BRPI1014574B1 - "Imitation-binding compounds, Pharmaceutical composition, Understanding them, Method for manufacturing and preparing the same, and Use of the pharmaceutical composition dialect" - Google Patents

Description

(54) Title: IMPACTING COMPOUNDS WITH REVERSE CONNECTION, PHARMACEUTICAL COMPOSITION UNDERSTANDING THE SAME, METHOD FOR MANUFACTURING AND PREPARING THE SAME AND USE OF THE DONE PHARMACEUTICAL COMPOSITION (51) lnt.CI .: C07D 487/04; C07D 403/02 (30) Unionist Priority: 4/15/2009 KR 10-2009-0032937 (73) Holder (s): JW PHARMACEUTICAL CORPORATION (72) Inventor (s): KYUNG-YUN JUNG; JAE UK CHUNG; MIN-WOOK JEONG; HEE-KYUNG JUNG; HYUN-JU LA; SANG-HO MA; YONG-SIL LEE (85) National Phase Start Date: 10/14/2011

1/51 “IMPELLING COMPOUNDS REVERSE CONNECTION, COMPOSITION

PHARMACEUTICAL UNDERSTANDING THE SAME, METHOD FOR MANUFACTURING

AND PREPARE THE SAME AND USE OF THAT PHARMACEUTICAL COMPOSITION ”

Technical Field [001] The present invention relates to new compounds of reverse linkers, a method of making them, and the use of these in the treatment of diseases, such as acute myeloid leukemia.

Background to the Technique [002] Random selection of molecules for possible activity as therapeutic agents has been conducted for many years and has resulted in numerous important drug discoveries. Recently, non-peptide compounds have been developed to further approximate the secondary structure of reverse bonds found in biologically active proteins or peptides. For example, U.S. patent No. 5,440,013, and PCT patent applications Nos. WO94 / 03494, W001 / 00210A1, and WO01 / 16135A2, all by Kahn, each describe conformationally limited, non-peptide compounds that mimic the secondary structure of reverse bonds. Furthermore, U.S. Patent Nos. 5,929,237 and 6,013,458, both by Kahn, describe conformationally limited compounds that mimic the secondary structure of reverse-binding regions of biologically active peptides and proteins. The synthesis and identification of conformationally limited reverse link mimics and their application to diseases have been well reviewed by Obrecht (Advances in Med. Chem., 4,1-68,1999).

[003] With the significant advances in the synthesis and identification of conformationally limited reverse linkers, techniques have been developed to synthesize and select library members of small molecules that mimic the secondary structure of peptides, in order to identify bioactive library members. In this way, attempts were made to look for compounds

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2/51 conformationally limited and highly bioactive compounds that mimic the secondary structure of reverse-binding regions of biologically active peptides and proteins. For example, reverse linkers, methods for making them and bioactivities of these are described in patent applications

PCT Nos. WO 04 / 093828A2, WO 05 / 116032A2, and WO 07 / 139346A1.

[004] Although a large number of reverse linkers have been manufactured, it has not been observed that many compounds have high bioactivity. Thus, efforts continue to manufacture compounds that can be applied to the treatment of diseases, such as cancer.

[005] Particularly, efforts were focused on developing compounds that strongly block the Wnt signaling pathway to effectively suppress the growth of acute myeloid leukemia (AML) cancer cells known to have an activated Wnt signaling pathway.

[006] Also, there is a need for methods to manufacture highly bioactive compounds on a mass scale if they are found.

Summary of the Invention

Technical problem [007] Thus, it is an objective of the present invention to provide new bioactive compounds, the use of these as therapeutic agents or prodrugs for cancer, in particular for acute myeloid leukemia, and a method to manufacture the same on a scale in pasta.

Technical Solution [008] In accordance with one aspect of this, the present invention provides new compounds, represented by the following chemical formula I:

[Chemical formula I]

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3/51

on what:

RaRa is a C1-C6 alkyl group, a C2-C6 alkenyl, or an alkynyl group

C2-C6;

Rb is an aryl group, a substituted aryl group, or -C (= O) Re where Re is a C1-C6 alkyl group, a C2-C6 alkenyl group, or a C2-C6 alkynyl group;

R _P is -H, -PO3H ² , -HPO3- Na ⁺ , -PO3 ² -Na ²⁺ , -PO3 ² -K2 ⁺ , -PO3 ² -Mg ²⁺ , -PO3 ² Ca ²⁺ , 'η, [ 009] The substituted arila can

be substituted acyl-aryl (as defined herein).

[0010] In one embodiment, in chemical formula I, Ra is a C1Οβ alkyl group or a C2-C6 alkenyl group; Rb is -C (= O) Re where Re is C1-C6 alkyl; and R _P is H, -PO3H2, -HPO3- Na ⁺ , or -PO ₃ ² -Na ²⁺ .

[0011] In another embodiment, in chemical formula I, Ra is methyl; Rb is (C = O) R _and where Re is C1-C6 alkyl; and Rp is -H.

[0012] In yet another modality, in chemical formula I, Ra is methyl; Rb is —C (= O) R _and where Re is C1-C6 alkyl; and Rp is -PO3H2, -HPO3- Na ⁺ , or -PO3 ² -Na ²⁺ .

[0013] In one aspect, the present description provides a pharmaceutical composition comprising a compound provided herein and a pharmaceutically acceptable excipient.

[0014] In another aspect, the present description provides a method for treating acute myeloid leukemia (AML) comprising administering to a patient

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4/51 with AML having an effective amount of the compound or composition provided herein. In certain embodiments, the method comprises injecting an effective amount of the compound or composition to a patient having AML.

[0015] In another aspect, the present description provides a method for making the compound provided herein, comprising the following sequential steps: (a) introducing an acyl group in indole-7-carbaldehyde by means of Friedel-Crafts acylation to provide 3-acyl-indole-7-carbaldehyde; (b) introducing an alkyl group and an aminoacetal group to 3-acyl-indol-7-carbaldehyde to provide a derivative of 1-alkyl-3-acyl-indole; (c) amide the 1-alkyl-3-acyl-indole derivative with Cbz-Tyrosine-OtBu stereoselectivity and 2- (1-allyl-4-benzylsemicarbazido) acetic acid to provide a reaction intermediate; (d) cyclizing the reaction intermediate in the presence of formic acid to provide a cyclic intermediate; and (e) phosphorylate the cyclic intermediate to provide a compound of the chemical formula (I). In certain embodiments, 2- (1-allyl-4-benzylsemicarbazido) acetic acid is synthesized by the following sequential steps: (1) adding TEA (triethylamine) to an ethylhydrazinoacetate solution to provide a reaction solution; (2) add allyl bromide to the reaction solution; and (3) add benzyl isocyanate. In certain additional embodiments, allyl bromide and benzyl isocyanate are added in a droplet manner.

[0016] In a related aspect, the present description provides a method for preparing a compound of the chemical formula (I), comprising: (a) converting

CHO

indol-7-carbaldehyde a ^R b, where Rb is an aryl group, a substituted aryl group, or -C (= O) R _and , where Re is a C1-C6 alkyl group, an alkenyl group

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C2-C6, or a C2-C6 alkynyl group; (b) convert

R _a

CHO

wherein R _a is a C1-C6 alkyl group, a C2-C6 alkenyl, or a R _b C2-C6 alkynyl group; (c) amide Eto

with stereoselectivity in the presence of Cbz-Tyrosine-OtBu and 2- (1-allyl-4-benzylsemicarbazido) acetic acid for R _b

Ot-Bu

Ν N NHBn

Η H (d) provide

Eto

Ot-Bu

// or 'N ^^ ^N ' N ^ NHBn Η H cyclize

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O-Rn in p, where Rp is -PO3H2, -HPO3- Na ⁺ , -PO3 ² Na ²⁺ , -PO3 ² -K2 ⁺ , -PO3 ² -Mg ²⁺ , -PO3 ² -Ca ²⁺ . In certain embodiments, Ra is methyl, Rb is -C (= O) R _e , and Re is methyl or cyclopropyl.

Advantageous Effect [0017] The new reverse binding mimics according to the present invention are observed to effectively inhibit the in vitro growth of AML cancer cells. Also, they are seen in tests of mouse models of acute myeloid leukemia to effectively inhibit tumor growth.

[0018] Without being bound by theory, it is believed that the leaving group (R _p ), also referred to as the functional prodrug group, is separated, the compounds of chemical formula I revolve in active forms. However, these active forms are difficult to prepare in an aqueous solution due to their poor solubility in water. In the prodrug forms, the compounds of chemical formula I according to the present invention are of high solubility and of high stability and are easy to be prepared as a preparation for injection.

[0019] Animal testing has shown that the compounds of the present invention have excellent pharmaceutical efficacy. This appears to be attributed to the rapid conversion of the compounds into their active forms shortly after the intravenous injection, and thus an increase in the initial drug concentration. In this way, the speed at which the prodrug compounds rotate in active forms has an influence on their medicinal efficacy, so that it is important to choose the functional group of prodrugs that allow ideal effects.

[0020] In a preferred embodiment, the functional prodrug groups

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7/51 are in the form of phosphate because phosphate prodrugs are converted faster in vivo to active forms than other prodrugs having other functional groups.

[0021] When the functional group of prodrugs is in the form of sodium salts, they are easy to prepare and have high water solubility. Furthermore, they are highly stable during storage at room temperature.

[0022] Usually, it is known that a suitable injection composition varies in pH from 4 to 9, and preferably has a pH that is close to human blood, 7.4. A composition that is strongly acidic or strongly basic is not preferred as an injection composition. In the case of a phosphate functional group, the final prodrugs of the present invention can be in the form of monosodium or disodium phosphate depending on the amount of sodium hydroxide. These compounds are advantageous for making a composition having pH values suitable for injection.

[0023] Furthermore, the manufacturing method according to the present invention allows the production not only of compounds of the chemical formula I, but also of imitators of reverse binding of it on an industrial scale.

Description of Drawings [0024] Figure 1 is a graph showing a correlation between changes in pH and the potential conducted during the final step of the method to manufacture the compound, in which 0.5 N NaOH is added in drops at 4- ( ((6S, 9aS) -1 (benzylcarbamoyl) -8 - ((3-acetyl-1-methyl-1 H-indol-7-yl) methyl) -2-allyl-octahydro-4,7-dioxo-1Hpyrazine [ 2,1-c] [1,2,4] triazin-6-yl) methyl) phenyl dihydrogen phosphate (Compound P2). In this graph, the horizontal axis represents the added amounts of NaOH. The first and second inflection points correspond to the beginning of monosodium and disodium production, respectively.

Best Mode

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8/51 [0025] Thus, one modality provides unprecedented reverse-binding mimics, represented by the following chemical formula I, which are used as therapeutic agents for cancer, in particular for acute myeloid leukemia.

[Chemical formula I]

on what

Rp can be any of the conventional functional groups that are available in prodrugs. Examples of functional groups include phosphate, carboxy, and C1-C6 alkylamino, and acylamino, such as -PO3H ² , -HPO3- Na ⁺ , -PO3 ² Na ²⁺ , -PO ₃ ² -K2 ⁺ , -PO3 ² -Mg ²⁺ , -PO ₃ ² -Ca ²⁺ o

ch ₃

O Ç ^H 3 ch ₃

N.

CH j

OU / X

VB ^CHa .

O

II —P-ORd [0026] Preferably, Rp is a phosphate functional group ( ^OR c) in which Rc and Rd are independently H, Na, Mg, Ca or K. Preferably, both Rc and Rd are H or Na, or one is Na while 0 the other is H.

[0027] R _p can also be -H, the resulting chemical structure in an active form of the corresponding prodrug as the functional group of the prodrug that is removed.

[0028] R _a is an alkyl group, an alkenyl group, or an alkynyl group; preferably a C1-C6 alkyl group, a C2-C6 alkenyl, or a C2-C6 alkynyl group; and more preferably a C1-C6 alkyl group.

[0029] Rb is an aryl group, a substituted aryl group, or -C (= O) Re where

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Re is a C1-C6 alkyl group, a C2-C6 alkenyl group, or a C2-C6 alkynyl group, and the substituted aryl group is an aryl substituted aryl group and preferably aryl substituted phenyl group.

[0030] The pro-drug compounds will transform into active forms in the body. When prodrugs have the phosphate functional group as a leaving group, the -POaRcRd group is rapidly cleaved by phosphatase and the prodrugs change in their active forms. At this point, Rp is changed to -H (a chemical structure in the active form as the functional group of the prodrug that left the structure).

[0031] As used herein, the term "alkyl" or "alkyl group" should include a straight, branched or cyclic hydrocarbon radical comprising carbon and hydrogen atoms, in which the carbon atoms are linked by simple bonds. In some embodiments, alkyl contains up to 20 carbons. In preferred embodiments, an alkyl can comprise one to six carbon atoms and be represented by "C1-C6 alkyl." An alkyl is attached to the rest of the molecule by a single bond. Examples of alkyls include, without limitation, methyl, ethyl, n-propyl, 1-methylethyl (iso-propyl), n-butyl, n-pentyl, n-hexyl, 1,1-dimethylethyl (t-butyl), 2, 2-dimethylpropyl (neo-pentyl), 3-methylhexyl, 2-methylhexyl, and the like. An alkyl may also be a monocyclic or bicyclic hydrocarbon ring radical, which may include fused or bonded ring systems. A cyclic alkyl is also referred to as cycloalkyl. In certain embodiments, a cycloalkyl may comprise three to six carbon atoms and may be represented by "C3-6 cycloalkyl." Examples of monocyclic cycloalkyl radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloeptyl, and cyclooctyl.

[0032] "Alkenyl" or "alkenyl group" refers to a straight, branched or cyclic hydrocarbon radical comprising carbon and hydrogen atoms, in which at least two carbon atoms are linked by a double bond. In some

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10/51 modalities, alkyl contains up to 20 carbons. In preferred embodiments, an alkenyl can comprise two to six carbon atoms and can be represented by "C2-C6 alkyl." An alkenyl is attached to the rest of the molecule by a single or double bond. Examples of alkenyls include, without limitation, ethylene, allyl, butenyl and the like.

[0033] "Alquinyl" or "alkynyl group" refers to a straight, branched or cyclic hydrocarbon radical comprising carbon and hydrogen atoms, in which at least two carbon atoms are linked by a triple bond. In some embodiments, alkyl contains up to 20 carbons. In preferred embodiments, an alkynyl can comprise two to six carbon atoms and can be represented by "C2-C6 alkynyl." An alkynyl is attached to the rest of the molecule by a single bond. Examples of alkynyls include, without limitation, ethynyl, 1-propynyl, or 2propynyl and the like.

[0034] Unless otherwise specifically stated in the specification, the term "alkyl" should include an alkyl having only carbon and hydrogen atoms, as well as "substituted alkyl," which refers to an alkyl radical in which one or more hydrogen atoms are replaced by one or more substituents independently selected from: acyl, alkoxy, aryl, cyano, cycloalkyl, halo, hydroxyl, nitro, -OC (O) -R ¹¹ , -N (R ¹¹ ) 2, -C (O) OR ¹¹ , -C (O) N (R ¹¹ ) 2, -N (R ¹¹ ) C (O) OR ¹¹ , -N (R ¹¹ ) C (O) R ¹¹ , -N (R ¹¹ ) S (O) tR ¹¹ (where t is 1 or 2), -S (O) tOR ¹¹ (where t is 1 or 2), -S (O) pR ¹¹ (where p is 0, 1 or 2), and -S (O) tN (R ¹¹ ) 2 (where t is 1 or 2) where each R ¹¹ is independently hydrogen, alkyl, aryl, as defined herein. The terms "alkenyl" and "alkynyl" are also defined as including "substituted alkenyl" and "substituted alkynyl," respectively.

[0035] "Aloxy" refers to a radical represented by the formula alkyl-O-, where alkyl is as defined herein. The alkyl portion can also be replaced by one or more halogen. An alkoxy can also be represented by the number of

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11/51 carbons in the alkyl group, for example, C1-6 alkoxy or C1-3 alkoxy.

[0036] "Acyl" refers to a radical represented by the formula R ¹² C (= O) -, where R ¹² is alkyl or aryl as defined herein. The alkyl or aryl can be optionally substituted by the substituents as described for an alkyl or an aryl group, respectively. Exemplary acyl groups include, without limitation, methylacyl (i.e., acetyl), phenylacyl, cyclopropylacil, and the like.

[0037] "Arila" refers to a radical derived from an aromatic monocyclic or bicyclic ring system removing a hydrogen atom from a carbon atom in the ring. The aromatic monocyclic or bicyclic hydrocarbon ring system comprises six to twelve carbon atoms (i.e., C6-12 aryl), where at least one of the rings in the ring system is completely unsaturated, that is, it contains an electron system π (4n + 2) delocalized, cyclic according to Hückel's theory. Examples of aryl radicals include, but are not limited to, phenyl and naphthyl. Unless otherwise specifically stated in the specification, the term "aryl" must include both aryl and "substituted aryl," which refers to an aryl radical in which one or more hydrogen atoms are replaced by one or more substituents independently selected from: acyl, alkoxy, aryl, cyano, cycloalkyl, halo, hydroxyl, nitro, -OC (O) -R ¹¹ , -N (R ¹¹ ) 2, -C (O) OR ¹¹ , -C (O) N (R ¹¹ ) 2, -N (R ¹¹ ) C (O) OR ¹¹ , -N (R ¹¹ ) C (O) R ¹¹ , -N (R ¹¹ ) S (O) tR ¹¹ (where t is 1 or 2), -S (O) tOR ¹¹ (where t is 1 or 2), -S (O) pR ¹¹ (where p is 0, 1 or 2), and -S (O) tN (R ¹¹ ) ₂ ( where t is 1 or 2) where each R ¹¹ is independently hydrogen, alkyl, aryl, as defined herein.

[0038] "Halo" refers to fluorine, chlorine, bromine and iodine.

[0039] The active form of the compounds is not suitable for I.V. injection due to their low solubility in an aqueous medium (for example, saline or water). The prodrug forms described here are suitable for I.V. injection due to their better solubility in the aqueous medium. In a preferred embodiment, a prodrug

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12/51 phosphate is used; and when one or two Na atoms are introduced into the phosphate fraction, the solubility is even better. To introduce Na atoms, sodium hydroxide is added (for example, in drops) to the phosphate compound at a specific pH value to perform replacement with one or two protons of the phosphate fraction with sodium ions.

Thus, an additional embodiment provides a pharmaceutical composition comprising a compound of the chemical formula (I) and a pharmaceutically acceptable excipient. The compounds or compositions of the present invention can be used in the treatment of AML in the manner described in more detail below.

[0041] The pharmaceutical composition of the present invention is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal or subcutaneous application may include the following components: a sterile diluent, such as water for injection, saline, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents, such as benzyl alcohol or methyl parabens; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylene diaminetetraacetic acid; buffers, such as acetates, citrates or phosphates and agents for adjusting tonicity, such as sodium chloride or dextrose.

[0042] In a preferred embodiment, the pharmaceutically acceptable excipient is suitable for use in I.V. administration, such as injection or infusion

I.V .. Suitable carriers for I.V. administration include physiological saline, bacteriostatic water, Cremophor EL ™ (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and must be fluid

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13/51 to the point where the syringe easily comes out. It must be stable in the conditions of manufacture and storage and must be preserved against the action of contamination by microorganisms, such as bacteria and fungi.

[0043] In other embodiments, oral compositions that generally include an inert diluent or an edible carrier are provided. Such compositions can be enclosed in gelatin capsules or tablets in the form of tablets. For the purpose of oral therapeutic administration, the compound described herein can be incorporated with excipients and used in the form of tablets, lozenges, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, in which the compound in the fluid carrier is applied orally and shaken and expectorated or ingested. Pharmaceutically compatible binding agents, and / or adjuvant materials can be included as part of the composition. Tablets, pills, capsules, lozenges and the like can contain any of the following ingredients, or compounds of a similar nature: a binder, such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient, such as starch or lactose, a disintegrating agent, such as alginic acid, Primogel, or corn starch; a lubricant, such as magnesium stearate or Sterotes; a glidant, such as colloidal silicon dioxide; a sweetening agent, such as sucrose or saccharin; or a flavoring agent, such as mint, methyl salicylate, or orange flavoring.

[0044] According to another aspect, the present description provides a method of treating diseases, particularly cancer, more particularly acute myeloid leukemia (AML) comprising administering to a cancer patient (e.g., a patient with AML) an amount effect of a compound of the chemical formula (I) or a pharmaceutical composition comprising the same. Example 23 provided below demonstrates that exemplary compounds of the present description are effective in treating AML in an animal model.

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14/51 [0045] Examples of the compounds of the chemical formula (I) are given in the table

1, below. Because the four compounds in Table 1 are different only in the fraction Rp that is H or phosphate functional group having the same NMR data, it is commonly given in Table 1 (Rp was not observed in the spectra of

1H NMR due to having been replaced by deuterium).

TABLE 1

At the. Cpd. M.W. NMR 1 rX ^h ÇàU ⁰ 736.69 1H NMR (500MHz, CDCI3) δ 8.43 (d, J = 4.8 Hz, 1H), 7.63 (s, 1H), ° XX ^ ° ^Na ΌΗ 7.38-7.35 (m, 2H), 7.317.30 (m, 1H), 7.29-7.21 (m, 2H), 7.00 (d, J = 4.8 Hz, 2H), 6.97 (d, J = 4.8 Hz, 1H), 6.69-6.65 (m, 2 ψςΑθν ° 758.67 ° XX ^ ° ^Na 3H), 5.87 (s, 1H), 5,555.44 (m, 3H), 5.34 (t, J = 4.6 Hz, 1H), 5.03 (d, 3 714.70 ^ • '' VOAílo yA ⁰ AA ° ^ TJ-P-OH J = 6.3 Hz, 1H), 4.87 (d, J = 9.0 Hz, 1H), 4.79 (d, J = 7.5 Hz, 1H), 4.42 (dd, Oh J = 9.0, 3.6 Hz, 1H), 4.29 4 634.72 (dd, J = 9.0, 3.6 Hz, 1H), 4.02 (s, 3H), 3.43 (d, J = 7.2 Hz, 1H), 3.38-3.33 ⁰ AA ^ OH (m, 3H), 3.27 (d, J = 7.2 Hz, 1H), 3.29-3.24 (m,

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1H), 3.18 (dd, J = 7.2, 2.4 Hz, 1H), 2.51 (s, 3H) 5 ÜMy ⁰ V / Ã ÇÇyU ⁰ 792.79 1H NMR (D2O, 300MHz) δ 7.27 (d, 2H, J = 8.4 Hz), 7.175 (d, 1H, ° XX ^ ° ^Na J = 7.2 Hz), 6.37-6.31 (m, 3H), 6.214 (d, 2H), 6 COy> γΧ 814.77 6.14-6.07 (m, 4H), 4.51-4.46 (dd, 2H, ΥΥ ₀ Μ ⁰ J = 10.8 Hz), 4.31 -4.04 ° XX Ã ° ^Na (dd, 2H, J = 14.7 Hz), ONa 3.39-3.34 (d, 1H, J = 8.4 Hz), 3.34-2.97 (dd, 2H, 7 CXx _r ° \ X N ^N N 770.81 J = 15.3, 15.3 Hz), 4.33 (dd, 1H, J = 15.3, 6.3 Hz), ° Ul 2.97 (s, 3H), 2.75 (d, | 1H), 2.49-2.05 (dd, 2H, OH J = 15.3Hz), 1.19 (s, 9H) 8 uuç ° \ ~ y N ' ^N Y * ^ N ^{/ X /} Ã ^{/ k} / Υ ^ ΑθΧΧ 690.83 ° XX

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16/51

Η

Νγ> 0

Ο '

Ό — P-ONa ΟΗ

762.72

784.71

Η

Ο — P-ONa ÓNa ^<5% ^ Ν ^Νχ 1 ^ 'Ν

Η

NyO

Ο '

740.74

Η

N _r O

Η / Ρ

660.76 ^ Οχσ £

Ο

ΟΗ

NMR 1Η (300ΜΗζ, CDCI3) δ 8.43 (d, J = 4.8 Hz, 1Η), 7.63 (s, 1H), 7.38-7.35 (m, 2H), 7.317.30 ( m, 1H), 7.29-7.21 (m, 2H), 7.00 (d, J = 4.8 Hz, 2H), 6.97 (d, J = 4.8 Hz, 1H), 6.69-6.65 (m, 3H), 5.87 (s, 1H), 5,555.44 (m, 3H), 5.34 (t, J = 4.6 Hz, 1H), 5.03 (d, J = 6.3 Hz, 1H), 4.87 (d, J = 9.0 Hz, 1H), 4.79 (d, J = 7.5 Hz, 1H), 4.42 (dd , J = 9.0, 3.6 Hz, 1H), 4.29 (dd, J = 9.0, 3.6 Hz, 1H), 4.02 (s, 3H), 3.43 (d, J = 7.2 Hz, 1H), 3.38-3.33 (m, 3H), 3.27 (d, J = 7.2 Hz, 1H), 3.29-3.24 (m, 1H ), 3.18 (dd, J = 7.2, 2.4 Hz, 1H), 1.28 (m, 1H), 0.63 (m, 2H), 0.38 (m, 2H)

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17/51

778.77

^{0 P} ~ ONa ONa

800.75

756.78

676.80

1H NMR (300MHz, CDCI3) δ 8.43 (d, J = 4.8 Hz, 1H), 7.63 (s, 1H),

7.38-7.35 (m, 2H), 7.317.30 (m, 1H), 7.29-7.21 (m, 2H), 7.00 (d, J = 4.8 Hz, 2H) , 6.97 (d, J = 4.8 Hz, 1H), 6.69-6.65 (m, 3H), 5.87 (s, 1H), 5,555.44 (m, 3H), 5, 34 (t, J = 4.6 Hz, 1H), 5.03 (d, J = 6.3 Hz, 1H), 4.87 (d, J = 9.0 Hz, 1H), 4.79 ( d, J = 7.5 Hz, 1H), 4.42 (dd, J = 9.0, 3.6 Hz, 1H), 4.29 (dd, J = 9.0, 3.6 Hz, 1H ), 4.02 (s, 3H), 3.43 (d, J = 7.2 Hz, 1H), 3.38-3.33 (m, 3H), 3.27 (d, J = 7, 2 Hz, 1H), 3.29-3.24 (m, 1H), 3.18 (dd, J = 7.2, 2.4 Hz, 1H), 2.51 (d, J = 5.0 Hz, 2H), 2.06 (m, 1H), 1.01 (d, J = 5.2 Hz, 6H)

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18/51

764.74

phono

ONa

786.72

742.76

662.78

1H NMR (300MHz, CDCI3) δ 8.43 (d, J = 4.8 Hz, 1H), 7.63 (s, 1H), 7.38-7.35 (m, 2H), 7.317.30 ( m, 1H), 7.29-7.21 (m, 2H), 7.00 (d, J = 4.8 Hz, 2H), 6.97 (d, J = 4.8 Hz, 1H), 6.69-6.65 (m, 3H), 5.87 (s, 1H), 5,555.44 (m, 3H), 5.34 (t, J = 4.6 Hz, 1H), 5.03 (d, J = 6.3 Hz, 1H), 4.87 (d, J = 9.0 Hz, 1H), 4.79 (d, J = 7.5 Hz, 1H), 4.42 (dd , J = 9.0, 3.6 Hz, 1H), 4.29 (dd, J = 9.0, 3.6 Hz, 1H), 4.02 (s, 3H), 3.43 (d, J = 7.2 Hz, 1H), 3.38-3.33 (m, 3H), 3.27 (d, J = 7.2 Hz, 1H), 3.29-3.24 (m, 1H ), 3.18 (dd, J = 7.2, 2.4 Hz, 1H), 2.51 (d, J = 5.0 Hz, 2H), 1.66 (m, 2H), 0.98 (t, J = 4.2 Hz, 3H)

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776.75

Ο — Ρ \ ONa

ONa

798.73

754.77

674.79

1H NMR (300MHz, CDCI3) δ 8.43 (d, J = 4.8 Hz, 1H), 7.63 (s, 1H),

7.38- 7.35 (m, 2H), 7.317.30 (m, 1H), 7.29-7.21 (m, 2H), 7.00 (d, J = 4.8 Hz, 2H), 6 , 97 (d, J = 4.8 Hz, 1H), 6.69-6.65 (m, 3H), 5.87 (s, 1H), 5,555.44 (m, 3H), 5.34 ( t, J = 4.6 Hz, 1H), 5.03 (d, J = 6.3 Hz, 1H), 4.87 (d, J = 9.0 Hz, 1H), 4.79 (d, J = 7.5 Hz, 1H), 4.42 (dd, J = 9.0, 3.6 Hz, 1H), 4.29 (dd, J = 9.0, 3.6 Hz, 1H), 4.02 (q, J = 4.8 Hz, 2H), 3.43 (d, J = 7.2 Hz, 1H),

3.38 - 3.33 (m, 3H), 3.27 (d, J = 7.2 Hz, 1H), 3.293.24 (m, 1H), 3.18 (dd, J = 7.2, 2, 4 Hz, 1H), 1.51 (t, J = 5 Hz, 3H), 1.28 (m, 1H), 0.63 (m, 2H), 0.38 (m, 2H)

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⁰ ^ ONa OH

788.76

\ —ON ONa ONa

810.74

766.78

686.80

1H NMR (300MHz, CDCI3) δ 8.43 (d, J = 4.8 Hz, 1H), 7.63 (s, 1H), 7.38-7.35 (m, 2H), 7.317.30 ( m, 1H), 7.29-7.21 (m, 2H), 7.00 (d, J = 4.8 Hz, 2H), 6.97 (d, J = 4.8 Hz, 1H), 6.69-6.65 (m, 3H), 5.87 (s, 1H), 5.83 (m, 1H), 5.55-5.44 (m, 3H), 5.34 (t, J = 4.6 Hz, 1H), 5.17 (m, 2H), 5.03 (d, J = 6.3 Hz, 1H), 4.87 (d, J = 9.0 Hz, 1H) , 4.79 (d, J = 7.5 Hz, 1H), 4.42 (dd, J = 9.0, 3.6 Hz, 1H), 4.29 (dd, J = 9.0, 3 , 6 Hz, 1H), 4.02 (m, 2H), 3.43 (d, J = 7.2 Hz, 1H), 3,383.33 (m, 3H), 3.27 (d, J = 7 , 2 Hz, 1H), 3.29-3.24 (m, 1H), 3.18 (dd, J = 7.2, 2.4 Hz, 1H), 1.28 (m, 1H), 0 63 (m, 2H), 0.38 (m, 2H)

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⁰ % ONa OH

778.77

O-P \ ONa

ONa

800.75

756.78

676.80

1H NMR (300MHz, CDCI3) δ 8.43 (d, J = 4.8 Hz, 1H), 7.63 (s, 1H), 7.38-7.35 (m, 2H), 7.317.30 ( m, 1H), 7.29-7.21 (m, 2H), 7.00 (d, J = 4.8 Hz, 2H), 6.97 (d, J = 4.8 Hz, 1H), 6.69-6.65 (m, 3H), 5.87 (s, 1H), 5,555.44 (m, 3H), 5.34 (t, J = 4.6 Hz, 1H), 5.03 (d, J = 6.3 Hz, 1H), 4.87 (d, J = 9.0 Hz, 1H), 4.79 (d, J = 7.5 Hz, 1H), 4.42 (dd , J = 9.0, 3.6 Hz, 1H), 4.29 (dd, J = 9.0, 3.6 Hz, 1H), 4.02 (s, 3H), 3.43 (d, J = 7.2 Hz, 1H), 3.38-3.33 (m, 3H), 3.27 (d, J = 7.2 Hz, 1H), 3.29-3.24 (m, 1H ), 3.18 (dd, J = 7.2, 2.4 Hz, 1H), 2.51 (d, J = 5.0 Hz, 2H), 1.62 (m, 2H), 1.33 (m, 2H), 0.96 (t, J = 4.0 Hz, 3H)

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22/51

ο

UxONa Ο'ΌΗ

750.71

u ^ ONa 0'ONa

772.69

728.73

648.75

1H NMR (300MHz, CDCI3) δ 8.43 (d, J = 4.8 Hz, 1H), 7.63 (s, 1H), 7.38-7.35 (m, 2H), 7.317.30 ( m, 1H), 7.29-7.21 (m, 2H), 7.00 (d, J = 4.8 Hz, 2H), 6.97 (d, J = 4.8 Hz, 1H), 6.69-6.65 (m, 3H), 5.87 (s, 1H), 5,555.44 (m, 3H), 5.34 (t, J = 4.6 Hz, 1H), 5.03 (d, J = 6.3 Hz, 1H), 4.87 (d, J = 9.0 Hz, 1H), 4.79 (d, J = 7.5 Hz, 1H), 4.42 (dd , J = 9.0, 3.6 Hz, 1H), 4.29 (dd, J = 9.0, 3.6 Hz, 1H), 4.02 (s, 3H), 3.43 (d, J = 7.2 Hz, 1H), 3.38-3.33 (m, 3H), 3.27 (d, J = 7.2 Hz, 1H), 3.29-3.24 (m, 1H ), 3.18 (dd, J = 7.2, 2.4 Hz, 1H), 2.44 (q, J = 4.1 Hz, 2H), 1.18 (t, J = 4.1 Hz , 3H)

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764.74

LxONa O 'ONa

786.72

742.76

662.78

1H NMR (300MHz, CDCI3) δ 8.43 (d, J = 4.8 Hz, 1H), 7.63 (s, 1H), 7.38-7.35 (m, 2H), 7.317.30 ( m, 1H), 7.29-7.21 (m, 2H), 7.00 (d, J = 4.8 Hz, 2H), 6.97 (d, J = 4.8 Hz, 1H), 6.69-6.65 (m, 3H), 5.87 (s, 1H), 5,555.44 (m, 3H), 5.34 (t, J = 4.6 Hz, 1H), 5.03 (d, J = 6.3 Hz, 1H), 4.87 (d, J = 9.0 Hz, 1H), 4.79 (d, J = 7.5 Hz, 1H), 4.42 (dd , J = 9.0, 3.6 Hz, 1H), 4.29 (dd, J = 9.0, 3.6 Hz, 1H), 3.85 (m, 2H), 3.43 (d, J = 7.2 Hz, 1H), 3.38-3.33 (m, 3H), 3.27 (d, J = 7.2 Hz, 1H), 3.29-3.24 (m, 1H ), 3.18 (dd, J = 7.2, 2.4 Hz, 1H), 2.51 (s, 3H), 1.81 (m, 2H), 0.96 (t, J = 4, 3 Hz, 3H)

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41 θΧ ° “to Y γγθυ ⁰ ° X.? - ° ^Na 820.85 1H NMR (D2O, 300MHz) δ 7.27 (d, 2H, J = 8.4 Hz), 7.175 (d, 1H, J = 7.2 Hz), 6.37-6.31 (m, 3H), 6.214 (d, 2H), 6.14-6.07 (m, 4H), 4.51-4.46 (dd, 2H, J = 10.8 Hz), 4.31 -4.04 (dd, 2H, J = 14.7 Hz), 3.39-3.34 (d, 1H, J = 8.4 Hz), 3.34-2.97 (dd, 2H, J = 15.3, 15.3 Hz), 4.33 (dd, 1H, J = 15.3, 6.3 Hz), 2.97 (m, 2H), 2.75 (d, 1H), 2.49-2.05 (dd, 2H, J = 15.3Hz), 1.92 (m, 2H), 1.20 (s, 9H), 1.01 (t, J = 4.3 Hz, 3H) 42 0o _r ° -Ύ ÇU U ^{0 0} Sí ^ l 0 || 1 ΐ / ONa ^ '' ^ 0 'ONa 842.83 43 CO _Y ° <ζ Y ⁰ ΎΊ j? .OH 798.86 44 X H? ~ -Y Y-A, U ° ° X. 718.88

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45 ÇCa ₀ U ^{0 0} o I / 1 ü- ^0Na ^^ 0 '''OH 862.93 1H NMR (D2O, 300MHz) δ 7.27 (d, 2H, J = 8.4 Hz), 7.175 (d, 1H, J = 7.2 Hz), 6.37-6.31 (m, 3H), 6.214 (d, 2H), 6.14-6.07 (m, 4H), 4.51-4.46 (dd, 2H, J = 10.8 Hz), 4.31 -4.04 (dd, 2H, J = 14.7 Hz), 3.39-3.34 (d, 1H, J = 8.4 Hz), 3.34-2.97 (dd, 2H, J = 15.3, 15.3 Hz), 4.33 (dd, 1H, J = 15.3, 6.3 Hz), 2.97 (m, 2H), 2.75 (d, 1H), 2.49-2.05 (dd, 2H, J = 15.3Hz), 1.77 - 1.30 (m, 8H), 1.20 (s, 9H) 0.98 (t, J = 5.0 Hz, 3H) 46 CO _r ° ÇaU ^{0 0} So θ l Jl k ^{0Na x} ONa 884.91 47 ^[M Al <-Ύ CCA nJ U ⁰ ₀ ⁰ o Ii ^0H OH 840.94 48 Al ^h / -y MOy ⁰ < _N A ° 760.96 49 (A ^H <W ^N y ° < _N N çUoU ° ⁰ ΎΊ ° II ^ ° - ^ ONa OH 804.80 1H NMR (300MHz, CDCI3) δ 8.43 (d, J = 4.8 Hz, 1H), 7.63 (s, 1H), 7.38-7.35 (m, 2H), 7.317.30 (m, 1H), 7.29-7.21

O

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50 826.78 (m, 2H), 7.00 (d, J = 4.8 Hz, 2H), 6.97 (d, J = 4.8 Hz, 1H), 6.69-6.65 (m, 3H), 5.87 (s, 1H), 5,555.44 (m, 3H), 5.34 (t, J = 4.6 Hz, 1H), 5.03 (d, J = 6.3 Hz, 1H), 4.87 (d, J = 9.0 Hz, 1H), 4.79 (d, J = 7.5 Hz, 1H), 4.42 (dd, J = 9.0, 3.6 Hz, 1H), 4.29 (dd, J = 9.0, 3.6 Hz, 1H), 4.02 (m, 2H), 3.43 (d, J = 7.2 Hz, 1H), 3.38-3.33 (m, 3H), 3.27 (d, J = 7.2 Hz, 1H), 3.29-3.24 (m, 1H), 3.18 (dd, J = 7.2, 2.4 Hz, 1H), 1.77 (m, 2H), 1.88 (m, 2H), 1.28 (m, 1H), 0.98 (t, J = 4.8 Hz, 3H), 0.63 (m, 2H), 0.38 (m, 2H) 51 Ol VV i> ÇU. / U ⁰ ° Vi ° ^ θ-ΧοΗ OH 782.82 52 ^H <ÇX V ° ° U 702.84 53 Qx _r ° \ ^ VfX0? γΧ V \ ⁰ 9 _z ^Na ^ q_P_q OH 812.78 1H NMR (CDCI3, 300MHz) δ 8.05 (d, 2H, J = 8.4 Hz), 7.91 (d, 1H, J = 7.2 Hz), 7.71 (d, 2H, J = 8.4 Hz), 7.40-7.20 (m,

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54 ° XX * P-0 ONa 834.76 4H), 7.16 (t, 1H, J = 7.2Hz), 7.05 (d, 2H, J = 8.4 Hz), 6.96 (d, 1H, J = 6.9 Hz), 6.69 (d, 2H, J = 8.4 Hz), 6.68 (m, 1H), 5.58-5.44 (m, 3H), 5.37 (t, 1H, J = 5.7 Hz), 5.03 (d, 1H, J = 10.8 Hz), 4.97 (d, 1H, J = 14.7 Hz), 4.81 (d, 1H, J = 17.1 Hz), 4.47 (dd, 1H, J = 15.3, 6.3 Hz), 4.33 (dd, 1H, J = 15.3, 6.3 Hz), 4.33 (s, 3H), 3.47-3.24 (m, 8H), 2.64 (s, 3H)

[0046] Methods known in the art can be used to determine the effectiveness of a compound provided herein in the treatment of cancer, such as AML. For example, the method described in example 23 can be used to estimate the anticancer activity of a given compound. Additional exemplary methods for estimating the activity of a compound in the treatment of AML include those described in Bishop et al., Blood 87: 1710-7, 1996; Bishop, Semin Oncol 24: 57-69, 1997; and Estey, Oncology 16: 343-52, 2002.

[0047] The compounds of the present description can be administered to a patient in need of him through various routes, such as oral, topical, transdermal or parenteral. In one embodiment, the compounds or compositions thereof are administered parenterally. The term “parenteral,” as used here

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28/51 used, includes subcutaneous, intravenous, intramuscular injection, intracisternal injections, and intravenous infusions. In preferred embodiments, the compounds or compositions are administered by injection, such as intravenous injections.

[0048] Toxicity and therapeutic efficacy of the compounds of the present description can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine LD50 (the lethal dose for 50% of the population) and ED50 (the therapeutically dose effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and can be expressed as the LD50 / ED50 ratio. Compounds that have high therapeutic indexes are preferred. Although compounds that have toxic side effects can be used, care must be taken to design a delivery system that targets such compounds to the affected tissue site in order to minimize potential damage to uninfected cells and thereby reduce side effects .

[0049] The data obtained from cell culture assays and animal studies can be used in the formulation of a dosage range for use in humans. The dosage of such compounds preferably falls within a range of circulating concentrations that include the ED50 with little or no toxicity. The in vitro cardiotoxicity of the compounds can be determined according to the method described in example 24 below. The dosage may vary in this range depending on the dosage form employed and the route of administration used. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC 50 (i.e., the concentration of the test compound that achieves half-maximum inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine

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29/51 useful doses in humans. Plasma levels can be measured, for example, by high performance liquid chromatography.

[0050] The effective dose depends on the type of disease, the composition used, the route of administration, the type of subject being treated, the physical characteristics of the specific subject under consideration for treatment, the concurrent medication and other factors than those versed in technology will realize. For example, to treat AML, a compound of the present description can be administered by injection or IV infusion in an amount between 0.5 mg / kg and 500 mg / kg (for example, 0.5 to 10 mg / kg, 10 to 100 mg / kg, about 100 to 500 mg / kg body weight) that can be administered as a single, daily, seminal, monthly dose, or at any appropriate interval. In certain embodiments, the compounds described can be used in the treatment of AML in a similar manner to that used for Ara-C.

[0051] In accordance with a further aspect of this, the present invention provides a method for making the reverse link mimics of the present invention on a mass scale. The method comprises the following sequential steps:

introducing an acyl group in indole-7-carbaldehyde, preferably by means of Friedel-Crafts acylation to provide 3-acyl-indole-7-carbaldehyde;

introducing an alkyl group and an aminoacetal group to 3-acyl-indol-7-carbaldehyde to provide a 1-alkyl-3-acyl-indole derivative;

amide the 1-alkyl-3-acyl-indole derivative with stereoselectivity with CbzTyr (OtBu) (i.e., (S) -2- (benzyloxycarbonylamino) -3- (4-tert-butoxyphenyl) propanoic acid) and 2- (1 -alyl-4-benzylsemicarbazide) acetic to provide a reaction intermediate;

cyclize the reaction intermediate in the presence of formic acid to provide a cyclic intermediate; and

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30/51 phosphorylate the cyclic intermediate.

[0052] In the previous method, 2- (1-allyl-4-benzylsemicarbazido) acetic acid can be prepared by the following sequential steps:

add TEA (triethylamine) to an ethylhydrazinoacetate solution to form a reaction solution;

add allyl bromide (for example, in drops) to the reaction solution; and add benzyl isocyanate (for example, in drops).

[0053] Representative compounds of the invention can be prepared from

7. formic acid (85%) 8. POCI ₃ , TEA

9. phosphorylation

10. lyophilization

[0054] In certain embodiments, Ra is methyl, Rb is -C (= O) R _e , and Re is methyl or cyclopropyl.

[0055] As seen here, the reaction scheme is directed to reverse bonded mimics, represented by chemical formula I.

[0056] The compounds according to the present invention are based on a pyrazine-triazinone framework, with four different functional groups attached to it. Due to their two chiral centers, the compounds must be synthesized stereoselectively.

[0057] An acyl group is introduced into the AA1 indole-7-carbaldehyde via

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31/51 of Friedel-Crafts acylation, followed by the introduction of alkyl and amino acid groups.

[0058] After the reaction of AA2 with the chiral compound (Cbz-Tyrosine-OtBu), the resulting intermediate is subjected to stereoselective amidation with PivCI (Pivaloyl chloride) and iBCF (isobutyl chloroformate) to make AA3 available. Thereafter, AA3 is cyclized with formic acid to obtain AA4, followed by phosphorylization, introduction of salt (adding Na to phosphate using 0.5 N NaOH) and lyophilization to synthesize highly pure pyrazine-triazone compounds, AA5.

Mode for the Invention [0059] A better understanding of the present invention can be obtained by means of the following examples which are presented to illustrate, but are not to be considered as limiting the present invention.

[0060] As shown here, the compounds of chemical formula I have anti-cancer activity.

[0061] The manufacturing method of the present invention is illustrated in detail as follows.

<Reaction scheme 1>

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[0062] In reaction scheme 1, side chain S3 can be prepared as shown below.

H tf

C ^ HuCINjO; Md, Wr: 154-6

Ally IbroTi itJií TEA

THF, rt 5h

EerkZylisocyúnetê rt, 3 min

CH _U N, O, Mol. Wt.:l5a.2

S-SH

O <0

Η H

CrHjiHjC-, Mol. Wt,: 231.36 S2 aq. KOH

O

€ _n H ₁₇ N ₅ 0j Mol. Wt .; 263 20

SS [0063] In the following, each step of the manufacturing method illustrated in reaction scheme 1 will be described in detail in examples 1 to 10.

EXAMPLE 1 [0064] Synthesis of S3

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33/51 2- (1-allyl-4-benzylsemicarbazido) acetic acid [0065] 67 g of ethyl hydrazinoacetate was dissolved in 673 ml of THF (tetrahydrofuran) and mixed with 121 ml of TEA (triethylamine). To this reaction mixture, 41 ml of allyl bromide were added in drops for 20 min. This solution was stirred for 5 hours and filtered. To the filtrate, drops of 53 mL of benzyl isocyanate were added for 15 min, followed by stirring for 30 min at room temperature. Thereafter, a solution of 48 g of KOH (potassium hydroxide) in 673 ml of distilled water was added in drops before stirring for 30 min. The separation of the layers was generated by adding 403 ml of MC (dichloromethane) and 269 ml of hexane and stirring. The aqueous solution was washed once with 201 ml of MC (dichloromethane). The aqueous solution was adjusted to a pH of 2 ~ 3 using 100 ml of concentrated HCI. After being stirred for 30 min, the pH-adjusted solution was extracted with 1009 ml of MC (dichloromethane). The layer of MC (dichloromethane) thus obtained was dehydrated with 269 g of Na2SO4, filtered and then concentrated in a vacuum. The concentrate is crystallized with 134 ml of EA (ethyl acetate) and 269 ml of hexane, followed by filtration. The solid thus obtained was plated in 134 ml of EA (ethyl acetate), filtered at 0 ^s C and dried in a vacuum to produce 40 g of S3 as a white solid (35% yield).

[0066JRMN 1H (500MHz, CDCI3) δ 10.84 (bs, 1H), δ 7.90 (s, 1H), δ 7.4-7.3 (m, 5H), δ 6.42 (t, J = 5.0 Hz, 1H), δ 5.85-5.72 (m, 1H), δ 5.28 (dd, J = 28.5, 2.0 Hz, 1 Η), δ 5.19 ( d, J = 17 Hz, 1 Η), δ 4.47-4.42 (m, 2H), δ 3.70 (dd, J = 40.0, 2.5 Hz, 1H).

EXAMPLE 2 [0067] Synthesis of P9 [0068] 3-Acetyl-1H-indole-7-carbaldehyde [0069) 23.5 ml of AcCI (acetyl chloride) were added dropwise to a solution of 55 g of AICI3 in 400 mL of MC (dichloromethane) with stirring. To this solution, a 40 g solution of

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34/51 broken (indole-7-carbaldehyde) in 400 ml of MC (dichloromethane). The temperature of the solution must be maintained at 0 ~ 5 ^s C upon addition and then naturally increased to room temperature. The progress of the reaction was monitored using thin layer chromatography (TLC) and high performance liquid chromatography. After the reaction was completed, the solution was subjected to separation of the layers with water. The organic layer thus formed was dried over MgSO4 (magnesium sulfate), filtered and then concentrated at 40 ^s C to give 41 g of P9 as a concentrated residue (80% yield).

EXAMPLE 3 [0070] Synthesis of P8 [0071] 3-Acetyl-1-methyl-1 H-indole-7-carbaldehyde [0072] 41 g of P9 was dissolved in 412 ml of DMF (dimethylformamide) and stirred. After the solution was cooled to 10 ^s C, 91 g of K2CO3 (potassium carbonate) were added to it, and 20 ml of honey (methyl iodide) was added in drops. The resulting solution naturally raised the temperature to room temperature and was stirred for 4 ~ 5 hrs. When the starting material was recognized to have disappeared, K2CO3 was filtered, followed by crystallization from hexane to give 35 g of P8 as a yellowish solid (80% yield).

EXAMPLE 4 [0073] Synthesis of P7 [0074] 1 - (7 - ((2,2-Diethoxyethylamino) methyl) -1-methyl-1 H-indol-3-yl) ethanone [0075] To a 35 g solution of P8 in 354 ml of MeOH (methanol) 3.5 ml of AcOH (acetic acid) were added. The solution was mixed with 33 ml of diethylacetal aminoacetaldehyde at room temperature and stirred for 3 ~ 4 hrs. After the solution was cooled to 10 ^s C, 3.3 g of the reducing agent NaBH4 (sodium borohydride) was slowly added to it. At this time, care must be taken due to the generation of hydrogen gas and exothermic reaction. The solution was stirred at

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35/51 room temperature for 1 hr. When the reaction was complete, 354 ml of EA (ethyl acetate) and 354 ml of distilled water were added in order to separate the layers. The organic layer thus formed was dried over 141 g of

MgSO4 (magnesium sulfate) and crystallized in hexane to provide 85 g of

P7 as a yellowish solid (80% yield).

[0076] 1H NMR (500MHz, CDCI3), δ 8.36 (d, J = 4.8 Hz, 1H), δ 7.61 (s, 1H), δ 7.17 (d, J = 4.2 Hz, 1 Η), δ 7.10 (d , J = 4.2 Hz, 1 Η), δ 4.58 (t, J = 3.3, 1 Η), δ 4.21 (s, 3H), δ 4.07 (s, 3H), δ 3.68 (m, 2H), δ 3.51 ( m, 2H), δ 2.82 (d, J = 3.3 Hz, 2H), δ 2.48 (s, 3H), δ 1.19 (t, J = 4.2 Hz, 6H).

EXAMPLE 5 [0077] Synthesis of P6 [0078] (S) -1 - (N - ((3-acetyl-1-methyl-1 H-indol-7-yl) methyl) -N- (2,2diethoxyethyl) carbamoyl ) Benzyl -2- (4-tert-butoxyphenyl) ethylcarbamate [0079] 85 g of Cbz-Tyr (OtBu) was dissolved in 449 ml of EA (ethyl acetate) with stirring. After the solution was cooled to 0 ~ 5 ^s C, 31 ml of NMM (Nmethylmorpholine) and 19 ml of pivaloyl chloride were added dropwise to it. The solution was stirred for 1 ~ 2 hrs and then 44.9 g of P7 was added to it at 0 ~ 5 ^s C. The solution was warmed to room temperature followed by stirring for 2 ~ 3 h. After the end of the reaction, distilled water was added to generate separation of the layers. The organic layer thus formed was washed with 898 ml of a 5% aqueous solution of citric acid and 898 ml of a 5% aqueous solution of NaHCO3 and then dried over 179 g of MgSO4 (magnesium sulfate) to be concentrated. 85 g of P6 were obtained as a residue (90% yield).

EXAMPLE 6

Synthesis of P5 [0080] (S) -3- (4-tert-butoxyphenyl) -N - ((3-acetyl-1-methyl-1 H-indol-7-yl) methyl) -2amino-N- (2 , 2-diethoxyethyl) propanamide

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36/51 [0081] Α 85 g of P6 in 853 ml of MeOH was added 8.5 g of 10% by weight of Pd / C. 16 g of ammonium formate were added and then refluxed for 2 hours. After completing the reaction, the solution was cooled to room temperature and Pd / C was filtered. The solution was concentrated before separating the layers with 853 ml of EA (ethyl acetate) and 1706 ml of distilled water. The organic layer thus formed was washed with 850 ml of a 5% aqueous solution of citric acid and 850 ml of a 5% aqueous solution of NaHCO3 and concentrated to give 56 g of P5 (90% yield).

EXAMPLE 7 [0082] Synthesis of P4 [0083] 40 g of side chain S3 was dissolved in 426 ml of EA (ethyl acetate) and cooled to -10 ^s C. To the solution, 41 ml of NMM (Nmethylmorpholine) was added in drops. and 20 ml of iBCF (iso-butylchloroformate) at the same temperature. The reaction mixture was stirred for 2 ~ 3 hours at -10 ^s C after which a solution of 56 g of P5 in 200 ml of EA (ethyl acetate) was added dropwise to it. The reaction mixture was warmed to room temperature and then stirred for 1 ~ 2 hours. When the reaction was over, EA (ethyl acetate) and 850 mL of distilled water were added to separate the layers. The organic layers thus formed were washed with 850 ml of a 5% aqueous solution of citric acid and 850 ml of a 5% aqueous solution of NaHCO3 and dried over 340 g of MgSO4 (magnesium sulfate) for concentration. 81 g of P4 was obtained as a concentrated residue (90% yield).

EXAMPLE 8 [0084] Synthesis of P3 [0085] (6S, 9aS) -6- (4-Hydroxybenzyl) -8 - ((3-acetyl-1-methyl-1H-indol-7-yl) methyl) -2allyl- N-benzyl-hexahydro-4,7-dioxo-2H-pyrazine [2,1-c] [1,2,4] triazine-1 (6H) -carboxamide [0086] 81 g of P4 were dissolved in 383 ml of 85% formic acid and

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37/51 heated to 50 ^s C. After being stirred for 1-2 hours at the same temperature, the solution was cooled to room temperature and mixed with acetone. This solution was adjusted to a pH of 4.0-4.2 by adding 5 N NaOH drops to form crude crystals. After cooling to 10 ~ 15 ^s C, the solid was filtered and completely dissolved in 767 ml of MeOH with heating. Precipitated crystals of slow cooling were filtered to provide P3 in the form of a pink crystal (40 g, 60% yield).

[0087] 1H NMR (500MHz, CDCl3) 8.43 (d, J = 4.8 Hz, 1H), 7.63 (s, 1H), 7.387.35 (m, 2H), 7.31-7, 30 (m, 1H), 7.29-7.21 (m, 2H), 7.00 (d, J = 4.8 Hz, 2H), 6.97 (d, J = 4.8 Hz, 1H ), 6.69-6.65 (m, 3H), 5.87 (s, 1H), 5.55-5.44 (m, 3H), 5.34 (t, J = 4.6 Hz, 1H), 5.03 (d, J = 6.3 Hz, 1H), 4.87 (d, J = 9.0 Hz, 1H), 4.79 (d, J = 7.5 Hz, 1H) , 4.42 (dd, J = 9.0, 3.6 Hz, 1H), 4.29 (dd, J = 9.0, 3.6 Hz, 1H), 4.02 (s, 3H), 3.43 (d, J = 7.2 Hz, 1H), 3.38-3.33 (m, 3H), 3.27 (d, J = 7.2 Hz, 1H), 3.29-3 , 24 (m, 1H), 3.18 (dd, J = 7.2, 2.4 Hz, 1H), 2.51 (s, 3H).

EXAMPLE 9 [0088] Synthesis of P2 [0089] 4 - (((6S, 9aS) -1 - (benzylcarbamoyl) -8 - ((3-acetyl-1 methyl-1 H-indol-7-yl) methyl) ) -2-allyl-octahydro-4,7-dioxo-1 H-pyrazine [2,1-c] [1,2,4] triazin-6yl) methyl) phenyl [0090] 40 g of P3 were dissolved in 217 ml of THF (tetrahydrofuran), cooled to 0 ~ 5 ^s C and mixed with 25 ml of POCI3. At the same temperature, 28 mL of TEA (triethylamine) was added in drops. Stirring for 1 h was followed by a slow addition of 87 mL of distilled water. 348 ml of a saturated aqueous solution of NaHCO3 was added to the solution which was then stirred for 30 min. After the solution was subjected to separation of the layers by adding 217 ml of EA (ethyl acetate), 217 ml of MC (methylene chloride) were added to the aqueous layer and then the pH was adjusted to 1-3 with 14 ml of HCI focused

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38/51 to separate the layers. The organic layer thus formed was dehydrated with

174 g of Na2SO4 (sodium sulfate) and concentrated in vacuo. The concentrate was crystallized from 130 ml of THF (tetrahydrofuran) and 435 ml of n-hexane, filtered and dried in vacuo to provide 40 g of P2 as a white solid (90% yield).

[0091] 1H NMR (500MHz, DMSO-d6) 8.27 (s, 1H), 8.16 (d, J = 7.5 Hz, 1H), 7.85 (t, J = 6.3 Hz, 1H), 7.34-7.29 (m, 3H), 7.22-7.01 (m, 9H), 6.79 (d, J = 6.9 Hz, 1H), 5.84-5 , 75 (m, 1H), 5.52 (dd, J = 8.1, 3.6 Hz, 1H), 5.38 (d, J = 15.6 Hz, 1H), 5.17-5, 13 (m, 1H), 5.09-5.03 (m, 2H), 4.90 (d, J = 15.6 Hz, 1H), 4.22 (d, J = 6.3 Hz, 2H ), 4.06 (s, 3H), 3.76-3.68 (m, 1H), 3.61-3.55 (m, 2H), 3.33-3.27 (m, 4H), 3.07-3.02 (m, 2H), 2.41 (s, 3H).

EXAMPLE 10 [0092] Synthesis of P1 [0093] 4 - ((((6S, 9aS) -1 - (benzylcarbamoyl) -8 - ((3-acetyl-1-methyl-1 H-indole-7yl) methyl) -2 -alyl-octahydro-4,7-dioxo-1 H-pyrazine [2,1-c] [1,2,4] triazin-6-yl) methyl) sodium phenyl hydrogen phosphate [0094] 40 g of dry P2 were dissolved in 2000 mL of distilled water with stirring. The solution was cooled to 0 ~ 5 ^s C, followed by adjusting its pH to 4.6-4.8 (130-110mV) slowly by adding a 0.1 N aqueous NaOH solution, and then lyophilized to make 40 g of P1 as a white solid (95% yield).

[0095] 1H NMR (300MHz, D2O) 7.86 (d, J = 7.8 Hz, 1H), 7.60 (s, 1H), 7.076.93 (m, 10H), 6.56 (d, J = 7.2 Hz, 1H), 5.39-5.32 (m, 2H), 5.09 (t, J = 5.4 Hz, 1H), 4.95 (d, J = 15.6 Hz, 1H), 4.70-4.53 (m, 2H), 4.14 (d, J = 15.6 Hz, 1H), 3.97 (d, J = 15.6 Hz, 1H), 3.57 (s, 3H), 3.56-3.49 (m, 1H), 3.30-2.81 (m, 6H), 2.84-2.81 (m, 1H), 2, 18 (s, 3H).

[0096] Another example of preparation for representative compounds

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<Reaction scheme 2>

l-k ^ o h ^ o

Η I2, KOH

N

J λ DMF, rt

K ₂ CO ₃ , CH ₃ I

DMF, rt, 1h

Mol. Wt .: 145.16 indole

Mol. Wt .: 271.05

Q10

Mol. Wt .: 285.08

Q9 aminoacetal NaBH ₄ , AcOH

MeOH, rt, 3 h

Cl6 ^ 23 ^ 2O2 Mol. Wt .: 402.27

Q8

FmocTyr (t-Bu) -OH HATU, DIPEA

CH2CI2 · laughs

piperidine

CH2CI2 · laughs

θ16 ^ 23 ^ 2θ2 Mol. Wt .: 402.27

Q8

C44H50IN3O6 Mol. Wt .: 843.79

CH2CI2 · laughs

S3

HATU, DIPEA

C2gH4QlNgO4 Mol. Wt .: 621.55

Q6 ° C, 30 min θ42 ^ 55 ^ θθθ

Mol. Wt .: 866.83

Q5 p-TsOH.H ₂ O, Toluene

Q4

THF, rt, 30 min

POCI3

TEA

Mol. Wt .: 790.80

Q2

Q3

O.INNaOH Lyophilzer H ₂ O, 5 ° C

Q1 [0097] The method illustrated in reaction scheme 2 is described in detail in examples 11 to 21.

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EXAMPLE 11 [0098] Synthesis of S3 (Side chain) [0099] S3 was obtained in the same way as in example 1.

EXAMPLE 12 [00100] Synthesis of Q10 [00101] 3-sludge-1 H-indole-7-carbaldehyde [00102] A solution of 24 g of I2 in 125 ml of DMF (dimethylformamide) was added to the starting material (indol- 7-carbaldehyde) and reacted with 5.3 g of KOH with stirring. The progress of the reaction was monitored with TLC. When the reaction was complete, 354 ml of EA (ethyl acetate) and 354 ml of distilled water were added to generate separation of the layers. The organic layer thus formed was washed with 10% aqueous Na2S2O3 solution, dried over Na2SÜ4 (sodium sulfate), filtered and concentrated at 40 ^s C to give Q10 as a concentrated residue.

[00103] 1H NMR (CDCI3, 300MHz) δ 10.3 (bs, 1H), 10.2 (s, 1H), 7.79 (d, 1H, J = 7.8 Hz), 7.75 (d , 1H, J = 7.2 Hz), 7.44 (d, 1H, J = 2.1 Hz), 7.37 (t, 1H, J = 7.2 Hz); m / z 272.14 [M + 1] +

EXAMPLE 13 [00104] Synthesis of Q9 [00105] 17 g of Q10 were dissolved in 100 ml of DMF (dimethylformamide) with stirring. The resulting solution was cooled to 10 ^s C and mixed with 18 g of K2CO3 (potassium carbonate). After 6 ml of honey (methyl iodide) was added dropwise to it, the solution was warmed to room temperature and stirred for 4 ~ 5 hours. When the starting material was found to have disappeared, K2CO3 was filtered, followed by crystallization from hexane to give Q9.

[00106] 1H NMR (CDCI3, 300MHz) δ 10.2 (s, 1H), 7.76 (td, 1H, J = 7.8, 1.2

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Ηζ), 7.31 (t, 1 Η, J = 7.8 Hz), 7.12 (s, 1H), 4.14 (s, 3H)

EXAMPLE 14 [00107] Synthesis of Q8 [00108] To a solution of 18 g of Q9 in 600 ml of MeOH (methanol) was added 0.4 ml of AcOH (acetic acid). At room temperature, 14 ml of diethylacetal aminoacetaldehyde was added to the solution, followed by stirring for 3 ~ 4 hours. The solution was cooled to 10 ^s C before 3.3 g of the reducing agent NaCNBH3 (sodium cyanoborohydride) was added slowly. At this time, care must be taken because hydrogen gas and value have been generated. After the reaction mixture was stirred at room temperature for 1 h, the progress of the reaction was monitored. When the reaction was complete, 354 ml of EA (ethyl acetate) and 354 ml of distilled water were used to separate the layers. The organic layer thus formed was dehydrated with 141 g of Na2SO4 (sodium sulfate) and crystallized from hexane to give Q8.

EXAMPLE 15 [00109] Synthesis of Q7 [00110] 27 g of Fmoc-Tyr (OtBu) were dissolved in 200 ml of MC (dichloromethane) with stirring. To this solution were added 23 g of HATU (O (7-azabenzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorphosphate) and 20 ml of DIPEA (diisopropylethylamine) at room temperature. The solution was stirred for 1 ~ 2 hours, mixed with 15.8 g of Q9 and further stirred for 2 ~ 3 hours. After completing the reaction, distilled water was added to separate the layers. The organic layer thus formed was washed with 898 ml of a 5% aqueous solution of citric acid and 898 ml of a 5% aqueous solution of NaHCO3, dehydrated with Na2SO4 (sodium sulfate), and concentrated to provide Q7 as a residue. focused.

EXAMPLE 16

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42/51 [00111] Synthesis of Q6 [00112] To a solution of 34 g of Q7 in 400 ml of MC (dichloromethane) was added 20 ml of piperidine. After the reaction was complete, the solution is concentrated, followed by separation of the layers with 400 ml of MC (dichloromethane) and 800 ml of distilled water. The organic layer thus formed was washed with 850 ml of a 5% aqueous solution of citric acid and 850 ml of a 5% aqueous solution of NaHCO3, and then concentrated to give Q6.

EXAMPLE 17 [00113] Synthesis of Q5 [00114] To a solution of 13 g of S3 in 400 ml of MC (dichloromethane) was added in drops 19 g of HATU (O- (7-azabenzotriazol-1-yl) -N, N, N ', N'tetramethyluronium hexafluorophosphate) and 16 mL of DIPEA (diisopropylethylamine) at room temperature. After the solution was stirred for 2 ~ 3 hours, a solution of 28 g of Q6 in 200 ml of MC (dichloromethane) was added dropwise to it. It was stirred at room temperature for 1 ~ 2 hours. When the reaction was complete, 200 ml of MC (dichloromethane) and 200 ml of distilled water were used to generate separation of the layers. The organic layer thus formed was washed with 200 ml of a 5% aqueous solution of citric acid and 200 ml of a 5% aqueous solution of NaHCO3 and dehydrated with 340 g of Na2SO4 (sodium sulfate) and then concentrated to make Q5 available as of a concentrated residue.

EXAMPLE 18 [00115] Synthesis of Q4 [00116] 289 mg of p-TsOH.H2O was added to a solution of 4 g of

Q5 in 100 ml of toluene which was then heated to 80 ^s C. The resulting solution was stirred at the same temperature for 30 min, cooled to room temperature and concentrated. Separation of the layers was generated with EA (ethyl acetate) and distilled water. The organic layer was washed with 200 ml of an aqueous solution of

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43/51 citric acid 5% and 200 ml of a 5% aqueous solution of NaHCO3 and dehydrated with 340 g of Na2SO4 (sodium sulfate) and then concentrated to give Q4 as a concentrated residue.

[00117] 1H NMR (CDCI3, 300MHz) δ 7.43-7.27 (m, 3H), 7.23-7.21 (m, 2H), 7.12 (t, 1H, J = 7.2Hz ), 7.08 (s, 1H), 7.05 (d, 2H, J = 7.8 Hz), 6.97 (d, 1H, J = 7.2 Hz), 6.90 (d, 2H , J = 8.4 Hz), 6.59 (t, 1H, J = 6.0 Hz), 5.62 (dd, 1H, J = 10.2, 4.8 Hz), 5.53-5 , 39 (m, 3H), 5.37 (t, 1H, J = 6.0 Hz), 5.02 (d, 1H, J = 10.2 Hz), 4.93 (d, 1H, J = 16.5 Hz), 4.77 (d, 1H, J = 17.1 Hz), 4.44 (dd, 1H, J = 15.0, 6.3 Hz), 4.32 (dd, 1H, J = 15.0, 6.0 Hz), 3.97 (s, 3H), 3.49-3.19 (m, 8H), 1.33 (s, 9H);

EXAMPLE 19 [00118] Synthesis of Q3 [00119] To a solution of 100 mg of Q4 in a mixture of 8 ml of 1,4dioxane and 4 ml of water was added to 33 mg of 4acetylbenzenoboronic acid, 41 mg of Na2CÜ3 (carbonate of sodium) and 15 mg of Pd (PPh3) 4 (tetraquistriphenylphosphinopalladium), followed by raising the temperature to 90 ^s C. After being stirred for 2 hours at the same temperature, the solution was cooled to room temperature and concentrated. EA (ethyl acetate) and distilled water were used to generate separation of the layers. The organic layer thus formed was dehydrated with Na2SO4 (sodium sulfate) for concentration. The concentrate was dissolved in MC (dichloromethane) to which 1 ml of TFA (trifluoroacetic acid) was then added in drops, followed by stirring at room temperature. After completing the reaction, the reaction mixture was washed with 10 ml of 5% aqueous NaHCO3 solution and dehydrated with Na2SO4 (sodium sulfate) to give Q3 as a concentrated residue.

[00120] 1H NMR (CDCI3, 300MHz) δ 8.05 (d, 2H, J = 8.4 Hz), 7.91 (d, 1H, J = 7.2 Hz), 7.71 (d, 2H , J = 8.4 Hz), 7.40-7.20 (m, 4H), 7.16 (t, 1H, J = 7.2Hz), 7.05 (d, 2H, J = 8.4 Hz), 6.96 (d, 1H, J = 6.9 Hz), 6.69 (d, 2H, J = 8.4 Hz), 6.68 (m, 1H),

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5.58-5.44 (m, 3H), 5.37 (t, 1H, J = 5.7 Hz), 5.03 (d, 1H, J = 10.8 Hz), 4.97 (d , 1H, J = 14.7 Hz), 4.81 (d, 1H, J = 17.1 Hz), 4.47 (dd, 1H, J = 15.3, 6.3 Hz), 4.33 (dd, 1H, J = 15.3, 6.3 Hz), 4.33 (s, 3H), 3.47-3.24 (m, 8H), 2.64 (s, 3H); m / z 711.56 [M + 1] +

EXAMPLE 20 [00121] Synthesis of Q2 [00122] A solution of 50 g of Q3 in 217 ml of THF (tetrahydrofuran) was cooled to 0 ~ 5 ^s C and mixed with 25 ml of POCb. At the same temperature, 28 mL of TEA (triethylamine) was added in drops to the solution, which was then stirred for 1 h. 87 mL of distilled water was added slowly. 348 mL of a saturated aqueous solution of NaHCO3 was added and the solution was stirred for 30 min. The addition of 217 mL of EA (ethyl acetate) resulted in the separation of the layers. To the aqueous layer were added 217 ml of MC (methylene chloride), followed by adjusting the pH of the solution to 1-3 with 14 ml of concentrated HCI. The organic layer thus formed was dehydrated with Na2SO4 (sodium sulfate) and concentrated in vacuo. The concentrate was crystallized from 130 ml of THF (tetrahydrofuran) and 435 ml of n-hexane and the solid was filtered and dried in a vacuum.

EXAMPLE 21 [00123] Synthesis of Q1 [00124] 44 g of dry Q2 were dissolved in 200 ml of distilled water with stirring. After cooling to 0 ~ 5 ^s C, 0.1 N NaOH was slowly added to adjust the pH of the solution to 4.6-4.8 (130-110 mV), followed by lyophilization to give Q1.

[00125] A detailed description will be given below of the effect of the prepared compounds.

EXAMPLE 22 [00126] The compounds were prepared in the form of prodrugs to improve their solubility. Phosphate can be introduced as a possible

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45/51 prodrug substitute that can exist in the form of either monosodium phosphate or disodium phosphate.

[00127] This prodrug was prepared by adding sodium hydroxide to P2, which was synthesized according to example 9. Both monosodium and disodium forms of the prodrug show a solubility of up to 400 mg / mL. Both forms have advantageous properties in the form of an I.V. injection composition in which a monosodium form has a pH of 4.45 and a disodium form has a pH of 7.62.

[00128] FIG. 1 graphically shows changes in pH and potential when 0.5 N NaOH is added dropwise to the compound of the present invention. In the graph, the horizontal axis represents the added amounts of sodium hydroxide. In the graph, the first and second inflection points correspond to the production time of the monosodium and disodium forms, respectively.

EXAMPLE 23 [00129] Anticancer activity in an animal model of acute myeloid leukemia (AML) [00130] Test materials were prepared in the form of prodrugs to increase the solubility of the compounds of interest. A functional phosphate group that can be either a monosodium or disodium form has been introduced as a substituting prodrug.

Compound A1 Compound A2

O

ONa

ONa

A3 compound

Compound A

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Compound B3

Compound C1 Compound C2

Compound C3

Compound C

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[00131 jReference material: Ara-C (Medically available drug to treat acute myeloid leukemia) [00132] The human AML cell line, MV4-11, was purchased (ATCC, USA) and grown at 37 ^S C in a 5% CO2 in Dulbecco's modified Iscove medium (GIBCO, cat # 21056) supplemented with 10% fetal bovine serum (GIBCO, cat # 25030-081). Female nude Balb / C mice (OrientBio, Sungnam-city, Korea), 5-6 weeks old, were acclimatized in the production room. Using a sterile syringe, a 1: 1 mixture of MV4-11: matrigel (v / v) cells was implanted in an amount of 5x106 / mouse below the axles of each of the mice. When a tumor was formed 2 weeks after implantation, the mice were divided into five (5) groups in such a way that a minimum deviation regarding the size of the tumor and body weight was obtained between the groups. The test materials were dissolved in physiological saline and intravenously injected in a dose of 10 ml / kg once a day and five times a week for two weeks (days of administration of the test materials, D1-D5, D8-D12). For a control, only physiological saline was used. The tumor size was determined as calculated by the following equation: Long axis x Short axis x Short axis / 2. The long and short axes of the tumor were measured in length using a digital caliper (Mitsutoyo, Japan). The anti-cancer activity of the test materials was numbered according to the following equation.

[00133] Tumor growth inhibition rate A (%) = 100 X [1- (bPetition 870180065902, of 07/30/2018, page 71/82

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a) / (Ref b-Ref a)] where a = the size of the average group tumor administered with medication on day 1 b = the size of the average group tumor administered with medication on day 12

Ref one = average tumor size of the control on day 1

Ref b = average tumor size of the control on day 12 [00134] When the average tumor size of the group administered with medication on day 12 was smaller than that of just before the administration of the test materials, it is indicated as Regression (> 100%). Rates of tumor growth inhibition of tumor growth of test materials are summarized in Table 2, below.

TABLE 2

Inhibition rate d and tumor growth Test material dose (mg / kg) Rate of inhibition of tumor growth Ara-C 50 77% Ara-C 25 66% A1 25 Regression (> 100%) Compound A2 25 Regression (> 100%) A3 compound 25 Regression (> 100%) Compound A 25 61% Compound B1 25 Regression (> 100%) Compound B2 25 Regression (> 100%) Compound B3 25 80% Compound B 25 49%

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Compound C1 25 Regression (> 100%) Compound C2 25 Regression (> 100%) Compound C3 25 70% Compound C 25 25%

[00135] Test results showed that all test compounds have inhibitory activity against tumor growth. In compounds A1-A3, B1-B3 and C1-C3 according to the present invention, tumor inhibition rates were measured to vary the regression from 70% (> 100%). In contrast, Ara-C, it was observed that a drug widely used for AML has a tumor inhibition rate of 66%. Taken together, the results demonstrate that the compounds of the present invention are highly inhibitory to tumor growth.

EXAMPLE 24 [00136] In vitro cardiotoxicity assay ·. Assay for inhibitory activity against hERG [00137] HEK293 was transfected with hERG cDNA (human related Ether-to-go-go gene) for 48 hours using Lipofectamine 2000 (Invitrogen, USA). The transfected HEK293 cells were cultured in modified Dulbecco's medium (MEM, Gibco, 1 L) supplemented with 10% FBS, sodium pyruvate (10 ml), penicillin / streptomycin (10 ml) and Zeocin (100 pg / ml, Invitrogen ) at 37 ^S C in 5% CO2. After being detached from the incubation vessels by trypsinization, HEK293 cells were placed in a chamber to record a patch clamp. A full cell patch clamp method was used to record K ⁺ hERG currents in HEK293 cells using the following intra / extracellular solutions. Thereafter, the effects of K ⁺ currents were observed with compounds applied outside the cells.

• intracellular solution: 100 mM K-aspartate, 25 mM KCI, 5 mM NaCI, 1 mM MgCI2, 4 mM Mg-ATP, 1,2-bis (o-aminophenoxy) ethane-N, N, N ', N' acid -tetraacetic

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50/51 (ΒΑΡΤΑ) 10 mM, 4- (2-hydroxyethyl) -1-piperazine ethanesulfonic acid (HEPES) 10 mM, normalized magnesium (NMG) were used to adjust the pH to 7.2;

• extracellular solution: 145 mM NaCl, 5 mM KCI, 10 mM glucose, 1 mM MgCI2, 2 mM CaCb, 10 mM HEPES, HCI were used to adjust the pH to 7.4.

The membrane potential was depolarized from -80 mV to +20 mV per 1,000 ms in a full cell patch clamp mode and then repolarized to -40 mV per 1,000 ms, during which the tail current of K ⁺ currents from hERG to have been recorded. In this regard, the concentrations of the compounds that are required for 50% inhibition of the current were represented as IC 50.

TABLE 3

Cardiotoxicity assay

Cpd. of test Cardiotoxicity (μΜ) (HERG inhibition activity test, IC50) A1 80 Compound A 14 Compound B1 18 Compound B2 25 Compound B3 20 Compound B 1.6

[00138] The risk of cardiotoxicity has been increased in many drugs. Some of them were withdrawn from the market due to their sudden death due to its cardiotoxicity. Cardiotoxicity of drugs is associated with the extension of QT intervals on electrocardiograms. In particular, it is known that most drugs that extend in QT intervals inhibit IKr channels (Bernard Fermini and Anthony A. Fossa, Nature Reviews Drug Discovery, 2003, 2, 439-447). The hERG channel shows the most important effect on cardiotoxicity

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51/51 between IKr channels. In this example, the risk of cardiotoxicity was assessed using mammalian cells that express human hERG channel, which are internationally recognized as a system (ICH guideline, S7B, Step4, 12, May, 2005). Although the pharmaceutical activity of the drug should be taken into account, a drug is rated as having a low risk of cardiotoxicity when its IC50 is 10 μΜ or more. In this assay, it was observed that most test compounds exceed this criterion. Having a higher IC50, compound A1 was evaluated as safer than compound A, and compounds Β1, B2 and B3 than compound B.

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Claims (11)

1. Composed of chemical formula I, CHARACTERIZED by the fact that it has the following structure: Chemical formula I where: Ra is an Ο-Οβ alkyl group; Rb is -C (= O) R _e , where Re is a C1-C6 alkyl group, a C2C6 alkenyl group or a C2-C6 alkynyl group; and R _P is -PO3H2, -HPO ₃ Na ⁺ , -PO3 ^{2 3} Na ²⁺ , -PO3 ² K2 ⁺ , -PO3 ² Mg ²⁺ , -PO3 ² Ca ²⁺ , 2. Composed, according to claim 1, CHARACTERIZED by the fact that: Ra is methyl, Rb is -C (= O) R _e , where Re is C1-C6 alkyl, and R _P is -PO3H2, - HPO ₃ Na ⁺ or -PO ₃ ² -Na ²⁺ . 3. Compound according to claim 1, CHARACTERIZED by the fact that the compound represented by chemical formula I is: 8- (3-Acetyl-1-methyl-1 H-indol-7-ylmethyl) -2-allyl 6- (4-hydroxybenzyl) -4,7-dioxo-hexahydro-pyrazine benzylamide [2,1 - c] [1,2,4] triazine-1-carboxylic acid, 2-Allyl-8- [3- (3,3-dimethyl-butyryl) -1-methyl-1 H-indole-7-ylmethyl] benzylamide] 6- (4-hydroxy-benzyl) -4,7-dioxo-hexahydro-pyrazine [2,1-c] [1,2,4] triazine-1-carboxylic acid, 2-Allyl-8- acid benzylamide (3 -cyclopropanocarbonyl-1-methyl-1 H-indole-7 Petition 870180065902, of 07/30/2018, page 76/82 2/5 ylmethyl) -6- (4-hydroxy-benzyl) -4,7-dioxo-hexahydro-pyrazine [2,1-c] [1,2,4] triazine-1 carboxylic acid, 2-Allyl benzylamide -6- (4-hydroxy-benzyl) -8- [1-methyl-3- (3-methyl-butyryl) 1 H-indol-7-ylmethyl] -4,7-dioxo-hexahydro-pyrazine [2,1 -c] [1,2,4] triazine-1-carboxylic acid, 2-Allyl-8- (3-butyryl-1-methyl-1 H-indol-7-ylmethyl) -6- (4-hydroxybenzyl) benzylamide ) -4,7-dioxo-hexahydro-pyrazine [2,1-c] [1,2,4] triazine-1-carboxylic acid, 2-Allyl-8- (3-cyclopropanecarbonyl-1-ethyl-1H) benzylamide -indole-7ylmethyl) -6- (4-hydroxy-benzyl) -4,7-dioxo-hexahydro-pyrazine [2,1-c] [1,2,4] triazine-1 carboxylic acid, 2-Allyl benzylamide -8- (1-allyl-3-cyclopropanecarbonyl-1H-indol-7ylmethyl) -6- (4-hydroxy-benzyl) -4,7-dioxo-hexahydro-pyrazine [2,1 -c] [1,2, 4] triazine-1 carboxylic acid, benzylamide of 2-Allyl-6- (4-hydroxy-benzyl) -8- (1-methyl-3-pentanoyl-1Hindol-7-ylmethyl) -4,7-dioxo-hexahydro- pyrazine [2,1-c] [1,2,4] triazine-1-carboxylic acid, 2-allyl-6- (4-hydroxy-benzyl) -8- (1-methyl-3-) benzylamide propionyl-1 Hindol-7-ylmethyl) -4,7-dioxo-hexahydro-pyrazine [2,1-c] [1,2,4] triazine-1-carboxylic acid, 8- (3-Acetyl-1) benzylamide -propyl-1 H-indol-7-ylmethyl) -2-ally-6- (4-hydroxybenzyl) -4,7-dioxohexahydro-pyrazine [2,1 -c] [1,2,4] triazine- 1-carboxylic acid, 2-Allyl-8- [3- (3,3-dimethyl-butyryl) -1-propyl-1 H-indol-7ylmethyl] -6- (4-hydroxy-benzyl) -4, benzylamide, 7-dioxo-hexahydro-pyrazine [2,1-c] [1,2,4] triazine-1 carboxylic acid, benzylamide 2-Allyl-8- [3- (3,3-dimethyl-butyryl) -1 - hexyl-1 H-indol-7-ylmethyl] 6- (4-hydroxy-benzyl) -4,7-dioxo-hexahydro-pyrazine [2,1-c] [1,2,4] triazine-1-carboxylic, or 2-Allyl-8- (1-butyl-3-cyclopropanecarbonyl-1H-indol-7ylmethyl) -6- (4-hydroxy-benzyl) -4,7-dioxo-hexahydro-pyrazine acid benzylamide [2,1 - c] [1,2,4] carboxylic triazine-1. 4. Pharmaceutical composition CHARACTERIZED by the fact that Petition 870180065902, of 07/30/2018, p. 77/82 3/5 comprises the compound as defined in any one of claims 1 to 3 and a pharmaceutically acceptable excipient. 5. Use of the pharmaceutical composition as defined in claim 4, CHARACTERIZED for what it is in the manufacture of a drug to treat acute myeloid leukemia (AML). 6. Use, according to claim 5, CHARACTERIZED by the fact that the pharmaceutical composition is formulated to be injected into the patient. 7. Method for making the compound as defined in claim 1, CHARACTERIZED by the fact that it comprises the following sequential steps: introducing an acyl group into the indole-7-carbaldehyde by means of Friedel-Crafts acylation to provide 3-acyl-indole-7-carbaldehyde; introducing an alkyl group and an aminoacetal group on 3-acyl-indole-7carbaldehyde to provide a 1-alkyl-3-acyl-indole derivative; amide the 1-alkyl-3-acyl-indole derivative with Cbz-TyrosineOtBu stereoselectivity and 2- (1-allyl-4-benzylsemicarbazido) acetic acid to provide a reaction intermediate; cyclize the reaction intermediate in the presence of formic acid to provide a cyclic intermediate; and phosphorylating the cyclic intermediate to provide a compound of chemical formula (I). 8. Method according to claim 7, CHARACTERIZED by the fact that 2- (1-allyl-4-benzylsemicarbazido) acetic acid is synthesized by the following sequential steps: add TEA (triethylamine) to an ethylhydrazinoacetate solution to provide a reaction solution; add allyl bromide to the reaction solution; and then add benzyl isocyanate. Petition 870180065902, of 07/30/2018, p. 78/82 4/5 9. Method according to claim 8, CHARACTERIZED by the fact that allyl bromide and benzyl isocyanate are added in a droplet manner. 10. Method for preparing a compound of chemical formula (I), CHARACTERIZED by the fact that it comprises: CHO convert indole-7-carbaldehyde to ^R b, where Rb is an aryl group, a substituted aryl group, or -C (= O) R _and , where Re is a C1-C6 alkyl group, a C2- alkenyl group C6 or a C2-C6 alkynyl group; convert, wherein R _a is a C1-C6 alkyl group, a C2-C6 alkenyl group or a C2-C6 alkynyl group; R _b with stereoselectivity in the presence of Cbz-Tyrosineamidar Eto o Petition 870180065902, of 07/30/2018, p. 79/82 5/5 oro EtO '^ ^N > ^ N ^x ^ ^N ' N ^ NHBn. .. II Η H cichzar o in the presence of formic acid H ΝγΟ N to provide R = Rh O H ^Ν γ ° 'N' ^N ^ NN OH R _a Rh Oh _a The converter ^{0 r} p, where RP is -ΡΟ3Η2> -HPO3-Na ⁺ , -POs ² -Na ²⁺ , PO3 ² -K2 ⁺ , - PO3 ² -Mg ²⁺ , - POs ² -Ca ²⁺ . 11. Method according to claim 10, CHARACTERIZED by the fact that Ra is methyl, Rb is -C (= O) R _e , and Re is methyl or cyclopropyl. Petition 870180065902, dc 07/30/2018, p. 80/82 1/1