KR20220118933A - Pharmaceutical composition comprising indoloquinazolidine alkaloid for treatment of cancer by targeting heat shock protein 70 - Google Patents

Abstract

The present invention relates to an anticancer pharmaceutical composition for targeting heat shock protein 70 containing indoloquinazolidine alkaloid. The indoloquinazolidine alkaloid of the present invention inhibits tumor growth and inhibits HSP70 protein expression and colony formation ability of cancer cells and inhibits tumor growth in cancer cell line heterologous graft and patient-derived cancer heterologous graft mouse models, and also inhibits growth of drug-resistant cancer cells such as pemetrexed, cisplatin, and paclitaxel. Therefore, the indoloquinazolidine alkaloid is expected to be widely used in prevention and treatment of various cancers.

Description

Pharmaceutical composition comprising indoloquinazolidine alkaloid for treatment of cancer by targeting heat shock protein 70

The present invention relates to a pharmaceutical composition for heat shock protein 70 target anticancer containing indoloquinazolidine alkaloid, and the like.

Cancer is a major cause of death, and early diagnosis and development of anticancer drugs that can effectively suppress cancer such as immunotherapy are being developed due to the development of diagnostic technology. It is essential to develop effective anticancer drugs because of the decrease and recurrence of cancer.

Lung cancer refers to a malignant tumor that occurs in the lungs, and may occur either in the lung itself (primary lung cancer) or by metastasis of cancer originating from other organs to the lungs. Primary lung cancer is classified into non-small cell lung cancer and small cell lung cancer based on the size and shape of cancer cells. Non-small cell lung cancer accounts for 80-85% of lung cancers. It is divided into squamous cell carcinoma, adenocarcinoma (adenocarcinoma), large cell carcinoma, adenosquamous cell carcinoma, sarcoma, carcinoid tumor, salivary gland carcinoma, and unclassified cancer. The rest, small cell lung cancer, has a high overall malignancy, and at the time of discovery, it has already metastasized to other organs or the contralateral lung, mediastinum (the space between the two lungs, where the heart, trachea, esophagus, aorta, etc. are located) through lymphatic vessels or blood vessels. there are often

Heat shock protein 70 (Hsp70) is overexpressed in several cancer cells and is known to mediate cancer cell survival and anticancer drug resistance. In particular, in recent papers, inhibition of Hsp70 has been shown to inhibit cancer initiation and metastasis through suppression of cancer initiating cells. Several Hsp70 inhibitors are under development, but most of them are in the preclinical stage, so there is a need to develop effective Hsp70 inhibitors.

Meanwhile, in previous studies, the anti-inflammatory, antiviral, hepatoprotective, anticancer, and cancer stem cell inhibitory effects of indoloquinazolidine alkaloids and representative compounds of this class, evodiamine and rutaecarpine, have been reported. In addition, the present inventors recently revealed the Hsp70 inhibitory effect as a new mechanism of the anticancer and cancer stem cell inhibitory action of evodiamine.

Domestic registered patent 10-18888744

The present invention has been devised to solve the problems in the prior art as described above, and the present inventors prepared a novel indoloquinazolidine alkaloid having anticancer activity, and inhibited the proliferation of cancer cells by treating it Through this, the preventive or therapeutic effect on cancer was confirmed, and the present invention was completed based on this.

Accordingly, it is an object of the present invention to provide an indoloquinazolidine alkaloid represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In Formula 1,

R ₁ and R ₂ are each independently hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, halogen, nitro (NO ₂ ), amide, —NH— aryl, hydroxy, or -COOR ₇ ;

R ₃ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, -CH ₂ -R ₈ , substituted or unsubstituted C ₁ -C ₆ acyl, substituted or unsubstituted aryl, -CO-R ₈ or benzyl ego,

R ₄ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₂ -C ₆ alkylalkenyl, substituted or unsubstituted C ₂ -C ₆ alkylalkynyl, -CH ₂ -R ₈ , substituted or unsubstituted aryl, propargyl, substituted or unsubstituted C ₁ -C ₆ acyl, or benzyl,

R ₅ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, halogen, nitro, or substituted or unsubstituted C ₁ -C ₆ alkoxy;

R ₆ is hydrogen, oxygen, or substituted or unsubstituted C ₁ -C ₆ alkyl,

R ₇ and R ₈ are each independently hydrogen, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted aryl;

X is C, N, O or S.)

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer comprising the indoloquinazolidine alkaloid or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide an anticancer adjuvant that enhances the anticancer efficacy of an anticancer agent comprising the indoloquinazolidine alkaloid or a pharmaceutically acceptable salt thereof as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the object of the present invention as described above, the present invention provides an indoloquinazolidine alkaloid represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In Formula 1,

R ₁ and R ₂ are each independently hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, halogen, nitro (NO ₂ ), amide, —NH— aryl, hydroxy, or -COOR ₇ ;

R ₃ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, -CH ₂ -R ₈ , substituted or unsubstituted C ₁ -C ₆ acyl, substituted or unsubstituted aryl, -CO-R ₈ or benzyl ego,

R ₄ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₂ -C ₆ alkylalkenyl, substituted or unsubstituted C ₂ -C ₆ alkylalkynyl, -CH ₂ -R ₈ , substituted or unsubstituted aryl, propargyl, substituted or unsubstituted C ₁ -C ₆ acyl, or benzyl,

R ₅ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, halogen, nitro, or substituted or unsubstituted C ₁ -C ₆ alkoxy;

R ₆ is hydrogen, oxygen, or substituted or unsubstituted C ₁ -C ₆ alkyl,

R ₇ and R ₈ are each independently hydrogen, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted aryl;

X is C, N, O or S.)

In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer comprising an indoloquinazolidine alkaloid or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a method for treating cancer comprising administering to an individual in need of cancer treatment a pharmaceutical composition comprising an indoloquinazolidine alkaloid or a pharmaceutically acceptable salt thereof as an active ingredient. do.

In addition, the present invention provides a cancer treatment use of a composition comprising an indoloquinazolidine alkaloid or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides the use of an indoloquinazolidine alkaloid or a pharmaceutically acceptable salt thereof for the production of a therapeutic agent for cancer.

,

, 또는 -COOR₇이고, In one embodiment of the present invention, the R ₁ and R ₂ are each independently hydrogen, methoxy, butoxy, methyl, hydroxy, F, Cl, nitro,

,

, or -COOR ₇ ,

R ₃ is hydrogen, methyl, ethyl, propyl, butyl, hexyl, benzyl, -CO-R ₈ or -CH ₂ -R ₈ ,

, 또는 -CH₂-R₈이고,R ₄ is hydrogen, methyl, benzyl, substituted or unsubstituted C ₂ -C ₅ alkenyl, substituted or unsubstituted C ₂ -C ₅ alkynyl, substituted or unsubstituted C ₂ -C ₅ alkylalkenyl, substituted or unsubstituted C ₂ -C ₅ alkyl alkynyl, cyclohexanecarbonyl,

, or -CH ₂ -R ₈ ,

R ₅ is hydrogen, methoxy, nitro, methyl, F, Cl, Br, or I;

R ₆ is hydrogen or oxygen,

R ₇ and R ₈ are each independently hydrogen, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted aryl, the substituted aryl is substituted with one or more selected from the group consisting of halogen, CN, C ₁ -C ₆ haloalkyl, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, nitro,

X may be N or O, but is not limited thereto.

In another embodiment of the present invention, the indoloquinazolidine alkaloid represented by Formula 1 may be one selected from the group consisting of the following compounds, but is not limited thereto.

In another embodiment of the present invention, the cancer is cervical cancer, lung cancer, pancreatic cancer, non-small cell lung cancer, liver cancer, colon cancer, colorectal cancer, bone cancer, skin cancer, head cancer, cervical cancer, skin melanoma, intraocular melanoma, uterine cancer ; , parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, bladder cancer, kidney cancer, ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma , may be one or more selected from the group consisting of a spinal cord tumor, a brainstem glioma, and a pituitary adenoma, but is not limited thereto.

In another embodiment of the present invention, the cancer may be lung cancer, but is not limited thereto.

In another embodiment of the present invention, the indoloquinazolidine alkaloid may reduce one or more selected from the group consisting of the number and volume of tumors, but is not limited thereto.

In another embodiment of the present invention, the indoloquinazolidine alkaloid may inhibit the expression of HSP70 protein, but is not limited thereto.

In another embodiment of the present invention, the indoloquinazolidine alkaloid may inhibit the expression of one or more proteins or genes selected from the group consisting of Akt, Src, and MEK, but is not limited thereto.

In another embodiment of the present invention, the composition may further include one or more selected from the group consisting of pemetrexed, cisplatin, and paclitaxel, but is not limited thereto.

In another embodiment of the present invention, the composition may be for preventing or treating lung cancer that has acquired resistance to an anticancer agent, but is not limited thereto.

In addition, the present invention provides an anticancer adjuvant that enhances the anticancer efficacy of an anticancer agent comprising the indoloquinazolidine alkaloid represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In one embodiment of the present invention, the anticancer agent may be one or more selected from the group consisting of pemetrexed, cisplatin, and paclitaxel, but is not limited thereto.

In addition, the present invention provides an anticancer agent comprising administering to an individual in need thereof an anticancer adjuvant comprising an indoloquinazolidine alkaloid represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. A method for enhancing or enhancing efficacy is provided.

In addition, the present invention provides an anticancer adjuvant use for improving or enhancing the anticancer efficacy of an anticancer agent of a composition comprising the indoloquinazolidine alkaloid represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides the use of the indoloquinazolidine alkaloid represented by Formula 1 or a pharmaceutically acceptable salt thereof for producing a medicament that improves or enhances the anticancer efficacy of an anticancer agent.

The indoloquinazolidine alkaloid of the present invention can inhibit tumor growth, inhibit HSP70 protein expression and colony formation ability of cancer cells, and inhibit tumor growth in cancer cell line xenografts and patient-derived cancer xenograft mouse models. In addition, it inhibits the growth of drug-resistant cancer cells such as pemetrexed, cisplatin, and paclitaxel, and is expected to be widely used in the prevention and treatment of various cancers.

1 shows the change in cell viability after 3 days of treatment with EV501-EV509 100 nM in H1299 lung cancer cells by MTT assay.
Figure 2 shows the change in cell viability after 3 days of treatment with EV507 at various concentrations in cancer cell lines of various origins by MTT assay (lung cancer cell line and chemotherapy-resistant lung cancer cell line) or crystal violet assay (remaining cell lines).
3 shows the change in colony-forming ability after 2 weeks of treatment with EV507 (50 nM) in cancer cell lines of various origins.
4 shows that the expression of Hsp70 and the expression of Hsp90/Hsp70 client protein are reduced by EV507 in H1299, H460, A549 and H226B lung cancer cell lines.
FIG. 5 shows that Hsp70 protein expression is reduced when EV507 (50 nM) is treated in H1299 lung cancer cells, HCT116 colorectal cancer cells, and UMSCC38 head and neck cancer cell lines.
6 shows that EV507 binds to the full-length Hsp70 and to the Hsp70 N-terminus.
7 shows the tumor growth inhibitory effect of EV507 on tumors derived from lung cancer cell lines or lung cancer patients transplanted into mice.
8 shows the effect of EV507 on the mouse survival rate increase in lung metastasis cancer allograft model.
9 is a view showing the change in cell viability was confirmed by MTT assay after 3 days of treatment with EV101-EV112 at various concentrations (0.5-10 μM) in H1299 cells.
Figure 10 shows the change in cell viability after 3 days of treatment with EV200-EV222 of various concentrations (0.5-10 μM) in H1299 cells was confirmed by MTT assay.
11 shows the change in cell viability was confirmed by MTT assay after 3 days of treatment with EV301-EV312 at various concentrations (0.5-10 μM) in H1299 cells.
12 shows the change in cell viability after 3 days of treatment with EV401-EV413 at various concentrations (0.5-10 μM) in H1299 cells by MTT assay.
13 shows the cell viability inhibitory effect of EV206 in various lung cancer cells and anticancer drug-resistant lung cancer cell lines.
14 shows the inhibitory effect of EV206 on anchorage-dependent colony formation in various lung cancer cells and anticancer drug-resistant lung cancer cell lines.
15 shows the cell viability inhibitory effect of EV408 in various lung cancer cells and anticancer drug-resistant lung cancer cell lines.
16 shows changes in adhesion-dependent colony-forming ability of EV408 in various lung cancer cells and anticancer drug-resistant lung cancer cell lines.
17 shows H1299 cells treated with up to 250 nM of evodiamine, EV206, and EV408 for 2 days, respectively, and the expression changes of Hsp70 and Hsp90/Hsp70 client proteins (Akt, Src, MEK) were confirmed by Western blot analysis.
18 shows the cancer cell survival inhibitory activity (MTT assay) of evodiamine derivatives EV501 to EV509.
19 shows the cancer cell survival inhibitory activity (Crystal violet assay) of evodiamine derivatives EV501, EV504, EV507, EV508, and EV509.
20 shows the colony formation inhibitory activity of evodiamine derivatives EV501 to EV509 on soft agar.
FIG. 21 shows the expression changes of Hsp70 and Hsp90/Hsp70 client proteins (Akt, and Src) of the evodiamine derivatives EV501 and EV507 by Western blot analysis.

Hereinafter, the present invention will be described in detail.

The present invention relates to an indoloquinazolidine alkaloid represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In Formula 1,

R ₁ and R ₂ are each independently hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, halogen, nitro (NO ₂ ), amide, —NH— aryl, hydroxy, or -COOR ₇ ;

R ₃ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, -CH ₂ -R ₈ , substituted or unsubstituted C ₁ -C ₆ acyl, substituted or unsubstituted aryl, -CO-R ₈ or benzyl ego,

R ₄ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₂ -C ₆ alkylalkenyl, substituted or unsubstituted C ₂ -C ₆ alkylalkynyl, -CH ₂ -R ₈ , substituted or unsubstituted aryl, propargyl, substituted or unsubstituted C ₁ -C ₆ acyl, or benzyl,

R ₅ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, halogen, nitro, or substituted or unsubstituted C ₁ -C ₆ alkoxy;

R ₆ is hydrogen, oxygen, or substituted or unsubstituted C ₁ -C ₆ alkyl,

R ₇ and R ₈ are each independently hydrogen, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted aryl;

X is C, N, O or S.)

In addition, the present invention relates to a pharmaceutical composition for preventing or treating cancer comprising an indoloquinazolidine alkaloid or a pharmaceutically acceptable salt thereof as an active ingredient.

,

, 또는 -COOR₇이고, In the present invention, the R ₁ and R ₂ are each independently hydrogen, methoxy, butoxy, methyl, hydroxy, F, Cl, nitro,

,

, or -COOR ₇ ,

R ₃ is hydrogen, methyl, ethyl, propyl, butyl, hexyl, benzyl, -CO-R ₈ or -CH ₂ -R ₈ ,

, 또는 -CH₂-R₈이고,R ₄ is hydrogen, methyl, benzyl, substituted or unsubstituted C ₂ -C ₅ alkenyl, substituted or unsubstituted C ₂ -C ₅ alkynyl, substituted or unsubstituted C ₂ -C ₅ alkylalkenyl, substituted or unsubstituted C ₂ -C ₅ alkyl alkynyl, cyclohexanecarbonyl,

, or -CH ₂ -R ₈ ,

R ₅ is hydrogen, methoxy, nitro, methyl, F, Cl, Br, or I;

R ₆ is hydrogen or oxygen,

R ₇ and R ₈ are each independently hydrogen, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted aryl, the substituted aryl is substituted with one or more selected from the group consisting of halogen, CN, C ₁ -C ₆ haloalkyl, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, nitro,

X may be N or O, but is not limited thereto.

As used herein, the term “C ₁ -C ₆ alkyl” refers to a monovalent alkyl group having 1 to 6 carbon atoms. The term includes functional groups such as methyl, ethyl, n -propyl, i -propyl, n -butyl, i -butyl, tert -butyl, n -hexyl and the like. Alkyl, and other substituents containing alkyl moieties described herein include both straight-chain and comminuted forms. Substituted C ₁ -C ₆ alkyl means that one or more of the hydrogen atoms is substituted with another substituent, the substituent is not limited, but halogen, N, O, S, or substituted or unsubstituted aryl, etc. include

As used herein, the term “halogen” may include fluoro (F), chloro (Cl), and bromo (Br) and iodine (I).

As used herein, the term “C ₁ -C ₆ alkoxy” refers to an —OR group, where R refers to “C ₁ -C ₆ alkyl”. Preferred alkoxy groups include, for example, methoxy, ethoxy, phenoxy, and the like.

As used herein, the term “aryl” refers to an unsaturated aromatic ring compound having 6 to 20 carbon atoms having a single ring (eg phenyl) or a plurality of condensed rings (eg naphthyl). The aryl may be selected from the group consisting of phenyl, naphthyl, anthryl and biaryl.

In the present invention, the indoloquinazolidine alkaloid may reduce one or more selected from the group consisting of the number and volume of tumors, but is not limited thereto.

In the present invention, the indoloquinazolidine alkaloid may inhibit the expression of HSP70 protein, but is not limited thereto. The HSP70 (heat shock protein 70, heat shock protein 70) is a kind of chaperone involved in the protein folding process for forming the complete structure of the protein. In several cancer cells, HSP70 is overexpressed and mediates cancer cell survival and anticancer drug resistance. The cancer is prevented or treated by inhibiting HSP70.

In the present invention, the indoloquinazolidine alkaloid may inhibit the expression of one or more proteins or genes selected from the group consisting of Akt, Src, and MEK, but is not limited thereto.

As used herein, the term “cancer” is a disease related to the regulation of cell death, and refers to a disease caused by excessive cell proliferation when the balance of normal apoptosis is disrupted. In some cases, these abnormal hyperproliferative cells may invade surrounding tissues and organs to form a mass, and may cause destruction or transformation of normal structures in the body. This condition is collectively called cancer.

In general, a tumor refers to an abnormally grown mass due to the autonomous overgrowth of body tissues, and tumors can be divided into benign tumors and malignant tumors. Malignant tumors grow very rapidly compared to benign tumors, and metastasis occurs while infiltrating the surrounding tissues, threatening life. Such malignant tumors are commonly referred to as 'cancer'.

In the present invention, the cancer is cervical cancer, lung cancer, pancreatic cancer, non-small cell lung cancer, liver cancer, colon cancer, colorectal cancer, bone cancer, skin cancer, head cancer, cervical cancer, skin melanoma, intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer , liver cancer, brain tumor, blood cancer, stomach cancer, perianal cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine adenocarcinoma, thyroid cancer, parathyroid cancer, parathyroid cancer Renal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, bladder cancer, kidney cancer, ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem It may be one or more selected from the group consisting of glioma and pituitary adenoma, but is not limited thereto.

In the present invention, the cancer may be drug-resistant cancer, but is not limited thereto. The drug-resistant cancer refers to a case in which drug-resistant cells occupy most of the cancer cells. This is progressed as most cancer masses become drug-resistant as cells with drug resistance selectively survive and proliferate. The agent may be pemetrexed, cisplatin or paclitaxel, but is not limited thereto.

In the present invention, the cancer may be lung cancer, specifically small cell lung cancer or non-small cell lung cancer, but is not limited thereto.

As used herein, the term “lung cancer” refers to a malignant tumor that occurs in the lung, and primary lung cancer that occurs in the lung itself is classified into non-small cell lung cancer and small cell lung cancer based on the size and shape of cancer cells.

As used herein, “non-small cell lung cancer (NSCLC)” refers to lung cancer in which the size of cancer cells is not small. It accounts for 80-85% of lung cancers, and includes squamous cell carcinoma, adenocarcinoma (adenocarcinoma), large cell carcinoma, adenosquamous cell carcinoma, sarcoma, carcinoid tumor, salivary gland carcinoma, and unclassified cancer. Non-small cell lung cancer can be caused by mutations, such as EGFR (epidermal growth factor receptor) mutated non-small cell lung cancer, KRAS (v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog) mutated non-small cell lung cancer, ALK (Anaplastic Lymphoma Kinase) mutated arsenic. and cell lung cancer.

The indoloquinazolidine alkaloid compound may have a non-aromatic double bond and one or more asymmetric centers. Thus, they may occur as racemates and racemic mixtures, single enantiomers, individual diastereomers, diastereomeric mixtures and cis- or trans-isomers. All such isomeric forms are contemplated.

The present invention may also include the pharmaceutically acceptable salt as an active ingredient. As used herein, the term "pharmaceutically acceptable salt" includes salts derived from pharmaceutically acceptable inorganic acids, organic acids, or bases.

Examples of suitable acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid , benzoic acid, malonic acid, gluconic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Acid addition salts can be prepared by conventional methods, for example, by dissolving the compound in an aqueous solution of an excess of acid, and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. It can also be prepared by heating an equimolar amount of the compound and an acid or alcohol in water and then evaporating the mixture to dryness, or by suction filtration of the precipitated salt.

Salts derived from suitable bases may include, but are not limited to, alkali metals such as sodium and potassium, alkaline earth metals such as magnesium, and ammonium. The alkali metal or alkaline earth metal salt can be obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and then evaporating and drying the filtrate. In this case, it is pharmaceutically suitable to prepare a sodium, potassium or calcium salt as the metal salt, and the corresponding silver salt can be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (eg, silver nitrate).

The content of the compound in the composition of the present invention can be appropriately adjusted according to the symptoms of the disease, the degree of progression of the symptoms, the condition of the patient, etc., for example, 0.0001 to 99.9% by weight, or 0.001 to 50% by weight based on the total weight of the composition. However, the present invention is not limited thereto. The content ratio is a value based on the dry amount from which the solvent is removed.

The pharmaceutical composition according to the present invention may further include suitable carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions. The excipient may be, for example, at least one selected from the group consisting of a diluent, a binder, a disintegrant, a lubricant, an adsorbent, a humectant, a film-coating material, and a controlled-release additive.

The pharmaceutical composition according to the present invention can be prepared according to a conventional method, respectively, in powders, granules, sustained-release granules, enteric granules, liquids, eye drops, elsilic, emulsions, suspensions, spirits, troches, fragrances, and limonaade. , tablets, sustained release tablets, enteric tablets, sublingual tablets, hard capsules, soft capsules, sustained release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, perfusates, Warnings, lotions, pasta, sprays, inhalants, patches, sterile injection solutions, or external preparations such as aerosols can be formulated and used, and the external preparations are creams, gels, patches, sprays, ointments, warning agents , lotion, liniment, pasta, or cataplasma.

Carriers, excipients and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used.

As additives for tablets, powders, granules, capsules, pills, and troches according to the present invention, corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, di-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, phosphoric acid Calcium monohydrogen, calcium sulfate, sodium chloride, sodium hydrogen carbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropylmethyl excipients such as cellulose (HPMC), HPMC 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, and Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose phthalate acetate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin , hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, purified shellac, starch powder, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, etc. Hydroxypropyl methylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, silicic anhydride, 1-hydroxy Propylcellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, disintegrants such as amylopectin, pectin, sodium polyphosphate, ethyl cellulose, sucrose, magnesium aluminum silicate, di-sorbitol solution, light anhydrous silicic acid; Calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodite, kaolin, petrolatum, sodium stearate, cacao fat, sodium salicylate, magnesium salicylate, polyethylene glycol 4000, 6000, liquid paraffin, hydrogenated soybean oil (Lubri) wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, silicic anhydride, higher fatty acid, higher alcohol, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, A lubricant such as sodium acetate, sodium oleate, dl-leucine, light anhydrous silicic acid; may be used.

As additives for the liquid formulation according to the present invention, water, diluted hydrochloric acid, diluted sulfuric acid, sodium citrate, monostearate sucrose, polyoxyethylene sorbitol fatty acid esters (Twinester), polyoxyethylene monoalkyl ethers, lanolin ethers, Lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc. can be used.

In the syrup according to the present invention, a sucrose solution, other sugars or sweeteners may be used, and if necessary, a fragrance, colorant, preservative, stabilizer, suspending agent, emulsifier, thickening agent, etc. may be used.

Purified water may be used in the emulsion according to the present invention, and if necessary, an emulsifier, preservative, stabilizer, fragrance, etc. may be used.

Suspension agents according to the present invention include acacia, tragacantha, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC 1828, HPMC 2906, HPMC 2910, etc. Agents may be used, and surfactants, preservatives, stabilizers, colorants, and fragrances may be used as needed.

The injection according to the present invention includes distilled water for injection, 0.9% sodium chloride injection, ring gel injection, dextrose injection, dextrose + sodium chloride injection, PEG (PEG), lactated ring gel injection, ethanol, propylene glycol, non-volatile oil-sesame oil , solvents such as cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; Solubilizing aids such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, tweens, nijeongtinamide, hexamine, and dimethylacetamide; Weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, buffers such as albumin, peptone, gum; isotonic agents such as sodium chloride; Stabilizers such as sodium bisulfite (NaHSO ₃ ) carbon dioxide gas, sodium metabisulfite (Na ₂ S ₂ O ₃ ), sodium sulfite (Na ₂ SO ₃ ), nitrogen gas (N ₂ ), ethylenediaminetetraacetic acid; sulphating agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate, acetone sodium bisulfite; analgesic agents such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; suspending agents such as SiMC sodium, sodium alginate, Tween 80, and aluminum monostearate.

The suppository according to the present invention includes cacao fat, lanolin, witepsol, polyethylene glycol, glycerogelatin, methyl cellulose, carboxymethyl cellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cacao butter + Cholesterol, Lecithin, Lanet Wax, Glycerol Monostearate, Tween or Span, Imhausen, Monolene (Propylene Glycol Monostearate), Glycerin, Adeps Solidus, Butyrum Tego -G), Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydroxote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydro Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium, A, AS, B, C, D, E, I, T, Massa-MF, Masupol, Masupol-15, Neosupostal-N, Paramound-B, Suposhiro (OSI, OSIX, A, B, C, D, H, L), Suppository IV type (AB, B, A, BC, BBG, E, BGF, C, D, 299), supostal (N, Es), Wecobi (W, R, S, M, Fs), tester triglyceride base (TG-95, MA, 57) and The same mechanism may be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose ) or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate talc are also used.

Liquid formulations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, drug activity, and type of the patient's disease; Sensitivity to the drug, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs and other factors well known in the medical field may be determined.

The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple. In consideration of all of the above factors, it is important to administer an amount capable of obtaining the maximum effect with a minimum amount without side effects, which can be easily determined by a person skilled in the art to which the present invention pertains.

The pharmaceutical composition of the present invention may be administered to an individual by various routes. All modes of administration can be envisaged, for example, oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal (intrathecal) injection, sublingual administration, buccal administration, rectal insertion, vaginal It can be administered according to internal insertion, ocular administration, ear administration, nasal administration, inhalation, spraying through the mouth or nose, skin administration, transdermal administration, and the like.

The pharmaceutical composition of the present invention is determined according to the type of drug as the active ingredient along with various related factors such as the disease to be treated, the route of administration, the patient's age, sex, weight, and the severity of the disease.

In the present invention, "individual" means a subject in need of treatment for a disease, and more specifically, human or non-human primates, mice, rats, dogs, cats, horses, cattle, etc. It may be a mammal of, but is not limited thereto.

In the present invention, "administration" means providing a predetermined composition of the present invention to a subject by any suitable method.

In the present invention, “prevention” means any action that suppresses or delays the onset of a target disease, and “treatment” means that the target disease and its metabolic abnormalities are improved or It means any action that is beneficially changed, and “improvement” means any action that reduces a parameter related to a desired disease, for example, the degree of a symptom by administration of the composition according to the present invention.

In addition, the present invention provides an anticancer adjuvant for enhancing the anticancer efficacy of an anticancer agent, comprising an indoloquinazolidine alkaloid or a pharmaceutically acceptable salt thereof as an active ingredient.

In the present invention, the cancer may be lung cancer, but is not limited thereto.

The anticancer adjuvant may enhance the efficacy of the anticancer agent. In addition, the anticancer adjuvant may be administered simultaneously or sequentially with the anticancer agent. When administered sequentially, the anticancer agent may be added after the anticancer adjuvant is injected, and the anticancer adjuvant may be added after the anticancer drug is injected, but is not limited thereto. The administration method may be changed to enhance the anticancer effect.

The anticancer agent may be pemetrexed, cisplatin, or paclitaxel, but is not limited thereto.

Since the present invention can apply various transformations and can have various embodiments, specific embodiments are illustrated in the drawings and described in detail in the detailed description. However, this is not intended to limit the present invention to specific embodiments, and it should be understood to include all modifications, equivalents and substitutes included in the spirit and scope of the present invention. In describing the present invention, if it is determined that a detailed description of a related known technology may obscure the gist of the present invention, the detailed description thereof will be omitted.

Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

synthesis example. Synthesis of Evodiamine Derivatives

Synthesis Example 1. General method of synthesis of Evodiamine (1) and an intermediate

N -formyltryptamine (3)

: Tryptamine ( 2 ) (1.00 g, 6.24 mmol) and ethyl formate (2.3 g, 31.210 mmol) were heated and stirred at 80°C under Ar substitution. After completion of the reaction, the mixture was concentrated under reduced pressure to remove excess ethyl formate, and the resulting mixture was purified by column chromatography (silica gel, MC:MeOH = 20:1 to 10:1) to form a brown oil of Compound 3 (1.2 g, 100 % yield) was obtained.

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.13 (br s, 2H), 7.61 (d, J = 8.3 Hz, 1H), 7.38-7.40 (m, 1H), 7.23 (td, J = 7.6, 1.4) Hz, 1H), 7.14 (t, J = 7.6 Hz, 1H), 7.06 (s, 1H), 5.58 (br s, 1H), 3.64-3.69 (m, 2H), 3.01 (t, J = 6.9 Hz, 2H); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 161.5, 136.5, 127.3, 122.4, 122.2, 119.5, 118.7, 112.4, 111.5, 42.1, 38.4, 27.4, 25.2; HRMS (FAB) m/z calcd for C ₁₁ H ₁₂ N ₂ O [M+H] ⁺ : 188.0950, found 188.0955.

3,4-dihydro-β-carboline (4)

: N -formyltryptamine ( 3 ) (1.1 g, 5.920 mmol) was dissolved in CH ₂ Cl ₂ (15.0 mL), and then POCl ₃ (1.7 mL, 17.770 mmol) was slowly added at 0°C. After stirring at 0° C. for 2 hours, the mixture was stirred at room temperature for 2 hours. After completion of the reaction, excess POCl ₃ and CH ₂ Cl ₂ were removed by concentration under reduced pressure, 1 M HCl (100 mL) was added to the obtained mixture, and then washed once with CH ₂ Cl ₂ (1 × 30 mL). The washed aqueous layer was adjusted to pH = 10 with 1 M NaOH at 0° C., and extracted three times with CH ₂ Cl ₂ (3 × 30 mL). The obtained organic layer was dried over MgSO ₄ , filtered, and concentrated under reduced pressure. Et ₂ O was added to the obtained mixture to precipitate a solid, and then filtered to obtain compound 4 (4.0 g, 64% yield) in the form of a yellow solid.

mp: 99-101°C; ¹ H-NMR (400 MHz, DMSO-D ₆ ) δ 11.32 (s, 1H), 8.36 (t, J = 2.3 Hz, 1H), 7.55 (d, J = 8.2 Hz, 1H), 7.41 (d, J) = 8.2 Hz, 1H), 7.17-7.21 (m, 1H), 7.04 (td, J = 7.5, 0.9 Hz, 1H), 3.78 (td, J = 8.7, 2.1 Hz, 2H), 2.80 (t, J = 8.7 Hz, 2H); ¹³ C-NMR (100 MHz, DMSO-D ₆ ) δ 151.6, 136.7, 128.4, 124.8, 123.7, 119.7, 119.6, 113.8, 112.5, 48.1, 18.7; HRMS (FAB) m/z calcd for C ₁₁ H ₁₁ N ₂ [M+H] ⁺ : 171.0917, found 171.0921.

Isatoic anhydride (6)

: Anthranilic acid ( 5 ) (1.1 g, 8.090 mmol) was dissolved in THF (15.0 mL), and then triphosgene (7.2 g, 24.280 mmol) was added at room temperature. Thereafter, the reaction temperature was raised to 50° C. and stirred for 3 hours. After completion of the reaction, ice water (50 mL) was added to the mixture, followed by filtration under reduced pressure to obtain Compound 6 (1.3 g, 95% yield) in the form of a white solid.

mp: 164-165° C.; ¹ H-NMR (400 MHz, DMSO-D ₆ ) δ 11.73 (s, 1H), 7.91 (dd, J = 7.8, 1.4 Hz, 1H), 7.72-7.76 (m, 1H), 7.23-7.27 (m, 1H), 7.15 (d, J = 7.8 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-D ₆ ) δ 159.9, 147.1, 141.4, 137.00, 129.00, 123.5, 115.4, 110.3; HRMS (FAB) m/z calcd for C ₈ H ₅ NO ₃ [M+H] ⁺ : 164.0269, found 164.0271.

N -methylisatoic anhydride (7)

: Isatoic anhydride ( 6 ) (200.0 mg, 1.226 mmol) was dissolved in DMAc (3.0 mL), DIPEA (0.64 mL, 3.678 mmol) and 1-iodomethane (522.1 mg, 3.678 mmol) were slowly added thereto, and then at 40°C. Stir overnight. Then, after completion of the reaction by adding H ₂ O to the mixture, the mixture was extracted several times with CH ₂ Cl ₂ . The collected organic layers were dried over MgSO ₄ , filtered, and concentrated under reduced pressure. After adding hexane to the obtained mixture to precipitate a solid, the mixture was filtered to obtain a solid compound 7 (1.7 g, 68% yield).

mp: 106-107 °C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.01 (dd, J = 7.8, 1.8 Hz, 1H), 7.86 (td, J = 7.9, 1.5 Hz, 1H), 7.45 (d, J = 8.3 Hz) , 1H), 7.34 (t, J = 7.6 Hz, 1H), 3.47 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 159.0, 147.8, 142.2, 137.2, 129.3, 123.6, 114.9, 111.6, 31.7; LRMS (FAB) m/z, 334 ([M+H] ⁺ ).

Evodiamine (1)

: 3,4-dihydro-β-carboline ( 4 ) (50.0 mg, 0.290 mmol) and N -methylisatoic anhydride ( 7 ) (46.1 mg, 0.260 mmol) were dissolved in CH ₂ Cl ₂ (0.7 mL) and stirred at 50° C. for 6 hours under Ar substitution. The resulting solid was filtered to obtain Compound 1 (67.2 mg, 85% yield) as a yellow solid.

mp: 302-303 °C; ¹ H-NMR (400 MHz, DMSO-D ₆ ) δ 11.07 (s, 1H), 7.79 (dd, J = 7.6, 1.6 Hz, 1H), 7.46-7.50 (m, 2H), 7.36 (d, J = 7.8 Hz, 1H), 6.94-7.13 (m, 4H), 6.13 (s, 1H), 4.61-4.65 (m, 1H), 3.17-3.24 (m, 1H), 2.90-2.95 (m, 1H), 2.88 (s, 3H), 2.79 (dd, J = 15.4, 4.8 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-D ₆ ) δ 164.3, 148.8, 136.5, 133.5, 130.6, 128.0, 126.00, 121.9, 120.3, 119.3, 118.9, 118.2, 117.5, 111.7, 111.5, 69.8, 40.9, 36.5, 19.5; HRMS (FAB) m/z calcd for C ₁₉ H ₁₈ N ₃ O [M+H] ⁺ : 304.1444, found 304.1434.

Synthesis Example 2. Synthesis of B-ring evodiamine derivatives (EV101~112)

<Synthesis method of EV101~112 derivatives>

<N General conditions for -alkylation>

: Evodiamine ( 1 ) (1.0 equiv.) was dissolved in DMF (0.1 M), NaH (2.0 equiv.) and an alkyl halide (2.0 equiv.) were sequentially added thereto, followed by stirring at room temperature overnight. After the reaction was terminated by adding H ₂ O (15 mL) to the reaction product, the aqueous layer was extracted three times with ethyl acetate (3 × 45 mL). The obtained organic layer was dried over MgSO ₄ , filtered, and concentrated under reduced pressure. The resulting mixture was purified by column chromatography (silica gel, hexane:ethyl acetate = 3:1 to 2:1) to obtain a solid compound EV101-112 .

EV101

: According to the general conditions of N -alkylation, EV101 (715.5 mg, 72% yield) in yellow solid form using Evodiamine ( 1 ) (872.7 mg, 2.877 mmol) and 1-chloropropane (451.9 mg, 5.754 mmol) was obtained. got it

mp: 168-170℃; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.96 (d, J = 7.8 Hz, 1H), 7.73-7.76 (m, 2H), 7.57 (t, J = 7.5 Hz, 1H), 7.43 (t) , J = 8.7 Hz, 2H), 7.13-7.17 (m, 1H), 7.01 (t, J = 7.6 Hz, 1H), 4.73-4.80 (m, 1H), 4.43-4.51 (m, 1H), 4.36 ( dd, J = 12.6, 4.8 Hz, 1H), 3.77 (q, J = 5.7 Hz, 1H), 3.37-3.43 (m, 1H), 2.72-2.85 (m, 2H), 1.92 (d, J = 5.5 Hz) , 3H), 1.78-1.88 (m, 1H), 1.41-1.54 (m, 1H), 1.05 (t, J = 7.4 Hz, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 167.2, 144.9, 136.5, 134.9, 132.5, 132.0, 129.6, 125.7, 124.5, 123.1, 122.2, 119.1, 118.8, 110.4, 109.9, 80.1, 46.4, 34.6, 27.9, 22.7, 21.7, 10.8; LRMS (FAB) m/z, 346 ([M+H] ⁺ ).

EV102

: According to the general conditions of N -alkylation, EV102 (412.2 mg, 97.4% yield) in the form of a pale yellow solid using Evodiamine ( 1 ) (300.0 mg, 0.989 mmol) and 2-chlorobenzyl chloride (318.5 mg, 1.978 mmol) ) was obtained.

mp: 242-244 °C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.17 (d, J = 6.9 Hz, 1H), 7.87 (d, J = 8.3 Hz, 2H), 7.53-7.71 (m, 6H), 7.13 (t) , J = 7.6 Hz, 1H), 7.01 (t, J = 7.8 Hz, 1H), 6.46 (d, J = 8.3 Hz, 1H), 5.03 (q, J = 5.5 Hz, 1H), 4.49-4.56 (m) , 1H), 3.43 (dd, J = 13.6, 5.7 Hz, 1H), 3.28 (d, J = 13.3 Hz, 1H), 3.01-3.08 (m, 1H), 2.81-2.85 (m, 1H), 2.52- 2.57 (m, 1H), 1.88 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 169.6, 143.2, 136.6, 132.1, 130.4, 130.2, 129.7, 128.7, 128.6, 124.6, 123.8, 123.00, 121.9, 119.6, 114.1, 112.9, 76.4, 58.8, 35.1, 23.35; LRMS (FAB) m/z, 428 ([M+H] ⁺ ).

EV103

: According to the general conditions of N -alkylation, EV103 (285.6 mg, 73% yield) in the form of a pale yellow solid was obtained using Evodiamine ( 1 ) (300.0 mg, 0.989 mmol) and benzyl bromide (338.3 mg, 1.978 mmol). got it

mp: 190-192°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.69 (d, J = 7.4 Hz, 1H), 7.63 (d, J = 7.8 Hz, 1H), 7.48 (q, J = 2.8 Hz, 1H), 7.43 (t, J = 7.4 Hz, 1H), 7.23-7.27 (m, 1H), 7.17-7.21 (m, 3H), 6.93-7.03 (m, 3H), 6.85-6.87 (m, 2H), 6.23 ( d, J = 17.0 Hz, 1H), 5.91 (d, J = 17.0 Hz, 1H), 4.41 (dd, J = 13.1, 3.4 Hz, 1H), 3.90 (q, J = 5.7 Hz, 1H), 3.46 3.53 (m, 1H), 2.79-2.90 (m, 2H), 1.89 (d, J = 5.5 Hz, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 167.3, 144.3, 138.3, 136.9, 135.6, 132.2, 131.6, 129.3, 128.2, 126.7, 126.0, 126.0, 124.8, 122.9, 122.4, 119.4, 118.9, 110.8, 110.4, 79.9, 48.7, 34.7, 27.9, 21.9; LRMS (FAB) m/z, 394 ([M+H] ⁺ ).

EV104

: EV104 (383 mg, 100 % yield) in the form of a pale yellow solid using Evodiamine ( 1 ) (300.0 mg, 0.989 mmol) and 1-iodohexane (419.4 mg, 1.978 mmol) according to the general conditions of N -alkylation got

mp: 66-68 ° C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.95 (d, J = 7.4 Hz, 1H), 7.71-7.75 (m, 2H), 7.58 (t, J = 7.4 Hz, 1H), 7.44 (d) , J = 7.8 Hz, 1H), 7.40 (d, J = 8.3 Hz, 1H), 7.14-7.18 (m, 1H), 7.01 (t, J = 7.1 Hz, 1H), 4.73-4.79 (m, 1H) , 4.50-4.58 (m, 1H), 4.36 (dd, J = 12.6, 4.8 Hz, 1H), 3.78 (q, J = 5.5 Hz, 1H), 3.40 (td, J = 12.1, 4.9 Hz, 1H), 2.71-2.84 (m, 2H), 1.91 (d, J = 6.0 Hz, 3H), 1.71-1.83 (m, 1H), 1.50 (s, 3H), 1.34 (d, J = 3.2 Hz, 4H), 0.91 (t, J = 6.9 Hz, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 167.2, 144.9, 136.5, 134.8, 132.5, 132.00, 129.6, 125.8, 124.5, 123.1, 122.2, 119.1, 118.6, 110.3, 110.0, 80.1, 45.0, 34.6, 31.0, 29.3, 27.7, 25.8, 22.0, 21.7, 13.9; LRMS (FAB) m/z, 388 ([M+H] ⁺ ).

EV105

: EV105 (385.5 mg, 91% yield) in the form of a yellow solid using Evodiamine ( 1 ) (300.0 mg, 0.989 mmol) and 3-chlorobenzyl chloride (630.0 mg, 1.978 mmol) according to the general conditions of N -alkylation got

mp: 168-170℃; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.90 (d, J = 7.8 Hz, 1H), 7.63 (d, J = 7.8 Hz, 1H), 7.52 (dd, J = 13.8, 7.8 Hz, 2H) ), 7.28-7.32 (m, 2H), 7.21 (t, J = 7.4 Hz, 4H), 7.12 (t, J = 7.4 Hz, 1H), 7.04 (s, 1H), 6.05 (s, 1H), 5.57 (q, J = 17.3 Hz, 2H), 4.66 (dd, J = 12.9, 4.1 Hz, 1H), 3.11-3.17 (m, 1H), 3.01 (d, J = 14.2 Hz, 1H), 2.83 (t, J = 10.6 Hz, 1H), 2.26 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.6, 140.8, 137.2, 133.1, 132.9, 130.3, 128.0, 127.1, 126.7, 125.4, 125.4, 123.6, 122.7, 119.6, 118.9, 113.0, 110.3, 46.1, 19.8; LRMS (FAB) m/z, 428 ([M+H] ⁺ ).

EV106

: EV106 (371.6 mg, 92% yield) in yellow solid form using Evodiamine ( 1 ) (300.0 mg, 0.989 mmol) and 4-methylbenzyl bromide (183.0 mg, 0.989 mmol) according to the general conditions of N -alkylation got

mp: 158-160°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.90 (d, J = 7.8 Hz, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.54 (t, J = 7.6 Hz, 1H), 7.48 (d, J = 7.8 Hz, 1H), 7.17-7.26 (m, 3H), 7.04-7.12 (m, 3H), 6.99 (d, J = 8.3 Hz, 2H), 5.98 (s, 1H), 5.52 (d, J = 3.4 Hz, 2H), 4.66 (dd, J = 12.9, 3.7 Hz, 1H), 3.13 (td, J = 12.3, 3.8 Hz, 1H), 3.01 (d, J = 14.7 Hz, 1H) , 2.78-2.85 (m, 1H), 2.34 (s, 3H), 2.21 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.5, 137.3, 136.3, 135.0, 133.0, 129.0, 128.0, 126.6, 125.4, 123.5, 122.8, 122.4, 119.4, 118.8, 112.8, 110.4, 46.4, 20.6, 19.8; HRMS (FAB) m/z calcd for C ₂₇ H ₂₆ N ₃ O [M+H] ⁺ : 408.2076, found 408.2078.

EV107

: EV107 (282.0 mg, 70% yield) in the form of a white solid using Evodiamine ( 1 ) (300.0 mg, 0.989 mmol) and 4-methoxybenzyl bromide (154.9 mg, 0.989 mmol) according to the general conditions of N -alkylation got

mp: 174-176°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.90 (d, J = 7.8 Hz, 1H), 7.61 (d, J = 7.8 Hz, 1H), 7.50-7.56 (m, 2H), 7.17-7.27 (m, 3H), 7.05-7.12 (m, 3H), 6.81 (d, J = 8.7 Hz, 2H), 6.01 (s, 1H), 5.49 (d, J = 3.2 Hz, 2H), 4.66 (dd, J = 12.9, 4.1 Hz, 1H), 3.66 (s, 3H), 3.12 (td, J = 12.2, 3.5 Hz, 1H), 3.00 (d, J = 14.7 Hz, 1H), 2.78-2.85 (m, 1H) ), 2.33 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.6, 158.4, 137.2, 133.0, 129.9, 128.1, 125.4, 122.4, 119.4, 118.8, 113.8, 112.7, 110.5, 55.0, 46.1, 19.8; LRMS (FAB) m/z, 424 ([M+H] ⁺ ).

EV108

: EV108 (128.4 mg, 30% yield) in the form of a yellow solid using Evodiamine ( 1 ) (300.0 mg, 0.989 mmol) and 4-chlorobenzyl chloride (159.3 mg, 0.989 mmol) according to the general conditions of N -alkylation got

mp: 202-204°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.86-7.91 (m, 1H), 7.62 (d, J = 7.8 Hz, 1H), 7.47-7.55 (m, 2H), 7.31 (t, J = 8.3 Hz, 2H), 7.20 (t, J = 7.1 Hz, 3H), 7.13 (q, J = 7.5 Hz, 3H), 6.02 (s, 1H), 5.56 (d, J = 5.5 Hz, 2H), 4.66 (dd, J = 12.9, 4.1 Hz, 1H), 3.14 (td, J = 12.3, 3.8 Hz, 1H), 3.01 (d, J = 14.7 Hz, 1H), 2.80-2.85 (m, 1H), 2.29 ( s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.6, 137.2, 133.0, 131.7, 128.5, 128.4, 128.0, 125.4, 122.6, 119.5, 118.9, 113.0, 110.3, 54.9, 46.0, 19.8; HRMS (FAB) m/z calcd for C ₂₆ H ₂₃ ClN ₃ O [M+H] ⁺ : 428.1530, found 428.1535.

EV109

: According to the general conditions of N -alkylation, EV109 (213.4 mg, 68% yield) in yellow solid form using Evodiamine ( 1 ) (300.0 mg, 0.989 mmol) and 1-iodomethane (140.4 mg, 0.989 mmol) was obtained. got it

mp: 176-178°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.13 (d, J = 7.9 Hz, 1H), 7.70-7.74 (m, 2H), 7.56 (t, J = 7.0 Hz, 1H), 7.43 (dd , J = 13.4, 7.9 Hz, 2H), 7.17 (td, J = 7.6, 1.2 Hz, 1H), 7.02 (t, J = 7.6 Hz, 1H), 4.36-4.40 (m, 1H), 4.19 (s, 3H), 3.79 (q, J = 5.7 Hz, 1H), 3.37-3.44 (m, 1H), 2.72-2.83 (m, 2H), 1.93 (d, J = 6.1 Hz, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 167.3, 145.2, 137.4, 135.0, 132.2, 132.2, 129.4, 125.3, 124.7, 123.1, 122.3, 119.1, 118.7, 109.9, 109.7, 79.7, 34.8, 32.5, 27.8, 21.7; LRMS (FAB) m/z, 320 ([M+H] ⁺ ).

EV110

: According to the general conditions of N -alkylation, EV110 (315.8 mg, 78.4% yield) in the form of a pale yellow solid was obtained using Evodiamine ( 1 ) (300.0 mg, 0.989 mmol) and benzoyl chloride (278.0 mg, 1.978 mmol). got it

mp: 233-235° C.; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.79 (t, J = 7.0 Hz, 3H), 7.71 (t, J = 3.1 Hz, 1H), 7.62 (t, J = 7.3 Hz, 1H), 7.51 (t, J = 7.6 Hz, 2H), 7.41-7.45 (m, 1H), 7.27-7.32 (m, 3H), 7.12 (t, J = 7.6 Hz, 1H), 7.03 (d, J = 7.9 Hz) , 1H), 5.66 (s, 1H), 4.66 (dd, J = 12.8, 4.9 Hz, 1H), 3.24 (td, J = 12.2, 3.7 Hz, 1H), 3.05 (d, J = 14.7 Hz, 1H) , 2.82-2.89 (m, 1H), 2.27 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 168.4, 163.4, 150.1, 137.1, 134.6, 133.1, 129.3, 128.8, 127.9, 127.2, 125.2, 123.5, 123.0, 122.7, 122.4, 121.1, 119.6, 114.1, 67.7, 37.7, 36.4, 20.0; LRMS (FAB) m/z, 334 ([M+H] ⁺ ), 307, 273.

EV111

: EV111 (334.5 mg, 81% yield) in the form of a pale yellow solid using Evodiamine ( 1 ) (300.0 mg, 0.989 mmol) and 4-cyanobenzyl bromide (387.7 mg, 1.978 mmol) according to the general conditions of N -alkylation ) was obtained.

mp: 207-209°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.89 (d, J = 7.3 Hz, 1H), 7.74 (d, J = 7.9 Hz, 2H), 7.64 (d, J = 7.9 Hz, 1H), 7.47-7.51 (m, 2H), 7.11-7.25 (m, 6H), 6.03 (s, 1H), 5.66 (dd, J = 28.4, 17.4 Hz, 2H), 4.66 (dd, J = 12.8, 3.7 Hz, 1H), 3.12-3.18 (m, 1H), 3.01 (d, J = 14.7 Hz, 1H), 2.85 (d, J = 9.8 Hz, 1H), 2.25 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.6, 144.1, 137.2, 133.0, 132.4, 128.0, 127.5, 125.5, 123.6, 122.7, 119.7, 118.9, 118.7, 113.2, 110.2, 109.9, 46.4, 19.8; LRMS (FAB) m/z, 421 ([M+H] ⁺ ).

EV112

: According to the general conditions of the N -alkylation, Evodiamine ( 1 ) (250.0 mg, 0.814 mmol) and cyclohexanecarbonyl chloride (241.6 mg, 1.648 mmol) were used to obtain EV112 (279.0 mg, 82% yield) in the form of a yellow solid. .

mp: 91-93°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.98 (d, J = 8.6 Hz, 1H), 7.92 (dd, J = 7.6, 1.5 Hz, 1H), 7.68 (d, J = 7.9 Hz, 1H) ), 7.52-7.57 (m, 1H), 7.39-7.43 (m, 1H), 7.32 (t, J = 7.3 Hz, 1H), 7.17-7.23 (m, 2H), 6.33 (s, 1H), 4.67 ( dd, J = 13.1, 4.0 Hz, 1H), 3.17-3.30 (m, 2H), 3.02-3.06 (m, 1H), 2.80 (tt, J = 12.8, 3.1 Hz, 1H), 2.29 (s, 3H) , 2.06 (d, J = 12.8 Hz, 1H), 1.89 (d, J = 12.2 Hz, 1H), 1.74-1.76 (m, 1H), 1.58-1.69 (m, 3H), 1.38-1.47 (m, 1H) ), 1.17-1.30 (m, 2H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 176.5, 163.4, 150.1, 136.4, 133.1, 128.3, 128.1, 127.2, 125.7, 123.4, 123.0, 123.0, 121.9, 121.4, 119.3, 114.9, 68.2, 44.7, 37.5, 35.8, 30.5, 28.2, 25.3, 25.2, 24.6, 20.0; LRMS (FAB) m/z, 413 ([M+H] ⁺ ).

Synthesis Example 3. Synthesis of D-ring evodiamine derivatives (EV201 to 222)

<EV201~220 Derivative Synthesis Method>

N -propargylisatoic anhydride (7ae)

: According to the above N -methylisatoic anhydride ( 7 ) synthesis conditions, using isatoic anhydride ( 6 ) (200.0 mg, 1.226 mmol) and propargyl bromide (437.5 mg, 3.678 mmol) as a beige solid compound 7ae (166.6 mg, 68 % yield) was obtained.

mp: 176-177°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.04 (d, J = 7.8 Hz, 1n), 7.91 (t, J = 7.8 Hz, 1H), 7.51 (d, J = 8.7 Hz, 1H), 7.38 (t, J = 7.6 Hz, 1H), 4.89 (s, 2H), 3.43 (s, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 158.5, 147.3, 140.5, 137.2, 129.6, 124.1, 115.0, 111.9, 77.6, 76.1, 34.4; LRMS (FAB) m/z, 202 ([M+H] ⁺ ).

N -allylisatoic anhydride (7af)

: According to the above N -methylisatoic anhydride ( 7 ) synthesis conditions, using isatoic anhydride ( 6 ) (200.0 mg, 1.226 mmol) and allyl bromide (445.0 mg, 3.678 mmol) as a beige solid compound 7af (106.0 mg, 50 % yield) was obtained.

mp: 127-128°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.02 (dd, J = 8.0, 1.1 Hz, 1H), 7.79-7.84 (m, 1H), 7.31-7.35 (m, 2H), 5.87-5.96 ( m, 1H), 5.31 (dq, J = 17.5, 1.7 Hz, 1H), 5.20 (dq, J = 10.6, 1.5 Hz, 1H), 4.67 (td, J = 3.2, 1.5 Hz, 2H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 159.0, 147.6, 141.3, 137.0, 131.2, 129.5, 123.6, 117.2, 115.2, 111.9, 46.5, 40.2, 39.9, 39.7, 39.5, 39.3, 39.1, 38.90; LRMS (FAB) m/z, 334 ([M+H] ⁺ ).

N -(2-methyl)allylisatoic anhydride (7aq)

: According to the synthesis conditions of N -methylisatoic anhydride ( 7 ), isatoic anhydride ( 6 ) (200.0 mg, 1.226 mmol) and 3-bromo-2-methylpropene (496.6 mg, 3.678 mmol) were used as a beige solid compound 7aq (151.6 mg, 57% yield) was obtained.

mp: 124-125°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.02 (dd, J = 7.7, 1.5 Hz, 1H), 7.78-7.82 (m, 1H), 7.31-7.35 (m, 1H), 7.24 (d, J = 8.6 Hz, 1H), 4.82-4.87 (m, 2H), 4.55 (s, 2H), 1.80 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 159.0, 147.8, 141.5, 138.2, 136.9, 129.4, 123.7, 115.4, 111.8, 111.0, 49.5, 19.8; LRMS (FAB) m/z, 334 ([M+H] ⁺ ).

EV201

: According to the above Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (200 mg, 1.175 mmol) and isatoic anhydride ( 6 ) (174.3 mg, 1.068 mmol) was used to form a white solid. Compound EV201 (153.0 mg, 50% yield) was obtained.

mp: 210-212°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.91 (s, 1H), 7.76 (dd, J = 7.8, 1.4 Hz, 1H), 7.51 (d, J = 7.8 Hz, 1H), 7.43 (d) , J = 8.3 Hz, 1H), 7.33-7.38 (m, 1H), 7.14 (td, J = 7.5, 1.1 Hz, 1H), 7.04 (td, J = 7.4, 1.0 Hz, 1H), 6.90 (s, 1H), 6.82-6.88 (m, 2H), 6.05 (d, J = 1.4 Hz, 1H), 4.80 (dd, J = 12.9, 3.7 Hz, 1H), 3.02 (td, J = 12.2, 4.7 Hz, 1H) ), 2.73-2.86 (m, 2H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.7, 147.3, 136.3, 133.3, 130.8, 128.2, 125.9, 121.9, 119.0, 118.6, 118.5, 116.1, 115.4, 115.3, 111.7, 109.3, 63.8, 38.7, 20.1; LRMS (FAB) m/z, 290 ([M+H] ⁺ ).

EV202

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (74.0 mg, 0.434 mmol) and N -benzylisatoic anhydride ( 7aa ) (100.0 mg, 0.395 mmol) were used as a yellow solid The form of compound EV202 (29.6 mg, 20% yield) was obtained.

mp: 251-252°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.16 (s, 1H), 7.73-7.75 (m, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.28-7.37 (m, 6H) , 7.15-7.25 (m, 1H), 7.09 (t, J = 7.3 Hz, 1H), 6.99 (t, J = 7.5 Hz, 1H), 6.84 (t, J = 8.0 Hz, 2H), 6.34 (s, 1H), 4.57-4.70 (m, 3H), 3.25 (td, J = 12.3, 4.8 Hz, 1H), 2.88-2.96 (m, 1H), 2.71 (dd, J = 15.6, 4.6 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.8, 146.7, 138.3, 136.3, 133.2, 131.6, 128.5, 128.3, 127.6, 127.2, 126.3, 122.0, 119.8, 119.0, 118.3, 117.3, 111.7, 111.4, 70.4, 52.9, 41.8, 19.3; LRMS (FAB) m/z, 380 ([M+H] ⁺ ).

EV203

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (39.0 mg, 0.229 mmol) and N -(3-chloro)benzylisatoic anhydride ( 7ab ) (60.0 mg, 0.209 mmol) was used to obtain compound EV203 (26.2 mg, 30% yield) in the form of a pale yellow solid.

mp: 230-232°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.21 (s, 1n), 7.78 (d, J = 7.8 Hz, 1H), 7.46 (d, J = 7.8 Hz, 1H), 7.21-7.37 (m) , 6H), 7.11 (t, J = 7.5 Hz, 1H), 7.00 (t, J = 7.3 Hz, 1H), 6.90 (t, J = 8.2 Hz, 2H), 6.34 (s, 1H), 4.54-4.64 (m, 3H), 3.25 (td, J = 12.1, 4.4 Hz, 1H), 2.87-2.95 (m, 1H), 2.75 (dd, J = 15.3, 3.9 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.7, 146.7, 141.0, 136.3, 133.3, 133.1, 131.1, 130.2, 128.3, 127.5, 127.2, 126.3, 126.2, 121.9, 120.4, 119.5, 119.0, 118.2, 117.6, 111.7, 111.6, 70.1, 52.3, 41.6, 19.3. LRMS (FAB) m/z, 414 ([M+H] ⁺ ).

EV204

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (711.4 mg, 4.180 mmol) and N -cyclohexanecarbonylisatoic anhydride ( 7ac ) (1.0 g, 3.800 mmol) were used to give a pale yellow color. Compound EV204 (237.1 mg, 14% yield) was obtained in solid form.

mp: 243-244° C.; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.16 (s, 1H), 7.82 (d, J = 7.8 Hz, 1H), 7.58 (t, J = 7.1 Hz, 2H), 7.34 (d, J) = 7.8 Hz, 1H), 7.31 (s, 1H), 7.24 (t, J = 3.9 Hz, 2H), 7.01-7.05 (m, 1H), 6.92-6.95 (m, 1H), 4.65 (dd, J = 13.0, 5.9 Hz, 1H), 3.64 (s, 1H), 2.91-3.04 (m, 2H), 2.58-2.67 (m, 1H), 2.08 (s, 1H), 1.79 (d, J = 11.9 Hz, 1H) ), 1.63 (d, J = 9.2 Hz, 4H), 1.37 (s, 2H), 1.21 (q, J = 12.4 Hz, 2H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 160.7, 147.4, 137.6, 135.7, 126.6, 122.2, 121.7, 118.9, 118.1, 111.5, 110.8, 117.0, 115.2, 111.7, 110.9, 71.0, 55.5, 42.5, 36.2, 29.7, 29.2, 25.4, 25.2, 24.8, 19.2; LRMS (FAB) m/z, 400 ([M+H] ⁺ ).

EV205

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (93.1 mg, 0.547 mmol) and N -propargylisatoic anhydride ( 7ad ) (100.0 mg, 0.497 mmol) were used as a yellow solid The form of compound EV205 (159.2 mg, 98% yield) was obtained.

mp: 182-184°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.26 (s, 1H), 7.90 (dd, J = 7.8, 1.4 Hz, 1H), 7.50-7.56 (m, 2H), 7.38 (d, J = 8.3 Hz, 1H), 7.31 (d, J = 7.4 Hz, 1H), 7.12-7.19 (m, 2H), 7.02-7.06 (m, 1H), 6.16 (s, 1H), 4.62 (ddd, J = 12.8) , 4.9, 2.0 Hz, 1H), 4.04 (dd, J = 18.2, 2.5 Hz, 1H), 3.61 (dd, J = 17.9, 2.3 Hz, 1H), 3.18-3.25 (m, 1H), 3.04 (t, J = 2.3 Hz, 1H), 2.84-2.98 (m, 2H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.5, 147.1, 136.8, 132.8, 129.0, 128.0, 125.8, 123.4, 122.9, 122.1, 121.3, 119.0, 118.5, 112.2, 111.7, 79.9, 75.2, 68.1, 19.5; HRMS (FAB) m/z calcd for C ₂₁ H ₁₈ N ₃ O [M+H] ⁺ : 328.1450, found 328.1456.

EV206

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (92.2 mg, 0.541 mmol) and N -allylisatoic anhydride ( 7ae ) (100.0 mg, 0.492 mmol) were used as a yellow solid The form of compound EV206 (125.1 mg, 84% yield) was obtained.

mp: 204-206°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.08 (s, 1H), 7.79 (d, J = 7.8 Hz, 1H), 7.42-7.47 (m, 2H), 7.35 (d, J = 7.8 Hz) , 1H), 7.11 (dd, J = 7.6, 5.3 Hz, 2H), 6.97 (td, J = 15.1, 7.8 Hz, 2H), 6.15 (s, 1H), 5.85 (dq, J = 22.4, 5.4 Hz, 1H), 5.02-5.07 (m, 2H), 4.62 (dd, J = 12.9, 5.1 Hz, 1H), 3.94 (d, 2H), 3.23 (td, J = 12.2, 4.6 Hz, 1H), 2.90-2.98 (m, 1H), 2.76 (dd, J = 15.6, 4.1 Hz, 1H). ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.5, 147.2, 136.4, 134.5, 133.1, 130.8, 128.0, 126.1, 121.8, 120.4, 120.0, 118.9, 118.5, 118.2, 117.1, 111.7, 111.4, 69.0, 52.6, 41.2, 19.3; HRMS (FAB) m/z calcd for C ₂₁ H ₂ ON ₃ O [M+H] ⁺ : 330.1606, found 330.1607.

EV207

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (35.1 mg, 0.206 mmol) and N - (4-methyl) benzylisatoic anhydride ( 7af ) (50.0 mg, 0.187 mmol) was used to obtain compound EV207 (45.1 mg, 61% yield) in the form of a yellow solid.

mp: 250-252°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.12 (s, 1H), 7.74 (dd, J = 8.0, 1.6 Hz, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.29-7.37 (m, 2H), 7.17 (d, J = 7.8 Hz, 2H), 7.08-7.12 (m, 3H), 6.97-7.01 (m, 1H), 6.83-6.87 (m, 2H), 6.31 (s, 1H) ), 4.51-4.63 (m, 3H), 3.24 (td, J = 12.2, 4.6 Hz, 1H), 2.88-2.96 (m, 1H), 2.72 (dd, J = 15.4, 4.8 Hz, 1H), 2.24 ( s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.8, 146.8, 136.3, 136.3, 135.1, 133.2, 131.6, 129.0, 128.1, 127.6, 126.3, 121.8, 119.8, 119.0, 118.9, 118.2, 117.4, 111.7, 111.4, 70.3, 52.6, 41.8, 20.7, 19.3. LRMS (FAB) m/z, 394 ([M+H] ⁺ ).

EV208

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (29.2 mg, 0.171 mmol) and N -(4-trifluoromethyl)benzylisatoic anhydride ( 7ag ) (50.0 mg, 0.156 mmol) were used to obtain compound EV208 (47.1 mg, 68% yield) in the form of a yellow solid.

mp: 207-209°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.14 (s, 1H), 7.78 (dd, J = 7.8, 1.4 Hz, 1H), 7.64 (d, J = 7.8 Hz, 2H), 7.47 (dd, J = 21.1, 7.8 Hz, 3H), 7.33-7.37 (m, 2H), 7.08-7.12 (m, 1H), 6.97-7.01 (m, 1H), 6.86-6.91 (m, 2H), 6.36 (s, 1H), 4.60-4.78 (m, 3H), 3.22-3.30 (m, 1H), 2.89-2.97 (m, 1H), 2.74 (dd, J = 15.6, 4.6 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.7, 146.7, 143.3, 136.3, 133.4, 131.2, 128.3, 128.2, 126.2, 125.3, 125.2, 121.9, 120.2, 119.2, 119.0, 118.2, 117.2, 111.7, 111.6 , 70.2, 52.3, 41.7, 19.3; LRMS (FAB) m/z, 448 ([M+H] ⁺ ).

EV209

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (34.5 mg, 0.203 mmol) and N - (4-fluoromethyl) benzylisatoic anhydride ( 7ah ) (50.0 mg, 0.184 mmol) was used to obtain compound EV209 (47.7 mg, 65% yield) in the form of a yellow solid.

mp: 224-226°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.16 (s, 1H), 7.77 (dd, J = 7.8, 1.4 Hz, 1H), 7.45 (d, J = 7.8 Hz, 1H), 7.27-7.37 (m, 4H), 7.08-7.13 (m, 2H), 6.98-7.02 (m, 1H), 6.85-6.91 (m, 2H), 6.32 (s, 1H), 4.50-4.62 (m, 3H), 3.21 -3.30 (m, 1H), 2.87-2.95 (m, 1H), 2.74 (dd, J = 15.4, 4.8 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.7, 146.8, 136.3, 134.4, 134.4, 133.2, 131.3, 129.6, 129.6, 128.2, 126.2, 121.9, 120.3, 119.5, 119.0, 118.2, 117.7, 115.3, 115.1, 111.7, 111.5, 70.1, 52.1, 41.6, 19.3; LRMS (FAB) m/z, 398 ([M+H] ⁺ ).

EV210

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (24.7 mg, 0.145 mmol) and N -(3-iodo)benzylisatoic anhydride ( 7ai ) (50.0 mg, 0.132 mmol) was used to obtain compound EV210 (34.8 mg, 52% yield) in the form of a pale yellow solid.

mp: 255-257°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.19 (s, 1H), 7.78 (dd, J = 8.0, 1.6 Hz, 1H), 7.57 (d, J = 7.4 Hz, 2H), 7.46 (d , J = 7.8 Hz, 1H), 7.36 (td, J = 7.7, 2.0 Hz, 2H), 7.25 (d, J = 7.8 Hz, 1H), 6.98-7.13 (m, 3H), 6.89-6.93 (m, 2H), 6.31 (s, 1H), 4.54-4.62 (m, 3H), 3.23 (td, J = 12.2, 4.3 Hz, 1H), 2.85-2.93 (m, 1H), 2.75 (dd, J = 15.4, 4.8 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.6, 146.8, 141.0, 136.3, 136.3, 135.9, 133.3, 131.0, 130.5, 128.2, 127.0, 126.2, 121.9, 120.4, 119.6, 119.0, 118.3, 117.7, 111.7, 111.6, 95.0, 70.0, 52.1, 41.5, 19.3; LRMS (FAB) m/z, 506 ([M+H] ⁺ ).

EV211

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (33.1 mg, 0.195 mmol) and N -(3-methoxy)benzylisatoic anhydride ( 7aj ) (50.0 mg, 0.177 mmol) was used to obtain compound EV211 (66.7 mg, 92% yield) in the form of a white solid.

mp: 212-214°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.16 (s, 1H), 7.76 (dd, J = 7.5, 1.4 Hz, 1H), 7.45 (d, J = 7.4 Hz, 1H), 7.31-7.37 (m, 2H), 7.20 (t, J = 7.7 Hz, 1H), 7.08-7.12 (m, 1H), 6.98-7.02 (m, 1H), 6.77-6.90 (m, 5H), 6.32 (s, 1H) ), 4.58-4.64 (m, 3H), 3.66 (s, 3H), 3.21-3.28 (m, 1H), 2.86-2.94 (m, 1H), 2.73 (dd, J = 15.6, 4.6 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.7, 159.3, 146.9, 139.9, 136.3, 133.2, 131.3, 129.5, 128.1, 126.2, 121.8, 119.9, 119.7, 119.1, 118.9, 118.2, 117.3, 113.2, 112.4, 111.7, 111.4, 70.1, 54.9, 52.7, 41.6, 19.3; LRMS (FAB) m/z, 410 ([M+H] ⁺ ).

EV212

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (33.1 mg, 0.195 mmol) and N -(4-methoxy)benzylisatoic anhydride ( 7ak ) (50.0 mg, 0.177 mmol) was used to obtain compound EV212 (24.1 mg, 33% yield) in the form of a white solid.

mp: 245-247°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.13 (s, 1H), 7.75 (dd, J = 8.3, 1.5 Hz, 1H), 7.45 (d, J = 8.0 Hz, 1H), 7.30-7.37 (m, 2H), 7.19 (d, J = 8.6 Hz, 2H), 7.10 (t, J = 7.1 Hz, 1H), 7.00 (t, J = 7.4 Hz, 1H), 6.83-6.88 (m, 4H) , 6.30 (s, 1H), 4.46-4.62 (m, 3H), 3.70 (s, 3H), 3.24 (td, J = 12.3, 4.5 Hz, 1H), 2.89-2.97 (m, 1H), 2.72 (dd , J = 15.3, 4.3 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.7, 158.4, 146.9, 137.0, 136.3, 133.1, 131.5, 129.9, 129.0, 128.1, 126.2, 121.8, 120.0, 119.3, 118.9, 118.2, 117.7, 114.1, 113.8, 111.7, 111.4, 70.1, 55.0, 52.3, 41.7, 19.3. LRMS (FAB) m/z, 410 ([M+H] ⁺ ).

EV213

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (35.1 mg, 0.206 mmol) and N -(2-methyl)benzylisatoic anhydride ( 7al ) (50.0 mg, 0.187 mmol) was used to obtain compound EV213 (49.7 mg, 68% yield) in the form of a pale yellow solid.

mp: 279-281°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.06 (s, 1H), 7.74 (dd, J = 7.7, 1.5 Hz, 1H), 7.42 (d, J = 8.0 Hz, 1H), 7.36 (d) , J = 8.0 Hz, 1H), 7.25-7.31 (m, 2H), 7.08-7.19 (m, 4H), 6.99 (t, J = 7.4 Hz, 1H), 6.79 (t, J = 7.7 Hz, 1H) , 6.66 (d, J = 8.6 Hz, 1H), 6.29 (s, 1H), 4.92 (d, J = 16.6 Hz, 1H), 4.65 (q, J = 6.3 Hz, 1H), 4.52 (d, J = 15.9 Hz, 1H), 3.26 (td, J = 12.3, 4.7 Hz, 1H), 2.96-3.04 (m, 1H), 2.68 (dd, J = 15.3, 4.9 Hz, 1H), 2.30 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 165.0, 146.4, 136.1, 136.0, 135.6, 133.4, 132.3, 130.3, 128.1, 127.1, 127.1, 126.4, 125.9, 121.8, 119.1, 119.0, 118.1, 117.8, 115.9, 111.6, 111.3, 70.5, 50.1, 42.4, 19.1, 18.9; LRMS (FAB) m/z, 394 ([M+H] ⁺ ).

EV214

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (32.6 mg, 0.191 mmol) and N - (4-chloro)benzylisatoic anhydride ( 7am ) (50.0 mg, 0.174 mmol) was used to obtain compound EV214 (48.3 mg, 67% yield) in the form of a pale yellow solid.

mp: 265-267°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.15 (s, 1H), 7.77 (dd, J = 8.0, 1.2 Hz, 1H), 7.45 (d, J = 8.0 Hz, 1H), 7.27-7.36 (m, 6H), 7.08-7.12 (m, 1H), 6.98-7.02 (m, 1H), 6.85-6.91 (m, 2H), 6.33 (s, 1H), 4.52-4.63 (m, 3H), 3.24 (td, J = 12.3, 4.5 Hz, 1H), 2.87-2.95 (m, 1H), 2.74 (dd, J = 15.3, 4.9 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.6, 146.7, 137.3, 136.3, 133.2, 131.7, 131.2, 129.5, 128.3, 128.2, 126.2, 121.9, 120.3, 119.4, 118.9, 118.2, 117.6, 111.7, 111.5, 70.1, 52.2, 41.6, 19.3; LRMS (FAB) m/z, 414 ([M+H] ⁺ ).

EV215

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (31.4 mg, 0.184 mmol) and N - (4-nitro) benzylisatoic anhydride ( 7an ) (50.0 mg, 0.168 mmol) was used to obtain compound EV215 (13.7 mg, 19% yield) in the form of a green-yellow solid.

mp: 228-230°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.14 (s, 1H), 8.12 (dd, J = 9.2, 2.5 Hz, 2H), 7.80 (dd, J = 7.7, 1.5 Hz, 1H), 7.54 (d, J = 8.6 Hz, 2H), 7.45 (d, J = 8.0 Hz, 1H), 7.31-7.38 (m, 2H), 7.07-7.10 (m, 1H), 6.89-7.01 (m, 3H), 6.36 (s, 1H), 4.61-4.76 (m, 3H), 3.24 (td, J = 12.3, 4.5 Hz, 1H), 2.89-2.97 (m, 1H), 2.76 (dd, J = 15.3, 4.9 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.6, 146.9, 146.6, 146.5, 136.3, 133.4, 130.8, 128.6, 128.3, 127.9, 126.1, 123.7, 123.4, 121.9, 120.7, 119.7, 119.0, 118.3, 117.6, 111.7, 111.6, 70.0, 52.3, 41.5, 19.3; LRMS (FAB) m/z, 425 ([M+H] ⁺ ).

EV216

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (35.1 mg, 0.206 mmol) and N- (3-methyl)benzylisatoic anhydride ( 7ao ) (50.0 mg, 0.187 mmol) was used to obtain compound EV216 (45 mg, 61% yield) in the form of a white solid.

mp: 243-245° C.; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.12 (s, 1H), 7.74-7.77 (m, 1H), 7.45 (d, J = 7.4 Hz, 1H), 7.30-7.37 (m, 2H) , 7.17 (t, J = 7.4 Hz, 1H), 6.98-7.12 (m, 5H), 6.86 (dd, J = 7.7, 6.4 Hz, 2H), 6.31 (s, 1H), 4.53-4.63 (m, 3H) ), 3.25 (td, J = 12.3, 4.5 Hz, 1H), 2.87-2.95 (m, 1H), 2.72 (dd, J = 15.3, 4.9 Hz, 1H), 2.23 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.7, 146.9, 138.1, 137.4, 136.3, 133.2, 131.4, 128.3, 128.2, 128.1, 127.8, 126.2, 124.6, 121.8, 119.8, 119.0, 118.9, 118.2, 117.2, 111.7, 111.5, 70.2, 52.8, 41.7, 21.0, 19.2; LRMS (FAB) m/z, 394 ([M+H] ⁺ ).

EV217

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (40.5 mg, 0.238 mmol) and N -(1-dimethyl)allylisatoic anhydride ( 7ap ) (50.0 mg, 0.216 mmol) was used to obtain compound EV217 (67 mg, 87% yield) in the form of a white solid.

mp: 219-221°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.10 (s, 1H), 7.79 (dd, J = 8.0, 1.2 Hz, 1H), 7.43-7.47 (m, 2H), 7.35 (d, J = 8.0 Hz, 1H), 7.10 (t, J = 7.7 Hz, 2H), 6.95-7.02 (m, 2H), 6.11 (s, 1H), 5.17 (t, J = 6.7 Hz, 1H), 4.58-4.63 ( m, 1H), 3.86 (d, J = 7.4 Hz, 2H), 3.19-3.26 (m, 1H), 2.75-2.91 (m, 2H), 1.57 (s, 3H), 1.33 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.4, 147.7, 136.4, 134.2, 133.0, 130.7, 128.0, 126.2, 121.8, 120.8, 120.7, 120.7, 119.0, 118.9, 118.1, 111.6, 111.4, 68.8, 47.8, 40.9, 40.2, 39.9, 39.7, 39.5, 39.3, 39.1, 38.9, 25.5, 19.4, 17.50; LRMS (FAB) m/z, 358 ([M+H] ⁺ ).

EV218

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (43.1 mg, 0.253 mmol) and N -(2-methyl)allylisatoic anhydride ( 7aq ) (50.0 mg, 0.230 mmol) was used to obtain compound EV218 (75.0 mg, 95% yield) in the form of a dark yellow solid.

mp: 181-183°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.94 (s, 1H), 7.72 (dd, J = 7.9, 1.8 Hz, 1H), 7.32-7.43 (m, 3H), 7.08 (t, J = 7.9 Hz, 1H), 6.98 (t, J = 7.9 Hz, 1H), 6.91 (d, J = 7.9 Hz, 1H), 6.80 (t, J = 7.0 Hz, 1H), 6.20 (s, 1H), 4.88 (d, J = 21.4 Hz, 2H), 4.62 (dd, J = 13.4, 5.5 Hz, 1H), 4.22 (d, J = 16.5 Hz, 1H), 3.94 (d, J = 16.5 Hz, 1H), 3.26 (td, J = 12.2, 4.9 Hz, 1H), 2.95-3.03 (m, 1H), 2.68 (dd, J = 15.6, 4.6 Hz, 1H), 1.72 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.9, 146.6, 141.3, 136.1, 133.3, 132.1, 128.0, 126.3, 121.7, 118.9, 118.8, 118.1, 117.4, 115.8, 112.3, 111.7, 111.2, 70.1, 54.9, 42.2, 20.0, 19.2; HRMS (FAB) m/z calcd for C ₂₂ H ₂₂ N ₃ O [M+H] ⁺ : 344.1763, found 344.1759.

EV219

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (43.5 mg, 0.256 mmol) and N -(1-methyl)propargylisatoic anhydride ( 7ar ) (50.0 mg, 0.232 mmol) was used to obtain compound EV219 (74.2 mg, 94% yield) in the form of a yellow solid.

mp: 195-198°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.20 (s, 1H), 7.88 (dd, J = 7.9, 1.2 Hz, 1H), 7.51-7.55 (m, 2H), 7.37 (d, J = 7.9 Hz, 1H), 7.26 (d, J = 7.9 Hz, 1H), 7.11-7.15 (m, 2H), 7.01-7.05 (m, 1H), 6.17 (s, 1H), 4.61-4.65 (m, 1H) ), 3.97 (dd, J = 17.7, 2.4 Hz, 1H), 3.68 (dd, J = 17.7, 2.4 Hz, 1H), 3.17-3.28 (m, 1H), 2.83-2.98 (m, 2H), 1.65 ( t, J = 2.1 Hz, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.6, 147.2, 136.7, 132.8, 129.4, 127.9, 125.9, 122.9, 122.5, 122.0, 120.8, 118.9, 118.4, 112.0, 111.7, 80.4, 75.2, 68.3, 40.2, 19.5, 3.1; LRMS (FAB) m/z, 342 ([M+H] ⁺ ).

EV220

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (33.6 mg, 0.197 mmol) and N -cinnamylisatoic anhydride ( 7as ) (50.0 mg, 0.179 mmol) were used to give a pale yellow color. Compound EV220 (13.7 mg, 19% yield) in solid form was obtained.

mp: 194-196° C.; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.18 (s, 1H), 7.80 (dd, J = 7.9, 1.2 Hz, 1H), 7.47 (d, J = 7.9 Hz, 1H), 7.35-7.44 (m, 2H), 7.25-7.33 (m, 5H), 7.17-7.20 (m, 1H), 7.11 (t, J = 7.6 Hz, 1H), 6.95-7.03 (m, 2H), 6.24-6.43 (m) , 2H), 6.21 (s, 1H), 4.62 (dd, J = 13.1, 4.6 Hz, 1H), 4.07 (ddd, J = 29.8, 15.7, 5.7 Hz, 2H), 3.25 (td, J = 12.2, 4.5) Hz, 1H), 2.88-2.96 (m, 1H), 2.77 (dd, J = 15.3, 4.3 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.5, 147.4, 136.4, 136.4, 133.2, 131.6, 130.7, 128.6, 128.0, 127.6, 126.3, 126.2, 126.1, 126.0, 121.9, 120.8, 120.5, 119.0, 118.9, 118.2, 111.7, 111.5, 68.9, 52.3, 41.1, 19.4; LRMS (FAB) m/z, 406 ([M+H] ⁺ ).

<EV221 derivative synthesis method>

: 3,4-dihydro-β-carboline ( 2 ) (189.0 mg, 1.110 mmol) was dissolved in CH ₂ Cl ₂ (3.0 mL, 0.4 M), and salicyl chloride (226.7 mg, 1.448 mmol) was slowly added under Ar substitution. After stirring at room temperature for 12 hours, the mixture was concentrated under reduced pressure to remove excess CH ₂ Cl ₂ . The resulting mixture was purified by column chromatography (silica gel, hexane:ethyl acetate = 3:1 to 2:1) to obtain compound EV221 (174.0 mg, 54% yield) in the form of a white solid.

mp: 113-115°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.52 (s, 1H) 7.91 (d, J = 7.6 Hz, 1H), 7.57-7.60 (m, 2H), 7.42 (d, J = 8.2 Hz, 1H), 7.13-7.27 (m, 3H), 7.07 (t, J = 7.7 Hz, 1H), 6.68 (s, 1H), 4.71-4.74 (m, 1H), 3.21-3.22 (m, 1H), 2.93 -3.02 (m, 2H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 162.6, 157.2, 137.4, 134.8, 128.5, 128.0, 125.7, 123.3, 123.1, 119.5, 119.3, 118.9, 117.0, 112.3, 112.1, 81.6, 20.3; LRMS (FAB) m/z, 291 ([M+H] ⁺ ).

<EV222 derivative synthesis method>

: Evodiamine ( 1 ) (300.0 mg, 0.989 mmol) was dissolved in THF (30.0 mL), and LiAlH ₄ (110.0 mg, 2.967 mmol) was added thereto. After stirring at room temperature for 12 hours, water (0.2 mL) was added to terminate the reaction. The mixture obtained by concentration under reduced pressure was purified by column chromatography (silica gel, hexane:ethyl acetate = 4:1 to 3:1) to obtain a white solid compound EV222 (182.4 mg, 64% yield).

mp: 162-164°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.22 (s, 1H), 7.45 (d, J = 7.6 Hz, 1H), 7.35 (d, J = 8.0 Hz, 1H), 7.06-7.15 (m) , 2H), 6.93-7.02 (m, 3H), 6.83 (td, J = 7.3, 1.0 Hz, 1H), 4.89 (s, 1H), 3.93 (s, 2H), 3.23-3.27 (m, 1H), 2.81-2.88 (m, 1H), 2.70-2.75 (m, 2H), 2.65 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 147.5, 136.5, 131.5, 126.9, 126.7, 126.2, 124.9, 121.2, 119.9, 119.1, 118.5, 118.8, 111.4, 109.5, 72.6, 55.0, 48.6, 38.0, 20.8; LRMS (FAB) m/z, 291 ([M+H] ⁺ ).

Synthesis Example 4. Synthesis of E-ring evodiamine derivatives (EV301 to 312)

<EV301~312 Derivative Synthesis Method>

5-Fluoroisatoic anhydride (6a)

: According to the isatoic anhydride ( 6 ) synthesis conditions, 5-fluoroanthranilic acid (300.0 mg, 1.932 mmol) was used to obtain compound 6a (350.0 mg, 99% yield) in the form of a white solid.

mp: 177-178°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.85 (d, J = 17.5 Hz, 1H), 7.62-7.70 (m, 2H), 7.19-7.24 (m, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 159.3, 159.2, 158.8, 156.4, 146.9, 138.2, 124.9, 124.7, 117.6, 117.6, 114.2, 113.9, 111.6, 111.5; LRMS (FAB) m/z, 182 ([M+H] ⁺ ).

5-Fluoro- N -methylisatoic anhydride (7ba)

: According to the N -methylisatoic anhydride ( 7 ) synthesis conditions, 6a (300.0 mg, 1.656 mmol) was used to obtain a beige solid compound 7ba (193.9 mg, 60% yield).

mp: 119-120°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.76 (td, J = 7.8, 2.6 Hz, 2H), 7.49-7.52 (m, 1H), 3.46 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 158.9, 158.3, 158.3, 156.5, 147.5, 139.1, 139.0, 124.7, 124.4, 117.5, 117.4, 114.7, 114.4, 113.0, 113.0, 32.0; LRMS (FAB) m/z, 196 ([M+H] ⁺ ).

EV301

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (96.0 mg, 0.564 mmol) and 5-fluoro- N -methyllisatoic anhydride ( 7ba ) (100.0 mg, 0.512 mmol) was used to obtain compound EV301 (125.0 mg, 76% yield) in the form of a pale yellow solid.

mp: 223-224°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.19 (s, 1H), 7.49-7.56 (m, 2H), 7.36-7.41 (m, 2H), 7.20 (q, J = 4.4 Hz, 1H) , 7.12 (td, J = 7.6, 1.3 Hz, 1H), 7.00-7.04 (m, 1H), 6.09 (s, 1H), 4.63 (dt, J = 12.6, 3.4 Hz, 1H), 3.17-3.25 (m) , 1H), 2.85-2.88 (m, 2H), 2.67 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.1, 163.1, 158.4, 156.0, 146.1, 136.7, 129.6, 125.8, 122.2, 122.2, 122.0, 121.9, 121.8, 120.7, 120.5, 118.9, 118.4, 113.6, 113.3, 111.7, 111.7, 69.2, 40.3, 36.6, 19.6; HRMS (FAB) m/z calcd for C ₁₉ H ₁₇ FN ₃ O [M+H] ⁺ : 322.1356, found 322.1362.

EV302

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (28.5 mg, 0.167 mmol) and 5-methoxy- N -methyllisatoic anhydride ( 7bb ) (31.6 mg, 0.152 mmol) was used to obtain compound EV302 (44.2 mg, 87% yield) in the form of a yellow solid.

mp: 222-223°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.25 (s, 1H), 7.51 (d, J = 8.2 Hz, 1H), 7.37 (t, J = 3.7 Hz, 2H), 7.10-7.18 (m) , 3H), 7.00-7.04 (m, 1H), 6.01 (s, 1H), 4.63-4.67 (m, 1H), 3.77 (s, 3H), 3.15-3.22 (m, 1H), 2.78-2.90 (m , 2H), 2.48 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.7, 155.0, 143.8, 136.8, 129.5, 125.8, 123.0, 122.9, 121.9, 120.6, 118.9, 118.4, 111.6, 111.6, 110.7, 69.0, 55.4, 36.8, 19.8; LRMS (FAB) m/z, 332 ([M+H] ⁺ ).

EV303

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (283.1 mg, 1.663 mmol) and 5-nitro- N -methylisatoic anhydride ( 7bc ) (336.0 mg, 1.512 mmol) was used to obtain compound EV303 (488.4 mg, 93% yield) in the form of a yellow solid.

mp: 290-292°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.90 (s, 1H), 8.45 (d, J = 2.8 Hz, 1H), 8.25 (dd, J = 9.2, 2.8 Hz, 1H), 7.43 (d , J = 7.8 Hz, 1H), 7.34 (d, J = 7.8 Hz, 1H), 6.98-7.12 (m, 3H), 6.51 (s, 1H), 4.65 (dd, J = 13.2, 5.9 Hz, 1H) , 3.46 (s, 3H), 3.30 (dd, J = 12.9, 5.1 Hz, 1H), 3.01-3.09 (m, 1H), 2.71 (dd, J = 15.4, 4.8 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 162.8, 151.2, 137.7, 136.1, 131.4, 129.3, 126.2, 124.1, 122.2, 119.2, 118.2, 114.1, 113.4, 111.8, 111.3, 71.3, 42.7, 37.2, 19.6; LRMS (FAB) m/z, 349 ([M+H] ⁺ ).

EV304

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (187.2 mg, 0.363 mmol) and 5-chloro- N -methylisatoic anhydride ( 7bd ) (69.9 mg, 0.330 mmol) were added. was used to obtain compound EV304 (83.9 mg, 75% yield) in the form of a yellow solid.

mp: 307-308°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.08 (s, 1H), 7.70 (d, J = 2.8 Hz, 1H), 7.51 (dd, J = 8.7, 2.3 Hz, 1H), 7.47 (d) , J = 7.8 Hz, 1H), 7.36 (d, J = 7.8 Hz, 1H), 7.07-7.13 (m, 2H), 6.99-7.03 (m, 1H), 6.17 (s, 1H), 4.61 (dd, J = 13.1, 4.4 Hz, 1H), 3.22 (td, J = 12.2, 4.6 Hz, 1H), 2.88-2.96 (m, 4H), 2.75-2.82 (m, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.2, 147.4, 136.5, 133.21, 130.3, 127.1, 126.0, 124.0, 122.0, 120.2, 119.2, 119.0, 118.3, 111.7, 111.6, 69.8, 41.2, 36.4, 19.4; HRMS (FAB) m/z calcd for C ₁₉ H ₁₇ ClN ₃ O [M+H] ⁺ : 338.1060, found 338.1065.

EV305

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (43.1 mg, 0.253 mmol) and 4-chloro- N -methylisatoic anhydride ( 7be ) (48.7 mg, 0.230 mmol) was used to obtain compound EV305 (28.0 mg, 36% yield) in the form of a pale yellow solid.

mp: 291-292° C.; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.94 (s, 1H), 7.71 (d, J = 8.3 Hz, 1H), 7.40 (dd, J = 37.2, 7.8 Hz, 2H), 7.08-7.12 (m, 1H), 6.98-7.01 (m, 2H), 6.87 (dd, J = 8.5, 2.1 Hz, 1H), 6.23 (s, 1H), 4.61 (dd, J = 13.1, 5.3 Hz, 1H), 3.19-3.28 (m, 1H), 3.11 (s, 3H), 2.92-3.00 (m, 1H), 2.73 (dd, J = 15.4, 4.8 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.8, 148.8, 138.4, 136.3, 131.2, 129.8, 126.1, 122.0, 119.0, 118.8, 118.2, 116.1, 114.7, 111.8, 111.5, 70.6, 41.8, 36.5, 19.3; LRMS (FAB) m/z, 338 ([M+H] ⁺ ).

EV306

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (43.1 mg, 0.253 mmol) and 5-bromo- N -methylisatoic anhydride ( 7bf ) (58.9 mg, 0.230 mmol) were added. was used to obtain compound EV306 (60.8 mg, 69% yield) in the form of a pale yellow solid.

mp: 200-201°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.06 (s, 1H), 7.82 (d, J = 2.3 Hz, 1H), 7.62 (dd, J = 8.7, 2.3 Hz, 1H), 7.47 (d) , J = 7.8 Hz, 1H), 7.36 (d, J = 7.8 Hz, 1H), 7.11 (td, J = 7.6, 1.1 Hz, 1H), 6.99-7.03 (m, 2H), 6.18 (s, 1H) , 4.61 (dd, J = 12.9, 4.6 Hz, 1H), 3.22 (td, J = 12.2, 4.6 Hz, 1H), 2.88-2.98 (m, 4H), 2.78 (dd, J = 15.6, 4.6 Hz, 1H) ); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.13, 147.58, 136.44, 135.97, 130.38, 130.03, 125.96, 121.99, 120.32, 119.17, 119.00, 118.28, 111.70, 111.57, 111.32, 69.86, 41.30, 36.86, 19.40; LRMS (FAB) m/z, 382 ([M+H] ⁺ ).

EV307

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (88.6 mg, 0.520 mmol) and 5-methyl- N -methylisatoic anhydride ( 7bg ) (90.4 mg, 0.473 mmol) were added. was used to obtain compound EV307 (49.5 mg, 33% yield) in the form of an orange solid.

mp: 245-247°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.14 (s, 1H), 7.64 (d, J = 1.8 Hz, 1H), 7.49 (d, J = 7.8 Hz, 1H), 7.30-7.37 (m) , 2H), 7.11 (td, J = 7.6, 1.1 Hz, 1H), 7.01 (t, J = 7.8 Hz, 2H), 6.05 (s, 1H), 4.61-4.66 (m, 1H), 3.14-3.22 ( m, 1H), 2.80-2.91 (m, 2H), 2.70 (s, 4H), 2.28 (s, 4H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.2, 147.2, 136.6, 134.1, 130.4, 130.2, 128.0, 125.9, 121.9, 120.4, 119.1, 118.9, 118.3, 111.7, 111.6, 69.4, 40.4, 36.6, 20.3, 19.6; HRMS (FAB) m/z calcd for C ₂₀ H ₂ ON ₃ O [M+H] ⁺ : 318.1606, found 318.1606.

EV308

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (71.7 mg, 0.421 mmol) and 6-chloro- N -methylisatoic anhydride ( 7bh ) (81.1 mg, 0.383 mmol) were added was used to obtain compound EV308 (83.9 mg, 65% yield) in the form of a pale yellow solid.

mp: 267-269°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.29 (s, 1H), 7.55 (d, J = 8.3 Hz, 1H), 7.38-7.45 (m, 2H), 7.10-7.18 (m, 3H) , 7.02-7.06 (m, 1H), 5.94 (s, 1H), 4.49 (dq, J = 12.8, 2.6 Hz, 1H), 3.25-3.31 (m, 1H), 2.81-2.94 (m, 2H), 2.65 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 161.3, 152.3, 136.8, 134.1, 132.8, 128.4, 125.6, 124.4, 122.2, 119.0, 118.7, 118.6, 111.9, 111.7, 67.4, 35.7, 19.7; LRMS (FAB) m/z, 338 ([M+H] ⁺ ).

EV309

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (67.8 mg, 0.398 mmol) and 6-methoxy- N -methylisatoic anhydride ( 7bi ) (75.0 mg, 0.362 mmol) was used to obtain compound EV309 (54.8 mg, 45 % yield) in the form of a yellow solid.

mp: 248-250°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.23 (s, 1H), 7.52 (d, J = 7.8 Hz, 1H), 7.36-7.41 (m, 2H), 7.13 (td, J = 7.6, 1.1 Hz, 1H), 7.01-7.05 (m, 1H), 6.73 (d, J = 8.3 Hz, 1H), 6.67 (d, J = 7.8 Hz, 1H), 5.83 (s, 1H), 4.44-4.49 ( m, 1H), 3.77 (s, 3H), 3.20-3.27 (m, 1H), 2.78-2.89 (m, 2H), 2.62 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 162.0, 160.5, 152.0, 136.8, 133.2, 129.1, 125.7, 122.0, 118.9, 118.5, 111.8, 111.7, 111.5, 110.7, 105.4, 67.7, 55.8, 35.8, 19.8; LRMS (FAB) m/z, 334 ([M+H] ⁺ ).

EV310

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (31.6 mg, 0.186 mmol) and 5-iodo- N -methylisatoic anhydride ( 7bj ) (51.2 mg, 0.169 mmol) were added. was used to obtain compound EV310 (40.4 mg, 56 % yield) in the form of a pale yellow solid.

mp: 293-295°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.03 (s, 1H), 7.98 (d, J = 2.3 Hz, 1H), 7.74 (dd, J = 8.7, 2.3 Hz, 1H), 7.46 (d , J = 7.8 Hz, 1H), 7.35 (d, J = 8.3 Hz, 1H), 7.09-7.13 (m, 1H), 6.99-7.02 (m, 1H), 6.87 (d, J = 8.7 Hz, 1H) , 6.17 (s, 1H), 4.60 (dd, J = 13.0, 4.5 Hz, 1H), 3.21 (td, J = 12.2, 4.7 Hz, 1H), 2.96 (s, 3H), 2.88-2.94 (m, 1H) ), 2.77 (dd, J = 15.6, 4.6 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.1, 147.8, 141.6, 136.4, 136.0, 130.6, 126.0, 122.0, 120.4, 119.1, 119.0, 118.3, 111.7, 111.5, 81.9, 69.9, 41.4, 36.3, 19.4; LRMS (FAB) m/z, 430 ([M+H] ⁺ ).

EV311

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (45.5 mg, 0.267 mmol) and 4-methoxy- N -methylisatoic anhydride ( 7bk ) (50.3 mg, 0.243 mmol) were added. Compound EV311 (44.2 mg, 55% yield) in the form of a pale yellow solid was obtained using

mp: 246-248°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.97 (s, 1H), 7.68 (d, J = 8.7 Hz, 1H), 7.45 (d, J = 7.8 Hz, 1H), 7.35 (d, J) = 8.3 Hz, 1H), 7.08-7.12 (m, 1H), 6.98-7.02 (m, 1H), 6.44-6.50 (m, 2H), 6.13 (s, 1H), 4.60 (dd, J = 12.9, 5.1) Hz, 1H), 3.81 (s, 3H), 3.18 (td, J = 12.2, 4.7 Hz, 1H), 3.00 (s, 3H), 2.88-2.96 (m, 1H), 2.73 (dd, J = 15.2, 4.6 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.4, 163.7, 150.0, 136.4, 131.3, 129.9, 126.1, 121.8, 118.9, 118.2, 111.7, 111.6, 111.4, 106.5, 100.4, 70.4, 55.4, 41.2, 36.5, 19.4; LRMS (FAB) m/z, 334 ([M+H] ⁺ ).

EV312

: According to the Evodiamine ( 1 ) synthesis conditions, 3,4-dihydro-β-carboline ( 4 ) (58.4 mg, 0.343 mmol) and 3-methyl- N -methylisatoic anhydride ( 7bl ) (59.6 mg, 0.312 mmol) was used to obtain compound EV312 (84.1 mg, 85% yield) in the form of an orange solid.

mp: 256-258°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.31 (s, 1H), 7.99-8.01 (m, 1H), 7.61 (d, J = 7.8 Hz, 1H), 7.41 (dd, J = 28.3, 7.6 Hz) , 2H), 7.27-7.30 (m, 1H), 7.13-7.23 (m, 3H), 5.88 (s, 1H), 4.90 (dq, J = 12.9, 2.5 Hz, 1H), 3.29-3.36 (m, 1H) ), 2.91-3.03 (m, 2H), 2.41 (s, 3H), 2.26 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.7, 149.1, 137.0, 134.4, 132.3, 128.8, 125.8, 125.8, 124.8, 124.5, 122.0, 118.8, 118.4, 111.7, 111.6, 68.8, 33.8, 19.8, 16.5; LRMS (FAB) m/z, 318 ([M+H] ⁺ ).

Synthesis Example 5. Synthesis of ring A evodiamine derivative (EV401~413 )

EV401

: Evodiamine ( 1 ) (100.0 mg, 0.330 mmol) was added with KNO ₃ (33.4 mg, 0.330 mmol) and concentrated H ₂ SO ₄ (1.0 mL, 0.33 M), followed by stirring at 0° C. overnight. After completion of the reaction, ice water (10 mL) was added to the mixture, and the resulting solid was filtered under reduced pressure to obtain a yellow solid compound EV401 (111.5 mg, 97% yield).

mp: 220-222°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.89 (s, 1H), 8.51 (s, 1H), 8.02 (dd, J = 9.0, 2.1 Hz, 1H), 7.79-7.87 (m, 1H) , 7.47-7.53 (m, 2H), 7.07 (d, J = 8.3 Hz, 1H), 6.98 (t, J = 7.6 Hz, 1H), 6.21 (s, 1H), 4.62-4.66 (m, 1H), 3.20-3.28 (m, 1H), 2.91-2.95 (m, 5H); ¹³ C-NMR (100 MHz, DMSO-D ₆ ) δ 164.2, 148.6, 140.6, 139.6, 134.9, 133.6, 128.0, 125.4, 120.4, 119.2, 117.7, 117.3, 115.6, 114.3, 112.1, 69.6, 40.7, 36.9, 19.2; HRMS (FAB) m/z calcd for C ₁₉ H ₁₇ N ₄ O ₃ [M+H] ⁺ : 349.1301, found 349.1294.

EV402

: KNO ₃ (66.8 mg, 0.660 mmol) and concentrated H ₂ SO ₄ (1.0 mL, 0.33 M) were added to Evodiamine ( 1 ) (100.0 mg, 0.330 mmol) at 0° C., followed by stirring at room temperature overnight. After completion of the reaction, ice water (10 mL) was added to the mixture, and the solid obtained by filtration under reduced pressure was purified by column chromatography (silica gel, hexane:ethyl acetate = 2:1 to 1:1), and the yellow solid compound EV402 (14.3 mg) , 11% yield) was obtained.

mp: 268-270°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 12.63 (s, 1H), 9.03 (d, J = 2.0 Hz, 1H), 8.80 (d, J = 2.0 Hz, 1H), 7.88 (dd, J) = 7.4, 1.3 Hz, 1H), 7.51-7.56 (m, 1H), 7.22 (d, J = 8.1 Hz, 1H), 7.11 (t, J = 7.2 Hz, 1H), 6.04 (s, 1H), 4.68 (dd, J = 12.1, 4.7 Hz, 1H), 3.10-3.21 (m, 2H), 2.85-2.92 (m, 1H), 2.63 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.6, 149.9, 139.5, 135.7, 133.2, 131.8, 131.3, 129.7, 128.2, 122.1, 121.8, 121.2, 119.7, 117.3, 114.4, 67.7, 40.4, 39.9, 39.7, 39.5, 39.3, 39.2, 39.1, 38.9, 36.0, 19.7; HRMS (FAB) m/z calcd for C ₁₉ H ₁₆ N ₅ O ₅ [M+H] ⁺ : 394.1151, found 394.1157.

5-1. Method for synthesizing EV403, EV405-407 derivatives

5-Fluoro- N- formyltryptamine (3b)

: According to the above N -formyltrypramine ( 3 ) synthesis conditions, using 5-fluorotryptamine (165.9 mg, 0.931 mmol) and ethyl formate (344.8 mg, 4.655 mmol) as a brown oil, compound 3b (192.0 mg, 99% yield) got

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.32 (s, 1H), 8.12 (s, 1H), 6.90-7.29 (m, 3H), 5.83 (d, J = 91.9 Hz, 1H), 3.61 (q) , J = 6.6 Hz, 2H), 2.89-2.95 (m, 2H); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 161.3, 124.0, 112.1, 112.0, 111.0, 110.8, 103.8, 103.6, 38.3, 25.3; LRMS (FAB) m/z, 303 ([M+H] ⁺ ), 273, 235. LRMS (FAB) m/z, 207 ([M+H] ⁺ ).

6-fluoro-4,9-dihydro-3 H -pyrido[3,4- b ]indole (4b)

: According to the above 3,4-Dihydro _- β-carboline ( 4 ) synthesis conditions, compound 4b (200.0 mg, 200.0 mg, 70% yield) was obtained.

mp: 150-152°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 9.19 (s, 1H), 7.90 (s, 1H), 7.26 (t, J = 6.7 Hz, 1H), 7.07 (dd, J = 9.2, 2.3 Hz, 1H) ), 6.95 (td, J = 9.0, 2.5 Hz, 1H), 4.18 (t, J = 8.5 Hz, 2H), 3.10 (t, J = 8.5 Hz, 2H); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 156.9, 151.4, 134.8, 129.6, 125.7, 116.5, 113.2, 100.8, 58.0, 49.6, 19.1; LRMS (FAB) m/z, 189 ([M+H] ⁺ ).

EV403

: Compound EV403 (56.6 mg, 60%) in the form of a pale yellow solid using N -methylisatoic anhydride ( 7 ) (50.1 mg, 0.283 mmol) and 4a (62.3 mg, 0.311 mmol) according to the Evodiamine ( 1 ) synthesis conditions yield) was obtained.

mp: 312-313°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.88 (s, 1H), 7.79 (dd, J = 7.8, 1.8 Hz, 1H), 7.45-7.49 (m, 1H), 7.24 (d, J = 8.7 Hz, 1H), 7.03 (d, J = 7.8 Hz, 1H), 6.93-6.97 (m, 2H), 6.74 (dd, J = 8.7, 2.3 Hz, 1H), 6.10 (s, 1H), 4.60- 4.64 (m, 1H), 3.75 (s, 3H), 3.19 (td, J = 12.2, 4.3 Hz, 1H), 2.84-2.92 (m, 4H), 2.75 (dd, J = 15.2, 4.6 Hz, 1H) ; ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.3, 153.4, 148.7, 133.5, 131.5, 131.3, 128.0, 126.3, 120.2, 119.2, 117.3, 112.4, 112.0, 111.3, 100.0, 69.9, 55.4, 41.0, 36.4, 19.6; LRMS (FAB) m/z, 334 ([M+H] ⁺ ).

EV405

: Compound EV405 (17.4 mg, 55%) in the form of a pale yellow solid using N -methylisatoic anhydride ( 7 ) (17.4 mg, 0.098 mmol) and 4b (20.3 mg, 1.108 mmol) according to the Evodiamine ( 1 ) synthesis conditions yield) was obtained.

mp: 313-314° C.; ¹ H-NMR (400 MHz, DMSO-d ₆ ); δ 11.15 (s, 1H), 7.77 (dd, J = 7.6, 1.5 Hz, 1H), 7.47-7.49 (m, 1H), 7.35 (d, J = 7.9 Hz, 1H), 7.22 (dd, J = 9.8 , 2.4 Hz, 1H), 7.04 (d, J = 7.9 Hz, 1H), 6.91-6.96 (m, 2H), 6.13 (s, 1H), 4.62 (dd, J = 12.8, 4.9 Hz, 1H), 3.20 (td, J = 12.2, 4.7 Hz, 1H), 2.91 (s, 3H), 2.70-2.77 (m, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.2, 148.6, 133.6, 133.0, 131.6, 128.0, 126.3, 123.6, 120.2, 118.9, 117.2, 114.0, 111.7, 110.7, 110.0, 69.9, 41.0, 36.6, 19.4; LRMS (FAB) m/z, 322 ([M+H] ⁺ ).

EV406

: Compound EV406 (16.1 mg, 19%) in the form of a pale yellow solid using N -methylisatoic anhydride ( 7 ) (47.3 mg, 0.267 mmol) and 4c (54.1 mg, 0.294 mmol) according to the Evodiamine ( 1 ) synthesis conditions yield) was obtained.

mp: 284-285°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.91 (s, 1H), 7.78 (dd, J = 7.8, 1.4 Hz, 1H), 7.47 (td, J = 7.7, 1.5 Hz, 1H), 7.24 (d, J = 8.3 Hz, 2H), 7.04 (d, J = 7.8 Hz, 1H), 6.92-6.97 (m, 2H), 6.10 (s, 1H), 4.62 (dd, J = 12.9, 4.6 Hz, 1H), 3.18 (td, J = 12.1, 4.4 Hz, 1H), 2.84-2.92 (m, 4H), 2.75 (dd, J = 15.2, 4.6 Hz, 1H), 2.36 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.3, 148.8, 134.9, 133.5, 130.7, 128.0, 127.4, 126.2, 123.5, 120.2, 119.2, 117.9, 117.4, 111.4, 111.0, 69.8, 40.9, 36.4, 21.2, 19.5; LRMS (FAB) m/z, 318 ([M+H] ⁺ ).

EV407

: Compound EV407 (18.5 mg, 39%) in the form of a pale yellow solid using N -methylisatoic anhydride ( 7 ) (24.6 mg, 0.139 mmol) and 4c (31.3 mg, 0.153 mmol) according to the Evodiamine ( 1 ) synthesis conditions yield) was obtained.

mp: 312-313°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.27 (s, 1H), 7.78 (d, J = 7.8 Hz, 1H), 7.53 (d, J = 2.1 Hz, 1H), 7.45-7.50 (m , 1H), 7.36 (d, J = 8.7 Hz, 1H), 7.10 (dd, J = 8.5, 2.1 Hz, 1H), 7.04 (d, J = 8.3 Hz, 1H), 6.95 (t, J = 7.6 Hz) , 1H), 6.15 (s, 1H), 4.61 (dd, J = 12.9, 5.1 Hz, 1H), 3.20 (td, J = 12.2, 4.9 Hz, 1H), 2.90 (s, 3H), 2.85-2.93 ( m, 1H), 2.77 (dd, J = 15.4, 4.4 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.2, 148.6, 134.9, 133.6, 132.8, 128.0, 127.2, 123.6, 121.8, 120.3, 119.0, 117.7, 117.4, 113.2, 111.4, 69.79, 40.9, 36.7, 19.3; LRMS (FAB) m/z, 338 ([M+H] ⁺ ).

5-2. EV408 Derivative Synthesis Method

General synthesis of intermediates 8 and 9

: Serotonin hydrochloride (100.0 mg, 0.470 mmol) and N -methylisatoic anhydride ( 7 ) (75.7 mg, 0.427 mmol) were dissolved in CH ₂ Cl ₂ (1.1 mL, 0.4 M), followed by triethylamine (43.3 mg, 0.427 mmol) added. Then, the temperature was raised to 50° C., and the mixture was stirred under reflux for 3 hours. After completion of the reaction, the mixture obtained by removing the excess CH ₂ Cl ₂ by concentration under reduced pressure was purified by column chromatography (silica gel, hexane:ethyl acetate = 2:1 to 1:1), and compound 8 in the form of a colorless oil (33.1 mg , 22% yield) and 9 (68.9 mg, 33 % yield) in the form of a white solid were obtained.

Intermediate 9 General Conditions for Hydrolysis

: Intermediate 9 (68.9 mg, 0.156 mmol) was dissolved in THF (2.3 mL, 0.06 M), LiOH (11.2 mg, 0.467 mmol) was dissolved in MeOH (2.3 mL, 0.06 M), and it was slowly added to the reaction mixture. After stirring at room temperature for 12 hours, the mixture was concentrated under reduced pressure to remove excess THF and MeOH. After the obtained mixture was dissolved in ethyl acetate (30 mL), the organic layer was washed with water 3 times (10 mL × 3). The aqueous layer was again extracted with ethyl acetate three times (10 mL × 3). The obtained organic layer was dried over MgSO ₄ , filtered, and concentrated under reduced pressure. The resulting mixture was purified by column chromatography (silica gel, hexane:ethyl acetate = 2:1 to 1:1) to obtain Compound 8 (48.2 mg, 99.9% yield) in the form of a colorless oil.

Intermediate 8

: ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.92 (d, J = 16.8 Hz, 1H), 7.39 (s, 1H), 7.27-7.31 (m, 1H), 7.20 (d, J = 8.7 Hz, 1H), 7.14 (dd, J = 7.8, 1.4 Hz, 1H), 7.00 (s, 3H), 6.78 (dd, J = 8.6, 2.4 Hz, 1H), 6.67 (d, J = 8.3 Hz, 1H), 6.52 (t, J = 7.5 Hz, 1H), 6.16 (s, 1H), 5.29 (d, J = 4.1 Hz, 1H), 3.67 (q, J = 6.4 Hz, 3H), 2.98 (t, J = 6.7) Hz, 3H), 2.85 (s, 4H); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 171.5, 170.3, 150.4, 149.9, 132.9, 131.6, 128.1, 127.4, 123.3, 115.8, 115.0, 112.4, 112.3, 112.1, 111.4, 103.3, 40.2, 29.9, 25.3; LRMS (FAB) m/z, 310 ([M+H] ⁺ ).

Intermediate 9

: mp : 81-83°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.18 (dd, J = 8.0, 1.6 Hz, 1H), 8.13 (s, 1H), 7.66 (d, J = 5.1 Hz, 1H), 7.44-7.48 (m) , 2H), 7.40 (d, J = 2.1 Hz, 1H), 7.37 (d, J = 8.7 Hz, 1H), 7.26-7.30 (m, 2H), 7.16 (dd, J = 7.8, 1.6 Hz, 1H) , 7.07 (d, J = 2.3 Hz, 1H), 7.01 (dd, J = 8.7, 2.1 Hz, 1H), 6.73 (d, J = 8.5 Hz, 1H), 6.66-6.70 (m, 1H), 6.62 6.64 (m, 1H), 6.50-6.54 (m, 1H), 6.11 (s, 1H), 3.71 (q, J = 6.4 Hz, 2H), 3.03 (t, J = 6.7 Hz, 2H), 2.92 (d) , J = 5.1 Hz, 3H), 2.84 (d, J = 4.4 Hz, 3H); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 170.0, 168.5, 152.7, 150.4, 144.3, 135.4, 134.4, 132.8, 132.2, 127.8, 127.3, 123.6, 116.9, 115.5, 114.8, 114.6, 113.4, 111.9, 111.3, 111.0, 109.3, 39.9, 29.9, 29.7, 25.4; LRMS (FAB) m/z, 443 ([M+H] ⁺ ).

EV408

: Intermediate 8 (1.0 equiv.) was dissolved in triehoxymethane-DMF (0.33 M), and then trifluoroacetic anhydride (1.0 equiv.) and DABCO (1.1 equiv.) were sequentially added. After raising the temperature of the reactant to 100 °C, the mixture was stirred under reflux for 6 hours. After completion of the reaction, the mixture was dissolved in ethyl acetate (30 mL), and the organic layer was washed three times (10 mL × 3) with water. The aqueous layer was again extracted with ethyl acetate three times (10 mL × 3). The obtained organic layer was dried over MgSO ₄ , filtered, and concentrated under reduced pressure. The resulting mixture was purified by column chromatography (silica gel, hexane:ethyl acetate = 2:1 to 1:1) to obtain compound EV408 (40.0 mg, 41% yield) in the form of a white solid.

mp: 276-277°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.71 (s, 1H), 8.70 (s, 1H), 7.78 (dd, J = 8.1, 1.3 Hz, 1H), 7.45-7.49 (m, 1H) , 7.14 (d, J = 8.8 Hz, 1H), 7.02 (d, J = 8.1 Hz, 1H), 6.94 (t, J = 7.4 Hz, 1H), 6.75 (d, J = 1.7 Hz, 1H), 6.62 (dd, J = 8.4, 2.4 Hz, 1H), 6.08 (s, 1H), 4.60 (dd, J = 12.8, 4.7 Hz, 1H), 3.18 (td, J = 12.1, 4.5 Hz, 1H), 2.90 ( s, 3H), 2.80-2.87 (m, 1H), 2.66 (dd, J = 15.5, 4.0 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.3, 150.7, 148.7, 133.5, 131.2, 130.9, 128.0, 126.7, 120.0, 119.0, 117.1, 112.1, 112.1, 110.6, 102.1, 70.0, 41.1, 36.4, 19.6; HRMS (FAB) m/z calcd for C ₁₉ H ₁₈ N ₃ O ₂ [M+H] ⁺ : 320.1399, found 320.1401.

5-3. Synthesis method of EV404, EV409~413 derivatives

O General conditions for -alkylation

: EV408 (1.0 equiv.) and K ₂ CO ₃ (1.2 equiv.) were dissolved in EtOH (0.05 M), and then an alkyl halide (1.2 equiv.) was added. After raising the temperature of the reaction product to 80 °C, the mixture was stirred under reflux overnight. After completion of the reaction, the mixture was concentrated under reduced pressure to remove excess EtOH. After the obtained mixture was dissolved in ethyl acetate (30 mL), the organic layer was washed with water 3 times (10 mL × 3). The aqueous layer was again extracted with ethyl acetate three times (10 mL × 3). The obtained organic layer was dried over MgSO ₄ , filtered, and concentrated under reduced pressure. The resulting mixture was purified by column chromato-graphy (silica gel, hexane:ethyl acetate = 3:1 to 1:1) to obtain solid compounds EV404 and EV409-413 .

EV404

: According to the general conditions of O -alkylation, EV408 (14.8 mg, 0.046 mmol) and benzyl bromide (9.4 mg, 0.055 mmol) were used to obtain compound EV404 (8.7 mg, 46% yield) in yellow solid form.

mp: 188-190°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.13 (s, 1H), 8.10 (m, 1H), 7.46 (m, 1H), 7.37 (t, J = 7.5 Hz, 1H), 7.30 (m , 1H), 7.17 (t, J = 7.5 Hz, 1H), 7.10 (m, 1H), 6.97 (dd, J = 8.5 Hz, 1H), 5.87 (s, 1H), 5.11 (s, 1H), 4.84 (m, 1H), 3.25 (m, 1H), 2.92 (m, 2H), 2.49 (s, 3H); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 164.8, 153.7, 150.7, 137.6, 133.1, 132.0, 129.2, 129.0, 128.6, 127.9, 127.6, 126.7, 124.0, 123.7, 122.3, 114.0, 113.5, 112.1, 102.5, 71.0, 69.0, 39.6, 37.3, 20.2; LRMS (FAB) m/z, 410 ([M+H] ⁺ ).

EV409

: According to the general conditions of O -alkylation, EV408 (32.2 mg, 0.101 mmol) and propargyl bromide (14.4 mg, 0.121 mmol) were used to obtain compound EV409 (31.7 mg, 88% yield) in yellow solid form.

mp: 170-172°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.19 (s, 1H), 8.11 (dd, J = 7.8, 1.4 Hz, 1H), 7.48 (td, J = 7.7, 1.5 Hz, 1H), 7.33 (t) , J = 8.2 Hz, 1H), 7.19-7.23 (m, 1H), 7.13 (q, J = 2.5 Hz, 2H), 6.98 (dd, J = 8.7, 2.3 Hz, 1H), 5.89 (s, 1H) , 4.87 (dt, J = 12.7, 3.7 Hz, 1H), 4.75 (d, J = 2.3 Hz, 2H), 3.22-3.30 (m, 1H), 2.92-2.95 (m, 2H), 2.51-2.53 (m) , 4H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.3, 151.3, 148.7, 133.5, 132.0, 131.6, 128.0, 126.2, 120.2, 119.2, 117.4, 112.5, 112.4, 111.4, 102.1, 80.0, 77.8, 69.8, 56.1, 41.1, 36.5, 19.6; LRMS (FAB) m/z, 360 ([M+H] ⁺ ).

EV410

: According to the general conditions of the O -alkylation, EV408 (20.0 mg, 0.063 mmol) and allyl bromide (11.4 mg, 0.094 mmol) were used to obtain a yellow solid compound EV410 (21.9 mg, 96.9% yield).

mp: 176-178°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.13 (s, 1H), 8.08 (d, J = 7.0 Hz, 1H), 7.44 (m, 1H), 7.26 (d, J = 6.5 Hz, 1H), 7.17 (t, J = 7.5 Hz, 1H), 7.10 (d, J = 8.0 Hz, 1H), 7.01 (s, 1H), 6.92 (dd, J = 8.0 Hz, 1H), 6.10 (m, 1H), 5.86 (s, 1H), 5.43 (dd, J = 18 Hz, 1H), 5.27 (dd, J = 10 Hz, 1H), 4.84 (m, 1H), 4.58 (d, J = 5.5 Hz, 2H), 3.26 (m, 1H), 2.90 (m, 2H), 2.49 (s, 3H); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 164.7, 153.4, 150.6, 133.8, 133.0, 131.9, 129.1, 129.0, 126.6, 124.0, 123.6, 122.2, 117.4, 113.9, 113.4, 112.0, 102.3, 69.8, 68.9, 39.6, 37.2, 20.1; LRMS (FAB) m/z, 358 ([M+H] ⁺ ).

EV411

: According to the general conditions of O -alkylation, EV408 (20.0 mg, 0.063 mmol) and benzoyl chloride (13.2 mg, 0.094 mmol) were used to obtain compound EV411 (19.2 mg, 72% yield) in the form of a yellow solid.

mp: 224-226°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.77 (s, 1H), 7.82 (s, 1H), 7.77 (dd, J = 7.8, 1.4 Hz, 1H), 7.44-7.48 (m, 1H) , 7.23 (s, 1H), 7.14 (d, J = 8.7 Hz, 1H), 7.01 (d, J = 7.8 Hz, 1H), 6.93 (t, J = 7.4 Hz, 1H), 6.75 (d, J = 1.8 Hz, 1H), 6.63 (dd, J = 8.7, 2.3 Hz, 1H), 6.08 (s, 1H), 4.60 (dd, J = 12.6, 4.8 Hz, 1H), 3.17 (td, J = 12.2, 4.4) Hz, 1H), 2.90 (s, 3H), 2.79-2.86 (m, 1H), 2.67 (td, J = 15.3, 4.3 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.2, 150.8, 148.6, 139.0, 133.5, 131.2, 130.9, 128.9, 127.9, 126.9, 126.7, 120.0, 118.9, 117.0, 112.1, 110.5, 102.1, 70.0, 41.1, 36.4, 28.2, 19.6; LRMS (FAB) m/z, 424 ([M+H] ⁺ ).

EV412

: According to the general conditions of O -alkylation, EV408 (20.0 mg, 0.063 mmol) and acetyl chloride (7.4 mg, 0.094 mmol) were used to obtain compound EV412 (12.5 mg, 55% yield) in the form of a yellow solid.

mp: 201-203° C.; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.29 (s, 1H), 8.09 (m, 1H), 7.46 (m, 1H), 7.26 (d, J = 2.0 Hz, 1H), 7.20 (m, 1H) ), 7.11 (d, J = 8.0 Hz, 1H), 6.94 (dd, J = 8.5 Hz, 1H), 5.87 (s, 1H), 5.11 (s, 1H), 4.82 (m, 1H), 3.25 (m) , 1H), 2.88 (m, 2H), 2.49 (s, 3H), 2.31 (s, 3H); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 170.5, 164.7, 150.5, 144.5, 134.5, 133.1, 129.8, 129.0, 126.6, 124.1, 123.6, 122.3, 117.1, 113.9, 111.8, 111.2, 68.8, 39.5, 37.3, 21.2, 20.1; LRMS (FAB) m/z, 362 ([M+H] ⁺ ).

EV413

: According to the general conditions of O -alkylation, EV408 (28.0 mg, 0.088 mmol) and 1-chloropropane (16.2 mg, 0.132 mmol) were used to obtain compound EV413 (14.0 mg, 44% yield) in the form of a yellow solid.

mp: 158-160°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 0.89-1.08 (t, J = 7.0 Hz, 3H), 1.84-1.86 (m, 2H), 2.50 (s, 3H), 2.92-2.93 (m, 2H) , 3.25-3.31 (m, 1H), 3.99 (t, J = 6.6 Hz, 2H), 4.84-4.88 (m, 1H), 5.90 (s, 1H), 6.92 (d, J = 8.8 Hz, 1H), 7.03 (s, 1H), 7.13 (d, J = 8.0 Hz, H), 7.20 (t, J = 7.7 Hz, 1H), 7.29 (d, J = 8.8 Hz, 1H), 7.48 (t, J = 8.0) Hz, 1H), 8.08 (s, 1H), 8.11 (d, J = 7.7 Hz, 1H). ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 164.3 153.4, 150.2, 132.6, 131.3, 128.5 (2C), 126.2, 123.4, 123.1, 121.7, 113.3, 112.9, 111.6, 101.4, 70.0, 68.4, 39.2, 36.7, 22.3, 19.7, 10.2; LRMS (FAB) m/z, 362 ([M+H] ⁺ ).

Synthesis Example 6. Synthesis of multi-substituted evodiamine derivatives (EV501 to 509)

<Synthesized evodiamine derivative EV501~509>

6-1. Synthesis method of EV501-506

N -formyltryptamine (1)

: Tryptamine (1.00 g, 6.24 mmol) and ethyl formate (2.3 g, 31.210 mmol) were heated and stirred at 80°C under Ar substitution. After completion of the reaction, the mixture was concentrated under reduced pressure to remove excess ethyl formate, and the resulting mixture was purified by column chromatography (silica gel, MC:MeOH = 20:1 to 10:1) to form a brown oil of Compound 1 (1.2 g, 100 % Yield) was obtained.

¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.13 (br s, 2H), 7.61 (d, J = 8.3 Hz, 1 H), 7.38-7.40 (m, 1H), 7.23 (td, J = 7.6, 1.4 Hz, 1H), 7.14 (t, J = 7.6 Hz, 1H), 7.06 (s, 1H), 5.58 (br s, 1H), 3.64-3.69 (m, 2H), 3.01 (t, J = 6.9 Hz) , 2H); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 161.5, 136.5, 127.3, 122.4, 122.2, 119.5, 118.7, 112.4, 111.5, 42.1, 38.4, 27.4, 25.2; HRMS (FAB) m/z calcd for C ₁₁ H ₁₂ N ₂ O [M+H] ⁺ : 188.0950, found 188.0955.

3,4-dihydro-β-carboline (2)

: N -formyltryptamine ( 1 ) (1.1 g, 5.920 mmol) was dissolved in CH ₂ Cl ₂ (15.0 mL), and then POCl ₃ (1.7 mL, 17.770 mmol) was slowly added at 0°C. After stirring at 0° C. for 2 hours, the mixture was stirred at room temperature for 2 hours. After completion of the reaction, excess POCl ₃ and CH ₂ Cl ₂ were removed by concentration under reduced pressure, 1 M HCl (100 mL) was added to the obtained mixture, and then washed once with CH ₂ Cl ₂ (1 × 30 mL). The washed aqueous layer was adjusted to pH = 10 with 1 M NaOH at 0° C., and extracted three times with CH ₂ Cl ₂ (3 × 30 mL). The obtained organic layer was dried over MgSO ₄ , filtered, and concentrated under reduced pressure. Et ₂ O was added to the resulting mixture to precipitate a solid, followed by filtration to obtain Compound 2 (4.0 g, 64% yield) in the form of a yellow solid.

mp: 99-101°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.32 (s, 1H), 8.36 (t, J = 2.3 Hz, 1H), 7.55 (d, J = 8.2 Hz, 1H), 7.41 (d, J) = 8.2 Hz, 1H), 7.17-7.21 (m, 1H), 7.04 (td, J = 7.5, 0.9 Hz, 1H), 3.78 (td, J = 8.7, 2.1 Hz, 2H), 2.80 (t, J = 8.7 Hz, 2H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 151.6, 136.7, 128.4, 124.8, 123.7, 119.7, 119.6, 113.8, 112.5, 48.1, 18.7; HRMS (FAB) m/z calcd for C ₁₁ H ₁₁ N ₂ [M+H] ⁺ : 171.0917, found 171.0921.

Isatoic anhydride (3)

: Anthranilic acid (1.1 g, 8.090 mmol) was dissolved in THF (15.0 mL), and then triphosgene (7.2 g, 24.280 mmol) was added at room temperature. Thereafter, the reaction temperature was raised to 50° C. and stirred for 3 hours. After completion of the reaction, ice water (50 mL) was added to the mixture and filtered under reduced pressure to obtain Compound 3 (1.3 g, 95% yield) in the form of a white solid.

mp: 164-165°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.73 (s, 1H), 7.91 (dd, J = 7.8, 1.4 Hz, 1H), 7.72-7.76 (m, 1H), 7.23-7.27 (m, 1H), 7.15 (d, J = 7.8 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 159.9, 147.1, 141.4, 137.00, 129.00, 123.5, 115.4, 110.3; HRMS (FAB) m/z calcd for C ₈ H ₅ NO ₃ [M+H] ⁺ : 164.0269, found 164.0271.

General alkylation conditions for isatoic anhydride

: Isatoic anhydride ( 3 ) (1.0 equiv.) was dissolved in DMAc (0.4 M), DIPEA (3.0 equiv.) and alkyl halide (3.0 equiv.) were slowly added thereto, followed by stirring at 40°C overnight. Then, after completion of the reaction by adding H ₂ O to the mixture, the mixture was extracted several times with CH ₂ Cl ₂ . The collected organic layers were dried over MgSO ₄ , filtered, and concentrated under reduced pressure. After adding hexane to the obtained mixture to precipitate a solid, it filtered and obtained 4a - 4c .

N -allyl isatoic anhydride (4a)

: According to the general alkylation conditions, isatoic anhydride ( 3 ) (200.0 mg, 1.226 mmol) and allyl bromide (445.0 mg, 3.678 mmol) were used to obtain a beige solid compound 4a (106.0 mg, 50% yield).

mp: 127-128°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.02 (dd, J = 8.0, 1.1 Hz, 1H), 7.79-7.84 (m, 1H), 7.31-7.35 (m, 2H), 5.87-5.96 ( m, 1H), 5.31 (dq, J = 17.5, 1.7 Hz, 1H), 5.20 (dq, J = 10.6, 1.5 Hz, 1H), 4.67 (td, J = 3.2, 1.5 Hz, 2H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 159.0, 147.6, 141.3, 137.0, 131.2, 129.5, 123.6, 117.2, 115.2, 111.9, 46.5, 40.2, 39.9, 39.7, 39.5, 39.3, 39.1, 38.90; LRMS (FAB) m/z, 334 ([M+H] ⁺ ).

N -(2-methyl)allyl isatoic anhydride (4b)

: Compound 4b (151.6 mg, 57%) in the form of a beige solid using isatoic anhydride ( 3 ) (200.0 mg, 1.226 mmol) and 3-bromo-2-methylpropene (496.6 mg, 3.678 mmol) according to the general alkylation conditions yield) was obtained.

mp: 124-125°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.02 (dd, J = 7.7, 1.5 Hz, 1H), 7.78-7.82 (m, 1H), 7.31-7.35 (m, 1H), 7.24 (d, J = 8.6 Hz, 1H), 4.82-4.87 (m, 2H), 4.55 (s, 2H), 1.80 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 159.0, 147.8, 141.5, 138.2, 136.9, 129.4, 123.7, 115.4, 111.8, 111.0, 49.5, 19.8; LRMS (FAB) m/z, 334 ([M+H] ⁺ ).

N -propargyl isatoic anhydride (4c)

: According to the general condition B of the above N -alkylation, using isatoic anhydride ( 6 ) (200.0 mg, 1.226 mmol) and propargyl bromide (437.5 mg, 3.678 mmol) as a beige solid compound 4c (166.6 mg, 68% yield) ) was obtained.

mp: 176-177°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.04 (d, J = 7.8 Hz, 1n), 7.91 (t, J = 7.8 Hz, 1H), 7.51 (d, J = 8.7 Hz, 1H), 7.38 (t, J = 7.6 Hz, 1H), 4.89 (s, 2H), 3.43 (s, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 158.5, 147.3, 140.5, 137.2, 129.6, 124.1, 115.0, 111.9, 77.6, 76.1, 34.4; LRMS (FAB) m/z, 202 ([M+H] ⁺ ).

General Condensation Conditions for Compound No. 5

: Dissolve N -alkyl isatoic anhydride ( 4 ) (1.0 equiv.) and 3,4-dihydro-β-carboline ( 2 ) (1.1 equiv.) in CH ₂ Cl ₂ (0.4 M) at 50°C under Ar substitution was stirred for 6 hours. The resulting solid was filtered to obtain 5a - 5c .

14 N -allyl evodiamine (5a)

: According to the general condensation reaction conditions, 3,4-dihydro-β-carboline ( 2 ) (92.2 mg, 0.541 mmol) and N -allyl isatoic anhydride ( 4a ) (100.0 mg, 0.492 mmol) were used to form a yellow solid. of compound 5a (125.1 mg, 84% yield) was obtained.

mp: 204-206°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.08 (s, 1H), 7.79 (d, J = 7.8 Hz, 1H), 7.42-7.47 (m, 2H), 7.35 (d, J = 7.8 Hz) , 1H), 7.11 (dd, J = 7.6, 5.3 Hz, 2H), 6.97 (td, J = 15.1, 7.8 Hz, 2H), 6.15 (s, 1H), 5.85 (dq, J = 22.4, 5.4 Hz, 1H), 5.02-5.07 (m, 2H), 4.62 (dd, J = 12.9, 5.1 Hz, 1H), 3.94 (d, 2H), 3.23 (td, J = 12.2, 4.6 Hz, 1H), 2.90-2.98 (m, 1H), 2.76 (dd, J = 15.6, 4.1 Hz, 1H). ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.5, 147.2, 136.4, 134.5, 133.1, 130.8, 128.0, 126.1, 121.8, 120.4, 120.0, 118.9, 118.5, 118.2, 117.1, 111.7, 111.4, 69.0, 52.6, 41.2, 19.3; HRMS (FAB) m/z calcd for C ₂₁ H ₂ ON ₃ O [M+H] ⁺ : 330.1606, found 330.1607.

14 N -(2-methyl)allyl evodiamine (5b)

: According to the general condensation reaction conditions, 3,4-dihydro-β-carboline ( 2 ) (43.1 mg, 0.253 mmol) and N -(2-methyl)allyl isatoic anhydride ( 4b ) (50.0 mg, 0.230 mmol) was used to obtain compound 5b (75.0 mg, 95% yield) in the form of a dark yellow solid.

mp: 181-183°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.94 (s, 1H), 7.72 (dd, J = 7.9, 1.8 Hz, 1H), 7.32-7.43 (m, 3H), 7.08 (t, J = 7.9 Hz, 1H), 6.98 (t, J = 7.9 Hz, 1H), 6.91 (d, J = 7.9 Hz, 1H), 6.80 (t, J = 7.0 Hz, 1H), 6.20 (s, 1H), 4.88 (d, J = 21.4 Hz, 2H), 4.62 (dd, J = 13.4, 5.5 Hz, 1H), 4.22 (d, J = 16.5 Hz, 1H), 3.94 (d, J = 16.5 Hz, 1H), 3.26 (td, J = 12.2, 4.9 Hz, 1H), 2.95-3.03 (m, 1H), 2.68 (dd, J = 15.6, 4.6 Hz, 1H), 1.72 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.9, 146.6, 141.3, 136.1, 133.3, 132.1, 128.0, 126.3, 121.7, 118.9, 118.8, 118.1, 117.4, 115.8, 112.3, 111.7, 111.2, 70.1, 54.9, 42.2, 20.0, 19.2; HRMS (FAB) m/z calcd for C ₂₂ H ₂₂ N ₃ O [M+H] ⁺ : 344.1763, found 344.1759.

14 N -propagyl evodiamine (5c)

: According to the general condensation reaction conditions, 3,4-dihydro-β-carboline ( 2 ) (93.1 mg, 0.547 mmol) and N -propargyl isatoic anhydride ( 4c ) (100.0 mg, 0.497 mmol) were used to form a yellow solid. of compound 5c (159.2 mg, 98% yield) was obtained.

mp: 182-184°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.26 (s, 1H), 7.90 (dd, J = 7.8, 1.4 Hz, 1H), 7.50-7.56 (m, 2H), 7.38 (d, J = 8.3 Hz, 1H), 7.31 (d, J = 7.4 Hz, 1H), 7.12-7.19 (m, 2H), 7.02-7.06 (m, 1H), 6.16 (s, 1H), 4.62 (ddd, J = 12.8) , 4.9, 2.0 Hz, 1H), 4.04 (dd, J = 18.2, 2.5 Hz, 1H), 3.61 (dd, J = 17.9, 2.3 Hz, 1H), 3.18-3.25 (m, 1H), 3.04 (t, J = 2.3 Hz, 1H), 2.84-2.98 (m, 2H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.5, 147.1, 136.8, 132.8, 129.0, 128.0, 125.8, 123.4, 122.9, 122.1, 121.3, 119.0, 118.5, 112.2, 111.7, 79.9, 75.2, 68.1, 19.5; HRMS (FAB) m/z calcd for C ₂₁ H ₁₈ N ₃ O [M+H] ⁺ : 328.1450, found 328.1456.

General Nitration Conditions for Evodiamine Derivatives

: KNO ₃ (1.0 equiv.) and concentrated H ₂ SO ₄ (0.33 M) were added to 14 N -alkyl evodiamine ( 5 ) (1.0 equiv.), followed by stirring at 0° C. overnight. After completion of the reaction, ice water was added to the mixture, and the resulting solid was filtered under reduced pressure to obtain EV501-503 .

EV501

: According to the general nitration conditions, 5a (200.0 mg, 0.607 mmol) and KNO ₃ (61.4 mg, 0.607 mmol) were used to obtain compound EV501 (83.8 mg, 37% yield) in the form of a yellow solid.

mp: 186-188°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.92 (s, 1H), 8.49 (d, J = 2.3 Hz, 1H), 8.01 (dd, J = 9.2, 2.3 Hz, 1H), 7.78 (dd , J = 7.8, 1.4 Hz, 1H), 7.43-7.51 (m, 2H), 7.12 (d, J = 8.3 Hz, 1H), 6.95 (t, J = 7.7 Hz, 1H), 6.22 (s, 1H) , 5.87 (ddd, J = 23.0, 10.1, 6.0 Hz, 1H), 5.04-5.10 (m, 2H), 4.63 (dd, J = 12.9, 4.6 Hz, 1H), 3.98 (d, J = 4.1 Hz, 2H) ), 3.26 (dd, J = 13.1, 4.8 Hz, 1H), 2.96-3.04 (m, 1H), 2.88 (dd, J = 15.6, 4.6 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.1, 147.9, 139.5, 136.2, 134.7, 133.0, 131.9, 131.1, 129.8, 128.1, 122.0, 121.6, 121.5, 120.1, 117.3, 116.9, 114.4, 67.6, 52.8, 19.4; HRMS (FAB) m/z calcd for C ₂₁ H ₁₉ N ₄ O ₃ [M+H] ⁺ : 375.1457, found 375.1459.

EV502

: According to the general nitration conditions, 5b (87.9 mg, 0.226 mmol) and KNO ₃ (22.9 mg, 0.226 mmol) were used to obtain compound EV502 (31.6 mg, 36% yield) in the form of a yellow solid.

mp: 261-262°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.94 (s, 1H), 8.52 (d, J = 2.3 Hz, 1H), 8.04 (dd, J = 9.2, 2.3 Hz, 1H), 7.71 (dd , J = 7.8, 1.4 Hz, 1H), 7.55 (m, 2H), 7.49-7.44 (m, 1H), 7.01 (t, J = 7.7 Hz, 1H), 6.95 (d, J = 4.1 Hz, 1H) , 6.11 (s, 1H), 4.98 (d, J = 4.1 Hz, 2H), 4.64 (dd, J = 13.8, 4.3 Hz, 1H), 4.06 (d, J = 4.6 Hz, 1H), 3.78 (d, J = 4.3 Hz, 1H), 3.21 (dt, J = 13.1, 4.8 Hz, 1H), 3.07-2.99 (m, 1H), 2.81 (dd, J = 15.8, 4.1 Hz, 1H), 1.86 (s, 3H) ); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 165.2, 147.4, 140.5, 139.2, 133.6, 131.9, 130.2, 127.7, 121.6, 119.2, 118.5, 117.2, 116.4, 115.4, 115.1, 114.0, 113.5, 72.6, 59.1, 20.4, 18.44; LRMS (FAB) m/z, 389 ([M+H] ⁺ ).

EV505

: According to the general nitration conditions, 5c (100.0 mg, 0.305 mmol) and KNO ₃ (30.9 mg, 0.305 mmol) were used to obtain compound EV505 (107.6 mg, 94.8% yield) in the form of a yellow solid.

mp: 164-166°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 12.10 (s, 1H), 8.57 (d, J = 2.0 Hz, 1H), 8.05 (dd, J = 8.9, 2.2 Hz, 1H), 7.89 (dd , J = 7.7, 1.3 Hz, 1H), 7.52-7.58 (m, 2H), 7.32 (d, J = 8.1 Hz, 1H), 7.16-7.20 (m, 1H), 6.24 (s, 1H), 4.60- 4.66 (m, 1H), 4.08 (dd, J = 18.1, 2.4 Hz, 1H), 3.69 (dd, J = 18.1, 2.4 Hz, 1H), 3.22-3.29 (m, 1H), 3.08 (t, J = 2.4 Hz, 1H), 3.00 (d, J = 4.7 Hz, 2H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.5, 146.9, 140.7, 139.9, 133.5, 133.1, 128.0, 125.3, 123.1, 121.4, 117.5, 115.9, 114.8, 112.2, 79.8, 75.5, 68.0, 19.3; HRMS (FAB) m/z calcd for C ₂₁ H ₁₇ N ₄ O ₃ [M+H] ⁺ : 373.1301, found 373.1305.

General dinitration conditions for Evodiamine derivatives

: KNO ₃ (2.0 equiv.) and concentrated H ₂ SO ₄ (0.33 M) were added to 14N -alkyl evodiamine ( 5 ) (1.0 equiv.) at 0° C., followed by stirring at room temperature overnight. After completion of the reaction, ice water was added to the mixture and filtered under reduced pressure. The solid obtained was purified by column chromatography (silica gel, hexane:ethyl acetate = 2:1 to 1:1) to obtain EV504-506 .

EV503

: According to the general dinitration conditions, 5a (200.0 mg, 0.607 mmol) and KNO ₃ (122.8 mg, 1.214 mmol) were used to obtain compound EV503 (28.8 mg, 11.3% yield) in the form of a yellow solid.

mp: 229-231°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 12.51 (s, 1H), 9.02 (d, J = 2.0 Hz, 1H), 8.82 (d, J = 2.0 Hz, 1H), 7.86 (dd, J) = 7.9, 1.5 Hz, 1H), 7.48-7.52 (m, 1H), 7.26 (d, J = 8.4 Hz, 1H), 7.07 (t, J = 7.4 Hz, 1H), 6.16 (s, 1H), 5.68 -5.78 (m, 1H), 4.86-4.91 (m, 2H), 4.66 (dd, J = 12.3, 4.9 Hz, 1H), 3.95 (dd, J = 16.3, 5.9 Hz, 1H), 3.63 (dd, J) = 16.3, 6.2 Hz, 1H), 3.12-3.23 (m, 2H), 2.89-2.97 (m, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.1, 147.9, 139.5, 136.2, 134.7, 133.0, 131.9, 131.1, 129.9, 128.1, 122.0, 121.6, 121.4, 120.1, 117.3, 116.9, 114.4, 67.6, 52.8, 19.4; HRMS (FAB) m/z calcd for C ₂₁ H ₁₈ N ₅ O ₈ [M+H] ⁺ : 420.1308, found 420.1301.

EV504

: According to the general dinitration conditions, 5b (211.0 mg, 0.614 mmol) and KNO ₃ (124.2 mg, 1.229 mmol) were used to obtain compound EV504 (34.6 mg, 13% yield) in the form of a yellow solid.

mp: 175-177°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 12.55 (s, 1H), 9.04 (d, J = 2.0 Hz, 1H), 8.84 (d, J = 2.0 Hz, 1H), 7.91 (dd, J) = 7.7, 1.7 Hz, 1H), 7.50-7.54 (m, 1H), 7.15 (t, J = 7.6 Hz, 1H), 7.07 (d, J = 7.7 Hz, 1H), 6.17 (s, 1H), 5.29 (dd, J = 30.7, 14.3 Hz, 2H), 5.07 (d, J = 16.8 Hz, 2H), 4.66-4.71 (m, 1H), 3.90 (d, J = 16.5 Hz, 1H), 3.63 (d, J = 16.5 Hz, 1H), 3.34-3.41 (m, 1H), 3.15-3.21 (m, 1H), 2.89-2.96 (m, 1H), 1.99 (s, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 165.3, 147.7, 140.6, 139.5, 133.7, 132.0, 130.3, 127.9, 121.8, 119.3, 118.7, 117.4, 116.7, 115.6, 115.3, 114.2, 113.7, 72.9, 59.8, 20.8, 18.7; LRMS (FAB) m/z, 434 ([M+H] ⁺ ).

EV506

: According to the general dinitration conditions, 5c (166.2 mg, 0.508 mmol) and KNO ₃ (102.6 mg, 1.015 mmol) were used to obtain compound EV506 (48.3 mg, 25% yield) in the form of a yellow solid.

mp: 232-234°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 12.53 (s, 1H), 12.11 (s, 1H), 9.07 (d, J = 2.0 Hz, 1H), 8.84 (d, J = 2.4 Hz, 1H) ), 7.87-7.92 (m, 3H), 7.52-7.59 (m, 4H), 7.37 (d, J = 8.1 Hz, 1H), 7.14-7.21 (m, 3H), 6.27-6.23 (1H), 4.64- 4.71 (m, 1H), 4.12 (dd, J = 14.3, 2.5 Hz, 1H), 3.60 (dd, J = 18.5, 2.4 Hz, 1H), 3.22-3.29 (m, 1H), 3.12 (t, J = 2.2 Hz, 1H), 3.00 (d, J = 7.4 Hz, 2H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.3, 147.1, 139.5, 135.7, 131.9, 131.4, 129.8, 128.0, 123.1, 122.1, 121.3, 117.6, 114.5, 112.1, 80.1, 75.6, 67.5, 40.1, 39.9, 39.7, 39.5, 39.3, 39.1, 38.9, 19.4; HRMS (FAB) m/z calcd for C ₂₁ H ₁₆ N ₅ O ₅ [M+H] ⁺ : 418.1151, found 418.1158.

6-2. Synthesis method of EV507-509

General Cleavage Conditions for Compound 6

: N -alkyl isatoic anhydride ( 4 ) (1.0 equiv.) and serotonin hydrochloride (1.1 equiv.) were dissolved in CH ₂ Cl ₂ (0.4 M), followed by addition of triethylamine (1.0 equiv.). Then, the temperature was raised to 50° C., and the mixture was stirred under reflux for 3 hours. After completion of the reaction, the excess CH ₂ Cl ₂ was removed by concentration under reduced pressure, and the resulting mixture was purified by column chromatography (silica gel, hexane:ethyl acetate = 2:1 to 1:1) to obtain the desired intermediate 6 and by-product 7 .

According to the general cleavage reaction conditions, serotonin hydrochloride (300.0 mg, 1.411 mmol) and N -allyl isatoic anhydride ( 4a ) (286.6 mg, 1.411 mmol) were used to form a colorless oil of Compound 6a (135.0 mg, 29% yield) and compound 7a (198.4 mg, 28% yield) in the form of a white solid was obtained.

2-(allylamino)- N -(2-(5-hydroxy-1) H -indol-3-yl)ethyl)benzamide (6a)

¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.91 (s, 1H), 7.70 (s, 1H), 7.14-7.16 (m, 1H), 7.04 (dd, J = 8.1, 2.0 Hz, 2H), 6.78 (dd, J = 8.6, 2.9 Hz, 1H), 6.67 (d, J = 8.7 Hz, 1H), 6.52 (t, J = 7.1 Hz, 1H), 6.13 (s, 1H), 5.91-6.00 (m, 1H), 5.28-5.33 (m, 1H), 5.17 (dd, J = 10.1, 1.3 Hz, 1H), 4.74 (s, 1H), 3.84 (d, J = 4.7 Hz, 2H), 3.70 (q, J) = 6.5 Hz, 2H), 3.02 (t, J = 6.7 Hz, 2H); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 171.4, 170.3, 149.8, 149.3, 132.7, 132.2, 127.9, 127.4, 123.3, 116.3, 115.1, 112.6, 112.2, 112.1, 111.5, 103.5, 77.5, 77.2, 76.8, 46.0, 45.7, 40.2, 25.4 ; LRMS (FAB) m/z, 336 ([M+H] ⁺ ).

3-(2-(2-(allylamino)benzamido)ethyl)-1 H -indol-5-yl 2-(allylamino)benzoate (7a)

mp: 67-69°C; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.19 (dd, J = 8.1, 1.3 Hz, 1H), 8.17 (s, 1H), 7.89 (t, J = 5.0 Hz, 1H), 7.73 (s, 1H) ), 7.36-7.45 (m, 3H), 7.23 (td, J = 7.9, 1.6 Hz, 1H), 7.16-7.17 (m, 1H), 7.06 (d, J = 2.0 Hz, 1H), 7.01 (dd, J = 8.6, 1.8 Hz, 1H), 6.62-6.73 (m, 3H), 6.52 (t, J = 8.1 Hz, 1H), 6.13 (t, J = 5.4 Hz, 1H), 5.88-5.98 (m, 2H) ), 5.26-5.32 (m, 2H), 5.13-5.18 (m, 2H), 3.85-3.90 (m, 2H), 3.80 (d, J = 5.4 Hz, 2H), 3.72 (q, J = 6.5 Hz, 2H), 3.04 (t, J = 6.7 Hz, 2H); ¹³ C-NMR (100 MHz, CDCl ₃ ) δ 170.0, 168.5, 151.7, 149.4, 144.3, 135.3, 135.0, 134.4, 132.7, 132.3, 127.8, 127.4, 123.7, 116.9, 116.4, 116.2, 115.6, 115.1, 115.0, 113.4, 112.1, 111.9, 111.8, 111.3, 109.4, 45.6, 45.4, 39.9, 25.4; LRMS (FAB) m/z, 495 ([M+H] ⁺ ).

General hydrolysis conditions for by-product 7

: By-product 7 (1.0 equiv.) was dissolved in THF (0.06 M), then LiOH (3.0 equiv.) was dissolved in MeOH (0.06 M) and slowly added to the reaction mixture. After stirring at room temperature for 12 hours, the mixture was concentrated under reduced pressure to remove excess THF and MeOH. After the obtained mixture was dissolved in ethyl acetate (30 mL), the organic layer was washed with water 3 times (10 mL × 3). The aqueous layer was again extracted with ethyl acetate three times (10 mL × 3). The obtained organic layer was dried over MgSO ₄ , filtered, and concentrated under reduced pressure. The obtained mixture was purified by column chromatography (silica gel, hexane:ethyl acetate = 2:1 to 1:1) to obtain the desired intermediate 6 quantitatively.

General conditions for cyclization of intermediate 6

: Intermediate 6 (1.0 equiv.) was dissolved in triehoxymethane-DMF (0.33 M), and then trifluoroacetic anhydride (1.0 equiv.) and DABCO (1.1 equiv.) were sequentially added. After raising the temperature of the reactant to 100 °C, the mixture was stirred under reflux for 6 hours. After completion of the reaction, the mixture was dissolved in ethyl acetate, and the organic layer was washed three times with water. The aqueous layer was again extracted with ethyl acetate three times. The obtained organic layer was dried over MgSO ₄ , filtered, and concentrated under reduced pressure. The obtained mixture was purified by column chromatography (silica gel, hexane:ethyl acetate = 2:1 to 1:1) to obtain EV507-509 .

EV507

According to the general cyclization conditions, 6a (135.0 mg, 0.402 mmol) was used to obtain compound EV507 (50.4 mg, 36% yield) in the form of a yellow solid.

mp: 234-236°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.74 (s, 1H), 8.70 (s, 1H), 7.77 (dd, J = 8.1, 1.3 Hz, 1H), 7.41-7.45 (m, 1H) , 7.13 (d, J = 8.8 Hz, 1H), 7.07 (d, J = 8.8 Hz, 1H), 6.92 (t, J = 7.1 Hz, 1H), 6.74 (d, J = 2.0 Hz, 1H), 6.61 (dd, J = 8.4, 2.4 Hz, 1H), 6.10 (s, 1H), 5.80-5.89 (m, 1H), 5.03-5.09 (m, 2H), 4.59 (dd, J = 13.1, 5.1 Hz, 1H) ), 3.94 (d, J = 4.7 Hz, 2H), 3.16-3.24 (m, 1H), 2.82-2.90 (m, 1H), 2.64 (dd, J = 15.2, 4.4 Hz, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 164.6, 150.7, 147.1, 134.5, 133.1, 131.4, 130.8, 128.0, 126.9, 120.2, 119.8, 118.2, 117.1, 112.1, 110.5, 102.1, 69.2, 52.5, 41.3, 19.4; HRMS (FAB) m/z calcd for C ₂₁ H ₂₀ N ₃ O ₂ [M+H] ⁺ : 346.1556, found 346.1558.

EV508

: According to the general cyclization reaction conditions, 6c (81.1 mg, 0.243 mmol) was used to obtain compound EV508 (56.2 mg, 67% yield) in the form of a yellow solid.

mp: 230-232°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.91 (s, 1H), 8.75 (s, 1H), 7.88 (dd, J = 8.1, 1.3 Hz, 1H), 7.51-7.55 (m, 1H) , 7.29 (d, J = 7.4 Hz, 1H), 7.15 (dd, J = 8.1, 6.0 Hz, 2H), 6.80 (d, J = 2.0 Hz, 1H), 6.65 (dd, J = 8.7, 2.0 Hz, 1H), 6.11 (s, 1H), 4.59 (td, J = 6.3, 4.8 Hz, 1H), 4.00-4.05 (m, 2H), 3.62 (dd, J = 18.1, 2.0 Hz, 1H), 3.19 (td) , J = 12.1, 4.1 Hz, 1H), 3.05 (t, J = 2.4 Hz, 1H), 2.83-2.90 (m, 1H), 2.72-2.76 (m, 1H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.5, 150.8, 147.1, 132.8, 131.2, 129.4, 128.0, 126.6, 123.3, 122.8, 121.2, 112.3, 112.1, 111.3, 102.3, 80.0, 75.2, 68.2, 40.2, 39.9, 39.7, 39.5, 39.3, 39.1, 39.0, 38.9, 19.6; HRMS (FAB) m/z calcd for C ₂₁ H ₁₈ N ₃ O ₂ [M+H] ⁺ : 344.1399, found 344.1407.

EV509

: According to the general cyclization reaction conditions, 6b (95.8 mg, 0.274 mmol) was used to obtain compound EV509 (32.7 mg, 33% yield) in the form of a yellow solid.

mp: 211-213°C; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.59 (s, 1H), 8.67 (s, 1H), 7.71 (dd, J = 7.8, 1.7 Hz, 1H), 7.34-7.38 (m, 1H) , 7.11 (d, J = 8.8 Hz, 1H), 6.89 (d, J = 8.1 Hz, 1H), 6.79 (t, J = 7.1 Hz, 1H), 6.70 (d, J = 2.0 Hz, 1H), 6.59 (dd, J = 8.4, 2.4 Hz, 1H), 6.14 (s, 1H), 4.89 (d, J = 22.2 Hz, 2H), 4.59 (q, J = 6.3 Hz, 1H), 4.21 (d, J = 16.2 Hz, 1H), 3.92 (d, J = 16.2 Hz, 1H), 3.23 (td, J = 12.1, 4.7 Hz, 1H), 2.87-2.95 (m, 1H), 2.57 (dd, J = 15.5, 4.7) Hz, 1H), 1.72 (s, 3H); ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 163.5, 150.8, 147.1, 132.8, 131.2, 129.4, 128.0, 126.6, 123.2, 122.8, 121.2, 112.3, 112.1, 111.3, 102.3, 80.0, 75.2, 68.2, 19.6; LRMS (FAB) m/z, 360 ([M+H] ⁺ ).

Example 1. Experimental method

1-1. cell culture

Human lung cancer cell line (H1299, H460, A549, H226B, PC9), breast cancer cell line (MDA-MB-231), liver cancer cell line (Hep3B), kidney cancer cell line (786-O), prostate cancer cell line (DU145, PC-3) and mouse lung cancer cell lines (Lewis Lung Carcinoma, LLC) were purchased from the American Type Culture Collection (ATCC) or obtained from Dr. Anderson Cancer Center, MD, USA. It was sold by John V. Heymach. The human head and neck cancer cell line (UMSCC38) was prepared by Dr. Anderson Cancer Center, MD, USA. It was sold by Jeffrey N. Myers. Human colorectal cancer cell lines (HCT116, HT-29) and glioblastoma cell lines (U87MG) were purchased from Professor Lee Sang-guk, College of Pharmacy, Seoul National University. Human renal cancer cell line (Caki-1) and gastric cancer cell line (SNU-638) were purchased from Korea Cell Line Bank. Chemotherapy-resistant cells [paclitaxel-resistant H460 cells (H460/PcR), paclitaxel-resistant H226B cells (H226B/PcR), cisplatin-resistant H1299 cells (H1299/CsR), pemetrexed-resistant H1299 cells (H1299/PmR)] and targeted anticancer drug-resistant cells [erlotinib-resistant PC9 cells (PC9/ER)] were constructed by treating the corresponding anticancer agent for at least 3 months. H1299, H460, H226B, PC9, Hep3B, 786-O, DU145, PC-3, HCT116, HT-29, SNU-638, chemotherapy-resistant cells and targeted anticancer-resistant cells contain 10% fetal bovine serum (FBS) and antibiotics RPMI 1640 medium containing the, MDA-MB-231, UMSCC38, U87MG, Caki-1 was cultured at 37 ^o C using DMEM medium containing 10% FBS and antibiotics under 5% CO ₂ conditions. Cells were passaged 1-2 times a week.

1-2. Cell viability assay

The cancer cell line was placed in a 96-well plate at the number of 2x10 ³ cells per well and cultured for 24 hours to attach the cells. After 3 days of treatment with the drug diluted in the medium, cell viability was evaluated using MTT assay or crystal violet (CB) assay. For MTT assay, the final concentration of MTT solution in PBS is 500 μg/mL, incubated for 2-4 hours, the medium is removed, and the resulting formazan is dissolved in 100% dimethyl sulfoxide (DMSO), followed by 570 nm absorbance was measured. For CB assay, cultured cells were fixed with 100% methanol, stained with 0.025% crystal violet solution, and the stained cells were dissolved in 1% SDS solution, and absorbance was measured at 570 nm. Cell viability was calculated by expressing the change in absorbance in the drug-treated group as a percentage compared to the vehicle (DMSO)-treated control group.

1-3. Assessment of anchorage-dependent colony formation

After attaching 100-250 cells per well to a 24-well or 6-well plate, the cancer cell lines were treated with drugs diluted in the medium and cultured for 2-3 weeks. The generated colonies were stained with crystal violet and counted. The effect of inhibiting the colony formation ability of the drug was calculated by expressing the change in the number of colonies in the drug-treated group as a percentage compared to the vehicle (DMSO)-treated control group.

1-4. Assessment of anchorage-independent, soft agar colony formation

Before the experiment, a 1% agar solution prepared with low-melting agar was put in a 24 well plate and hardened at room temperature to prepare a bottom agar, and the cells were diluted with a medium so that 2 × 10 ³ cells were seeded per well in a 24 well plate. Then, it was mixed with 1% agar solution to make a final 0.4% agar solution, and then put into a plate containing bottom agar and hardened at room temperature. A drug-containing medium was placed on the cells mixed with agar, and cultured at 37° C., 5% CO ₂ conditions for 1-3 weeks, and the resulting colonies were stained with 250-500 μg/mL MTT solution and counted. The effect of inhibiting the colony formation ability of the drug was calculated by expressing the change in the number of colonies in the drug-treated group as a percentage compared to the vehicle (DMSO)-treated control group.

1-5. Western blot analysis

RIPA lysis buffer [50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM EDTA, 0.25% sodium deoxycholate, 1% Triton X-100, 100 mM NaF, 5 mM Na ₃ VO ₄ , 1 mM PMSF, 1 μg/mL aprotinin, 1 μg/mL leupeptin, and 1 μg/mL pepstatin], incubated on ice for 10 minutes, and centrifuged at 13,000 rpm for 15 minutes to extract proteins. After quantifying the protein by BCA assay, the same amount of protein was separated by 8-10% SDS-PAGE. After the separated protein was transferred to the PVDF membrane, the membrane was incubated in 5% non-fat milk powder solution or 3% BSA solution diluted in TBST for 1 hour at room temperature to block the membrane area where the protein was not transferred. Membrane was incubated at 4 ^o C for 12 hours or more in a primary antibody solution diluted in 3% BSA at a ratio of 1:1,000. As the primary antibody, an anti-Hsp70 or anti-actin antibody was used. The anti-Hsp70 antibody was purchased from Enzo Life Science, and the actin antibody was purchased from Santa Cruz Biotechnology. After incubation, the membrane was washed for 1 hour while exchanging TBST several times, and incubated with a secondary antibody (GeneTex) diluted in 5% skim milk powder solution at a ratio of 1:5,000 for 1 hour at room temperature. After incubation, the membrane was washed for 1 hour while exchanging TBST several times, and after treatment with an enhanced chemiluminescence (ECL) solution, Hsp70 or Actin protein expression was detected using LAS4000.

1-6. Drug affinity responsive target stability (DARTS)

35 μg of Hsp70 full length, Hsp70 N-terminal, and Hsp70 C-terminal proteins were treated with vehicle (DMSO) or EV507 up to 10 μM (final DMSO concentration: 1%) at 4 ^o C for 30 minutes, followed by proteinase K (proteinase K: protein = 1:100) was treated at room temperature for 15 minutes. Treated with 5x SDS-PAGE sample buffer, boiled at 95 ^o C for 5 minutes to complete the reaction, and then electrophoresed with 8% SDS-PAGE. After the protein was transferred to the PVDF membrane, the Western blot analysis method described above was performed.

1-7. animal testing

Tumors were generated by transplanting H460 cells (5x10 ⁶ cells/spot) or lung cancer patient-derived tumors into the flanks of 6-week-old NOD/SCID mice, or by transplanting LLC-Luc cells (5x10 ⁵ cells/spot) into the flanks of 8-week-old C57BL/6 mice. did. When the tumor volume reached 50-100 mm ³ , the mice were divided into a control group and an experimental group (more than 6 mice were assigned to each group), and a vehicle (10% DMSO and 40% PEG400 solution prepared in sterile distilled water) was applied to the control group and the experimental group EV507 (20 mg/kg) was orally administered 6 times a week. The tumor size was measured at least 3-4 times a week by measuring the long axis and short axis of the tumor, and the toxicity of the drug was evaluated by measuring the body weight at least once a week. In addition, survival curves were determined by monitoring the survival of C57BL/6 mice transplanted with LLC-Luc cells. Tumor volume was calculated as (short) ² x long x 0.5.

1-8. Statistical processing

All results were expressed as mean ± standard deviation, and statistical treatment was performed by Student's t -test or Mann-Whitney test when there were two groups, and one-way ANOVA when there were three or more groups. The significance of survival analysis was evaluated through the log-rank test. Statistical significance analysis was performed using Graphpad Prism (version 9). When the P value was less than 0.05, statistical significance was evaluated.

Example 2. Confirmation of the effect of inhibiting cancer cell survival rate and colony formation of the evodiamine derivative EV507

The cancer cell survival inhibitory effect of 100 nM of EV compounds (EV501-EV509) on H1299 cells was evaluated.

As a result, as shown in FIG. 1 , EV507, the compound showing the best efficacy, was discovered, and it was confirmed that EV507 was significantly superior in the effect of inhibiting cancer cell survival than evodiamine and other evodiamine derivatives.

In addition, as shown in FIGS. 2 and 3 , EV507 showed a concentration-dependent inhibition of cancer cell survival against several cancer cells of different origins, and at a concentration as low as 50 nM, the formation of colonies under adhesion-dependent conditions was significantly reduced. suppressed.

Example 3. Confirmation of the inhibitory effect of EV507 on Hsp70 and client protein expression

The effect of EV507 on Hsp70 and related client proteins was confirmed.

As shown in FIGS. 4 and 5 , it was confirmed that the expression of Hsp70 protein and Hsp70/Hsp90 client protein (Akt, Src, and MEK) was remarkably reduced when EV507 was treated in several lung cancer cells. This effect was also shown in HCT116, a colorectal cancer cell line, and UMSCC38, a head and neck cancer cell line.

In addition, as shown in FIG. 6 , it was confirmed that EV507 binds to Hsp70 protein, particularly Hsp70 N-terminal, through DARTS.

Example 4. Anti-cancer effect of EV507

The antitumor effect of EV507 was confirmed in H460 and lung cancer patient-derived tumor transplantation mouse models.

As shown in FIG. 7 , EV507 significantly inhibited tumor growth compared to the control group.

In addition, it is known that in the LLC-Luc transplant allograft model, which spontaneously generates lung metastases, mouse death appears due to the proliferation of LLC-Luc transplanted tumors and the formation of lung metastases. Accordingly, EV507 was administered to the allograft model to confirm the inhibitory effect of EV507 on mouse death according to tumor growth.

As shown in FIG. 8 , it was confirmed that mouse death was significantly reduced by EV507 treatment, confirming the anticancer and cancer malignancy inhibitory effects of EV507.

Example 5. Confirmation of Lung Cancer Treatment Effect of Various Ebodiamine Derivatives

Based on the anticancer effect of EV507 confirmed in the above Example, the H1299 lung cancer cell killing effect of various evodiamine derivatives was confirmed.

As shown in FIGS. 9 to 12 , it was confirmed that various evodiamine derivatives in addition to EV507 exhibited anticancer effects on lung cancer cells.

In particular, lung cancer cells with excellent EV205, EV206, EV217, EV218, EV219, EV301, EV302, EV303, EV304, EV306, EV307, EV308, EV309, EV310, EV401, EV402, EV403, EV406, EV407, EV408, EV411, and EV413 lung cancer cells. It was confirmed that it exhibits a killing effect.

Example 6. Confirmation of lung cancer treatment effect of EV206

Based on Example 5, the effect of EV206 on various lung cancer cells and anticancer drug-resistant lung cancer cell lines was confirmed.

As shown in FIG. 13 , it was confirmed that EV206 exhibited excellent apoptosis effects on various lung cancer cell lines and anticancer drug-resistant lung cancer cell lines.

As shown in FIG. 14 , EV206 significantly inhibited colony formation under adhesion-dependent conditions at a concentration as low as 0.5 μM.

Example 7. Confirmation of lung cancer treatment effect of EV408

Based on Example 5, the effect of EV408 on various lung cancer cells and anticancer drug-resistant lung cancer cell lines was confirmed.

As shown in FIG. 15 , it was confirmed that EV408 exhibited an excellent apoptosis effect on various lung cancer cell lines and anticancer drug-resistant lung cancer cell lines.

As shown in FIG. 16 , EV408 significantly inhibited colony formation under adhesion-dependent conditions at a concentration as low as 0.5 μM.

Example 8. Confirmation of the inhibitory effect of EV206 and EV408 on Hsp70 and client protein expression

In the same manner as in Example 3, the effect of EV507 on Hsp70 and related client proteins was confirmed.

As shown in FIG. 17 , it was confirmed that when lung cancer cells were treated with EV206 and EV408, the expression of Hsp70 protein and Hsp70/Hsp90 client protein (Akt, Src, and MEK) was remarkably reduced.

Example 9. Confirmation of lung cancer treatment effect of EV508 and EV509

As shown in FIG. 18 , among the developed evodiamine derivatives, EV501, EV507, EV508, and EV509 exhibited excellent anticancer efficacy in inhibiting the survival of lung cancer cells by 65% or more at the lowest concentration of 0.5 μM, and EV502, EV503, EV506 also At 0.5 μM, the survival of lung cancer cells was inhibited by more than 50%.

In addition, as shown in FIG. 19 , it was reconfirmed that the cancer cell survival inhibitory activity of the major evodiamine derivatives EV501, EV504, EV508, and EV509 was as good as that of EV507 using crystal violet assay.

Example 10. EV501, EV507-EV509 Anchor-Independent Colony Formation Inhibitory Effect

The effect of evodiamine derivatives EV501 and EV507~EV509 on the colony-forming ability of lung cancer cells in the adhesion-dependent culture condition of soft agar was confirmed.

As shown in FIG. 20 , it was confirmed that EV501, EV508, and EV509 effectively inhibited the adhesion-dependent colony formation of lung cancer cells like EV507.

Example 11. Confirmation of Hsp70 inhibitory activity of EV501

The effects of EV501 and EV507, one of the major evodiamine derivatives, on Hsp70 and Hsp70/Hsp90 client proteins were evaluated.

As shown in FIG. 21 , it was confirmed that the evodiamine derivative EV501 reduced the expression of Hsp70 and the Hsp70/Hsp90 client proteins Src and Akt like EV507.

The above description of the present invention is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (15)

An indoloquinazolidine alkaloid represented by the following formula (1) or a pharmaceutically acceptable salt thereof.
[Formula 1]

(In Formula 1,
R ₁ and R ₂ are each independently hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, halogen, nitro (NO ₂ ), amide, —NH— aryl, hydroxy, or -COOR ₇ ;
R ₃ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, -CH ₂ -R ₈ , substituted or unsubstituted C ₁ -C ₆ acyl, substituted or unsubstituted aryl, -CO-R ₈ or benzyl ego,
R ₄ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₂ -C ₆ alkylalkenyl, substituted or unsubstituted C ₂ -C ₆ alkylalkynyl, -CH ₂ -R ₈ , substituted or unsubstituted aryl, propargyl, substituted or unsubstituted C ₁ -C ₆ acyl, or benzyl,
R ₅ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, halogen, nitro, or substituted or unsubstituted C ₁ -C ₆ alkoxy;
R ₆ is hydrogen, oxygen, or substituted or unsubstituted C ₁ -C ₆ alkyl,
R ₇ and R ₈ are each independently hydrogen, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted aryl;
X is C, N, O or S.)
According to claim 1,
The R ₁ and R ₂ are each independently hydrogen, methoxy, butoxy, methyl, hydroxy, F, Cl, nitro,

,

, or -COOR ₇ ,
R ₃ is hydrogen, methyl, ethyl, propyl, butyl, hexyl, benzyl, -CO-R ₈ or -CH ₂ -R ₈ ,
R ₄ is hydrogen, methyl, benzyl, substituted or unsubstituted C ₂ -C ₅ alkenyl, substituted or unsubstituted C ₂ -C ₅ alkynyl, substituted or unsubstituted C ₂ -C ₅ alkylalkenyl, substituted or unsubstituted C ₂ -C ₅ alkyl alkynyl, cyclohexanecarbonyl,

, or -CH ₂ -R ₈ ,
R ₅ is hydrogen, methoxy, nitro, methyl, F, Cl, Br, or I;
R ₆ is hydrogen or oxygen,
R ₇ and R ₈ are each independently hydrogen, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted aryl, the substituted aryl is substituted with one or more selected from the group consisting of halogen, CN, C ₁ -C ₆ haloalkyl, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, nitro,
X is N, or O, characterized in that the indoloquinazolidine alkaloid or a pharmaceutically acceptable salt thereof.
According to claim 1,
The indoloquinazolidine alkaloid represented by Formula 1 is an indoloquinazolidine alkaloid or a pharmaceutically acceptable salt thereof, characterized in that one selected from the group consisting of the following compounds.

A pharmaceutical composition for the prevention or treatment of cancer comprising an indoloquinazolidine alkaloid represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

(In Formula 1,
R ₁ and R ₂ are each independently hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, halogen, nitro (NO ₂ ), amide, —NH— aryl, hydroxy, or -COOR ₇ ;
R ₃ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, -CH ₂ -R ₈ , substituted or unsubstituted C ₁ -C ₆ acyl, substituted or unsubstituted aryl, -CO-R ₈ or benzyl ego,
R ₄ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₂ -C ₆ alkylalkenyl, substituted or unsubstituted C ₂ -C ₆ alkylalkynyl, -CH ₂ -R ₈ , substituted or unsubstituted aryl, propargyl, substituted or unsubstituted C ₁ -C ₆ acyl, or benzyl,
R ₅ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, halogen, nitro, or substituted or unsubstituted C ₁ -C ₆ alkoxy;
R ₆ is hydrogen, oxygen, or substituted or unsubstituted C ₁ -C ₆ alkyl,
R ₇ and R ₈ are each independently hydrogen, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted aryl,
X is C, N, O or S.)
5. The method of claim 4,
The R ₁ and R ₂ are each independently hydrogen, methoxy, butoxy, methyl, hydroxy, F, Cl, nitro,

,

, or -COOR ₇ ,
R ₃ is hydrogen, methyl, ethyl, propyl, butyl, hexyl, benzyl, -CO-R ₈ or -CH ₂ -R ₈ ,
R ₄ is hydrogen, methyl, benzyl, substituted or unsubstituted C ₂ -C ₅ alkenyl, substituted or unsubstituted C ₂ -C ₅ alkynyl, substituted or unsubstituted C ₂ -C ₅ alkylalkenyl, substituted or unsubstituted C ₂ -C ₅ alkyl alkynyl, cyclohexanecarbonyl,

, or -CH ₂ -R ₈ ,
R ₅ is hydrogen, methoxy, nitro, methyl, F, Cl, Br, or I;
R ₆ is hydrogen or oxygen,
R ₇ and R ₈ are each independently hydrogen, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted aryl, the substituted aryl is substituted with one or more selected from the group consisting of halogen, CN, C ₁ -C ₆ haloalkyl, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, nitro,
X is N, or O, the prevention or treatment of cancer pharmaceutical composition.
5. The method of claim 4,
The indoloquinazolidine alkaloid represented by Formula 1 is a pharmaceutical composition for the prevention or treatment of cancer, characterized in that one selected from the group consisting of the following compounds.

5. The method of claim 4,
The cancer is cervical cancer, lung cancer, pancreatic cancer, non-small cell lung cancer, liver cancer, colon cancer, colorectal cancer, bone cancer, skin cancer, head cancer, cervical cancer, skin melanoma, intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, liver cancer, brain tumor , blood cancer, gastric cancer, perianal cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine adenocarcinoma, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma , urethral cancer, penile cancer, prostate cancer, bladder cancer, kidney cancer, ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma and pituitary gland A pharmaceutical composition for the prevention or treatment of cancer, characterized in that at least one selected from the group consisting of adenoma.
5. The method of claim 4,
The cancer is a pharmaceutical composition for the prevention or treatment of cancer, characterized in that lung cancer.
5. The method of claim 4,
The indoloquinazolidine alkaloid is a pharmaceutical composition for the prevention or treatment of cancer, characterized in that it reduces at least one selected from the group consisting of the number and volume of tumors.
5. The method of claim 4,
The indoloquinazolidine alkaloid is a pharmaceutical composition for preventing or treating cancer, characterized in that it suppresses the expression of HSP70 protein.
5. The method of claim 4,
The indoloquinazolidine alkaloid is a pharmaceutical composition for the prevention or treatment of cancer, characterized in that it inhibits the expression of one or more proteins selected from the group consisting of Akt, Src, and MEK.
5. The method of claim 4,
The composition is a pharmaceutical composition for the prevention or treatment of cancer, characterized in that it further comprises one or more selected from the group consisting of pemetrexed, cisplatin, and paclitaxel.
5. The method of claim 4,
The composition is a pharmaceutical composition for preventing or treating cancer, characterized in that it is for the prevention or treatment of lung cancer that has acquired resistance to an anticancer agent.
An anticancer adjuvant for enhancing the anticancer efficacy of an anticancer agent comprising an indoloquinazolidine alkaloid represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

(In Formula 1,
R ₁ and R ₂ are each independently hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, halogen, nitro (NO ₂ ), amide, —NH— aryl, hydroxy, or -COOR ₇ ;
R ₃ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, -CH ₂ -R ₈ , substituted or unsubstituted C ₁ -C ₆ acyl, substituted or unsubstituted aryl, -CO-R ₈ or benzyl ego,
R ₄ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₂ -C ₆ alkylalkenyl, substituted or unsubstituted C ₂ -C ₆ alkylalkynyl, -CH ₂ -R ₈ , substituted or unsubstituted aryl, propargyl, substituted or unsubstituted C ₁ -C ₆ acyl, or benzyl,
R ₅ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, halogen, nitro, or substituted or unsubstituted C ₁ -C ₆ alkoxy;
R ₆ is hydrogen, oxygen, or substituted or unsubstituted C ₁ -C ₆ alkyl,
R ₇ and R ₈ are each independently hydrogen, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₁ -C ₆ alkyl, or substituted or unsubstituted aryl;
X is C, N, O or S.)
15. The method of claim 14,
The anticancer agent, characterized in that at least one selected from the group consisting of pemetrexed, cisplatin, and paclitaxel, an anticancer adjuvant.

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