TW202323254A - Polycyclic pyridotriazine derivative - Google Patents

Abstract

The present inventon provides the compound represented by the following formula (I). wherein ring A is a substituted or unsubstituted heterocycle; ring B is a benzene ring or a pyridine ring; Q is -NHC(O)- or a 5-membered aromatic heterocycle; R1 is each independently halogen, alkyl, haloalkyl, alkyloxy, cyano or haloalkyloxy; R<SP>2a</SP> and R<SP>2b</SP> are each independently a hydrogen atom, alkyl or haloalkyl; R3 is alkyl or haloalkyl; R4 is a hydrogen atom or alkyl; and n is an integer of 1 to 3.

Description

polycyclic pyrido triderivatives

衍生物及含有該衍生物的醫藥，特別是抗HIV藥物。 The present invention relates to a novel compound with antiviral effect, more specifically, a polycyclic pyridotridioide with HIV integrase (integrase) inhibitory activity

Derivatives and medicines containing the derivatives, especially anti-HIV medicines.

Among viruses, human immunodeficiency virus (Human Immunodeficiency Virus, hereinafter abbreviated as HIV), which is one of retroviruses, is known as a cause of acquired immunodeficiency syndrome (hereinafter abbreviated as AIDS). As a therapeutic agent for this AIDS, at present, various guidelines recommend integrase inhibitors (for example, dolutegravir (Dolutegravir) etc.) as the main agent to naive patients with different drug resistance profiles ( Resistance profile) is a combination of two nucleic acid-based reverse transcriptase inhibitors (for example, ABC+3TC, FTC+TAF, etc.). Compared with the initial treatment drugs, satisfaction has been improved due to its strong efficacy and high safety. On the other hand, due to the emergence of safe drugs and good prognosis, it is recommended to start treatment once HIV infection is known, and the average life expectancy of HIV-infected patients is also close to that of healthy people, which leads to long-term medication. Due to long-term medication, when the side effects of nucleic acid-based reverse transcriptase inhibitors appear or once drug-resistant viruses appear, there is no easy treatment afterwards, so there are measures not to use nucleic acid-based reverse transcriptase inhibitors. Therefore, expect Dual-agent therapy consisting of two primary agents is well established, and it is desirable to develop primary agents that can be combined with integrase inhibitors. Furthermore, in order to improve the patient's quality of life (Quality of Life, QOL) such as improving fatigue caused by long-term medication and enjoying daily life better, it is expected to develop therapeutic drugs with longer intervals between medications, that is, for example, every interval A continuous injection that can complete the treatment with one injection over a month.

To meet this need, the integrase inhibitor Cabotegravir is being developed as a continuous injection in Ph3 (Phase 3). Furthermore, non-nucleic acid-based reverse transcriptase inhibitor Rilpivirin is also being prescribed as a continuous injection, with the aim of establishing a therapeutic method using the two doses. However, these drugs have to be injected every one to two months and are injected in 3 to 4 places with pain. Therefore, in order to further improve the QOL of patients, it is desired to develop a drug with lower dosage and less pain, which can be completed once every three months.

Integrase inhibitors have been marketed as first-generation oral agents Raltegravir and Elvitegravir; and second-generation dolutegravir. Drug-resistant mutations will not occur when dolutegravir is used in newly diagnosed patients. Addition of a further resistance mutation renders dolutegravir ineffective. Therefore, it is desirable to develop inhibitors with a higher barrier to resistance than dolutegravir.

衍生物。 Furthermore, as one of anti-HIV drugs having an integrase inhibitory effect, bicyclic or more bicyclic carbamidylpyridone derivatives are known (Patent Documents 1 to 29). Among these documents, it is described in Patent Document 3 that carbamoyl three

derivative.

衍生物。再者，專利文獻5記載了縮合三環性的吡啶并吡

衍生物和縮合三環性的吡啶并三

衍生物。 Further, one of the anti-HIV drugs with integrase inhibitory effect is known to have pyridone derivatives with heterocyclic side chains (patent documents 5, 8, 9, 12, 13, 19, 23, 24, 27, 30 to 33). Among these documents, Patent Document 9 describes a condensed tricyclic pyridopyridine

derivative. Furthermore, Patent Document 5 describes a condensed tricyclic pyridopyridine

Derivatives and condensed tricyclic pyridotri

derivative.

[Prior Art Literature]

[Patent Document]

Patent Document 1: International Publication No. WO2006/088173

Patent Document 2: International Publication No. WO2006/116764

Patent Document 3: International Publication No. WO2007/049675

Patent Document 4: International Publication No. WO2011/129095

Patent Document 5: International Publication No. WO2014/099586

Patent Document 6: International Publication No. WO2014/100323

Patent Document 7: International Publication No. WO2014/104279

Patent Document 8: International Publication No. WO2014/183532

Patent Document 9: International Publication No. WO2014/200880

Patent Document 10: International Publication No. WO2015/039348

Patent Document 11: International Publication No. WO2015/048363

Patent Document 12: International Publication No. WO2015/089847

Patent Document 13: International Publication No. WO2015/095258

Patent Document 14: International Publication No. WO2015/006731

Patent Document 15: International Publication No. WO2015/006733

Patent Document 16: International Publication No. WO2015/19967

Patent Document 17: International Publication No. WO2016/090545

Patent Document 18: International Publication No. WO2016/094198

Patent Document 19: International Publication No. WO2016/094197

Patent Document 20: International Publication No. WO2016/106237

Patent Document 21: International Publication No. WO2016/154527

Patent Document 22: International Publication No. WO2016/161382

Patent Document 23: International Publication No. WO2016/187788

Patent Document 24: International Publication No. WO2016/191239

Patent Document 25: International Publication No. WO2017/087256

Patent Document 26: International Publication No. WO2017/087257

Patent Document 27: International Publication No. WO2017/106071

Patent Document 28: International Publication No. WO2017/113288

Patent Document 29: International Publication No. WO2017/116928

Patent Document 30: International Publication No. WO2005/016927

Patent Document 31: International Publication No. WO2011/105590

Patent Document 32: International Publication No. WO2013/054862

Patent Document 33: International Publication No. WO2016/027879

The object of the present invention is to provide a novel compound with a high drug resistance barrier and sustained integrase inhibitory activity.

衍生物，該衍生物具有高抗藥性屏障的整合酶抑制作用。進一步發現本發明化合物及含有該等的醫藥係可用作為抗病毒藥(例如，抗反轉錄病毒藥、抗HIV藥、抗HTLV-1(Human T cell leukemia virus type 1：人類T細胞白血病病毒1型)藥、抗FIV(Feline immunodeficiency virus：貓AIDS病毒)藥、抗SIV(Simian immunodeficiency virus：猿AIDS病毒)藥)，特別是抗HIV藥、抗AIDS藥或與其相關疾病的治療藥等，遂而完成如下所示之本發明。 As a result of in-depth examination, the present inventors have found a novel pyridotri

Derivatives that have integrase inhibition with a high resistance barrier. It was further found that the compounds of the present invention and pharmaceuticals containing them can be used as antiviral drugs (for example, antiretroviral drugs, anti-HIV drugs, anti-HTLV-1 (Human T cell leukemia virus type 1: Human T cell leukemia virus type 1) ) drugs, anti-FIV (Feline immunodeficiency virus: feline AIDS virus) drugs, anti-SIV (Simian immunodeficiency virus: ape AIDS virus) drugs), especially anti-HIV drugs, anti-AIDS drugs or therapeutic drugs for related diseases, etc., and then The present invention as shown below was accomplished.

The present invention provides the inventions shown below.

[1] A compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

(where,

Ring A is the following rings:

X1 is CR ^9a R ^9b or O;

R ^5a , R ^5b , R ^6a , R ^6b , R ^7a and R ^7b are each independently hydrogen, alkyl, alkoxy or alkoxyalkyl;

R ^5a and R ^6a , or R ^6a and R ^7a together with adjacent atoms can form an aromatic carbocycle that can be substituted by halogen, a non-aromatic carbocycle with 3 to 6 members that can be substituted by halogen or can be substituted by halogen 4 to 6-membered non-aromatic heterocyclic ring (except, when forming an aromatic carbocyclic ring, R ^5b and R ^6b , or R ^6b and R ^7b together form a bond);

R ^5b and R ^6b can form a bond together;

R ^8a , R ^8b , R ^9a , R ^9b , R ^10a , R ^10b , R ^11a and R ^11b are each independently hydrogen, alkyl, alkoxy or alkoxyalkyl;

R ^8a and R ^10a can form C1-C3 crosslinks together;

R ^10a and R ^11a can form a 5-membered non-aromatic carbocyclic ring together with adjacent atoms;

R ^9a and R ^9b can form a 4-membered non-aromatic carbocyclic ring or a 5-membered non-aromatic heterocyclic ring together with adjacent atoms;

R ^8a and R ^9a can form a bond together;

Ring B is a benzene ring or a pyridine ring;

Q is -NHC(O)- or a 5-membered aromatic heterocycle;

Each R1 is independently halogen, alkyl, haloalkyl, alkoxy, cyano, or haloalkoxy;

R ^2a and R ^2b are each independently hydrogen, alkyl or haloalkyl;

R ³ is alkyl or haloalkyl;

^R4 is hydrogen or alkyl; and

n is an integer from 1 to 3)

[2] The compound according to [1] or a pharmaceutically acceptable salt thereof, wherein R ³ is an alkyl group.

[3] The compound according to [1] or [2] or a pharmaceutically acceptable salt thereof, wherein R ¹ are each independently halogen.

[4] The compound according to any one of [1] to [3] or a pharmaceutically acceptable salt thereof, wherein R ^2a is hydrogen, and R ^2b is hydrogen or an alkyl group.

[5] The compound according to any one of [1] to [4] or a pharmaceutically acceptable salt thereof, wherein Q is -NHC(O)-.

[6] The compound according to any one of [1] to [4], or a pharmaceutically acceptable salt thereof, wherein Q is a 5-membered aromatic heterocycle.

[7] The compound as described in [1] or a pharmaceutically acceptable salt thereof, which is selected from compounds I-3, I-7, I-11, I-16, I-23, I-24 and I Groups of -32.

[8] The compound as described in [1] or a pharmaceutically acceptable salt thereof, which is selected from compounds II-1, II-4, II-5, II-13, II-14, II-16, II -19, II-21, II-23, II-26, II-31, II-34, II-36, II-38, II-41, II-43, II-45, II-47, II-49 , II-52, II-56, II-58, II-62, II-63, II-64, II-68, II-89, II-92, II-93, II-95, II-96, II -98, II-106, II-109, II-110, II-112, II-113, II-117, II-118, II-125, II-127, II-128, II-129, II-130 , II-131, II-132, II-136, II-138, II-139, II-142, II-143, II-144, II-147, II-148, II-149, II-150, II The group consisting of -151, II-155, II-156 and II-157.

[9] A pharmaceutical composition comprising the compound described in any one of [1] to [8] or a pharmaceutically acceptable salt thereof.

[10] The pharmaceutical composition according to [9], which is an anti-HIV agent.

[11] The pharmaceutical composition as described in [9], which is an HIV integrase inhibitor.

[12] An anti-HIV agent comprising the compound described in any one of [1] to [8] or a pharmaceutically acceptable salt thereof.

[13] Use of the compound described in any one of [1] to [8] or a pharmaceutically acceptable salt thereof for producing a therapeutic agent and/or a preventive agent for HIV infection.

[14] The compound according to any one of [1] to [8], or a pharmaceutically acceptable salt thereof, for use in the treatment and/or prevention of HIV infection.

The present invention further provides a method for preventing or treating HIV infection, the method comprising administering an effective amount of the above compound to humans.

The present invention further provides the above compounds for use as anti-HIV agents.

The compound of the present invention has integrase inhibitory activity and/or cell proliferation inhibitory activity to virus, especially HIV or its drug-resistant virus. Therefore, it is useful for the prevention or treatment of various integrase-related diseases or viral infections (for example, AIDS). Preferably, the compounds of the invention are useful as persistent integrase inhibitors. In addition, it is also excellent in drug resistance spectrum in which new HIV drug-resistant viruses are not easily generated. More preferably, the compounds of the present invention also have prophylactic or therapeutic effects on HIV drug-resistant viruses. Still more preferably, the compounds of the present invention have small clearance, long half-life in vivo, excellent solubility or metabolic stability, etc., and can be used as cytotoxicity or side effects (for example, CYP inhibition, mutagenicity, electrocardiogram QT interval prolongation, arrhythmia) less doubts about medicines.

Hereinafter, the meaning of each term used in this specification will be explained. Unless otherwise specified, each term has the same meaning when used alone or in combination with other terms.

The phrase "consisting of" means having only the constituent elements.

The term "contains" means not limiting constituent elements, and not excluding unrecited elements.

"Halogen" includes fluorine atom, chlorine atom, bromine atom and iodine atom. Particularly preferred are fluorine atoms and chlorine atoms.

"Alkyl" refers to a straight or branched hydrocarbon group having 1 to 15 carbons, preferably 1 to 10 carbons, more preferably 1 to 6 carbons, more preferably 1 to 4 carbons. For example: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, second butyl, third butyl, n-pentyl, isopentyl, neopentyl, n-hexyl , isohexyl, n-heptyl, isoheptyl, n-octyl, isooctyl, n-nonyl, n-decyl, etc.

Preferable examples of "alkyl" include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, second-butyl, third-butyl, and n-pentyl. Even better implementations may include, for example: methyl, ethyl, n-propyl, isopropyl, and tert-butyl.

The "aromatic carbocyclic group" means a monocyclic cyclic aromatic hydrocarbon group. Examples include phenyl.

A preferred embodiment of the "aromatic carbocyclic group" includes phenyl.

The "non-aromatic carbocyclic group" means a monocyclic cyclic saturated hydrocarbon group or a cyclic non-aromatic unsaturated hydrocarbon group.

The monocyclic non-aromatic carbocyclic group preferably has 3 to 16 carbons, more preferably 3 to 6 carbons. For example: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, Cyclodecyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclohexadienyl and the like.

The "aromatic heterocyclic group" means an aromatic ring group having one or more five-membered heteroatoms arbitrarily selected from O, S, and N in the ring.

唑基、

二唑基、異噻唑基、噻唑基、噻二唑基等。 The 5-membered aromatic heterocyclic group can be exemplified by: pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl,

Azolyl,

Oxadiazolyl, isothiazolyl, thiazolyl, thiadiazolyl, etc.

"Non-aromatic heterocyclic group" means a monocyclic non-aromatic ring group having one or more identical or different heteroatoms arbitrarily selected from O, S and N in the ring.

The monocyclic non-aromatic heterocyclic group is preferably 3 to 8 members, more preferably 4 to 6 members.

基、

啉基(morpholinyl)、N-

啉基(morpholino)、硫代

啉基(thiomorpholinyl)、硫代N-

啉基(thiomorpholino)、二氫吡啶基、四氫吡啶基、四氫吡喃基、二氫

基(dihydroxazinyl)、四氫嗒

基(tetrahydropyridazinyl)、六氫嘧啶基、二

基、噻吩基(thienyl)、噻

基等。7員非芳芳族雜環基可舉例如：六氫氮呯基(azepinyl)、四氫二氮呯基、氧雜環庚烷基。 Examples of the 3-membered non-aromatic heterocyclic group include thioethylene group, oxirane group and aziridinyl group. Examples of the 4-membered non-aromatic heterocyclic group include oxetanyl and azetidinyl. The 5-membered non-aromatic heterocyclic group can be exemplified by: oxirane, tetrahydrothiazolyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidine, pyrazolinyl, Tetrahydrofuryl, dihydrothiazolyl, tetrahydroisoquinolinethiazolyl, dioxolanyl, dioxolenyl, tetrahydrothienyl, and the like. The 6-membered non-aromatic heterocyclic group can be exemplified as: dioxanyl, thianyl, piperidinyl, piperidine

base,

Morpholinyl, N-

Morpholino, Thio

Linyl (thiomorpholinyl), thio N-

thiomorpholino, dihydropyridyl, tetrahydropyridyl, tetrahydropyranyl, dihydro

base (dihydroxazinyl), tetrahydroxazinyl

base (tetrahydropyridazinyl), hexahydropyrimidinyl, two

base, thienyl (thienyl), thiophene

Base etc. Examples of the 7-membered non-aromatic aromatic heterocyclic group include: azepinyl, tetrahydrodiazepinyl, and oxepinyl.

"Aromatic carbocycle", "non-aromatic carbocycle", "aromatic heterocycle" and "non-aromatic heterocycle" respectively mean Rings of "cyclic group", "aromatic heterocyclic group" and "non-aromatic heterocyclic group".

Preferable aspects of each symbol in the compound represented by formula (I) are as follows. The compound represented by formula (I) can illustrate all combinations of the specific examples shown below.

A ring can enumerate the following rings:

One of the better forms of the A ring is the following ring:

The most preferable aspect of ring A is the above (a) or (b-1).

The B ring may, for example, be a benzene ring or a pyridine ring.

A preferred embodiment of the B ring is a benzene ring.

Each of R ¹ independently includes halogen, alkyl, haloalkyl, alkoxy, cyano, or haloalkoxy.

One of the preferred embodiments of R ¹ is halogen, alkyl or haloalkyl.

A further preferred embodiment of ^R1 is halogen.

R ^2a and R ^2b each independently include hydrogen, alkyl or haloalkyl.

One of the preferred aspects of R ^2a and R ^2b is hydrogen.

One of the preferred embodiments of R ^2a is hydrogen.

A preferred embodiment of R ^2b is hydrogen or methyl, and a further preferred embodiment is hydrogen.

Examples of R ³ include alkyl or haloalkyl.

A preferred aspect of R ³ is an alkyl group.

R ⁴ can be, for example: hydrogen or alkyl.

A preferred embodiment of R ⁴ is hydrogen or methyl, and a further preferred embodiment is hydrogen.

R ^5a , R ^5b , R ^6a , R ^6b , R ^7a and R ^7b each independently include hydrogen, alkyl, alkoxy or alkoxyalkyl.

A preferred embodiment of R ^5a is hydrogen or an alkyl group, and a further preferred embodiment is hydrogen.

A preferred embodiment of R ^5b is hydrogen or an alkyl group, and a further preferred embodiment is hydrogen.

One of the preferred embodiments of R ^6a is hydrogen, alkyl or alkoxyalkyl, and another preferred embodiment is hydrogen.

One of the preferred aspects of R ^6b is hydrogen.

A preferred embodiment of R ^7a is hydrogen, an alkyl group or an alkoxyalkyl group, and a further preferred embodiment is an alkoxyalkyl group.

One of the preferred embodiments of R ^7b is hydrogen.

R ^5a and R ^6a , or R ^6a and R ^7a can form, together with adjacent atoms, an aromatic carbocyclic ring that can be substituted by halogen, a 3- to 6-membered non-aromatic carbocyclic ring that can be substituted by halogen, or a carbocyclic ring that can be substituted by halogen 4 to 6-membered non-aromatic heterocycle (provided that when forming an aromatic carbocycle, R ^5b and R ^6b , or R ^6b and R ^7b together form a bond).

R ^5b and R ^6b may together form a bond.

R ^8a , R ^8b , R ^9a , R ^9b , R ^10a , R ^10b , R ^11a , and R ^11b each independently include hydrogen, alkyl, alkoxy, or alkoxyalkyl.

One of the preferred embodiments of R ^8a is hydrogen or alkyl, and another preferred embodiment is hydrogen.

One of the preferred embodiments of R ^8b is hydrogen or alkyl, and another preferred embodiment is hydrogen.

One of the preferred embodiments of R ^9a is hydrogen, alkyl or alkoxyalkyl.

A preferred embodiment of R ^9b is hydrogen or an alkyl group, and another preferred embodiment is hydrogen.

A preferred embodiment of R ¹⁰ is hydrogen, alkyl or alkoxy, and a further preferred embodiment is hydrogen.

One of the preferred embodiments of R ^10b is hydrogen.

A preferred embodiment of R ^11a is hydrogen or an alkyl group, and a further preferred embodiment is hydrogen.

One of the preferred embodiments of R ^11b is hydrogen.

R ^8a and R ^10a can together form a C1-C3 crosslink, preferably a C1-C2 crosslink.

R ^10a and R ^11a may form a 5-membered non-aromatic carbocyclic ring together with adjacent atoms.

R ^9a and R ^9b may form, together with adjacent atoms, a 4-membered non-aromatic carbocyclic ring or a 5-membered non-aromatic heterocyclic ring.

R ^8a and R ^9a may together form a bond.

The integer of 1-3 is mentioned as n.

One of the preferable aspects of n is an integer of 2-3.

One of the best forms of n is an integer of 1-2.

Q includes -NHC(O)- or a 5-membered aromatic heterocyclic ring.

One of the preferred forms of Q is -NHC(O)- (the bond on the left is bonded to CR ^2a R ^2b ).

One of the other preferred embodiments of Q is a 5-membered aromatic heterocycle.

One of the other preferred forms of Q is the following ring (the bond on the left is bonded to CR ^2a R ^2b );

One of the best forms of Q is the ring shown in (1) above.

衍生物，而具有優異的抗藥性譜、體內動態及安全性優異的特點。 The compound of the present invention is characterized in that ring A is fixed in a specific three-dimensional structure in the formula (I), so it has the characteristics of excellent drug resistance spectrum, in vivo dynamics and excellent safety. Furthermore, the compound of the present invention is characterized in that in formula (I), it becomes an optically active tricyclic or higher pyridotricyclic

Derivatives, which have excellent drug resistance spectrum, in vivo dynamics and excellent safety characteristics.

The compounds of the present invention are not limited to a specific isomer unless otherwise stated, and include all possible isomers (e.g., keto-enol isomers, imine-enamine isomers, diastereomers compounds, optical isomers, rotamers, etc.), racemates or mixtures thereof.

Pharmaceutically acceptable salts of the compounds of the present invention include, for example: the compounds of the present invention and alkali metals (for example, lithium, sodium, potassium, etc.), alkaline earth metals (for example, calcium, barium, etc.), magnesium, transition metals (for example, zinc , iron, etc.), ammonia, organic bases (for example, trimethylamine, triethylamine, dicyclohexylamine, ethanolamine, diethanolamine, triethanolamine, meglumine (Meglumine), ethylenediamine, pyridine, picoline, quinine phyline, etc.) and amino acid salts; or with inorganic acids (such as hydrochloric acid, sulfuric acid, nitric acid, carbonic acid, hydrobromic acid, phosphoric acid, hydroiodic acid, etc.), and organic acids (such as formic acid, acetic acid, propionic acid, Trifluoroacetic acid, citric acid, lactic acid, tartaric acid, oxalic acid, maleic acid, fumaric acid, mandelic acid, glutaric acid, malic acid, benzoic acid, phthalic acid, ascorbic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, etc.) salts, etc. Such salts can be formed by commonly performed methods.

The compounds of the present invention or pharmaceutically acceptable salts thereof may form solvates (for example, hydrates, etc.), co-crystals and/or crystal polymorphs, and the present invention also includes such various solvates, co-crystals and Polymorphic crystals. "Solvate" refers to a compound of the present invention that can to coordinate with any number of solvent molecules (eg, water molecules, etc.). When the compound of the present invention or a pharmaceutically acceptable salt thereof is left to stand in the air, water may be absorbed and adsorbed water may be attached or a hydrate may be formed. Furthermore, the compound of the present invention or a pharmaceutically acceptable salt thereof may be recrystallized to form polymorphic crystals. "Co-crystal" means that the compound or salt of the present invention and the counter molecule exist in the same crystal lattice, and can form together with any number of counter molecules.

The compound of the present invention or a pharmaceutically acceptable salt thereof may form a prodrug, and the present invention also includes such various prodrugs. A prodrug is a derivative of the compound of the present invention having a chemically or metabolically decomposable base, and is a compound that becomes a pharmaceutically active compound of the present invention by solvolysis or in vivo under physiological conditions. Prodrugs include compounds that are converted into compounds represented by formula (I) by enzyme oxidation, reduction, hydrolysis, etc. under physiological conditions in vivo, and compounds that are converted into compounds represented by formula (I) by gastric acid hydrolysis, etc. compound etc. Methods for selecting appropriate prodrug derivatives and production methods are described in, for example, "Design of Prodrugs, Elsevier, Amsterdam, 1985". There are instances where a prodrug is itself active.

When the compound represented by the formula (I) or a pharmaceutically acceptable salt thereof has a hydroxyl group, it can be exemplified as a prodrug of an acyloxy derivative or a sulfonyloxy derivative, and the prodrug is suitably prepared by a compound having a hydroxyl group. Acyl halides, appropriate acid anhydrides, appropriate sulfonyl chlorides, appropriate sulfonic anhydrides, and mixed anhydrides are reacted or produced by using a condensing agent. Examples include CH ₃ COO-, C ₂ H ₅ COO-, tert-BuCOO-, C ₁₅ H ₃₁ COO-, PhCOO-, (m-NaOOCPh)COO-, NaOOCCH ₂ CH ₂ COO-, CH ₃ CH( NH ₂ )COO-, CH ₂ N(CH ₃ ) ₂ COO-, CH ₃ SO ₃ -, CH ₃ CH ₂ SO ₃ -, CF ₃ SO ₃ -, CH ₂ FSO ₃ -, CF ₃ CH ₂ SO ₃ - , p-CH ₃ O-PhSO ₃ -, PhSO ₃ -, p-CH ₃ PhSO ₃ -.

(Method for producing the compound of the present invention)

The compound of the present invention can be produced, for example, by the general synthetic method shown below. Extraction, purification, and the like may be performed as long as they can be performed by ordinary organic chemistry experiments.

The compounds of the present invention can be synthesized by referring to methods known in the art.

(Preparation method 1) When Q is -NHC(O)-

(In the formula, P ¹ is a hydroxyl protecting group; P ² is an amino protecting group; R and R' are carboxyl protecting groups; Z ¹ is CR ^5a R ^5b or CR ^8a R ^8b ; m is an integer from 2 to 3, m When it is 2, -(Z) ₂ -system-(CR ^7a R ^7b -CR ^6a R ^6b )-, when m is 3, -(Z) ₃ -system-(CR ^11a R ^11b -CR ^10a R ^10b -CR ^9a R ^9b )-, or -(CR ^11a R ^11b -CR ^10a R ^10b -O)-; Hal is a halogen; P ¹ , P ² , R and R' are available as long as they are available and described in Protective Groups in Organic Synthesis, The method in Theodora W Greene (John Wiley & Sons) etc. and the protection and/or deprotection group get final product, for example, P ¹ series aromatic carbocycloalkyl etc., P ² series alkoxycarbonyl etc., R and R ' is an alkyl group, etc.; other symbols have the same meaning as above)

step 1

Compound a1 can be obtained by adding a commercially available or prepared compound a by a known method to a general deprotection reaction of a carboxyl protecting group.

step 2

啉、吡啶、DIEA等3級胺，在10℃至60℃，較佳為在20℃至40℃，反應0.1小時至24小時，較佳為1小時至12小時，藉此，可以獲得化合物a3。 In the presence of solvents such as DMF, DMA, NMP, THF, chloroform, and dichloromethane, add condensing agents such as HATU, WSC‧HCl, and PyBOP to compound a1, and then add commercially available or prepared compounds by known methods a2 and triethylamine, N-methyl

tertiary amines such as morphine, pyridine, and DIEA, react at 10°C to 60°C, preferably at 20°C to 40°C, for 0.1 hour to 24 hours, preferably 1 hour to 12 hours, thereby obtaining compound a3 .

step 3

In the presence of THF, methanol, ethanol, chloroform, dichloromethane, THF and other solvents, add compound a4 to compound a3, and react at 60°C to 120°C, preferably at 80°C to 100°C, for 0.5 hours to 24 hours , preferably 1 hour to 12 hours, whereby compound a5 can be obtained.

step 4

Compound a6 can be obtained by adding compound a5 to a general deprotection reaction of an amine protecting group.

step 5

In the presence of solvents such as dichloromethane, dichloroethane, chloroform, methanol, ethanol, toluene, DMF, DMA, THF, etc., add commercially available or prepared compound a7 and acetic acid, Acids such as p-toluenesulfonic acid and methanesulfonic acid are reacted at 20°C to 130°C, preferably at 20°C to 100°C, for 0.1 hour to 24 hours, preferably 1 hour to 12 hours, whereby the compound can be obtained a8.

step 6

In the presence of DMF, DMA, NMP, THF and other solvents, add bases such as cesium carbonate or potassium carbonate and salts such as sodium iodide or potassium iodide to compound a8, at 0°C to 60°C, preferably at 0°C to 40°C , react for 0.1 hour to 24 hours, preferably 1 hour to 12 hours, whereby compound a9 can be obtained.

step 7

Compound a9 can be split into a10 by chiral SFC.

Step 8

Compound a10 is subjected to a general deprotection reaction of a hydroxyl protecting group, whereby compound Ia can be obtained.

(Recipe 2)

(In the formula, each symbol has the same meaning as above)

step 1

In the presence of solvents such as DMF, DMA, NMP, and THF, bases such as cesium carbonate, potassium carbonate, and triethylamine are added to compound a5, and salts such as sodium iodide or potassium iodide are added when Hal is chlorine, and commercially available or available Compound b1 prepared by a known method is reacted at 0°C to 60°C, preferably at 20°C to 40°C, for 0.1 hour to 24 hours, preferably 1 hour to 12 hours, whereby the compound can be obtained b2.

step 2

Compound b3 can be obtained by adding compound b2 to a general deprotection reaction of acetal.

step 3

In the presence of dichloromethane, dichloroethane, chloroform, acetonitrile, methanol, ethanol, toluene, DMF, DMA, THF and other solvents, add acetic acid, p-toluenesulfonic acid, methanesulfonic acid, trifluoroacetic acid to compound b3 Waiting for acid, at 20°C to 130°C, preferably 80°C to 120°C, reacting for 0.1 to 24 hours, preferably 1 hour to 12 hours, whereby compound a9 can be obtained.

step 4

Compound Ia can be synthesized according to Steps 7 and 8 of Preparation Method 1.

(Recipe 3)

(In the formula, each symbol has the same meaning as above)

step 1

In the presence of solvents such as THF and toluene, add commercially available or prepared compound c1 to compound a5, and then add DEAD/PPh ₃ , DIAD/PPh ₃ , DMEAD/PPh ₃ , ADDP/n -Mitsunobu such as Bu ₃ P, at 0°C to 100°C, preferably at 20°C to 80°C, react for 0.1 hour to 24 hours, preferably 1 hour to 12 hours, thereby, can obtain Compound c2.

step 2

Compound c3 can be obtained by adding compound c2 to a general oxidative cracking reaction of olefins. For example, using _ozonolysis or _{K2O4O4 / NaIO4} , etc.

step 3

Compound a9 can be obtained by reacting compound c3 under the same conditions as in Step 3 of Preparation method 2.

step 4

Compound Ia can be synthesized according to Steps 7 and 8 of Preparation Method 1.

(Recipe 4)

(In the formula, each symbol has the same meaning as above)

step 1

Compound d2 can be obtained by reacting Compound a5 and Compound d1 under the same conditions as in Step 1 of Preparation Method 3.

step 2

Compound d3 can be obtained by a general deprotection reaction in which compound d2 is attached to a hydroxyl protecting group.

step 3

Compound d3 is subjected to a general oxidation reaction of a hydroxyl group, whereby compound d4 can be obtained.

step 4

Compound a9 can be obtained by reacting compound d4 under the same conditions as in Step 3 of Preparation method 2.

step 5

Compound Ia can be synthesized according to Steps 7 and 8 of Preparation Method 1.

(Recipe 5)

(In the formula, each symbol has the same meaning as above)

step 1

Compound e2 can be obtained by reacting compound a5 and compound e1 under the same conditions as in step 5 of Production method 1.

step 2

In the presence of solvents such as DMF, DMA, NMP, THF, etc., add a base such as cesium carbonate or potassium carbonate to compound e2, and react at 0°C to 60°C, preferably at 0°C to 40°C, for 0.1 hours to 24 hours , preferably 1 hour to 12 hours, whereby compound e3 can be obtained.

step 3

A general deprotection reaction of a hydroxyl protecting group is carried out, whereby compound e4 can be obtained.

step 4

In the presence of solvents such as THF and toluene, add DEAD/PPh ₃ , DIAD/PPh ₃ , DMEAD/PPh ₃ , ADDP/n-Bu ₃ P and other Mitsunobu reagents (Mitsunobu) to compound e4. °C, preferably at 0°C to 80°C, and react for 1 hour to 24 hours, preferably 1 hour to 12 hours, whereby compound a9 can be obtained.

step 5

Compound Ia can be synthesized according to Steps 7 and 8 of Preparation Method 1.

(Preparation method 6) When Q is a 5-membered aromatic heterocycle

(wherein, Q is a 5-membered aromatic heterocyclic ring; other symbols have the same meaning as above)

step 1

啉、吡啶、二異丙基乙胺等3級胺，在10℃至60℃，較佳為在20℃至40℃，反應0.1小時至24小時，較佳為1小時至12小時，藉此，可以獲得化合物f1。 In the presence of solvents such as DMF, DMA, NMP, THF, chloroform, and dichloromethane, condensing agents such as HATU, WSC‧HCl, and PyBOP are added to compound f that is commercially available or can be prepared by known methods, and then added Compound a4, which is commercially available or can be prepared by known methods, and triethylamine, N-methyl

tertiary amines such as phenoline, pyridine, and diisopropylethylamine, at 10°C to 60°C, preferably at 20°C to 40°C, react for 0.1 hour to 24 hours, preferably 1 hour to 12 hours, thereby , Compound f1 can be obtained.

step 2

In the presence of solvents such as DMF, DMA, and NMP, add commercially available or prepared compound f2, acetic acid, pyridinium p-toluenesulfonate, p-toluenesulfonic acid, methanesulfonic acid, etc. to compound f1 The acid is reacted at 20°C to 120°C, preferably at 60°C to 100°C, for 0.1 hour to 24 hours, preferably 1 hour to 12 hours, whereby compound f3 can be obtained.

step 3

Compound f4 can be obtained by a known general deprotection reaction of adding compound f3 to an amino protecting group.

step 4

Compound f5 can be obtained by reacting compound f4 under the same conditions as those described in Preparations 1 to 5.

step 5

In solvents such as dichloromethane, dichloroethane, acetonitrile or DMF, add bromine, NBS, NCS, NIS and other halogenating reagents to compound f5, when Hal is bromine, at -30°C to 50°C, preferably at -30°C to 50°C -10°C to 20°C, react for 0.1 hour to 10 hours, preferably 0.5 hour to 2 hours, thereby obtaining compound f6. When Hal is chlorine or iodine, compound f6 can be obtained by reacting at 10°C to 150°C, preferably at 60°C to 120°C, for 0.5 hours to 24 hours, preferably 1 hour to 6 hours.

step 7

、DMF、DME、THF、DMSO等溶劑或混合溶劑中，於化合物f6中添加Pd(PPh₃)₄、Pd(OAc)₂、Pd(PPh₃)₂Cl₂、Pd(dppf)₂Cl₂或Pd(dtbpf)等鈀催化劑；乙酸鉀、乙酸鈉、碳酸鉀或磷酸鉀等鹼；及雙聯(頻哪醇)硼酸酯(Bis(pinacolato)diboron)，在氮環境下，在0℃至150℃，較佳為在60℃至120℃，反應0.5小時至24小時，較佳為1小時至12小時，藉此，可以獲得化合物f7。 in two

, DMF, DME, THF, DMSO and other solvents or mixed solvents, add Pd(PPh ₃ ) ₄ , Pd(OAc) ₂ , Pd(PPh ₃ ) ₂ Cl ₂ , Pd(dppf) ₂ Cl ₂ or Palladium catalysts such as Pd(dtbpf); bases such as potassium acetate, sodium acetate, potassium carbonate or potassium phosphate; 150°C, preferably 60°C to 120°C, react for 0.5 hours to 24 hours, preferably 1 hour to 12 hours, thereby obtaining compound f7.

Step 8

、DMF、DME、THF、水等溶劑或混合溶劑中，於化合物f7中添加Pd(PPh₃)₄、Pd(OAc)₂、Pd(PPh₃)₂Cl₂、Pd(dppf)₂或Pd(dtbpf)等鈀催化劑；碳酸鉀、碳酸鈉、碳酸銫或磷酸鉀等鹼；及市售或藉由已知的方法調製的化合物f8，在氮環境中，在0℃至150℃，較佳為在60℃至120℃，反應0.5小時至24小時，較佳為1小時至12小時，藉此，可以獲得化合物f9。 in two

, DMF, DME, THF, water and other solvents or mixed solvents, add Pd(PPh ₃ ) ₄ , Pd(OAc) ₂ , Pd(PPh ₃ ) ₂ Cl ₂ , Pd(dppf) ₂ or Pd( dtbpf) and other palladium catalysts; potassium carbonate, sodium carbonate, cesium carbonate or potassium phosphate and other bases; and commercially available or compound f8 prepared by known methods, in a nitrogen environment, at 0 ° C to 150 ° C, preferably React at 60°C to 120°C for 0.5 hours to 24 hours, preferably 1 hour to 12 hours, whereby compound f9 can be obtained.

step 9

Compound Ib can be synthesized according to Steps 7 and 8 of Preparation Method 1.

(Recipe 7)

(In the formula, each symbol has the same meaning as above)

step 1

In the presence of dichloromethane, dichloroethane, chloroform, DMF, DMA, NMP, THF and other solvents, it is prepared by adding triethylamine or diisopropylethylamine and ethyl chloroformate to compound g After acetyl chloride, add commercially available or compound g1 that can be prepared by known methods, and react at 0°C to 60°C, preferably at 0°C to 20°C, for 0.1 hour to 24 hours, preferably 1 hours to 12 hours, whereby compound g2 can be obtained.

step 2

、DMF、DMA、THF等溶劑的存在下，使化合物g2與T3P、三氟乙酸、磷酸、鹽酸、硫酸、氫溴酸等酸作用，在20℃至130℃，較佳為60℃至100℃，反應0.1小時至24小時，較佳為1小時至12小時，藉此，可以獲得化合物g3。 In ethyl acetate, dichloromethane, dichloroethane, chloroform, di

In the presence of solvents such as , DMF, DMA, THF, compound g2 reacts with acids such as T3P, trifluoroacetic acid, phosphoric acid, hydrochloric acid, sulfuric acid, hydrobromic acid, at 20°C to 130°C, preferably 60°C to 100°C , react for 0.1 hour to 24 hours, preferably 1 hour to 12 hours, whereby compound g3 can be obtained.

step 3

Compound Ic can be synthesized according to Steps 7 and 8 of Preparation Method 1.

The compounds of the present invention obtained above can be further chemically modified to synthesize other compounds. Furthermore, in the above reaction, when there is a reactive functional group (for example, OH, COOH, NH ₂ ) in the side chain moiety, it can be protected before the reaction or deprotected after the reaction as needed.

Protecting groups (amino group protecting group, hydroxyl protecting group, etc.) include, for example: ethoxycarbonyl, tert-butoxycarbonyl, acetyl, benzyl, and the like described in Protective Groups in Organic Synthesis (Protective Groups in Organic Synthesis ), T.W. Green, John Wiley & Sons Inc. (1991) et al. The method for introducing and removing the protecting group can utilize methods commonly used in organic synthetic chemistry [for example, see Protective Groups in Organic Synthesis (Protective Groups in Organic Synthesis), written by T.W. Greene, John Wiley & Sons Inc. (1991)] and the like. recorded method or based on such income. Furthermore, the conversion of the functional groups contained in each substituent can be performed by known methods [for example, Comprehensive Organic Transformations (Comprehensive Organic Transformations), written by R.C.Larock (1989), etc.] in addition to the above-mentioned production methods. The present invention Some of the compounds can be further transformed into new derivatives by using this compound as a synthetic intermediate. Intermediates and target compounds in the above-mentioned production methods can be purified by common purification methods in organic synthetic chemistry, for example, It can be isolated and purified by adding to neutralization, filtration, extraction, washing, drying, concentration, recrystallization, various chromatography, etc. Furthermore, the intermediate may be provided in the subsequent reaction without being particularly purified.

The compounds of the present invention are useful as medicines such as antiviral drugs. The compound of the present invention has significant inhibitory effect on virus integrase. Therefore, the compound of the present invention can be expected to have prophylactic or therapeutic effects on various diseases caused by viruses that produce at least integrase upon infection in animal cells, and can be used, for example, as an antiretrovirus (for example, HIV-1, HIV- 2. Integrase inhibitors of HTLV-1, SIV, FIV, etc.), and can be used as anti-HIV drugs. Even better compounds, in terms of in vivo dynamics, have characteristics such as high blood concentration, long duration of effect, and/or significant tissue metastasis. Furthermore, preferred compounds are safe from the viewpoint of side effects (eg, inhibition of CYP enzymes, mutagenicity, prolongation of QT interval in electrocardiogram, arrhythmia).

Furthermore, the compound of the present invention can also be used in combination therapy with anti-HIV drugs having different action mechanisms such as reverse transcriptase inhibitors, protease inhibitors, and/or invasion inhibitors.

Furthermore, the above-mentioned use includes not only the use as an anti-HIV mixture, but also use as a combination agent for increasing the anti-HIV activity of other anti-HIV drugs such as cocktail therapy.

Furthermore, in the field of gene therapy, the compound of the present invention can also be used to prevent the infection of the retroviral vector from spreading beyond the target tissue when HIV or MLV-based retroviral vectors are used. In particular, when an infection carrier such as a cell is returned to the body in a test tube, unnecessary infection in the body can be prevented by administering the compound of the present invention in advance.

The pharmaceutical composition of the present invention can be administered by either oral or parenteral methods. The methods of parenteral administration include transdermal, subcutaneous, intravenous, intraarterial, intramuscular, intraperitoneal, transmucosal, inhalation, nasal, eye drops, ear drops, and vaginal administration.

In the case of oral administration, solid preparations for internal use (for example, tablets, powders, granules, capsules, pills, films, etc.), liquid preparations for internal use (for example, suspensions, emulsions, etc.) , elixirs, syrups, lemonades, alcoholic preparations, aromatic water preparations, extractants, decoctions, tinctures, etc.) and other commonly used dosage forms may be administered. Tablets can be sugar-coated tablets, film-coated tablets, enteric-coated tablets, sustained-release tablets, lozenges, sublingual tablets, buccal tablets, chewable tablets or orally disintegrating tablets; powders and granules can be dry syrup , the capsules can be soft capsules, microcapsules or sustained-release capsules.

In the case of parenteral administration, injections, spot drops, external preparations (for example, eye drops, nasal drops, ear drops, aerosols, inhalants, emulsions, injections, coatings, etc.) can be administered as appropriate. , gargle, colon cleanser, ointment, plaster, jelly, cream, patch, poultice, external powder, suppository, etc.) and any other commonly used dosage forms. The injection agent may be an emulsion of O/W, W/O, O/W/O, W/O/W type, or the like.

The effective amount of the compound of the present invention is mixed with various pharmaceutical additives such as excipients, binders, disintegrants, lubricants, etc. that are suitable for the dosage form as required to prepare a pharmaceutical composition. Furthermore, by appropriately changing the effective amount, dosage form, and/or various pharmaceutical additives of the compound of the present invention, it can be used as a pharmaceutical composition for children, the elderly, severe patients, or surgery. For example, the pharmaceutical composition for children can be used for neonates (less than 4 weeks after birth), infants (4 weeks after birth to less than 1 year old), infants (over 1 year old and under 7 years old), children (over 7 years old and under 15 years old) or patients aged 15 to 18 years. For example, the pharmaceutical composition for the elderly can be administered to patients over 65 years old.

The dose of the pharmaceutical composition of the present invention is set in consideration of the patient's age, body weight, type or degree of disease, and route of administration, but it is usually 0.05 to 100 mg/kg/day for oral administration, preferably In the range of 0.1 to 10 mg/kg/day. In the case of parenteral administration, it varies greatly depending on the route of administration, but it is usually in the range of 0.005 to 10 mg/kg/day, preferably 0.01 to 1 mg/kg/day. The pharmaceutical composition can be administered once a day to once a month or once every three months.

[Example]

Examples are shown below.

(abbreviation)

ADDP: 1,1'-(Azodicarbonyl)dipiperide

Bn: benzyl

DEAD: Diethyl azodicarboxylate

DIAD: Diisopropyl azodicarboxylate

DIEA: N,N-Diisopropylethylamine

DMA: Dimethylacetamide

DME: Dimethoxyethane

DMEAD: Di-2-methoxyethyl azodicarboxylate

DMF: Dimethylformamide

DMSO: Dimethylsulfone

HATU: O-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate

NBS: N-Bromosuccinimide

NCS: N-chlorosuccinimide

NIS: N-iodosuccinimide

NMP: N-Methylpyrrolidone

PyBOP: (Benzotriazol-1-yloxy)tripyrrolidinylphosphonium hexafluorophosphate

TBAF: Tetrabutylammonium fluoride

THF: Tetrahydrofuran

WSC‧HCl: 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride

The NMR analysis obtained in each example was carried out at 300 MHz or 400 MHz and measured using DMSO-d ₆ and CDCl ₃ . In addition, when displaying NMR data, there may be cases where not all measured peaks are described.

In the examples, "No." means the compound number, "Structure" means the chemical structure, and "MS" means the molecular weight in LC/MS (liquid chromatography/mass spectrometry).

(measurement conditions)

(A) Column: ACQUITY UPLC (registered trademark) BEH C18 (1.7μm i.d.2.1x 50mm) (waters)

Flow rate: 0.8mL/min; UV detection wavelength: 254nm;

Mobile phase: [A] is an aqueous solution containing 0.1% formic acid, [B] is an acetonitrile solution containing 0.1% formic acid

After a linear gradient of 5% to 100% solvent [B] for 3.5 minutes, 100% solvent [B] was maintained for 0.5 minutes.

(B) Column: Shim-pack XR-ODS (2.2μm, i.d.50 x 3.0mm) (Shimadzu)

Flow rate: 1.6mL/min; UV detection wavelength: 254nm;

Mobile phase: [A] is an aqueous solution containing 0.1% formic acid, [B] is an acetonitrile solution containing 0.1% formic acid

Gradient: Run a linear gradient from 10% to 100% solvent [B] for 3 min, hold 100% solvent [B] for 0.5 min.

(C) Column: Shim-pack XR-ODS (2.2μm, i.d.50 x 3.0mm) (Shimadzu)

Flow rate: 1.6mL/min; UV detection wavelength: 254nm;

Mobile phase: [A] is an aqueous solution containing 0.1% formic acid, [B] is an acetonitrile solution containing 0.1% formic acid

Gradient: A linear gradient of 10%-100% solvent [B] was performed for 8 minutes and maintained at 100% solvent [B] for 0.5 minutes.

Example 1

step 1

Add 2mol/L methanol solution of ethylamine (17.9ml, 35.9mmol) to Compound 1 (1.50g, 3.59mmol), and stir at 100°C for 1 hour under microwave irradiation. After distilling off the solvent of the reaction liquid under reduced pressure, dilute hydrochloric acid was added to make it acidic, and it extracted with ethyl acetate. After drying the organic layer with sodium sulfate, the solvent was distilled off. The resulting residue was purified by silica gel column chromatography (chloroform-methanol) to obtain compound 2 (1.15 g, yield 74%).

1H-NMR (CDCL ₃ )δ: 14.53(s,1H), 8.64(brs,1H), 8.46(s,1H), 7.37(m,5H), 6.57(brs,1H), 5.38(s,2H) ,3.24(dt,J=14.0,6.6Hz,2H),1.45(s,9H),1.02(t,J=7.3Hz,4H).

step 2

Compound 2 (9.59g, 22.2mmol) was dissolved in dichloromethane (180ml), and (2,4-difluorophenyl)methylamine (4.77g, 33.3mmol), PyBOP (13.9g, 26.7mmol) and DIEA (11.7ml, 66.7mmol) and stirred at room temperature for 18 hours. The reaction liquid was washed with water and saturated brine, the organic layer was dried over sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel column chromatography (chloroform-methanol) to obtain compound 3 (11.5 g, yield 93%).

1H-NMR(CDCL ₃ )δ: 10.20(t,J=5.8Hz,1H),8.54(brs,1H),8.49(s,1H),7.38(m,5H),6.87-6.79(m,2H) ,6.61(t,J=5.5Hz,1H),5.28(s,2H),4.64(d,J=5.9Hz,2H),3.18(ddt,J=18.8,10.2,3.8Hz,3H),1.83- 1.80(m,1H),1.43(s,9H),0.99(t,J=7.3Hz,3H).

step 3

(57.5ml)，添加4mol/L鹽酸/二

溶液(300ml)，並在室溫攪拌4小時。將反應液的溶劑減壓餾除後，添加飽和碳酸鈉水溶液，並用氯仿-甲醇萃取。將有機層用硫酸鈉乾燥 後，餾除溶劑所得到的粗產物用二異丙醚固體化，得到化合物4(7.80g，產率83%)。 Compound 3 (11.5 g, 9.54 mmol) was dissolved in di

(57.5ml), add 4mol/L hydrochloric acid/two

solution (300ml) and stirred at room temperature for 4 hours. After distilling off the solvent of the reaction liquid under reduced pressure, a saturated aqueous sodium carbonate solution was added, followed by extraction with chloroform-methanol. After the organic layer was dried over sodium sulfate, the solvent was distilled off, and the obtained crude product was solidified with diisopropyl ether to obtain compound 4 (7.80 g, yield 83%).

1H-NMR (CDCL ₃ )δ: 10.33(s,1H), 8.60(s,1H), 7.39(m,5H), 6.83(m,3H), 5.82(s,2H), 5.26(s,2H) ,4.64(d,J=5.8Hz,2H),3.28-3.21(m,2H),1.02(t,J=7.3Hz,3H).

step 4

Compound 4 (200 mg, 0.438 mmol) was dissolved in dichloromethane (4 ml), compound 5 (111 mg, 0.920 mmol) and acetic acid (catalytic amount) were added, and stirred at room temperature for 19 hours. After the reaction solution was concentrated under reduced pressure, it was purified by silica gel column chromatography (chloroform-methanol) to obtain compound 6 (265 mg, yield 100%).

MS: m/z=559[M+H]+

step 5

Compound 6 (245 mg, 0.438 mmol) was dissolved in DMF (5 ml), cesium carbonate (428 mg, 1.31 mmol) was added at 0° C. and stirred at room temperature for 18 hours. Dilute hydrochloric acid was added to the reaction solution, and extracted with ethyl acetate. The organic layer was washed with water, dried over sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel column chromatography (chloroform-methanol) to obtain a racemic compound (139 mg, yield 60%).

The obtained racemic compound was optically resolved by SFC to obtain Compound 7.

Column: CHIRALPAK IA/SFC (5μm, i.d.250 x 20mm)

Flow rate: 30mL/min

UV detection wavelength: 250nm

Preparation conditions: maintain the composition ratio of MeOH/CO ₂ =45/55, and send liquid for 21 minutes.

1H-NMR (CDCL ₃ )δ: 10.46(s,1H), 8.51(s,1H), 7.58(m,2H), 7.34(m,4H), 6.81(m,2H), 5.41(d,J= 10.4Hz,1H),5.26(d,J=10.4Hz,1H),4.91(s,1H),4.64(m,2H),4.39(dd,J=14.3,7.2Hz,1H),3.18-2.88( m,3H),2.24(d,J=14.7Hz,1H),2.00(m,1H),1.85(m,2H),1.72(d,J=13.6Hz,1H),1.38(m,1H), 1.16(t,J=7.1Hz,3H).

step 6

Compound 7 (44.0 mg, 0.0840 mmol) was dissolved in DMF (0.88 ml), lithium chloride (35.7 mg, 0.842 mmol) was added, and stirred at 90° C. for 1.5 hours. Water was added to the reaction liquid, acidified with 10% citric acid aqueous solution, and extracted with ethyl acetate. The organic layer was washed with water, dried over sodium sulfate, and the solvent was distilled off. The obtained crude product was solidified with diethyl ether to obtain compound I-23 (19 mg, yield 52%).

1H-NMR(CDCl ₃ )δ: 11.98(s,1H),10.42(s,1H),8.46(s,1H),7.36(dd,J=15.2,8.6Hz,1H),6.83-6.77(m, 2H)),5.06(s,1H),4.64(m,2H),4.35(td,J=14.2,6.9Hz,1H),3.20-3.09(m,2H),3.00(d,J=10.8Hz, 1H),2.31(d,J=15.4Hz,1H),2.06(m,1H),1.89(m,2H),1.76(m,1H),1.42-1.36(m,1H),1.24(t,J =7.1Hz,4H).

Example 2

step 1

Under a nitrogen atmosphere, a THF (7.0 mL) solution of compound 8 (1.3 mL, 11.1 mmol) was added dropwise to a THF (3.0 mL) solution of magnesium (322 mg, 13.3 mmol), and stirred at room temperature for 30 minutes. The reaction solution was cooled to 0°C, copper iodide (210 mg, 1.1 mmol) was added, a solution of compound 9 (1.2 mL, 16.6 mmol) in THF (6.0 mL) was added dropwise, and the temperature was raised to room temperature and stirred for 2 hours. Saturated ammonium chloride aqueous solution was added to the reaction solution, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The obtained residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain compound 10 (192 mg, yield 11%).

1H-NMR(CDCl ₃ )δ: 4.86(t,J=4.8Hz,1H),3.99-3.96(m,2H),3.90-3.79(m,3H),1.72-1.67(m,2H),1.55- 1.48(m,4H),1.36(d,J=4.5Hz,1H),1.20(d,J=6.3Hz,3H).

step 2

In compound 11 (334mg, 0.60mmol) in THF (2.0mL) solution, add compound 10 (192.2mg, 1.2mmol), triphenylphosphine (315mg, 1.2mmol) and azodicarboxylic acid bis (2-methane oxyethyl ester) (281 mg, 1.0 mmol) and stirred at room temperature for 1 hour. Water was added to the reaction liquid, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The resulting residue was roughly purified by silica gel column chromatography (hexane-ethyl acetate).

MS: m/z=699[M+H]+

step 3

To a solution of the crude product (100 mg) obtained in Step 2 in acetonitrile (1.0 mL), p-toluenesulfonic acid hydrate (45.1 mg, 0.242 mmol) was added, followed by heating under reflux for 210 minutes. The reaction liquid was cooled to room temperature, added with saturated aqueous sodium bicarbonate solution, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The obtained residue was dissolved in DMF (1.0 mL), cesium carbonate (140 mg, 0.43 mmol) and benzyl bromide (34.1 μL, 0.29 mmol) were added, and the mixture was stirred at room temperature for 3 hours. Water was added to the reaction liquid, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel column chromatography (chloroform-methanol) to obtain Compound 13 (65.1 mg).

MS: m/z=537[M+H]+

step 4

The same reaction as in Step 6 of Example 1 was carried out to obtain compound I-31 (31 mg, yield 57%).

1H-NMR (CDCl ₃ )δ: 11.93(s,1H), 10.40(s,1H), 8.39(s,1H), 7.40-7.34(m,1H), 6.84-6.77(m,2H), 511- 5.09(m,1H),4.64(d,J=5.8Hz,2H),4.40-4.31(m,1H),3.27-3.21(m,1H),3.13-3.06(m,1H),2.32-2.28( m,1H),2.12-2.04(m,1H),1.86-1.83(m,1H),1.79-1.75(m,1H),1.63-1,48(m,2H),1.21(t,J=7.2 Hz,3H),0.89(d,J=6.3Hz,3H).

Example 3

step 1

Potassium carbonate (261 mg, 1.89 mmol) and 4-bromobutene (147 mg, 0.943 mmol) were added to a solution of compound 11 (352 mg, 0.629 mmol) in DMF (3.5 ml), and reacted overnight at room temperature. Water was added to the reaction liquid, and extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off.

MS: m/z=611[M+H]+

step 2

溶液(3.15ml)，並在室溫攪拌2小時。將飽和碳酸氫鈉水添加至反應液中，並用乙酸乙酯萃取。將有機層用飽和食鹽水洗淨，用無水硫酸鈉乾燥後，餾除溶劑。 In the crude product gained in step 1, add 4mol/L hydrochloric acid/di

solution (3.15ml) and stirred at room temperature for 2 hours. Saturated sodium bicarbonate water was added to the reaction liquid, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off.

MS: m/z=511[M+H]+

step 3

The crude product obtained in step 2, acrolein (102 mg, 1.83 mmol), p-toluenesulfonic acid hydrate (11.6 mg, 0.061 mmol) were dissolved in dichloroethane (9.6 mL), and stirred at 100 ° C for 6 hours hour. After cooling the reaction solution to room temperature, water and saturated sodium bicarbonate water were added, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain Compound 16 (115 mg).

MS: m/z=549[M+H]+

step 4

Compound 16 (66.4 mg, 0.121 mmol), Hoveyda-Grubbs second generation catalyst (60 mg, 0.139 mmol) were dissolved in dichloromethane (10 mL), and heated to reflux for 6 hours. The solvent of the reaction solution was distilled off, and the obtained residue was roughly purified by silica gel column chromatography (ethyl acetate-methanol).

MS: m/z=521[M+H]+

step 5

Compound 17 obtained in step 4 was optically resolved by SFC to obtain compound 18.

Column: CHIRALPAK IC/SFC (5μm, i.d.250 x 20mm)

Flow rate: 20mL/min

UV detection wavelength: 220nm

Preparation conditions: maintain the composition ratio of MeOH/CO ₂ =70/30, and feed the solution for 21 minutes.

step 6

The same reaction as in step 6 of Example 1 was carried out to obtain compound II-65 (11 mg, yield 74%).

1H-NMR(CDCl ₃ )δ: 11.93(s,1H),10.42(t,J=5.6Hz,1H),8.50(s,1H),7.40-7.33(m,1H),6.84-6.77(m, 2H),6.28-6.24(m,1H),5.96-5.91(m,1H),5.32(d,J=5.2Hz,1H),4.68(dd,J=15.2,6.0Hz,1H),4.61( dd,J=15.6,6.0Hz,1H),3.83(dt,J=21.2,7.2Hz,1H),3.53(dt,J=20.8,6.8Hz,1H),3.39(td,J=11.2,4.4Hz ,1H),3.04(dd,J=10.8,6.8Hz,1H),2.77-2.68(m,1H),2.35(dt,J=18.8,4.8Hz,1H),1.23(t,J=7.2Hz, 3H).

Example 4

step 1

In compound 11 (326mg, 0.59mmol), compound 19 (87mg, 0.77mmol) and triphenylphosphine (307mg, 1.18mmol) in THF (3.5mL) solution, add azodicarboxylic acid bis-2- Methoxyethyl ester (274mg, 1.18mmol), let stand at room temperature for 12 hours. Water was added to the reaction liquid, and extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain compound 20 (293 mg, yield 77%).

MS: m/z=653[M+H]+

step 2

(3.4mL)、水(2.3mL)，並在0℃添加2,6-二甲吡啶(0.10mL)、過碘酸氫鈉(282mg，1.32mmol)、 鋨酸鉀(VI)二水合物(8.0mg，0.02mmol)，並花費5小時由0℃升溫至室溫。將反應液用矽藻土(Celite)(註冊商標)過濾，添加10%硫代硫酸鈉水溶液，並用乙酸乙酯萃取。將有機層用水、飽和食鹽水洗淨，用無水硫酸鈉乾燥後，餾除溶劑。所得之殘渣藉由矽膠管柱層析(己烷-乙酸乙酯)精製，得到化合物21(223mg，產率78%)。 Compound 20 (287mg, 0.44mmol) was suspended in two

(3.4mL), water (2.3mL), and added 2,6-lutidine (0.10mL), sodium hydrogen periodate (282mg, 1.32mmol), potassium osmate (VI) dihydrate ( 8.0mg, 0.02mmol), and spent 5 hours warming from 0°C to room temperature. The reaction liquid was filtered through Celite (registered trademark), 10% aqueous sodium thiosulfate solution was added, and extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain compound 21 (223 mg, yield 78%).

MS: m/z=655[M+H]+

step 3

溶液(1.47ml)，並在室溫攪拌2小時。餾除溶劑，將所得之粗產物溶解在甲苯(2.0ml)中，添加觸媒量的乙酸，在90℃攪拌2小時。將飽和碳酸氫鈉水添加反應溶液，並用乙酸乙酯萃取。將有機層用水、飽和食鹽水洗淨，用硫酸鈉乾燥後，餾除溶劑。所得之殘渣藉由矽膠管柱層析精製，得到非鏡像異構物混合物。將所得之非鏡像異構物混合物藉由SFC進行光學分割，得到化合物22(69mg，產率44%)。 Compound 21 (192mg, 0.29mmol) was dissolved in 4mol/L hydrochloric acid/di

solution (1.47ml) and stirred at room temperature for 2 hours. The solvent was distilled off, the obtained crude product was dissolved in toluene (2.0 ml), a catalytic amount of acetic acid was added, and the mixture was stirred at 90° C. for 2 hours. Saturated sodium bicarbonate water was added to the reaction solution, and extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel column chromatography to obtain a mixture of diastereomers. The resulting mixture of diastereomers was optically resolved by SFC to obtain compound 22 (69 mg, yield 44%).

Column: Two CHIRALPAK IC/SFC (5μm, i.d.250 x 20mm) used in series

Flow rate: 20mL/min

UV detection wavelength: 220nm

Preparation conditions: maintain the composition ratio of MeOH/CO ₂ =65/35, and feed the solution for 35 minutes.

MS: m/z=537[M+H]+

step 4

The same reaction as step 6 of Example 1 was carried out to obtain compound II-34.

MS: m/z=447[M+H]+

Example 5

step 1

In a solution of compound 23 (1.59g, 12.2mmol) in DMF (16.0mL), add imidazole (0.998g, 14.66mmol) and tert-butyldimethylsilyl chloride (1.84g, 12.21mmol) at 0°C, Stir at room temperature for 3 hours. Aqueous saturated ammonium chloride solution was added to the reaction liquid, followed by extraction with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The obtained residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain compound 24 (1.39 g, yield 47%).

1 H-NMR (CDCl ₃ ) δ: 3.47-3.55 (m, 4H), 2.09-2.15 (m, 2H), 1.88-1.95 (s, 1H), 1.65-1.79 (m, 2H), 1.32-1.42 ( m,2H),0.88-0.89(m,1H),0.85(s,9H),0.039(s,6H).

step 2

In compound 24 (400mg, 0.164mmol), compound 11 (700mg, 1.26mmol) and triphenylphosphine (660mg, 2.52mmol) in THF (7mL) solution, add azodicarboxylic acid bis-2-methyl Oxyethyl ester (589 mg, 2.52 mmol), and allowed to stand at room temperature for 12 hours. add water into the reaction solution, and extracted with ethyl acetate. The organic layer was washed with water, dried over anhydrous sodium sulfate, and the solvent was distilled off. The resulting residue was roughly purified by silica gel column chromatography (hexane-ethyl acetate).

step 3

To a THF (10.0 mL) solution of compound 25 (1.06 g, 1.35 mmol) was added 1 mol/L TBAF/THF solution (1.63 ml, 1.63 mmol), and stirred at room temperature for 12 hours. Aqueous saturated ammonium chloride solution was added to the reaction liquid, followed by extraction with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain compound 26 (720 mg, yield 80%).

MS: m/z=669[M+H]+

step 4

To a solution of compound 26 (720 mg, 1.08 mmol) in dichloromethane (8.0 mL) was added Dess-Martin periodinane at 0° C. and stirred at room temperature for 1 hour. A 10% aqueous sodium thiosulfate solution and a saturated aqueous sodium bicarbonate solution were added to the reaction liquid, followed by extraction with chloroform. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain compound 27 (393 mg, yield 55%).

MS: m/z=667[M+H]+

step 5

A solution of compound 27 (393 mg, 0.59 mmol) in acetonitrile (8.0 mL) was heated and stirred at 60° C. for 80 minutes. Saturated aqueous sodium bicarbonate solution was added to the reaction liquid, followed by extraction with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The resulting crude product was dissolved in DMF (4.0 mL), and cesium carbonate (576 mg, 1.77 mmol), Toluene bromide (0.21 mL, 1.77 mmol), and stirred at room temperature overnight. Water was added to the reaction liquid, and extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel column chromatography (hexane-ethyl acetate), and optically resolved by SFC to obtain compound 28 (89 mg, yield 28%).

Column: Two CHIRALPAK IC/SFC (5μm, i.d.250 x 20mm) used in series

Flow rate: 20mL/min

UV detection wavelength: 220nm

Preparation conditions: maintain the composition ratio of MeOH/CO ₂ =75/25, and feed the solution for 45 minutes.

MS: m/z=549[M+H]+

step 6

The same reaction as in Step 6 of Example 1 was carried out to obtain Compound II-1 (11 mg, yield 74%).

MS: m/z=459[M+H]+

Example 6

step 1

Potassium carbonate (2.02g, 14.6mmol) and 2-(4-bromobutyl)-1,3-dioxolane (2.53ml, 16.7 mmol), and react overnight at room temperature. The reaction solution was neutralized with 1mol/L hydrochloric acid and extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over sodium sulfate, and the solvent was distilled off to obtain Compound 29.

MS: m/z=688[M+H]+

step 2

A 2 mol/L sodium hydroxide aqueous solution (17.38 ml, 139 mmol) was added to a THF (47.8 mL) solution of compound 29, and stirred at room temperature for 2 hours. The reaction liquid was neutralized by adding 2mol/L hydrochloric acid gradually, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over sodium sulfate, and the solvent was distilled off to obtain Compound 30.

MS: m/z=560[M+H]+

step 3

(5mL)溶液中添加4mol/L氯化氫(1,4-二

溶液，34.8ml，139mmol)，在室溫攪拌1小時。餾除反應液的溶劑，添加甲苯再次餾除，得到化合物31。 1,4-bis in compound 30

(5mL) solution was added 4mol/L hydrogen chloride (1,4-di

solution, 34.8ml, 139mmol), stirred at room temperature for 1 hour. The solvent of the reaction solution was distilled off, and toluene was added to distill off again to obtain Compound 31.

MS: m/z=416[M+H]+

step 4

A few drops of acetic acid were added to a solution of compound 31 in toluene (50 mL), followed by stirring at 110° C. for 30 minutes. The solvent of the reaction solution was distilled off, and the resulting residue was solidified with ethanol/isopropyl ether to obtain Compound 32 (2.44 g, 88% yield over 4 steps).

¹ H-NMR(CDCl ₃ )δ: 15.1(s,1H), 8.48(s,1H), 7.57-7.55(m,2H), 7.36-7.29(m,3H), 5.53(d,J=10.4Hz ,1H),5.36(d,J=10.4Hz,1H),4.93-4.91(m,1H),4.20(td,J=21.6,7.2Hz,1H),3.24-3.02(m,3H),2.28- 1.73(m,5H),1.41-1.31(m,1H),1.18(t,J=7.2Hz,3H).

step 5

In compound 32 (300mg, 0.755mmol) in dichloromethane (3ml) solution, add triethylamine (0.419ml, 3.02mmol) and ethyl chloroformate (90.0mg, 0.830mmol) at 0 ℃, stir at room temperature 30 minutes. Compound 33 (216 mg, 0.906 mmol) was added to the reaction liquid, and stirred at room temperature for 1 hour. The reaction solution was concentrated, and the resulting residue was purified by silica gel column chromatography (chloroform-methanol) to obtain compound 34 (466 mg, yield 100%).

MS: m/z=582[M+H]+

step 6

To a solution of compound 34 (439mg, 0.755mmol) in ethyl acetate (6ml) was added 50% T3P/ethyl acetate solution (2.25ml, 7,55mmol), and stirred at 100°C for 1 hour. A saturated aqueous sodium bicarbonate solution was added to the reaction liquid, followed by extraction with ethyl acetate. The organic layer was washed with water, dried over sodium sulfate, and the solvent was distilled off, and the resulting residue was purified by silica gel column chromatography (chloroform-methanol) to obtain a racemic compound. The obtained racemic compound was optically resolved by SFC to obtain compound 35.

Column: CHIRALPAK IA/SFC (5μm, i.d.250 x 20mm)

Flow rate: 20mL/min

UV detection wavelength: 220nm

Preparation conditions: maintain the composition ratio of MeOH/CO 2 =65/35, and feed the solution for 25 minutes.

1H-NMR(CDCl3)δ: 8.80(s,1H),7.60(m,2H),7.34-7.27(m,4H),6.85(t,J=8.9Hz,2H),5.55(d,J=10.3 Hz,1H),5.33(d,J=10.4Hz,1H),4.97(m,1H),4.46(s,2H),4.40(m,1H),3.23(m,1H),3.10-3.03(m ,2H),2.24(m,1H),2.02(m,1H),1.89(m,2H),1.72(m,1H),1.42(m,1H),1.17(t,J=7.2Hz,3H) .

step 7

By the same method as Step 6 of Example 1, compound I-11 (63 mg, yield 68%) was obtained.

1H-NMR (CDCl3) δ: 12.04(s,1H), 8.73(s,1H), 7.31(m,1H), 6.84(t, J=8.6Hz, 2H), 5.14(s,1H), 4.45( s,2H),4.36(m,1H),3.26-3.04(m,3H),2.33(d,J=14.9Hz,1H),2.08(t,J=14.7Hz,1H),1.91(m,3H ),1.42(m,1H),1.24(t,J=7.2Hz,3H).

Example 7

step 1

A dichloromethane (10 mL) solution of compound 36 (1.0 g, 2.8 mmol) synthesized in the same manner as in Example 1 was cooled to 0° C., NBS (0.56 g, 3.1 mmol) was added, and stirred overnight at room temperature. The solvent of the reaction solution was distilled off, and the resulting residue was purified by silica gel column chromatography (chloroform-methanol). to obtain the racemic compound. The obtained racemic compound was optically resolved by SFC to obtain compound 37.

Column: CHIRALPAK IB/SFC (5μm, i.d.250 x 20mm)

Flow rate: 30mL/min

UV detection wavelength: 220nm

Preparation conditions: maintain the composition ratio of MeOH/CO ₂ =35/65, and feed the solution for 21 minutes.

1H-NMR(CDCl ₃ )δ: 7.83(s,1H),7.64(d,J=7.0Hz,2H),7.34-7.27(m,3H),5.48(d,J=10.3Hz,1H),5.26 (d,J=10.3Hz,1H),4.92-4.90(m,1H),4.43-4.39(m,1H),3.20-3.14(m,1H),3.05-2.98(m,2H),2.26-2.22 (m,1H),1.97-1.94(m,1H),1.84-1.82(m,2H),1.73-1.69(m,1H),1.41-1.39(m,1H),1.16(t,J=7.2Hz ,3H).

step 2

Compound 37 (250 mg, 0.58 mmol) was dissolved in toluene, compound 38 (183 mg, 0.87 mmol), Pd(OAc) ₂ (13.0 mg, 0.06 mmol), 2-dicyclohexylphosphino-2′-(N, N-diamino)biphenyl (46mg, 0.12mmol) and cesium carbonate (565mg, 1.7mmol) were sealed and stirred at 140°C for 2 hours. After cooling to room temperature, the insoluble matter was removed by filtration with celite, and the solvent was distilled off. The obtained residue was roughly purified by silica gel column chromatography (hexane-ethyl acetate), and purified by reverse phase to obtain compound 39 (40 mg, yield 12%).

1H-NMR(CDCl3)δ: 8.63(s,1H),7.64-7.61(m,3H),7.34-7.19(m,4H),6.83-6.79(m,2H),5.58(d,J=10.3Hz ,1H),5.33(d,J=10.3Hz,1H),4.96-4.94(m,1H),4.44-4.40(m,1H),4.19(s,2H),3.22-3.18(m,1H), 3.09-3.02(m,2H),2.27-2.23(m,1H),2.01-1.97(m,1H),1.86-1.84(m,2H),1.74-1.70(m,1H),1.43-1.40(m ,1H),1.17(t,J=7.0Hz,3H).

step 3

By the same method as Step 6 of Example 1, compound I-2 (22 mg, yield 67%) was obtained.

1H-NMR (CDCl ₃ )δ: 11.81 (brs, 1H), 8.59 (s, 1H), 7.54 (s, 1H), 7.20-7.18 (m, 1H), 6.83-6.78 (m, 2H), 5.11- 5.09(m,1H),4.40-4.31(m,1H),4.18(s,2H),3.24-3.02(m,3H),2.34-2.29(m,1H),2.09-2.01(m,1H), 1.90-1.85(m,2H),1.78-1.74(m,1H),1.46-1.36(m,1H),1.23(t,J=7.0Hz,3H).

Example 8

step 1

To a solution of compound 40 (528 mg, 2.10 mmol) in THF (5.0 mL), sodium hydride (60 wt%, 135 mg, 3.38 mmol) was added at 0°C, and stirred at 0°C for 10 minutes. Compound 41 (500 mg, 2.415 mmol) was added to the reaction solution, and the temperature was raised to room temperature to react overnight. Water was added to the reaction liquid, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The obtained residue was purified by silica gel column chromatography (hexane-ethyl acetate) to obtain compound 42 (517 mg, yield 67%).

MS: m/z=321[M+H]+

step 2

溶液(1.8mL)，在90℃反應7小時。將水添加至反應液中，並用乙酸乙酯萃取。將有機層用飽和食鹽水洗淨，用無水硫酸鈉乾燥後，餾除溶劑。所得之殘渣透過矽膠柱層析(乙酸乙酯-甲醇)粗精製。 Compound 42 (89 mg, 0.278 mmol), compound 37 (60 mg, 0.139 mmol), cesium carbonate (68 mg, 0.208 mmol), tetrakis(triphenylphosphine) palladium (16 mg, 0.014 mmol) were dissolved in di

solution (1.8 mL), reacted at 90°C for 7 hours. Water was added to the reaction liquid, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. The resulting residue was crudely purified by silica gel column chromatography (ethyl acetate-methanol).

MS: m/z=546[M+H]+

step 3

Compound II-100 was obtained by the same method as step 6 of Example 1.

1H-NMR (CDCl ₃ )δ: 8.66(s,1H), 7.77(s,1H), 7.72(s,1H), 7.20-7.15(m,1H), 6.86-6.80(m,2H), 5.35( s,2H),5.08(s,1H),4.40-4.30(m,1H),3.20-2.95(m,3H),2.35-2.25(m,1H),2.01-1.40(m,6H),1.22( t,J=7.2Hz,3H).

Using the above-mentioned general synthesis method or the synthesis method described in the Examples, the following compounds were synthesized in the same manner.

[Table 1]

[Table 2]

[table 3]

[Table 4]

[table 5]

[Table 6]

[Table 7]

[Table 8]

[Table 9]

[Table 10]

[Table 11]

The physical data of each compound are shown below.

[Table 12]

Hereinafter, biological test examples of the compounds of the present invention are described.

Test Example 1: Anti-HIV Activity

A stepwise dilution series of the test samples was prepared in a 96-well microtiter plate (50 μL/well). After dispensing 2.5×10 ⁵ cells/mL of MT-4 cell suspension at 100 μL/well each into the plate into which the test sample was injected, HIV virus liquid was dispensed at 50 μL/well each. Mix with a pan mixer and incubate for 4 days in a _CO incubator. 30 μL of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) solution was dispensed into each well. React for 1 hour in a _CO incubator. 150 [mu]L of supernatant was removed from each well without aspirating the cells. Add 150 μL of cell lysate and mix well with a pan mixer until all cells are lysed. The absorbance of the mixed plate was measured at two wavelengths of 560nm/690nm with a microplate detector. The 50% HIV inhibitory concentration (EC50) was determined from the concentration dependence curve using the four parameter logistic curve fitting model shown below.

y=A+((BA)/(1+(C/x) ^D ))

A = minimum value of inhibition rate (negative control, 0%)

B = maximum value of inhibition rate (positive control, 100%)

C = compound concentration at the turning point

D = slope coefficient

x = compound concentration

y=inhibition rate (%)

(result)

[Table 13]

From the above test results, it can be known that the compounds of the present invention are useful as HIV medicines because they exhibit high anti-HIV activity.

Test Example 2: Drug Resistance Evaluation Test

A stepwise dilution series of the test samples was prepared in a 96-well microtiter plate (50 μL/well). After dispensing 2.5×10 ⁵ cells/mL of HeLa-CD4 cell suspension at 100 μL/well each into the plate containing the test sample, HIV virus liquid (wild strain and mutant strain) was dispensed at 50 μL/well each. Mix with a pan mixer and incubate for 3 days in a _CO incubator. The culture supernatant of each well was removed by suction, and 100 μL of the cell lysis buffer in the reporter assay kit was dispensed, followed by freezing in a freezer (-80° C.). The plate frozen in the freezer was thawed at room temperature, mixed with a plate mixer, and centrifuged at 1,200 rpm for 5 minutes. Dispense 20 μL of the supernatant from each well into a 96-well microplate (BLACK), and then inject 100 μl of the chemiluminescent reagent in the reporter assay kit. After reacting at room temperature for about 1 hour, use MicroBeta TRILUX Measure the amount of luminescence. The 50% HIV inhibitory concentration (EC50) was determined from the concentration dependence curve using the four parameter logistic curve fitting model shown below.

y=A+((BA)/(1+(C/x) ^D ))

A = minimum value of inhibition rate (negative control, 0%)

B = maximum value of inhibition rate (positive control, 100%)

C = compound concentration at the turning point

D = slope coefficient

x = compound concentration

y=inhibition rate (%)

Furthermore, the degree of drug resistance (fold difference (FC)) of each mutant strain was calculated according to the following calculation formula.

FC=EC50 of mutant strain/EC50 of wild strain

(result)

FC relative to mutant strain 1 (E138K/G140S/Q148H/N155H) and FC relative to mutant strain 2 (E92Q/E138T/G140S/Q148H) are shown in the table.

[Table 14]

FC relative to mutant strain 3 (E92Q/E138K/G140S/Q148H)

Compound I-032: 7.7

Compound I-011: 7.7

FC relative to mutant strain 4 (T97A/E138T/G140S/Q148H)

Compound I-032:10

Compound I-011: 3.2

From the above test results, it can be seen that the compound of the present invention has a high barrier to drug resistance and prevents the generation of HIV drug-resistant viruses.

Test Example 3: CYP Inhibition Test

Using commercially available pooled human liver microsomes (Pooled human liver microsomes), the 7-ethoxyresorufin that belongs to the typical substrate metabolic reaction of the major human CYP5 molecular species (CYP1A2, 2C9, 2C19, 2D6, 3A4) was adjacent to - Deethylation (CYP1A2), methyl-hydroxylation of tolbutamide (CYP2C9), 4'-hydroxylation of Mephenytoin (CYP2C19), ortho-de-hydroxylation of dextromethorphan Methylation (CYP2D6) and hydroxylation of Terfenadine (CYP3A4) were used as indicators to evaluate the degree of inhibition of the production of various metabolites by the compound of the present invention.

The reaction conditions are as follows, substrate: ethoxy resorufin (CYP1A2) 0.5 μmol/L, tolbutamide (CYP2C9) 100 μmol/L, S-mephenytoin (CYP2C19) 50 μmol/L, dextromethorphan ( CYP2D6) 5 μmol/L, tefenadine (CYP3A4) 1 μmol/L; Reaction time: 15 minutes; Reaction temperature: 37°C; Enzyme: mixed human liver microsome 0.2mg protein/mL; Compound concentration of the present invention: 1,5 , 10, 20 μmol/L (4 points).

In the 50mmol/L Hepes buffer in a 96-well plate, each of the five substrates, human liver microsomes and the compound of the present invention was added according to the above composition, and the coenzyme NADPH was added to start the metabolic reaction as an indicator. After reacting at 37° C. for 15 minutes, the reaction was terminated by adding methanol/acetonitrile=1/1 (V/V) solution. After centrifugation at 3000rpm for 15 minutes, use a fluorescent multi-label counter or LC/MS/MS to quantify resorufin (CYP1A2 metabolite) in the supernatant. LC/MS/MS quantification of tolbutamide hydroxide (CYP2C9 metabolite), mephenytoin 4'-hydroxide (CYP2C19 metabolite), dextromethorphan (CYP2D6 metabolite), terfenadine alcoholic body (CYP3A4 metabolites).

In the reaction solution, instead of the compound of the present invention, only DMSO, which is a solvent for dissolving the compound, was added as a control group (100%), and the residual activity (%), concentration and inhibition rate were calculated, and the inverse The estimated IC ₅₀ was calculated.

Test Example 4: CYP3A4(MDZ) MBI test

This is a test for evaluating the mechanism-based inhibition (MBI) ability of the compound of the present invention for the inhibition of CYP3A4 by enhancing the inhibitory effect due to the metabolic reaction of the compound of the present invention. CYP3A4 inhibition was assessed using pooled human liver microsomes using 1-hydroxylation of midazolam (Midazolam, MDZ) as an indicator.

The reaction conditions are as follows, substrate: MDZ 10 μmol/L; pre-reaction time: 0 or 30 minutes; substrate metabolism reaction time: 2 minutes; reaction temperature: 37°C; mixed human liver microsomes: 0.5 mg/mL during pre-reaction , 0.05 mg/mL during the reaction (10-fold dilution); the concentration of the compound of the present invention during the pre-reaction: 1, 5, 10, 20 μmol/L (4 points) or 0.83, 5, 10, 20 μmol/L (4 points ).

In the K-Pi buffer (pH 7.4) as a pre-reaction solution in a 96-well plate, add and mix human liver microsomes and the compound solution of the present invention according to the above-mentioned pre-reaction composition, and then transfer a part of it to another Dilute the 96-well plate to 1/10 with the K-Pi buffer containing the matrix, add NADPH which is a coenzyme, and start the reaction as an indicator (no pre-reaction: pre-incubation for 0 minutes), after the predetermined time of reaction, by The reaction was terminated by adding methanol/acetonitrile=1/1 (V/V) solution. Furthermore, NADPH is added to the remaining pre-reaction solution to start the pre-reaction (there is a pre-reaction: pre-incubation 30 minutes), and after the pre-reaction predetermined time, a part of it was transferred to another plate, and diluted to 1/10 with the K-Pi buffer containing the matrix, and the reaction as an indicator was started. After reacting for a predetermined time, the reaction was terminated by adding methanol/acetonitrile=1/1 (V/V) solution. After centrifuging each plate for the index reaction at 3000 rpm for 15 minutes, 1-midazolam hydroxide in the supernatant was quantified by LC/MS/MS.

In the reaction solution, instead of the compound of the present invention, only DMSO, which is a solvent for dissolving the compound, was added as a control group (100%), and the residual activity (%) when the compound of the present invention was added at various concentrations was calculated, and the concentration and the concentration were used. Inhibition rate, IC was calculated by inverse estimation in logarithmic mode. The IC of pre-incubation for 0 minutes/IC of pre-incubation for 30 minutes was used as the shifted IC value. If the shifted IC is 1.5 or more, it is positive (+), and if the shifted IC is 1.0 or less, it is negative ( -).

(result)

Compound I-032: (-)

Compound II-058: (-)

Compound II-117: (-)

Compound II-130: (-)

Test example 5: BA test

Oral Absorbability Review Experimental Materials and Methods

(1) Animals used: Rats were used.

(2) Breeding conditions: Rats were allowed to ingest solid feed and sterilized tap water freely.

(3) Dosage amount and group setting: Oral administration and intravenous administration with a predetermined dosage. Grouping is set as follows. (Dosage varies according to each compound)

Oral administration of 2 to 60 μmol/kg or 1 to 30 mg/kg (n=2 to 3)

Intravenous administration of 1 to 30 μmol/kg or 0.5 to 10 mg/kg (n=2 to 3)

(4) Preparation of administration solution: Oral administration is administered as a solution or suspension. Intravenous administration is administered after solubilization.

(5) Administration method: Oral administration is forcibly administered into the stomach by an oral probe. Intravenous administration was via the tail vein via a syringe with a needle attached.

(6) Evaluation items: blood was collected over time, and the concentration of the compound of the present invention in plasma was measured using LC/MS/MS.

(7) Statistical analysis: Aiming at the change of the concentration of the compound of the present invention in plasma, the area under the concentration-time curve (AUC) in plasma was calculated by the moment analysis method, and the results were determined from the oral administration group and the intravenous administration group. The bioavailability (BA) of the compounds of the present invention was calculated from the dose ratio and AUC ratio of the groups.

Test Example 6: Clearance Evaluation Test

Experimental materials and methods

(1) Animals used: Rats were used.

(2) Breeding conditions: Rats were allowed to ingest solid feed and sterile tap water freely.

(3) Dosage and grouping setting: the predetermined dosage is administered intravenously. Grouping is set as follows.

Intravenous administration of 1 μmol/kg (n=2)

(4) Preparation of dosing solution: Solubilized in a solvent of dimethylsulfoxide/propylene glycol=1/1, and then administered.

(5) Administration method: administration from the tail vein by a syringe with a needle attached.

(6) Evaluation items: blood was collected over time, and the concentration of the compound of the present invention in plasma was measured using LC/MS/MS.

(7) Statistical analysis: With regard to the concentration transition of the compound of the present invention in plasma, the systemic clearance rate (CLtot) and the disappearance half-life (t1/2) were calculated by the momentum analysis method.

(result)

Compound I-032: 0.111 mL/min/kg, 12.3 hr

Compound II-023: 0.102mL/min/kg, 26.7hr

Compound II-104: 0.0226mL/min/kg, 35.4hr

Compound II-110: 0.0364mL/min/kg, 23.6hr

From the above test results, it can be seen that the compounds of the present invention can be used as persistent integrase inhibitors due to their low clearance rate and long disappearance half-life.

Test Example 7: Metabolic Stability Test

The commercially available mixed human liver microsomes were reacted with the compound of the present invention for a certain period of time, the residual rate was calculated by comparing the reacted sample with the unreacted sample, and the degree of the compound of the present invention being metabolized in the liver was evaluated.

In 0.2 mL of buffer (Tris-HCl 50 mmol/L, pH 7.4, potassium chloride 150 mmol/L, magnesium chloride 10 mmol/L) containing 0.5 mg protein/mL of human liver microsomes, in the presence of 1 mmol/L NADPH, the The reaction was carried out at 37° C. for 0 or 30 minutes (oxidation reaction). After the reaction, 50 μL of the reaction solution was added to 100 μL of a methanol/acetonitrile=1/1 (v/v) solution, mixed, and centrifuged at 3000 rpm for 15 minutes. The compound of the present invention in the centrifuged supernatant was quantified by LC/MS/MS or solid phase extraction (SPE)/MS, and the amount of the compound at 0 minutes of reaction was taken as 100% to calculate the residual amount of the compound of the present invention after the reaction.

(Results) The residual rate at a compound concentration of 0.5 μmol/L is shown in the table below.

[Table 15]

Test Example 8: Fluctuation Ames Test

The mutagenicity of the compounds of the invention was evaluated.

Inoculate 20 μL of frozen-preserved Salmonella typhimurium (Salmonella typhimurium strain TA98, TA100 strain) in 10 mL of liquid nutrient medium (2.5% Oxoid nutrient broth No.2), and culture at 37°C for 10 hours before shaking. Centrifuge 7.70 to 8.00 mL of the bacterial liquid of the TA98 strain (2000×g, 10 minutes) and remove the culture liquid. Suspend the bacteria in Micro F buffer with the same capacity as the bacteria solution used for centrifugation (K ₂ HPO ₄ : 3.5g/L, KH ₂ PO ₄ : 1g/L, (NH ₄ ) ₂ SO ₄ : 1g/L, lemon Natrate trisodium dihydrate: 0.25g/L, MgSO ₄ .7H ₂ O: 0.1g/L), and added to 120mL exposure medium (Containing biotin: 8μg/mL, histidine: 0.2 μg/mL, glucose: 8 mg/mL in MicroF buffer). For the TA100 strain, 3.10 to 3.42 mL of the bacterial liquid was added to 120 to 130 mL of the exposure medium to prepare a test bacterial liquid. 12 μL of the DMSO solution of the compound of the present invention (several stages of dilution from the highest dosage of 50 mg/mL with a common ratio of 2 to 3 times), DMSO as a negative control group, and DMSO as a positive control group were placed under non-metabolic activation conditions, 4-nitroquinoline-1-oxide DMSO solution at 50 μg/mL for TA98 strain, 2-(2-furyl)-3-(5-nitro-2- Furanyl) acrylamide DMSO solution, 40 μg/mL 2-aminoanthracene DMSO solution for TA98 strain, 20 μg/mL 2-aminoanthracene DMSO solution for TA 100 strain under metabolic activation conditions, and 588 μL of the test bacteria solution was mixed (under the condition of metabolic activation, the test bacteria solution was a mixture of 498 μL and 90 μL of S9 mix), and incubated at 37°C for 90 minutes with shaking. Combine 460 μL of bacterial solution exposed to the compound of the present invention with indicator medium (containing biotin: 8 μg/mL, histidine: 0.2 μg/mL, glucose: 8 mg/mL, bromocresol purple: 37.5 μg/mL) MicroF buffer) 2300 μL was mixed, and 50 μl each was dispensed into 48 wells of a microplate, and cultured statically at 37° C. for 3 days. The wells containing bacteria that acquired the ability to proliferate due to the mutation of the amino acid (histidine) synthesis enzyme gene changed from purple to yellow due to pH changes, so the bacteria that changed color to yellow in 48 wells per 1 dose were counted Proliferate wells, compare to negative control and evaluate. Those with negative mutagenicity are expressed as (-), and those with positive lines are expressed as (+).

Test Example 9: hERG test

For the purpose of risk assessment of QT interval in electrocardiogram of the compound of the present invention, using CHO cells expressing the human ether-a-go-go related gene (hERG) channel, the delayed rectification that the compound of the present invention plays an important role in the process of ventricular repolarization was examined. Effect of K ⁺ current (I _Kr ).

乙磺酸、4-(2-羥乙基)-1-哌

乙磺酸)：10mmol/L，pH=7.4)作為媒介，將媒介及溶解有目標濃度的本發明化合物溶解之細胞外液分別在室溫條件下適用於細胞7分鐘以上。由所得的I_Kr使用解析軟體(QPatch Assay軟體；Sophion Bioscience A/S)，將保持膜電位中的電流值作為基準計測最大尾電流的絕對值。進一步，將本發明化合物適用後的最大尾電流相對於媒介適用後的最大尾電流作為抑制率算出，評估本發明化合物對I_Kr的影響。 Using the automatic patch clamp system (QPatch; Sophion Bioscience A/S) to maintain the cell membrane potential at -80mV through the whole-cell patch clamp method, and after applying a leak potential of -50mV, a +20mV depletion I _Kr evoked by 2 s of polarizing stimulation followed by 2 s of repolarizing stimulation at -50 mV was recorded. Adjust dimethylsulfoxide to 0.1% extracellular fluid (NaCl: 145mmol/L, KCl: 4mmol/L, CaCl2: 2mmol/L, MgCl2: 1mmol/L, glucose: 10mmol/L, HEPES (4- (2-Hydroxyethyl)-1-piperene

Ethylsulfonic acid, 4-(2-hydroxyethyl)-1-piperene

Ethylsulfonic acid): 10mmol/L, pH=7.4) was used as a medium, and the medium and the extracellular fluid dissolved with the compound of the present invention dissolved in the target concentration were applied to the cells at room temperature for more than 7 minutes. Using analysis software (QPatch Assay software; Sophion Bioscience A/S) from the obtained I _Kr , the absolute value of the maximum tail current was measured with the current value in the membrane potential maintained as a reference. Furthermore, the maximum tail current after application of the compound of the present invention was calculated as the inhibition rate relative to the maximum tail current after application of the vehicle, and the effect of the compound of the present invention on I _Kr was evaluated.

Test Example 10: Solubility Test

The solubility of the compounds of the present invention was determined with the addition of 1% DMSO. Prepare a 10 mmol/L compound solution with DMSO. 2 µL of the compound solution of the present invention was added to 198 µL of the JP-1 solution and the JP-2 solution, respectively. After shaking at room temperature for 1 hour, the mixture was filtered by suction. Dilute the filtrate 10 or 100 times with methanol/water=1/1(V/V) or acetonitrile/methanol/water=1/1/2(V/V/V), and use LC by absolute calibration curve method /MS or solid phase extraction (SPE)/MS to determine the concentration in the filtrate.

The composition of JP-1 liquid is as follows.

Add water to 2.0g of sodium chloride and 7.0mL of hydrochloric acid to make up to 1000mL.

The composition of JP-2 is as follows.

Dissolve 3.40 g of potassium dihydrogen phosphate and 3.55 g of anhydrous disodium hydrogen phosphate in water to make 1000 mL, and add 1 volume of water to 1 volume.

Test Example 11: Powder Solubility Test

Put an appropriate amount of the compound of the present invention into an appropriate container, and add 200 μL of JP-1 solution (2.0 g of sodium chloride, 7.0 mL of hydrochloric acid to make 1000 mL with water), JP-2 solution (with phosphoric acid Potassium dihydrogen 3.40g and anhydrous disodium hydrogen phosphate 3.55g were dissolved in water to make 1000mL, and water was added to 1 volume), sodium taurocholate (TCA)/JP-2 liquid (add to TCA 1.08g JP-2 liquid and make 100mL) 20mmol/L. After adding the test solution and dissolving the entire amount, the compound of the present invention is appropriately added. After sealing and shaking at 37° C. for 1 hour, it was filtered, and 100 μL of methanol was added to 100 μL of each filtrate to perform a 2-fold dilution. Dilution ratios were changed as needed. Seal and shake after confirming that there are no air bubbles and precipitates. Compounds of the invention were quantified by HPLC by the absolute calibration curve method.

Test Example 12: Angle's Test

Salmonella typhimurium (Salmonella typhimurium) TA98 strain, TA100 strain, TA1535 strain, TA1537 strain and Escherichia coli (Escherichia coli) WP2uvrA strain were used as test strains, and the mutagenicity of the compound of the present invention was evaluated by Angle's test. In 0.1mL of the DMSO solution of the compound of the present invention, mix 0.5mL of S9 mix under the condition of metabolic activation, mix 0.5mL of phosphate buffer saline and 0.1mL of the test bacteria solution under the condition of non-metabolic activation, and then mix it with histidine and Multilayers of biotin or tryptophan were overlayed together with 2 mL of soft agar on minimal glucose agar plates. At the same time, for the negative control substance (DMSO) and the positive control substance (2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, sodium azide, 9-aminoacridine Pyridine or 2-aminoanthracene) is also carried out in the same way. After culturing at 37° C. for 48 hours, count the revertant colonies that appeared and compare them with the negative control group for evaluation. The number of back mutant colonies increases in a concentration-dependent manner, and when the number of colonies is more than twice that of the negative control group, it is judged as positive (+).

Test Example 13: Nav Test

For the purpose of assessing the risk of arrhythmia induced by the compounds of the present invention, HEK cells expressing a potential-gated sodium channel (Voltage gated sodium channel, Nav1.5 channel) encoding the SCN5A gene were used to examine the effects of the compounds of the present invention on myocardial depolarization. The role of Na ⁺ current (I _Na ), which plays an important role in the process.

乙磺酸、4-(2-羥乙基)-1-哌

乙磺酸)：10mmol/L、TEA(四乙基氫氧化銨)：10mmol/L，pH=7.4)作為媒介，將媒介及溶解有目標濃度的本發明化合物的細胞外液分別在室溫條件下適用於細胞5分鐘以上。由所得的I_Na使用分析軟體(QPatch Assay軟體；Sophion Bioscience A/S)，以保持膜電位中的電流值作為基準計測最大峰值電流的絕對值。進一步，算出本發明化合物適用時的最大峰值電流相對於媒介適用時的最大峰值電流的比率，以評估本發明化合物對I_Na的影響。 Using the automatic patch clamp system (QPatch; Sophion Bioscience A/S) to maintain the cell membrane potential at -100mV through the whole-cell patch clamp method, record the I _Na induced by depolarizing stimulation of -10mV for 20ms . Dimethylsulfone was adjusted to 0.3% extracellular fluid (NaCl: 145mmol/L, KCl: 4mmol/L, CaCl ₂ : 2mmol/L, MgCl ₂ : 1mmol/L, glucose: 10mmol/L, HEPES (4 -(2-Hydroxyethyl)-1-piperene

Ethylsulfonic acid, 4-(2-hydroxyethyl)-1-piperene

Ethylsulfonic acid): 10mmol/L, TEA (tetraethylammonium hydroxide): 10mmol/L, pH=7.4) as the medium, the medium and the extracellular fluid of the compound of the present invention dissolved with the target concentration were respectively at room temperature Next apply to cells for 5 min more. Using analysis software (QPatch Assay software; Sophion Bioscience A/S) from the obtained I _Na , the absolute value of the maximum peak current was measured with the current value in the membrane potential maintained as a reference. Furthermore, the ratio of the maximum peak current when the compound of the present invention was applied to the maximum peak current when the compound was applied was calculated to evaluate the effect of the compound of the present invention on _Ina .

(result)

Compound I-007 103%

Compound I-011 102%

Compound I-022 97.1%

Compound I-023 101%

Compound I-032 92.1%

Compound II-026 79%

Compound II-098 96.7%

Compound II-106 109%

Compound II-109 93.3%

Compound II-110 89.3%

Compound II-117 88.8%

Compound II-118 86.2%

Compound II-120 78.8%

Compound II-158 90.7%

From the above results, it can be seen that a significant increase in current was not observed, and the compound of the present invention has low suspicion of arrhythmia caused by an increase in Na current.

Test Example 14: Anti-HIV Activity Evaluation Test Using Peripheral Blood Mononuclear Cell (PBMC) of Healthy Persons

A stepwise dilution series of test samples was prepared in a 96-well microtiter plate (50 μL/well). Mix 2.5×10 ⁵ cells/well of PBMCs stimulated with Phytohemagglutinin (PHA) and HIV virus liquid according to the number of holes required, and react at 37°C for 1 hour. After the reaction, the cell suspension was centrifuged and the supernatant was discarded, and the infected cells were dispersed into the culture solution of the required number of wells at 150 μL/well, and 150 μL/well were dispensed into the 96-well microplate containing the test sample. Mix with a pan mixer and incubate for 4 days in a _CO incubator. The reverse transcriptase activity in the culture medium was measured. The 90% HIV inhibitory concentration (EC90) was determined from the concentration dependence curve using the four parameter logistic curve fitting model shown below.

y=A+((BA)/(1+(C/x) ^D ))

A = minimum value of inhibition rate (negative control, 0%)

B = maximum value of inhibition rate (positive control, 100%)

C = compound concentration at the turning point

D = slope coefficient

x = compound concentration

y=inhibition rate (%)

(result)

Compound I-007 1.0nM

Compound II-026 0.73nM

Compound II-045 3.3nM

Compound II-109 1.7nM

Test Example 15: Anti-HIV Activity Evaluation Test in the Presence of Human Serum Protein

A stepwise dilution series of test samples was prepared in a 96-well microtiter plate (50 μL/well). 100 μL/well of a human serum protein solution (human serum protein concentration: 50%) was dispensed into the disk containing the test sample, and left to stand at room temperature for 1 hour. The culture solution was dispensed at 100 μL/well for each plate system in the absence of serum. Mix 3.0×10 ⁵ cells/well of MT-4 cells and 3 μL/well of HIV virus solution according to the required number of wells, and react at 37°C for 1 hour. After the reaction, centrifuge the cell suspension and discard the supernatant, disperse the infected cells at 50 μL/well into the culture solution of the required number of wells, and inject 50 μL each into the 96-well microplate containing the test sample and human serum albumin /well (final human serum protein concentration: 25%). Mix with a pan mixer and incubate for 4 days in a _CO incubator. 30 μL of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) solution was dispensed into each well. React for 1 hour in a _CO incubator. 150 [mu]L of supernatant was removed from each well without aspirating the cells. Add 150 μL of cell lysate and mix well with a pan mixer until all cells are lysed. The absorbance of the mixed plate was measured at two wavelengths of 560nm/690nm with a microplate detector. The 50% HIV inhibitory concentration (EC50) was determined from the concentration dependence curve using the four parameter logistic curve fitting model shown below.

y=A+((BA)/(1+(C/x) ^D ))

A = minimum value of inhibition rate (negative control, 0%)

B = maximum value of inhibition rate (positive control, 100%)

C = compound concentration at the turning point

D = slope coefficient

x = compound concentration

y=inhibition rate (%)

In addition, potency shift (PS) was calculated according to the following calculation formula. In addition, the PS line was taken as 100% of the extrapolated value of human serum protein concentration.

PS=4×(EC50 in the presence of 25% human serum albumin/EC50 in the absence of human serum albumin)

(result)

The PS in the presence of human serum albumin is shown in the table (100% extrapolated values).

Compound I-007 116

Compound II-026 364

Compound II-045 236

Compound II-109 56

Preparation example

The compound of the present invention can be administered by any previous route, especially enterally, such as orally, such as in the form of tablets or capsules; or parenterally, such as in the form of injection or suspension; topically, such as in the form of emulsion , gel, ointment or cream form; nasal form or suppository form as a pharmaceutical composition for administration. The pharmaceutical composition containing the compound of the present invention in free form or pharmaceutically acceptable salt form and at least one pharmaceutically acceptable carrier or diluent can be produced by mixing, granulating or film coating using conventional methods. For example, compositions for oral administration can be made into tablets, granules, and capsules containing excipients, disintegrants, binders, lubricants, etc., and active ingredients. In addition, the composition for injection can be made into a solution or a suspension, which can also be sterilized, and can also contain preservatives, stabilizers, buffers and the like.

[Industrial availability]

The compound of the present invention has integrase inhibitory activity and/or cell proliferation inhibitory activity against viruses, especially HIV. Therefore, it can be used for the prevention or treatment of various diseases or viral infections (such as AIDS) in which integrase is involved.

Claims (10)

A compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

In the formula, Ring A is any one of the following rings,

X1 is CR ^9a R ^9b or O; R ^5a , R ^5b , R ^6a , R ^6b , R ^7a and R ^7b are each independently hydrogen, alkyl, alkoxy or alkoxyalkyl; R ^5a and R ^6a , or R ^6a and R ^7a together with adjacent atoms can form an aromatic carbocycle which can be substituted by halogen, a non-aromatic carbocycle with 3 to 6 members which can be substituted by halogen or which can be substituted by halogen Substituted non-aromatic heterocycles with 4 to 6 members, except that when forming an aromatic carbocycle, R ^5b and R ^6b , or R ^6b and R ^7b together form a bond; R ^5b and R ^6b can form a bond together; R ^8a , R ^8b , R ^9a , R ^9b , R ^10a , R ^10b , R ^11a and R ^11b are each independently hydrogen, alkyl, alkoxy or alkoxyalkyl; R ^8a and R ^10a can form C1-C3 crosslinks together; R ^10a and R ^11a can form a 5-membered non-aromatic carbocyclic ring together with adjacent atoms; R ^9a and R ^9b can form a 4-membered non-aromatic carbocyclic ring or a 5-membered non-aromatic heterocyclic ring together with adjacent atoms; R ^8a and R ^9a can form a bond together; Ring B is a benzene ring; Q is any one of the following rings;

^Each R is independently halogen, alkyl, haloalkyl, alkoxy, cyano, or haloalkoxy; R ^2a and R ^2b are each independently hydrogen, alkyl or haloalkyl; R ³ is alkyl or haloalkyl; ^R4 is hydrogen or alkyl; and n is an integer of 1 to 3. The compound or a pharmaceutically acceptable salt thereof as described in item 1 of the patent claims, wherein R ³ is an alkyl group. The compound or a pharmaceutically acceptable salt thereof as described in claim 1, wherein R ¹ are each independently halogen. The compound or a pharmaceutically acceptable salt thereof as described in item 1 of the patent claims, wherein R ^2a is hydrogen, and R ^2b is hydrogen or an alkyl group. The compound or the pharmaceutically acceptable salt thereof as described in item 1 of the scope of the patent application, wherein, Q is a ring represented by the following formula

The compound or the pharmaceutically acceptable salt thereof as described in item 1 of the scope of the patent application, wherein R ^5a , R ^5b , R ^6a , R ^6b , R ^7a and R ^7b are each independently hydrogen, alkyl, or alkoxy or alkoxyalkyl; and R ^8a , R ^8b , R ^9a , R ^9b , R ^10a , R ^10b , R ^11a , and R ^11b are each independently hydrogen, alkyl, alkoxy, or alkoxyalkyl. The compound or the pharmaceutically acceptable salt thereof as described in item 1 of the scope of the patent application, wherein R ^5a , R ^5b , R ^6a , R ^6b , R ^7a and R ^7b are each independently hydrogen, alkyl, or alkoxy or alkoxyalkyl; R ^8a , R ^8b , R ^9a , R ^9b , R ^10a , R ^10b , R ^11a and R ^11b are each independently hydrogen, alkyl, alkoxy or alkoxyalkyl; ^Each R is independently halogen; R ^2a is hydrogen, and R ^2b is hydrogen or alkyl; and R ³ is an alkyl group. The compound described in Item 1 of the patent application or the pharmaceutically acceptable salt thereof, the compound is selected from the group consisting of the following compounds:

A pharmaceutical composition, which contains the compound described in any one of items 1 to 8 of the patent application or its pharmaceutically acceptable salt, and at least one pharmaceutically acceptable carrier and diluent. A use of the compound described in any one of items 1 to 8 of the patent application or a pharmaceutically acceptable salt thereof, which is used to manufacture a therapeutic agent and/or a preventive agent for HIV infection.