Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 52: Preparation of (R)/(S)-(1-oxo-1-((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)but-2-yl)carbamate tert-butyl ester (57)

Using reaction e, compound 4 (0.42 g, 2.08 mmol), NMM (0.57 ml, 5.21 mmol), IBCF (0.35 ml, 2.71 mmol) and compound 19 (0.66 g, 2.19 mmol) were reacted to synthesize a white powder compound 57, (R)/(S)-(1-oxo-1-((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)but-2-yl)carbamate tert-butyl ester (0.92 g, 98%).

_Rf = 0.16 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.70 (s, ArH), 7.57 (d, J=8.0Hz, ArH), 7.29-7.33 (m, ArH), 6.15 (s, C(O)NH), 5.00 (s, Boc-NH), 3.94-3.99 (m, Chiral-H), 3.53-3.66 (m, NHCH ₂ CH ₂ ), 2.90 (t, J=7.0Hz, NHCH ₂ CH ₂ ), 1.58-1.90 (m, CH ₂ CH ₃ ), 1.45 (s, Boc), 0.93 (t, J=7.5Hz, CH ₂ CH ₃ ).

Preparation Example 53: Preparation of (R)/(S)-(1-oxo-1-((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)pent-2-yl)carbamate tert-butyl ester (58)

Using reaction e, compound 5 (0.55 g, 2.53 mmol), NMM (0.69 ml, 6.31 mmol), IBCF (0.43 ml, 3.28 mmol) and compound 19 (0.80 g, 2.65 mmol) were reacted to synthesize a white powder compound 58, (R)/(S)-(1-oxo-1-((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)pent-2-yl)carbamate tert-butyl ester (1.05 g, 89%).

_Rf = 0.20 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 300MHz) 7.67 (s, ArH), 7.41 (dd, J=8.1Hz, 63.6Hz, ArH), 6.36 (s, C(O)NH), 5.05-5.07 (m, Boc-NH), 4.00-4.05 (m, Chiral-H), 3.45-3.66 (m, NHCH ₂ CH ₂ ), 2.87 (t, J=7.1Hz, NHCH ₂ CH ₂ ), 1.50-1.83 (m, CH ₂ CH ₂ CH ₃ ), 1.42 (s, Boc), 1.28-1.39 (m, CH ₂ CH ₂ CH ₃ ), 0.90 (t, J=7.2Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 54: Preparation of (R)/(S)-(1-oxo-1-((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)carbamate tert-butyl ester (59)

Using reaction e, compound 6 (0.76 g, 3.16 mmol), NMM (0.87 ml, 7.89 mmol), IBCF (0.53 ml, 4.10 mmol) and compound 19 (1.00 g, 3.31 mmol) were reacted to synthesize a white powder compound 59, (R)/(S)-(1-oxo-1-((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)carbamate tert-butyl ester (1.34 g, 89%).

_Rf = 0.13 (EtOAc 1: n-hexane)3;

* ^1H NMR(DMSO-d ₆ , 400MHz) 7.87 (d, J=8.2Hz, ArH, C(O)NH), 7.80 (d, J=8.5Hz, ArH), 7.66 (d, J=8.1Hz, ArH), 7.35 (d, J=8.1Hz, ArH), 6.73 (d, J=8.2Hz, Boc-NH), 3.83 (q, J=5.6Hz, 8.4Hz, Chiral-H), 3.24-3.43 (m, NHCH ₂ CH ₂ ), 2.77 (t, J=7.0Hz, NHCH ₂ CH ₂ ), 1.37-1.52 (m, CH ₂ CH ₂ CH ₂ CH ₃ , Boc), 1.15-1.20 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.80 (t, J=6.8Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 55: Preparation of (R)/(S)-(1-oxo-1-((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)but-2-yl)carbamate tert-butyl ester (60)

Using reaction e, compound 4 (0.49 g, 2.40 mmol), NMM (0.66 ml, 6.00 mmol), IBCF (0.41 ml, 3.12 mmol) and compound 20 (0.80 g, 2.52 mmol) were reacted to synthesize a white powder compound 60, (R)/(S)-(1-oxo-1-((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)but-2-yl)carbamate tert-butyl ester (0.94 g, 84%).

_Rf = 0.14 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.68-7.73 (m, ArH), 7.51-7.62 (m, ArH), 7.29-7.33 (m, ArH), 6.17 (d, J=5.6Hz, C(O)NH), 5.01 (s, NH), 3.95-3.98 (m, Chiral-H), 3.51-3.67 (m, NHCH ₂ CH ₂ ), 2.87-2.92 (m, NHCH ₂ CH ₂ ), 1.56-1.92 (m, CH ₂ CH ₃ ), 1.45 (s, Boc), 0.93 (t, J=7.4Hz, CH ₂ CH ₃ ).

Preparation Example 56: Preparation of (R)/(S)-(1-oxo-1-((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)pent-2-yl)carbamate tert-butyl ester (61)

Using reaction e, compound 5 (0.33 g, 1.50 mmol), NMM (0.41 ml, 3.75 mmol), IBCF (0.25 ml, 1.95 mmol) and compound 20 (0.50 g, 1.57 mmol) were reacted to synthesize a white powder compound 61, (R)/(S)-(1-oxo-1-((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)pent-2-yl)carbamate tert-butyl ester (0.70 g, 98%).

_Rf = 0.12 (EtOAc 1: n-hexane)3);

¹ H NMR (DMSO-d ₆ , 400MHz) 7.88 (t, J=5.6Hz, C(O)NH), 7.74-7.77 (m, ArH), 7.59 (d, J=8.2Hz, ArH), 7.44 (d, J=8.1Hz, ArH), 7 .32 (d, J=8.2Hz, ArH), 6.74 (d, J=8.2Hz, Boc-NH), 4.86 (q, J=5.6Hz, 8.2Hz, Chiral-H), 3.26-3.40 (m, NHCH ₂ CH ₂ ), 2.76 (t, J=7.0Hz, NHCH ₂ CH ₂ ), 1.37-1.52 (m, CH ₂ CH ₂ CH ₃ , Boc), 1.17-1.25 (m, CH ₂ CH ₂ CH ₃ ), 0.81 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 57: Preparation of (R)/(S)-(1-oxo-1-((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)hex-2-yl)carbamate tert-butyl ester (62)

Using reaction e, compound 6 (0.50 g, 2.18 mmol), NMM (0.60 ml, 5.45 mmol), IBCF (0.37 ml, 2.83 mmol) and compound 20 (0.72 g, 2.29 mmol) were reacted to synthesize a white powder compound 62, (R)/(S)-(1-oxo-1-((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)hex-2-yl)tert-butyl carbamate (0.95 g, 88%).

_Rf = 0.13 (EtOAc 1: n-hexane)3;

[462] ¹ H NMR (CDCl ₃ , 400MHz) 7.70-7.73 (m, ArH), 7.57 (d, J = 8.1Hz, ArH), 7.32 (d, J = 8.1Hz, ArH), 6.15 (s , C(O)NH), 4.95 (s, Boc-NH), 4.00 (q, J=6.4Hz, 7.2Hz, Chiral-H), 3.53-3.65 (m, NHCH ₂ CH ₂ ), 2.90 (t, J=7.0Hz, NHCH ₂ CH ₂ ), 1.52-1.85 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.45 (s, Boc), 1.31-1.34 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.90 (t, J=6.9Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 58: Preparation of (R)/(S)-(1-((2-(3’,4’-difluoro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxopent-2-yl)carbamate tert-butyl ester (63)

Using reaction e, compound 5 (0.50 g, 2.29 mmol), NMM (0.70 ml, 6.40 mmol), IBCF (0.39 ml, 2.97 mmol) and compound 21 (0.80 g, 2.40 mmol) were reacted to synthesize a white powder compound 63, (R)/(S)-(1-((2-(3’,4’-difluoro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxopent-2-yl)tert-butyl carbamate (0.97 g, 98%).

_Rf = 0.10 (EtOAc 1: n-hexane)3);

¹ H NMR (CDCl ₃ , 400MHz) 7.46 (d, J=7.9Hz, ArH), 7.33-7.38 (m, ArH), 7.17-7.27 (m, ArH), 6.11 (s, C(O)NH), 4.92 (s, Boc-NH), 3.96-4.01 (m, Chiral-H), 3.47-3.63 (m, NHCH ₂ CH ₂ ), 2.85 (t, J=7.0Hz, NHCH ₂ CH ₂ ), 1.48-1.82 (m, CH ₂ CH ₂ CH ₃ ), 1.42 (s, Boc), 1.26-1.36 (m, CH ₂ CH ₂ CH ₃ ), 0.90 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 59: Preparation of (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxobut-2-yl)(methyl)carbamate tert-butyl ester (64)

Using reaction e, compound 7 (0.82 g, 3.78 mmol), NMM (1.04 ml, 9.44 mmol), IBCF (0.64 ml, 4.91 mmol) and compound 18 (1.20 g, 3.97 mmol) were reacted to synthesize an oily compound 64, (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxobut-2-yl)(methyl)carbamate tert-butyl ester (1.38 g, 79%).

_Rf = 0.22 (EtOAc 1: n-hexane)3);

¹ H NMR (CDCl ₃ , 400MHz) 7.66 (d, J=2.0Hz, ArH), 7.49-7.53 (m, ArH), 7.41 (dd, J=2.1Hz, 6.3Hz, ArH), 7.27-7.29 (m ArH), 6.26 (s, C(O)NH), 4.44 (s, Chiral-H), 3.55-3.64 (m, NHCH ₂ CH ₂ ), 2.87 (t, J=6.7Hz, NHCH ₂ CH ₂ ), 2.70 (s, NCH ₃ ), 1.61-1.98 (m, CH ₂ CH ₃ ), 1.45 (s, Boc), 0.88 (t, J=7.4Hz, CH ₂ CH ₃ ).

Preparation Example 60: Preparation of (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxopent-2-yl)(methyl)carbamate tert-butyl ester (65)

Using reaction e, compound 8 (0.35 g, 1.51 mmol), NMM (0.33 ml, 3.03 mmol), IBCF (0.26 ml, 1.97 mmol), and compound 18 (0.48 g, 1.59 mmol) were reacted to synthesize an oily compound 65, (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxopent-2-yl)(methyl)carbamate tert-butyl ester (0.69 g, 95%).

_Rf = 0.33 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.46-7.64 (m, ArH), 7.35-7.40 (m, ArH), 7.25-7.27 (m, ArH), 5.95-6.25 (m, C(O)NH), 4.51 (s, Chiral-H), 3.53-3.60 (m, NHCH ₂ CH ₂ ), 2.84 (t, J=6.6Hz, NHCH ₂ CH ₂ ), 2.67 (s, NCH ₃ ), 1.57-1.90 (m, CH ₂ CH ₂ CH ₃ ), 1.42 (s, Boc), 1.22-1.32 (m, CH ₂ CH ₂ CH ₃ ), 0.95 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 61: Preparation of (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxohex-2-yl)(methyl)carbamate tert-butyl ester (66)

Using reaction e, compound 9 (0.29 g, 1.20 mmol), NMM (0.33 ml, 2.99 mmol), IBCF (0.20 ml, 0.16 mmol) and compound 18 (0.38 g, 1.26 mmol) were reacted to synthesize an oily compound 66, (R)/(S)-(1-((2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)amino)-1-oxohex-2-yl)(methyl)carbamate tert-butyl ester (0.60 g, 97%).

_Rf = 0.30 (EtOAc 1: n-hexane)3;

¹ H NMR (DMSO-d ₆ , 400MHz) 7.89 (d, J=1.9Hz, ArH, C(O)NH), 7.62-7.71 (m, ArH), 7.30 (d, J=8.1Hz, ArH), 4.27-4.47 (m, Chiral-H), 3.32-3.35 (m, NHCH ₂ CH ₂ ), 2.77 (t, J=6.5Hz, NHCH ₂ CH ₂ ), 2.67 (s, NCH ₃ ), 1.53-1.71 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.39 (s, Boc), 1.12-1.28 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.84 (s, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 62: Preparation of (R)/(S)-(1-oxo-1((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)but-2-yl)carbamate tert-butyl methyl ester (67)

Using reaction e, compound 7 (0.62 g, 2.84 mmol), NMM (0.87 ml, 7.95 mmol), IBCF (0.48 ml, 3.69 mmol) and compound 19 (0.90 g, 2.98 mmol) were reacted to synthesize a white powder compound 67, (R)/(S)-(1-oxo-1((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)but-2-yl)carbamate tert-butyl methyl ester (0.89 g, 67%).

_Rf = 0.15 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.70 (t, J=9.7Hz, ArH), 7.56 (d, J=7.9Hz, ArH), 7.28-7.32 (m, ArH), 5.94-6.22 (m, C(O)NH), 4.45 (s, Chiral-H), 3.57-3.64 (m, NHCH ₂ CH ₂ ), 2.89 (d, J=6.1Hz, NHCH ₂ CH ₂ ), 2.70 (s, NCH ₃ ), 1.61-1.99 (m, CH ₂ CH ₃ ), 1.45 (s, Boc), 0.89 (t, J=7.3Hz, CH ₂ CH ₃ ).

Preparation Example 63: Preparation of (R)/(S)-(1-oxo-1((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)pent-2-yl)carbamate tert-butyl methyl ester (68)

Using reaction e, compound 8 (0.58 g, 2.53 mmol), NMM (0.69 ml, 6.31 mmol), IBCF (0.43 ml, 3.28 mmol) and compound 19 (0.80 g, 2.65 mmol) were reacted to synthesize an oily compound 68, (R)/(S)-(1-oxo-1((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)pent-2-yl)carbamate tert-butyl methyl ester (1.19 g, 98%).

_Rf = 0.26 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 300MHz) 7.64-7.70 (m, ArH), 7.41 (dd, J=8.1Hz, 67.4Hz, ArH), 5.95-6.21 (s, C(O)NH), 4.52 (s, Chiral-H), 3.54-3.63 (m, NHCH ₂ CH ₂ ), 2.86 (t, J=6.8Hz, NHCH ₂ CH ₂ ), 2.67 (s, NCH ₃ ), 1.55-1.91 (m, CH ₂ CH ₂ CH ₃ ), 1.42 (s, Boc), 1.23-1.27 (m, CH ₂ CH ₂ CH ₃ ), 0.88-0.95 (m, CH ₂ CH ₂ CH ₃ ).

Preparation Example 64: Preparation of (R)/(S)-((1-oxo-1((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)hex-2-yl)carbamate tert-butyl methyl ester (69)

Using reaction e, compound 9 (0.70 g, 2.84 mmol), NMM (0.87 ml, 7.95 mmol), IBCF (0.48 ml, 3.69 mmol) and compound 19 (0.90 g, 2.98 mmol) were reacted to synthesize a yellow powder, compound 69, (R)/(S)-((1-oxo-1((2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)amino)hex-2-yl)tert-butyl methyl carbamate (0.88 g, 63%).

_Rf = 0.32 (EtOAc 1: n-hexane)3;

¹ H NMR (DMSO-d ₆ , 400MHz) 7.87 (d, J=8.3Hz, ArH, C(O)NH), 7.80 (d, J=8.4Hz, ArH), 7.66 (d, J=8.2Hz, ArH), 7.33 (d, J=8.1Hz, ArH), 4.27-4.47 (m, Chiral-H), 3.32-3.33 (m, NHCH ₂ CH ₂ ), 2.78 (t, J=6.8Hz, NHCH ₂ CH ₂ ), 2.66 (s, NCH ₃ ), 1.53-1.71 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.39 (s, Boc), 1.12-1.28 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.84-0.89(m, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 65: Preparation of (R)/(S)-(1-oxo-1((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)but-2-yl)carbamate tert-butyl methyl ester (70)

Using reaction e, compound 7 (0.65 g, 3.00 mmol), NMM (0.92 ml, 8.99 mmol), IBCF (0.51 ml, 3.90 mmol) and compound 20 (1.00 g, 3.15 mmol) were reacted to synthesize an oily compound 70, (R)/(S)-(1-oxo-1((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)but-2-yl)carbamate tert-butyl methyl ester (1.00 g, 69%).

_Rf = 0.19 (EtOAc 1: n-hexane)3;

¹ H NMR (CDCl ₃ , 400MHz) 7.51-7.70 (m, ArH), 7.29 (t, J=3.7Hz, ArH), 5.98-6.26 (m, C(O)NH), 4.45 (s, Chiral-H), 3.59-3.60 (m, NHCH ₂ CH ₂ ), 2.87 (s, NHCH ₂ CH ₂ ), 2.70 (d, J=3.2Hz, NCH ₃ ), 1.62-2.07 (m, CH ₂ CH ₃ ), 1.45 (s, Boc), 0.90 (m, CH ₂ CH ₃ ).

Preparation Example 66: Preparation of (R)/(S)-(1-oxo-1((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)pent-2-yl)carbamate tert-butyl methyl ester (71)

Using reaction e, compound 8 (0.25 g, 1.08 mmol), NMM (0.30 ml, 2.69 mmol), IBCF (0.18 ml, 1.40 mmol) and compound 20 (0.36 g, 1.13 mmol) were reacted to synthesize a white powder compound 71, (R)/(S)-(1-oxo-1((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)pent-2-yl)carbamate tert-butyl methyl ester (0.32 g, 60%).

_Rf = 0.18 (EtOAc 1: n-hexane)3;

¹ H NMR (DMSO-d ₆ , 400MHz) 7.89 (s, C(O)NH), 7.76 (d, J=8.4Hz, ArH), 7.59 (d, J=7.8Hz, ArH), 7.44 (d, J=8.2Hz, ArH), 7.30 (d, J=7.8Hz, ArH), 4.30-4.49 (m, Chiral-H), 3.28-3.40 (m, NHCH ₂ CH ₂ ), 2.77 (s, NHCH ₂ CH ₂ ), 2.66 (s, NCH ₃ ), 1.53-1.90 (m, CH ₂ CH ₂ CH ₃ ), 1.39 (s, Boc), 1.16-1.23 (m, CH ₂ CH ₂ CH ₃ ), 0.88-0.89 (m, CH ₂ CH ₂ CH ₃ )

Preparation Example 67: Preparation of (R)/(S)-(1-oxo-1((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)hex-2-yl)carbamate tert-butyl methyl ester (72)

Using reaction e, compound 9 (0.74 g, 3.00 mmol), NMM (0.92 ml, 8.99 mmol), IBCF (0.51 ml, 3.90 mmol) and compound 20 (1.00 g, 3.15 mmol) were reacted to synthesize an oily compound 72, (R)/(S)-methyl(1-oxo-1((2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)amino)hex-2-yl)carbamate tert-butyl methyl ester (0.92 g, 60%).

_Rf = 0.29 (EtOAc 1: n-hexane)3;

¹ H NMR (DMSO-d ₆ , 400MHz) 7.88 (s, C(O)NH), 7.74-7.77 (m, ArH), 7.59 (d, J=8.2Hz, ArH), 7.44 (d, J= 8.0Hz, ArH), 7.30 (d, J=8.1Hz, ArH), 4.28-4.47 (m, Chiral-H), 3.30-3.34 (m, NHCH ₂ CH ₂ ), 2.77 (t, J=6.7Hz, NHCH ₂ CH ₂ ), 2.66 (s, NCH ₃ ), 1.53-1.72 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.39 (s, Boc), 1.12-1.29 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.84-0.90 (m, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 68: Preparation of (R)/(S)-2-amino-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)butyramide hydrochloride (73)

Compound 73, (R)/(S)-2-amino-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)butyramide hydrochloride (0.52 g, 91%), was synthesized by reacting compound 54 (0.66 g, 1.47 mmol) with 4.0 M HCl (2.20 ml, 8.81 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.66 (t, J=5.3Hz, C(O)NH), 8.23 (s, NH ₃ ), 7.92 (d, J=1.9Hz, ArH), 7.65-7.72 (m, ArH), 7.35 (d, J=8.1Hz, ArH), 3.67 (t, J=6.0Hz, Chiral-H), 3.31-3.55 (m, NHCH ₂ CH ₂ ), 2.79-2.83 (m, NHCH ₂ CH ₂ ), 1.67-1.74 (m, CH ₂ CH ₃ ), 0.79 (t, J=7.4Hz, CH ₂ CH ₃ ).

Preparation Example 69: Preparation of (R)/(S)-2-amino-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)pentanamide hydrochloride (74)

Using reaction f, compound 55 (0.95 g, 2.04 mmol) was reacted with 4.0 M HCl (3.06 ml, 12.2 mmol in dioxane) to synthesize compound 74, (R)/(S)-2-amino-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)pentanamide hydrochloride (0.66 g, 80%), which was a yellow powder.

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.82 (t, J=5.3Hz, C(O)NH), 8.34 (s, NH ₃ ), 7.64-7.90 (m, ArH), 7.35-7.38 (d, J=8.1Hz, ArH), 3.73 (t, J=6.3Hz, Chiral-H), 3.29-3.57 (m, NHCH ₂ CH ₂ ), 2.82-2.85 (m, NHCH ₂ CH ₂ ), 1.60-1.67 (m, CH ₂ CH ₂ CH ₃ ), 1.14-1.22 (m, CH ₂ CH ₂ CH ₃ ), 0.80 (t, J=7.1Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 70: Preparation of (R)/(S)-2-amino-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)hexamethylene amide hydrochloride (75)

Compound 75, (R)/(S)-2-amino-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)hexamethylene hydrochloride (0.60 g, 92%), was synthesized by reacting compound 56 (0.75 g, 1.56 mmol) with 4.0 M HCl (2.35 ml, 9.39 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.60 (t, J=5.5Hz, C(O)NH), 8.18 (s, NH ₃ ), 7.91 (d, J=2.0Hz, ArH), 7.65-7.72 (m, ArH), 7.35 (d, J=8.2Hz, ArH), 3.66 (t, J=6.2Hz, Chiral-H), 3.28-3.58 (m, NHCH ₂ CH ₂ ), 2.81-2.85 (m, NHCH ₂ CH ₂ ), 1.59-1.64 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.14-1.23 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.79 (t, J=6.8Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 71: Preparation of (R)/(S)-2-amino-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)butyramide hydrochloride (76)

Compound 76, (R)/(S)-2-amino-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)butyramide hydrochloride (0.75 g, 96%), was synthesized by reacting compound 57 (0.90 g, 2.00 mmol) with 4.0 M HCl (3.00 ml, 12.0 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.62 (s, C(O)NH), 8.17 (s, NH ₃ ), 7.88 (d, J=8.2Hz, ArH), 7.81 (d, J=8.4Hz, ArH), 7.68 (d, J=8.3Hz, ArH), 7.38 (d, J=8.1Hz, ArH), 3.66 (t, J=6.1Hz, Chiral-H), 3.30-3.57 (m, NHCH ₂ CH ₂ ), 2.82 (t, J=7.0Hz, NHCH ₂ CH ₂ ), 1.67-1.74 (m, CH ₂ CH ₃ ), 0.79 (t, J=7.5Hz, CH ₂ CH ₃ ).

Preparation Example 72: Preparation of (R)/(S)-2-amino-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)pentanamide hydrochloride (77)

Compound 77, (R)/(S)-2-amino-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)pentanamide hydrochloride (0.75 g, 85%), was synthesized by reacting compound 58 (1.03 g, 2.22 mmol) with 4.0 M HCl (5.56 ml, 22.2 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 300MHz) 8.81 (t, J=5.3Hz, C(O)NH), 8.35 (s, NH ₃ ), 7.84 (dd, J=8.2Hz, 15.7Hz, ArH), 7.54 (dd, J=8.1Hz, 73.4Hz, ArH), 3.71-3.75 (t, J=6.3Hz, Chiral-H), 3.30-3.60 (m, NH ₂ CH ₂ CH ₂ ), 2.85 (t, J=6.3Hz, NHCH ₂ CH ₂ ), 1.61-1.68 (m, CH ₂ CH ₂ CH ₃ ), 1.15-1.22 (m, CH ₂ CH ₂ CH ₃ ), 0.80 (t, J=7.2Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 73: Preparation of (R)/(S)-2-amino-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)hexamethylene amide hydrochloride (78)

Compound 78, a white powder, was synthesized by reacting compound 59 (1.32 g, 2.76 mmol) with 4.0 M HCl (4.14 ml, 16.6 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.69 (t, J=4.9Hz, C(O)NH), 8.26 (s, NH ₃ ), 7.88 (d, J=8.2Hz, ArH), 7.81 (d, J=8.3Hz, ArH), 7.68 (d, J=8.1Hz, ArH), 7.39 (d, J=8.0Hz, ArH), 3.69 (t, J=6.2Hz, Chiral-H), 3.29-3.59 (m, NHCH ₂ CH ₂ ), 2.80-2.89 (m, NHCH ₂ CH ₂ ), 1.61-1.67 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.14-1.23 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.79 (t, J=6.8Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 74: Preparation of (R)/(S)-2-amino-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)butyramide hydrochloride (79)

Compound 79, (R)/(S)-2-amino-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)butyramide hydrochloride (0.43 g, 54%), was synthesized by reacting compound 60 (0.91 g, 1.95 mmol) with 4.0 M HCl (2.92 ml, 11.7 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.67 (s, C(O)NH), 8.24 (s, NH ₃ ), 7.35-7.89 (m, ArH), 3.67 (d, J=4.3Hz, Chiral-H), 3.30-3.55 (m, NHCH ₂ CH ₂ ), 2.80-2.85 (m, NHCH ₂ CH ₂ ), 1.68-1.75 (m, CH ₂ CH ₃ ), 0.80 (t, J=7.5Hz, CH ₂ CH ₃ ).

Preparation Example 75: Preparation of (R)/(S)-2-amino-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)pentanamide hydrochloride (80)

Compound 80, a white powder, was synthesized by reacting compound 61 (0.70 g, 1.45 mmol) with 4.0 M HCl (2.18 ml, 8.70 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.71 (t, J=5.4Hz, C(O)NH), 8.28 (s, NH ₃ ), 7.77 (d, J=8.8Hz, ArH), 7.62 (d, J=8.2Hz, ArH), 7.35-7.45 (m, ArH), 3.70 (t, J=6.4Hz, Chiral-H), 3.28-3.57 (m, NHCH ₂ CH ₂ ), 2.80-2.86 (m, NHCH ₂ CH ₂ ), 1.61-1.65 (m, CH ₂ CH ₂ CH ₃ ), 1.13-1.24 (m, CH ₂ CH ₂ CH ₃ ), 0.80 (t, J=7.4Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 76: Preparation of (R)/(S)-2-amino-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)hexamethylene amide hydrochloride (81)

Compound 81, a white powder, was synthesized by reacting compound 62 (0.79 g, 1.59 mmol) with 4.0 M HCl (2.38 ml, 9.54 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.66 (t, J=5.4Hz, C(O)NH), 8.23 (s, NH ₃ ), 7.88 (d, J=8.2Hz, ArH), 7.81 (d, J=8.4Hz, ArH), 7.68 (d, J=8.2Hz, ArH), 7.39 (d, J=8.2Hz, ArH), 3.68 (t, J=6.4Hz, Chiral-H), 3.29-3.59 (m, NHCH ₂ CH ₂ ), 2.81-2.87 (m, NHCH ₂ CH ₂ ), 1.60-1.66 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.13-1.23 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.79 (t, J=7.0Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 77: Preparation of (R)/(S)-2-amino-N-(2-(3’,4’-difluoro-[1,1’-biphenyl]-4-yl)ethyl)pentanamide hydrochloride (82)

Compound 62 (0.94 g, 2.17 mmol) was reacted with 4.0 M HCl (3.26 ml, 13.0 mmol in dioxane) to synthesize a white powder, compound 82, (R)/(S)-2-amino-N-(2-(3’,4’-difluoro-[1,1’-biphenyl]-4-yl)ethyl)pentanamide hydrochloride (0.68 g, 85%).

R _f =0.00 (EtOAc 9: aetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.55 (s, C(O)NH), 8.10 (s, NH ₃ ), 7.72-7.77 (m, ArH), 7.62 (d, J=8.1Hz, ArH), 7.50-7.53 (m, ArH), 7.33 (d, J=8.1Hz, ArH), 3.65 (t, J=5.9Hz, Chiral-H), 3.29-3.57 (m, NHCH ₂ CH ₂ ), 2.78-2.84 (m, NHCH ₂ CH ₂ ), 1.58 (q, J=6.9Hz, 8.8Hz, CH ₂ CH ₂ CH ₃ ), 1.14-1.20 (m, CH ₂ CH ₂ CH ₃ ), 0.80 (t, J=7.3Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 78: Preparation of (R)/(S)-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)butyramide hydrochloride (83)

Compound 83, (R)/(S)-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)butyramide hydrochloride (1.07 g, 94%), was synthesized by reacting compound 64 (1.31 g, 2.81 mmol) with 4.0 M HCl (4.20 ml, 16.9 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.79 (s, C(O)NH, NH ₂ ), 7.92 (d, J=2.0Hz, ArH), 7.65-7.72 (m, ArH), 7.36 (d, J=8.1Hz, ArH), 3.59 (t, J=6.4Hz, Chiral-H), 3.39-3.53 (m, NHCH ₂ CH ₂ ), 2.83 (t, J=6.8Hz, NHCH ₂ CH ₂ ), 2.38 (t, J=6.1Hz, NCH ₃ ), 1.68-1.80 (m, CH ₂ CH ₃ ), 0.77 (t, J=7.5Hz, CH ₂ CH ₃ ).

Preparation Example 79: Preparation of (R)/(S)-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)pentanamide hydrochloride (84)

Compound 84, (R)/(S)-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)pentanamide hydrochloride (0.33 g, 57%), was synthesized by reacting compound 65 (0.65 g, 1.36 mmol) with 4.0 M HCl (2.03 ml, 8.13 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 9.26 (s, NH ₂ ), 8.96 (t, J=5.3Hz, C(O)NH), 7.90-7.91 (m, ArH), 7.64-7.71 (m, ArH), 7.36 (d, J=8.1Hz, ArH), 3.64-3.68 (m, Chiral-H), 3.42-3.56 (m, NHCH ₂ CH ₂ ), 2.84 (t, J=6.8Hz, NHCH ₂ CH ₂ ), 2.35 (s, NCH ₃ ), 1.60-1.77 (m, CH ₂ CH ₂ CH ₃ ), 1.09-1.16 (m, CH ₂ CH ₂ CH ₃ ), 0.80 (t, J=7.2Hz, CH _{2 CH2CH3 )} .

Preparation Example 80: Preparation of (R)/(S)-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)hexamethylene amide hydrochloride (85)

Compound 85, (R)/(S)-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)hexamethylene hydrochloride (0.38 g, 75%), was synthesized by reacting compound 66 (0.57 g, 1.16 mmol) with 4.0 M HCl (1.74 ml, 6.95 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.70-9.36 (s, NH ₂ ), 8.80 (t, J=5.4Hz, C(O)NH), 7.91 (d, J=2.0Hz, ArH), 7.65-7.72 (m, ArH), 7.35 (d, J=8.2Hz, ArH), 3.55 (t, J=6.6Hz, Chiral-H), 3.38-3.52 (m, NHCH ₂ CH ₂ ), 2.83 (t, J=6.8Hz, NHCH ₂ CH ₂ ), 2.35 (s, NCH ₃ ), 1.60-1.76 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.05-1.23 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.78 (t, J=7.2Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 81: Preparation of (R)/(S)-2-(methylamino)-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)butyramide hydrochloride (86)

Compound 86, (R)/(S)-2-(methylamino)-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)butyramide hydrochloride (0.65 g, 88%), was synthesized by reacting compound 67 (0.86 g, 1.84 mmol) with 4.0 M HCl (2.77 ml, 11.1 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 9.04 (s, NH ₂ ), 8.79 (s, C(O)NH), 7.87 (d, J=8.3Hz, ArH), 7.80 (d, J=8.4Hz, ArH), 7.68 (d, J=8.2Hz, ArH), 7.39 (d, J=8.2Hz, ArH), 3.59 (t, J=2.5Hz, 4.9Hz, Chiral-H), 3.40-3.56 (m, NHCH ₂ CH ₂ ), 2.84 (t, J = 6.9 Hz, NHCH ₂ CH ₂ ), 2.37 (s, NCH ₃ ), 1.67-1.85 (m, CH ₂ CH ₃ ), 0.77 (t, J = 7.5 Hz, CH ₂ CH ₃ ).

Preparation Example 82: Preparation of (R)/(S)-2-(methylamino)-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)pentanamide hydrochloride (87)

Compound 68 (1.17 g, 2.45 mmol) was reacted with 4.0 M HCl (3.67 ml, 14.7 mmol in dioxane) to synthesize a white powder, compound 87, (R)/(S)-2-(methylamino)-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)pentanamide hydrochloride (81%).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.93-9.79 (m, NH ₂ ), 9.04 (t, J=5.2Hz, C(O)NH), 7.84 (dd, J=8.3Hz, 15.1Hz, ArH), 7.54 (dd, J=8.0Hz, 73.4Hz, ArH), 3.68-3.72 (m, Chiral-H), 3.42-3.58 (m, NHCH ₂ CH ₂ ), 2.87 (t, J=6.7Hz, NHCH ₂ CH ₂ ), 2.36 (s, NCH ₃ ), 1.64-1.82 (m, CH ₂ CH ₂ CH ₃ ), 1.09-1.22 (m, CH ₂ CH ₂ CH ₃ ), 0.81 (t, J=7.1Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 83: Preparation of (R)/(S)-2-(methylamino)-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)hexamethylene amide hydrochloride (88)

Compound 88, a white powder, was synthesized by reacting compound 69 (0.85 g, 1.72 mmol) with 4.0 M HCl (2.60 ml, 10.3 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 9.00 (s, NH ₂ ), 8.75 (t, J=5.4Hz, C(O)NH), 7.87 (d, J=8.3Hz, ArH), 7.81 (d, J=8.4Hz, ArH), 7.68 (d , J=8.2Hz, ArH), 7.38 (d, J=8.2Hz, ArH), 3.57-3.61 (m, Chiral-H), 3.40-3.55 (m, NHCH ₂ CH ₂ ), 2.84 (t, J=6.8Hz, NHCH ₂ CH ₂ ), 2.36 (s, NCH ₃ ), 1.60-1.75 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.06-1.24 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.78 (t, J=7.2Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 84: Preparation of (R)/(S)-2-(methylamino)-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)butyramide hydrochloride (89)

Compound 70 (0.97 g, 2.01 mmol) was reacted with 4.0 M HCl (3.00 ml, 12.1 mmol in dioxane) to synthesize a yellow powder, compound 89, (R)/(S)-2-(methylamino)-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)butyramide hydrochloride (0.73 g, 88%).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 9.00 (s, NH ₂ ), 8.77 (s, C(O)NH), 7.77 (d, J=8.7Hz, ArH), 7.61 (d, J=8.2Hz, ArH), 7.44 (d, J=8 .2Hz, ArH), 7.35 (d, J=8.1Hz, ArH), 3.57-3.60 (m, Chiral-H), 3.39-3.55 (m, NHCH ₂ CH ₂ ), 2.82 (t, J = 6.9 Hz, NHCH ₂ CH ₂ ), 2.37 (s, NCH ₃ ), 1.67-1.83 (m, CH ₂ CH ₃ ), 0.77 (t, J = 7.5 Hz, CH ₂ CH ₃ ).

Preparation Example 85: Preparation of (R)/(S)-2-(methylamino)-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)pentanamide hydrochloride (90)

Compound 90, a white powder, was synthesized by reacting compound 71 (0.83 g, 1.67 mmol) with 4.0 M HCl (2.51 ml, 10.0 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.85-9.19 (m, NH ₂ ), 8.75 (s, C(O)NH), 7.76 (d, J=7.7Hz, ArH), 7.61 (d, J=7.5Hz, ArH), 7.44 (d, J=8.0Hz, ArH), 7.35 (d, J=7.4Hz, ArH), 3.60 (s, Chiral-H), 3.40-3.56 (m, NHCH ₂ CH ₂ ), 2.83 (t, J=6.5Hz, NHCH ₂ CH ₂ ), 2.36 (s, NCH ₃ ), 1.60-1.66 (m, CH ₂ CH ₂ CH ₃ ), 1.09-1.17 (m, CH ₂ CH ₂ CH ₃ ), 0.80 (t, J=7.0Hz, CH ₂ CH ₂ CH ₃ ).

Preparation Example 86: Preparation of (R)/(S)-2-(methylamino)-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)hexamethylene amide hydrochloride (91)

Compound 91, (R)/(S)-2-(methylamino)-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)hexamamide hydrochloride (0.66 g, 86%), was synthesized by reacting compound 72 (0.89 g, 1.74 mmol) with 4.0 M HCl (2.61 ml, 10.4 mmol in dioxane).

_Rf = 0.00 (EtOAc 9: acetone 1);

¹ H NMR (DMSO-d ₆ , 400MHz) 8.95 (s, NH ₂ ), 8.72 (s, C(O)NH), 7.76 (d, J=8.7Hz, ArH), 7.61 (d, J=8.1Hz, ArH), 7.44 (d, J=8 .4Hz, ArH), 7.35 (d, J=8.1Hz, ArH), 3.57-3.60 (m, Chiral-H), 3.29-3.56 (m, NHCH ₂ CH ₂ ), 2.83 (t, J=6.8Hz, NHCH ₂ CH ₂ ), 2.37 (s, NCH ₃ ), 1.59-1.74 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 1.08-1.24 (m, CH ₂ CH ₂ CH ₂ CH ₃ ), 0.78 (t, J=7.2Hz, CH ₂ CH ₂ CH ₂ CH ₃ ).

Preparation Example 87: Preparation of (R)/(S)-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(dimethylamino)pentanamide (92)

Compound 92,(R)/(S)-N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(dimethylamino)pentanamide was synthesized by reacting 74 (1.0 equivalent), triethylamine (6.0 equivalent), formaldehyde (1.05 equivalent) and palladium catalyst (0.2 equivalent) using reaction formula g.

¹ H NMR (DMSO-d ₆ , 400MHz) 8.96 (t, J=5.3Hz, C(O)NH), 7.90-7.91 (m, ArH), 7.64-7.71 (m, ArH), 7.36 (d, J=8.1Hz, ArH), 3.64-3.68 (m, Chiral-H), 3.42-3.56 (m, NHCH ₂ CH ₂ ), 2.82-2.85 (m, NHCH ₂ CH ₂ ), 2.35 (s, N(CH ₃ ) 2), 1.60-1.77 (m, CH ₂ CH ₂ CH ₃ ), 1.09-1.16 (m, CH ₂ CH ₂ CH ₃ ), 0.80 (t, J=7.2Hz, CH ₂ CH ₂ CH ₃ ).

Implementation of the invention

Example

Example 1: Analysis of antifungal activity against human pathogenic fungi

To determine the degree of antifungal activity against opportunistic fungi of the compounds synthesized in the above preparation examples, in vitro antifungal susceptibility testing was performed according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI). In this example, using *Cryptococcus neoformans*, *Candida albicans*, *Candida glabrata*, and *Aspergillus fumigatus* as targets, the minimum inhibitory concentration (MIC) at which the compounds could inhibit fungal growth was measured and is shown in Table 1. MIC values (μg/ml) are expressed as range values. Specifically, Candida species and Cryptococcus neoformans were cultured on Sabouraud dextrose agar (SDA) (Sigma-Aldrich) solid medium for 24 hours and 48 hours, respectively. Single colonies that grew after culture were dissolved in 0.85% physiological saline to prepare a cell suspension with a concentration of 1 cell/ml to 5 × ^10⁶ cells/ml. This cell suspension was then diluted 2000-fold in RPMI 1640 liquid medium (Sigma-Aldrich) to prepare a final concentration of 5 × ^10² cells/ml to 2.5 × ^10³ cells/ml, and then aliquoted at 195 μl/well into 96-well plates. Then, the compounds synthesized according to the above preparation examples were diluted 2-fold to prepare final concentrations of 128 μg/ml, 64 μg/ml, 32 μg/ml, 16 μg/ml, 8 μg/ml, 4 μg/ml, 2 μg/ml, 1 μg/ml, and 0.5 μg/ml, respectively. Each concentration was then treated with 5 μl per well to prepare a final 200 μl/well test cell culture. Candida species treated with the compounds were cultured for 48 hours, and Cryptococcus neoformans were cultured at 35°C for 72 hours. The bottom of the 96-well plates was visually observed to confirm growth, thus determining the MIC. As a positive control, amphotericin B (AMB), a well-known representative antifungal agent with high toxicity to humans, was used. The results are shown in Tables 1 to 3.

Table 1

Table 2

Table 3

Example 2: Analysis of the antifungal activity of compound 74

To confirm the antifungal activity of compound 74, the minimum inhibitory concentration (MIC) for inhibiting fungal growth was determined according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI). Each strain was spread onto YPD solid medium (Sigma-Aldrich) to achieve a final concentration of 2.5 × ^10³ microconidia for *T. rubrum* and *T. menagrophytes*, and 2.5 × ^10³ cfu/ml for *Candida albicans*, *Candida glabrata*, and *Cryptococcus neoformans*. After preparation, 195 μl of each strain was aliquoted into 96-well plates. Compound 74 was diluted 2-fold to prepare final concentrations of 128 μg/ml, 64 μg/ml, 32 μg/ml, 16 μg/ml, 8 μg/ml, 4 μg/ml, 2 μg/ml, 1 μg/ml, and 0.5 μg/ml, respectively. Each concentration was then treated with 5 μl per well to prepare a final concentration of 200 μl per well. Candida species treated with the compound were cultured for 48 hours, and Cryptococcus neoformans and Trichophyton species were cultured at 35°C for 72 hours. The bottom of the 96-well plate was visually observed to confirm growth, thus determining the MIC (μg/ml). The results are shown in Table 4.

Table 4

Example 3: Confirmation of the rapid-acting effect of compound 74 compared with commercially available competing drugs

Confirmation of the rapid-acting effect of compound 74 compared with competing drugs in the market.

To confirm the rapid-acting effect of compound 74, the growth of *Candida albicans* was assessed by treating it with the same concentration of compound 74 and commercially available drugs such as efinaconazole, tavaborole, terbinafine, or cilopirox for the same duration. *Candida albicans* was prepared at a concentration of 2.5 × ^10³ cfu/ml, and 195 μl/well was divided into 96-well plates. Then, compound 74 and the aforementioned commercially available control drugs were treated at 50 μg/ml and 100 μg/ml, respectively, for 30 minutes, and then spread onto YPD solid medium (Sigma-Aldrich) to confirm the number of single colonies. The results are shown in Figure 1. This confirms that compound 74 has a superior rapid-acting effect compared to commercially available competing drugs.

Example 4: Confirmation of the antifungal efficacy of compound 74 compared to commercially available drugs

To confirm the antifungal effect of compound 74, *Trichophyton rubrum* was treated with 0.5, 1, 2, 4, 8, and 16 times the MIC concentration of compound 74 at predetermined times, along with commercially available drugs such as efinaconazole, amorolfine, terbinafine, cilopirox, and amphotericin B. *Trichophyton rubrum* was prepared at a concentration of 6 × ^10³ cfu/ml, and 195 μl was aliquoted into 96-well plates. Then, compound 74 and competing drugs were treated with 5 μl of each drug at concentrations of 0.5, 1, 2, 4, 8, and 16 times their respective MIC concentrations, adjusted to a final concentration of 200 μl/well, and treated for 2, 4, 6, 12, 24, 48, 72, and 120 hours. Samples were then spread onto YPD solid medium at each time point to confirm the number of single colonies formed. The results are shown in Figure 2. Figure 2 confirms that, compared with commercially available drugs, the MIC concentration of compound 74 is the same as its fungicidal concentration, indicating that it has a fungicidal effect at low concentrations compared to other drugs.

Example 5: Confirmation of the biofilm removal effect of compound 74 compared with commercially available drugs

To confirm the rapid-acting effect of compound 74, Candida albicans was treated with caspofungin, amphotericin B, and efinaconazole (common drugs) at concentrations of 4 μg/ml, 8 μg/ml, 16 μg/ml, and ³² μg/ml to confirm the growth of Candida albicans. Candida albicans was prepared at a concentration of 2.5 × 10³ cfu/ml and cultured in 96-well plates for 90 minutes to form a biofilm. Compound 74 and the aforementioned control drugs were then treated, and growth of Candida albicans was confirmed after 24 hours. The results are shown in Figure 3. Figure 3 confirms that the effect of inhibiting the growth of Candida albicans by disrupting the biofilm is significantly superior to that of the commercially available control drugs.

Example 6: Antibacterial Activity Analysis

The antibacterial activity of the compounds synthesized in this invention against Gram-negative or Gram-positive bacteria and multidrug-resistant bacteria was determined. MIC testing was performed according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI). Escherichia coli (−), Pseudomonas aeruginosa (−), and Salmonella enterica (−) were used as Gram-negative bacteria. Bacillus cereus (+), Bacillus subtilis (+), Bacillus coagulans (+), Listeria monocytogenes (+), Micrococcus luteus (+), Propionibacterium acnes (+), Staphylococcus epidermidis (+), and Staphylococcus aureus (+) were used as Gram-positive bacteria. Methicillin-resistant Staphylococcus aureus (MRSA) was used as a multidrug-resistant bacterium. The OD ₆₀₀ of single colonies formed in the solid medium used to culture the bacteria of the above-mentioned experimental subjects was adjusted to 0.1. These colonies were then diluted 100-fold in MHB liquid medium (Sigma-Aldrich) to a final OD ₆₀₀ of 0.001, and 100 μl/well was aliquoted into 96-well plates. The compound of the present invention was then diluted 2-fold to prepare final concentrations of 64 μg/ml, 32 μg/ml, 16 μg/ml, 8 μg/ml, 4 μg/ml, 2 μg/ml, 1 μg/ml, and 0.5 μg/ml, respectively, and treated with 100 μl/well to achieve a final concentration of 200 μl/well. Each strain was then incubated at 37°C for 24 hours, and the MIC (μg/ml) was determined visually and by OD _600. Norfloxacin and vancomycin were used as positive controls. The results are shown in Tables 5 to 8.

Table 5

Table 6

Table 7

Table 8

Example 7: Anti-inflammatory activity analysis.

The anti-inflammatory effect of the compounds synthesized in this invention was confirmed by inducing interleukin-6 (IL-6) in RAW264.7 cells. RAW264.7 cells (from a Korean cell line bank) used as mouse macrophages were seeded in 12-well plates at an initial density of 5 × ^10⁵ cells/well and cultured at 37°C for 16–24 hours. The culture medium containing the RAW264.7 cells was then treated with the compounds synthesized in this invention to achieve final concentrations of 0 μg/ml or 8 μg/ml, and cultured at 37°C for 1 hour. Then, lipopolysaccharide (LPS, Sigma-Aldrich, USA) was treated to achieve a final concentration of 1 μg/ml, and the cells were cultured at 37°C for 24 hours to induce IL-6. The amount of IL-6 generated in RAW264.7 cells was then confirmed using an ELISA kit purchased from Komabiotech, according to the manufacturer's instructions. The results are shown in Table 9. In Table 9, the amount of IL-6 is expressed as the ratio (%) of the amount induced by LPS to the amount reduced by treatment with the compounds of the present invention.

Table 9

Industrial availability

This invention relates to novel compounds and their uses, which can be used in the preparation of antifungal and antimicrobial compositions, and can be applied to the development of pharmaceutical compositions for antifungal and antifungal prevention and treatment.

Claims (21)

1. A compound, its stereoisomers, or pharmaceutically acceptable salts thereof, characterized in that, As shown in the following chemical formula 1: Chemical Formula 1: In the above chemical formula 1, n is 0, 1, 2, 3, 4, or 5. _R1 , _R2 , and _R3 are each independently the same or different. _R1 and _R2 are each independently selected from hydrogen, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, _C1-7 alkyl, hydroxyl, halogen, halo- _C1-7 alkyl, _C1-7 alkoxy, and halo- _C1-7 alkoxy. _R3 is independently selected from hydrogen, cyclopropyl, _C1-7 alkyl, hydroxyl, halogen, halo- _C1-7 alkyl, _C1-7 alkoxy, and halo- _C1-7 alkoxy. X represents m identical or different substituents selected from the group consisting of halogen groups, halogenated _C1-7 alkyl groups, and halogenated _C1-7 alkoxy groups, where m is an integer from 1 to 5. The conditions are: when n is 0 or 1 and _R1 and _R2 are hydrogen, _R3 is not a _C1-3 alkyl group; when n is 0, _R3 is not hydrogen. 2. The compound, its stereoisomer, or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, _R1 and _R2 are each independently H or methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, cyclobutyl, n-pentyl, cyclopentyl, n-hexyl or cyclohexyl. _R3 above is methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl. 3. The compound, its stereoisomers, or pharmaceutically acceptable salts thereof according to claim 1, characterized in that X is one or two identical or different substituents from the group consisting of fluorine, chlorine, trifluoromethyl and trifluoromethoxy. 4. The compound, its stereoisomers, or pharmaceutically acceptable salts thereof according to claim 3, characterized in that X is p-fluorine, m-fluorine, p-, m-difluoro, p-chloro, m-chloro, p-, m-dichloro, p-trifluoromethyl, or p-trifluoromethoxy. 5. The compound, its stereoisomer, or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, The above compounds are as follows: 1) 2-Amino-N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)butyramide; 2) 2-Amino-N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)pentanamide; 3) 2-Amino-N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)hexamethylenediamide; 4) 2-Amino-N-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)pentanamide; 5) N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)-2-(methylamino)butyramide; 6) N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)-2-(methylamino)pentanamide; 7) N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)-2-(methylamino)hexamamide; 8) N-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)-2-(dimethylamino)pentanamide; 9) 2-Amino-N-((3',4'-dichloro-[1,1'-biphenyl]-4-yl)methyl)pentanamide; 10) 2-Amino-N-((3',4'-dichloro-[1,1'-biphenyl]-4-yl)methyl)hexamamide; 11) 2-Amino-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)pentanamide; 12) 2-Amino-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)hexamamide; 13) 2-Amino-N-((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methyl)pentanamide; 14) 2-Amino-N-((4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)methyl)hexamamide; 15)N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)-2-(methylamino)pentanamide; 16) 2-(methylamino)-N-((4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)methyl)pentanamide; 17) N-((3’,4’-dichloro-[1,1’-biphenyl]-4-yl)methyl)-2-(dimethylamino)pentanamide; 18) 2-Amino-N-(2-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)ethyl)butyramide; 19) 2-Amino-N-(2-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)ethyl)pentanamide; 20) 2-Amino-N-(2-(3',4'-dichloro-[1,1'-biphenyl]-4-yl)ethyl)hexamamide; 21) 2-Amino-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)butyramide; 22) 2-Amino-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)pentanamide; 23) 2-Amino-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)hexamamide; 24) 2-Amino-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)butyramide; 25) 2-Amino-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)pentanamide; 26) 2-Amino-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)hexamamide; 27) 2-Amino-N-(2-(3',4'-difluoro-[1,1'-biphenyl]-4-yl)ethyl)pentanamide; 28)N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)butyramide; 29)N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)pentanamide; 30)N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(methylamino)hexamamide; 31) 2-(methylamino)-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)butyramide; 32)2-(methylamino)-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)pentanamide; 33) 2-(methylamino)-N-(2-(4’-(trifluoromethyl)-[1,1’-biphenyl]-4-yl)ethyl)hexamamide; 34) 2-(methylamino)-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)butyramide; 35) 2-(methylamino)-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)pentanamide; 36) 2-(methylamino)-N-(2-(4'-(trifluoromethoxy)-[1,1'-biphenyl]-4-yl)ethyl)hexamamide; Or 37)N-(2-(3’,4’-dichloro-[1,1’-biphenyl]-4-yl)ethyl)-2-(dimethylamino)pentanamide. 6. A method for preparing a compound, its stereoisomer, or a pharmaceutically acceptable salt thereof, as described in any one of claims 1 to 5, characterized in that, include: The first step involves reacting the aminoalkyl acid derivative compound protected by the tert-butoxycarbonyl protecting group shown in Formula 2 with the biphenyl derivative compound containing a primary amine group shown in Formula 3 to form a peptide bond; and The second step involves reacting the compound obtained in the first step with an acid to remove the tert-butyloxycarbonyl protecting group. Chemical Formula 1: Chemical formula 2: Chemical formula 3: In the above chemical formula, n is 0, 1, 2, 3, 4, or 5. _R1 , _R2 , and _R3 are each independently the same or different. _R1 and _R2 are each independently selected from hydrogen, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, _C1-7 alkyl, hydroxyl, halogen, halo- _C1-7 alkyl, _C1-7 alkoxy, and halo- _C1-7 alkoxy. _R3 is independently selected from hydrogen, cyclopropyl, _C1-7 alkyl, hydroxyl, halogen, halo- _C1-7 alkyl, _C1-7 alkoxy, and halo- _C1-7 alkoxy. X is selected from the group consisting of halogen groups, halogenated _C1-7 alkyl groups and halogenated _C1-7 alkoxy groups, where m is an integer from 1 to 5. 7. The method for preparing the compound, its stereoisomer, or a pharmaceutically acceptable salt thereof according to claim 6, characterized in that, The amino acid derivative compound shown in Formula 4 was prepared by reacting it with di-tert-butyl dicarbonate, which is protected by a tert-butyloxycarbonyl protecting group. Chemical formula 4: In the above chemical formula, _R1 ' is hydrogen. _R2 and _R3 are each independently the same or different, and are each independently selected from hydrogen, _C1-7 alkyl, hydroxyl, halogen, halogenated _C1-7 alkyl, _C1-7 alkoxy and halogenated _C1-7 alkoxy. 8. A method for preparing the compound, its stereoisomer, or a pharmaceutically acceptable salt thereof according to claim 7, characterized in that _R2 in the compound of formula 4 is hydrogen, and the compound of formula 4 is subjected to an alkylation step by reacting with a haloalkane in the presence of a base. 9. A method for preparing the compound, its stereoisomers, or pharmaceutically acceptable salts thereof according to claim 6, characterized in that, The _C0-2 alkylamine derivative shown in Formula 5, with a terminal substituted halophenyl group, is reacted with di-tert-butyl dicarbonate to introduce a tert-butyloxycarbonyl protecting group into the amine group. This is followed by reaction with a phenylboronic acid derivative shown in Formula 6, and then reaction with an acid to remove the tert-butyloxycarbonyl protecting group, thereby preparing the biphenyl derivative compound shown in Formula 3 containing a primary amine group. Chemical formula 5: Chemical formula 6: In the above chemical formula, X’ is a halogen. X is selected from the group consisting of halogen groups, halogenated _C1-7 alkyl groups and halogenated _C1-7 alkoxy groups, where m is an integer from 1 to 5. 10. A method for preparing the compound, its stereoisomer, or a pharmaceutically acceptable salt thereof according to claim 9, characterized in that the reaction with the above-mentioned phenylboronic acid derivative is achieved by a cross-linking reaction based on a metal catalyst in the presence of a base. 11. A method for preparing the compound, its stereoisomers, or pharmaceutically acceptable salts thereof according to claim 6, characterized in that the first step is achieved by an anhydrous coupling reaction in an organic solvent in the presence of N-methylmorpholine and isobutyl chloroformate. 12. A method for preparing the compound, its stereoisomers, or pharmaceutically acceptable salts thereof according to claim 6, characterized in that, after the second step, a third step is further included to alkylate the amine to form a tertiary amine, so as to obtain a compound in which both _R1 and _R2 are alkyl groups. 13. An antifungal composition, characterized in that it comprises the compound, its stereoisomer, or a pharmaceutically acceptable salt thereof as the active ingredient, according to any one of claims 1 to 5. 14. The antifungal composition according to claim 13, characterized in that it has antifungal activity against Cryptococcus neoformans, Candida albicans, Candida glabrata or Aspergillus fumigatus. 15. A pharmaceutical composition for treating or preventing fungal infections, characterized in that it comprises, as an active ingredient, any one of claims 1 to 5, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. 16. The pharmaceutical composition for treating or preventing fungal infections according to claim 15, wherein the fungal infection is caused by infection with Cryptococcus neoformans, Candida albicans, Candida glabrata, or Aspergillus fumigatus. 17. The pharmaceutical composition for treating or preventing fungal infections according to claim 15, characterized in that it further comprises a pharmaceutically acceptable carrier, diluent, or excipient. 18. An antibacterial composition, characterized in that it comprises, as an active ingredient, any one of claims 1 to 5, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. 19. The antibacterial composition according to claim 18, characterized in that it has antibacterial activity against Gram-negative bacteria, Gram-positive bacteria or multidrug-resistant bacteria. 20. A pharmaceutical composition for treating or preventing bacterial infections, characterized in that it comprises, as an active ingredient, a compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as any one of claims 1 to 5. 21. A pharmaceutical composition for treating or preventing anti-inflammatory diseases, characterized in that it comprises the compound, its stereoisomer, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient, as described in any one of claims 1 to 5.