JP6710376B2 - HIV infection inhibitor - Google Patents

Description

The present invention relates to a novel compound having an HIV infection inhibitory effect. More specifically, the invention relates to novel CD4 mimic compounds capable of blocking the entry of HIV cells into host cells.

Human immunodeficiency virus (HIV) is known as a virus that causes acquired immunodeficiency syndrome (AIDS). HIV is not transmitted through the air, but is transmitted through three main routes: sexual transmission, blood transmission, and mother-to-child transmission, and there is no risk of air transmission. To date, approximately 78 million people worldwide have been infected with HIV, of which 39 million have died of AIDS-related diseases. Currently, although chemotherapy with drugs has been successful in suppressing AIDS, the cure of AIDS has not yet been achieved.

HIV includes HIV-1 and HIV-2, and HIV-1 is further classified into AK subtypes. HIV-1 is common in the Western Hemisphere, Europe, Asia, and central/south/eastern Africa, and HIV-2 is common in western Africa. Because HIV-2 is less infectious than HIV-1, which has many cases of infection, and the epidemic is limited in some areas, development of anti-HIV drugs and AIDS vaccine is mainly targeted at HIV-1. ..

HIV-1 is a type of retrovirus. The mature virus is spherical with a diameter of 100-110 nm, and has a nucleus (capsid) containing two copies of the single-stranded RNA genome, reverse transcriptase, integrase, etc., and an envelope surrounding it (Fig. 1 ). On the surface of HIV-1, coat proteins gp120 and gp41 are present in the form of trimers, and these are CD4, CXCR4, which are present on human CD4-positive T cells, helper T cells and macrophages. It binds specifically to CCR5 and plays an important role in the entry of HIV into host cells.

Currently, clinically used anti-HIV drugs are mainly enzyme inhibitors such as "reverse transcriptase inhibitors", "protease inhibitors" and "integrase inhibitors" that inhibit the action of HIV-specific enzymes. Is becoming Highly active anti-retroviral therapy (HAART), in which multiple drugs are administered in combination, has been established, but few invasion inhibitors block the entry of HIV into cells and it is an effective membrane fusion inhibitor. Only the clinical application of enfuvirtide and maraviroc as a CCR5 inhibitor. However, these drugs, while capable of suppressing the growth of the virus but not killing them, require regular lifelong drug administration. Therefore, the risk of side effect accumulation due to long-term administration and high treatment cost are problems. Furthermore, HIV easily mutates, making the emergence of drug-resistant viruses a major problem and making vaccine development difficult.

The first step in the process of HIV-1 entry into host cells is the interaction of the HIV-1 coat protein gp120 with the host cell surface protein CD4 (first receptor). Along with this interaction, the structure of gp120 changes significantly, exposing a region called the V3 loop. Next, the interaction of this V3 loop with the second receptor (coreceptor, CCR5 or CXCR4) exposes gp41 on the surface, penetrates the host cell membrane, and enters the host cell through membrane fusion ( (Fig. 2).

In 2005, by screening HIV-1 syncytium formation inhibition, a low molecular weight compound NBD-556 having an action of inhibiting the invasion was reported (Non-Patent Document 1). NBD-556 also attracted attention as a small molecule CD4 mimetic compound because it can bind to the Phe43-cavity of gp120, which is the interaction site of CD4, and induce structural changes in gp120. 2-4). However, NBD-556 has problems such as low anti-HIV activity, high cytotoxicity, and low water solubility.

Thereafter, structure-activity relationship studies using NBD-556 as a lead compound have been actively conducted (Non-Patent Documents 5 to 8).

Qian, Z. et al., Virology 339, 213-225 (2005) Schon, A. et al., Biochemistry 45, 10973-10980 (2006) Madani, N. et al., Structure 16, 1689-1701 (2008) Hillel, H. et al., Plos Pathogens 5, e1000360 (2009) Yamada, Y. et al., Bioorg. Med. Chem. Lett. 20, 354-358 (2010) Narumi, T. et al., Bioorg. Med. Chem. 19, 6735-6742 (2011) Nguyen, W. et al., Bioorg. Med. Chem. Lett. 22, 7106-7109 (2012) Narumi, T. et al., Bioorg. Med. Chem. 21, 2518-2526 (2013)

The group of the present inventors has so far synthesized a compound HAR-171 having two cyclohexyl groups by modifying the piperidine ring site of NBD-556, and this compound has lower cytotoxicity than NBD-556. , And found to have stronger anti-HIV activity (Narumi, T. et al., Bioorg. Med. Chem. 19, 6735-6742 (2011)). HAR-171 is a compound synthesized on the basis of the co-crystal structure of NBD-556 and gp120, intended to interact with the two amino acids Val430 and Asp368 existing near the entrance of the Phe43-cavity. However, in the co-crystal structure of NBD-556 and the molecular modeling analysis of HAR-171 and gp120, hydrophobic interaction with Val430 was confirmed, but no significant interaction with Asp368 was observed.

The present inventors, as a strategy for inhibiting HIV-1 entry into cells, act on the Phe43-cavity, which is the binding site of the first receptor CD4, to inhibit the interaction between gp120 and CD4. We investigated the acquisition of effective low molecular weight CD4 mimic compounds.

This inhibition strategy not only has a mechanism of action different from conventional ones, but also aims to develop new drugs that suppress HIV-1 resistance to HIV-1 in a sustained and effective manner.

Based on the above-mentioned molecular design strategy considering the interaction with Val430 and Asp368 existing near the entrance of Phe43-cavity, the present inventors have an even stronger anti-HIV activity and change the structure of gp120. We aimed to create a new CD4 mimic derivative capable of inducing CD4. Specifically, with the intention of forming a hydrophobic interaction with Val430 and an electrostatic interaction with Asp368, a monocyclohexyl type compound in which one cyclohexyl group was introduced into the piperidine ring of NBD-556 was used. We designed and synthesized various novel derivatives with enhanced properties, evaluated their anti-HIV activity, cytotoxicity, gp120 structural change inducing ability, and used the integrated computational chemistry system Molecular Operating Environment (MOE, Chemical Computing Group Inc.). The interaction mode was predicted by the docking simulation.

As a result, the cyclohexyl group of the monocyclohexyl type compound has a hydrophobic interaction with Val430, and the compound having a guanidino group introduced therein has a compound that electrostatically interacts with the carboxy group of the Asp368 side chain. In the above, it was possible to obtain a compound having an anti-HIV activity superior to that of a conventionally known compound and having low cytotoxicity. Furthermore, it was also found that the obtained compound produces a synergistic anti-HIV effect when used in combination with a neutralizing antibody that specifically recognizes the V3 loop exposed in the middle of the HIV entry mechanism.

［式中、
XはCl、Br、及びFから選ばれるハロゲン原子であり、
Aは炭素数1〜5のアルキレン基であり、
Bは、下記の式(II)〜(IV)：

（式中、R¹及びR²はそれぞれ独立して、カルボニル基を含んでも良い炭素数1〜5のアルキレン基である）、

（式中、R³はカルボニル基を含んでも良い炭素数1〜5のアルキレン基である）、及び

から選択される基である。］
で示される化合物又はその塩。
［２］XがClであり、Aがエチレンである、［１］記載の化合物又はその塩。
［３］R¹及びR²が共にエチレンである、［１］もしくは［２］記載の化合物又はその塩。
［４］R³が-CO-(CH₂)_n-(式中、nは1〜4である)である、［１］もしくは［２］記載の化合物又はその塩。
［５］［１］〜［４］のいずれか記載の化合物又はその塩を有効成分として含む、HIV感染阻害剤。
［６］［５］記載のHIV感染阻害剤を含む、HIV感染の治療又は予防のための医薬組成物。
［７］抗-HIV抗体と組み合わせて投与されることを特徴とする、［５］記載のHIV感染阻害剤、又は［６］記載の医薬組成物。
［８］抗-HIV抗体が、HIV-1表面上のV3ループに対して特異的な中和抗体である、［７］記載のHIV感染阻害剤、又は医薬組成物。 That is, the present invention is as follows.
[1] General formula (I):

[In the formula,
X is a halogen atom selected from Cl, Br, and F,
A is an alkylene group having 1 to 5 carbon atoms,
B is represented by the following formulas (II) to (IV):

(In the formula, R ¹ and R ² are each independently an alkylene group having 1 to 5 carbon atoms which may include a carbonyl group),

(In the formula, R ³ is an alkylene group having 1 to 5 carbon atoms which may include a carbonyl group), and

Is a group selected from. ]
Or a salt thereof.
[2] The compound or salt thereof according to [1], wherein X is Cl and A is ethylene.
[3] The compound or salt thereof according to [1] or [2], wherein R ¹ and R ² are both ethylene.
[4] R ³ is -CO- (CH _{2) n} - (wherein, n is 1 to 4) is, [1] or [2] or a salt thereof, wherein.
[5] An HIV infection inhibitor containing the compound or salt thereof according to any one of [1] to [4] as an active ingredient.
[6] A pharmaceutical composition for treating or preventing HIV infection, which comprises the HIV infection inhibitor according to [5].
[7] The HIV infection inhibitor according to [5] or the pharmaceutical composition according to [6], which is administered in combination with an anti-HIV antibody.
[8] The HIV infection inhibitor or pharmaceutical composition according to [7], wherein the anti-HIV antibody is a neutralizing antibody specific for the V3 loop on the surface of HIV-1.

The present specification includes the disclosure content of Japanese Patent Application No. 2015-105723, which is the basis of priority of the present application.

INDUSTRIAL APPLICABILITY The present invention can provide a novel HIV infection inhibitor that has high anti-HIV activity and low cytotoxicity, and can bring about a synergistic anti-HIV activity when used in combination with a neutralizing antibody. The compound of the present invention has higher hydrophilicity compared to NBD-556 and is suitable for administration in an aqueous preparation such as an injection. Further, the HIV infection inhibitor of the present invention can be widely applied to HIV, which is known to cause mutations at a high frequency.

1 schematically shows the structure of HIV-1. 1 schematically shows the mechanism of HIV-1 entry into host cells. 7 shows the activity enhancing effect when the compound of the present invention is used in combination with the neutralizing antibody KD-247. A. YIR-819 (Compound 23), B. YIR-802 (Compound 29), C. YIR-821 (Compound 26), D. YIR-329 (Compound 11), E. NBD-556 5 shows the results of docking simulation of the compound of the present invention and HIV-1 Phe43-cavity. A. NBD-556, B. YIR-329 (Compound 11), C. YIR-821 (Compound 26) The result of the docking simulation of the compound of the present invention (YIR-819, compound 23) and Phe43-cavity of HIV-1 is shown.

Hereinafter, the present invention will be described in detail.

で示される化合物又はその塩を提供する。As described above, the present invention has the general formula (I):

And a salt thereof.

In the above formula, X is a halogen atom selected from Cl, Br, and F. The halogen atom X is preferably in the meta or para position with respect to the other substituent. Particularly preferably, X is Cl.

In the above formula, A is an alkylene group having 1 to 5 carbon atoms. Preferably, A is an alkylene group having 2 to 4 carbon atoms, particularly preferably ethylene.

（式中、R¹及びR²はそれぞれ独立して、カルボニル基を含んでも良い炭素数1〜5のアルキレン基である）
を有する基である。 In one embodiment of the invention, in the above formula, B is of formula (II):

(In the formula, R ¹ and R ² are each independently an alkylene group having 1 to 5 carbon atoms and optionally containing a carbonyl group)
Is a group having

Thus, in one embodiment, preferred compounds are compounds of formula (I) or salts thereof, wherein X is Cl, A is ethylene and B has the structure of formula (II).

In this embodiment, a particularly preferred compound is a compound of formula (I) or salt thereof, wherein X is Cl, A is ethylene, and R ¹ and R ² are both ethylene, for example, in the structure below: It is Compound 29 (YIR-802) shown.

（式中、R³はカルボニル基を含んでも良い炭素数1〜5のアルキレン基である）
を有する基である。 In another embodiment of the present invention, wherein B is of formula (III):

(In the formula, R ³ is an alkylene group having 1 to 5 carbon atoms which may contain a carbonyl group.)
Is a group having

Thus, in this embodiment, the preferred compound is a compound of formula (I) or salt thereof, wherein X is Cl, A is ethylene and B has the structure of formula (III).

In this embodiment, particularly preferred compounds, X is Cl, A is ethylene, R ³ is -CO- (CH _{2) n} - is (wherein, n is 1 to 4), wherein A compound of (I) or a salt thereof, for example, compound 23 (YIR-819), compound 26 (YIR-821), and compound 30 (YIR-913) shown by the following structures.

を有する基である。 In another embodiment of the present invention, wherein B is of formula (IV):

Is a group having

Thus, in this embodiment, preferred compounds are compounds of formula (I) or salts thereof, wherein X is Cl, A is ethylene and B has the structure of formula (IV), for example the structure of Is compound 20 (YIR-737).

The compound of general formula (I) can form a salt with an inorganic or organic base at the guanidino group moiety. The salt is not particularly limited, but a pharmaceutically acceptable salt is preferable, for example, hydrochloride, sulfate, phosphate, succinate, fumarate, mesylate, tosylate, bromide. Hydrochloride, acetate, trifluoroacetate and the like can be preferably used.

The synthetic procedure of the compound of the present invention will be specifically described below. Incidentally, for each new compound obtained, the compound name and the compound number given by the group of the present inventors are described together, but in the present specification, the same compound for both a certain compound and its salt The name and compound number may be used.

[1. Synthesis of monocyclohexyl type CD4 mimic derivative]
In NBD-556 and HAR-171, docking simulation results showed that no significant interaction was observed on one side of the hydrophobic functional group at the piperidine ring site. It was also considered that the presence of the hydrophobic functional groups on both sides increases the steric hindrance around the piperidine nitrogen atom and may interfere with the interaction with Asp368. Therefore, the present inventors first designed and synthesized a monocyclohexyl-type CD4 mimic derivative having a cyclohexyl group on only one side of the piperidine ring.

[1.1 Synthesis of monocyclohexyl 4-aminopiperidine]
Scheme 1-3 below shows a method for synthesizing monocyclohexyl 4-aminopiperidine.

Starting from compound 1 (phthalimide) as a starting material, it is treated with methyl vinyl ketone in the presence of a base to give compound 2 (Michael adduct), followed by acetal protection of the carbonyl group, followed by hydrazine decomposition to give compound 4 (ring). (Scheme 1).

Next, cyclohexanone is allowed to act on compound 4 to form compound 5 (imine), and then BF ₃ ·OEt ₂ complex is added as a Lewis acid and reacted under microwave irradiation to give compound 6 (cyclized compound). ). Then, by deprotecting the acetal group under acidic conditions, 2-cyclohexyl-4-oxopiperidine (Compound 7) was obtained (Scheme 2).

Then, the compound 7 was benzylated, followed by reductive amination with PMBNH ₂ to give the compound 9, followed by oxidative cleavage with ceric ammonium nitrate to give the desired 2-cyclohexyl-4-aminopiperidine ( Compound 10) was obtained (Scheme 3).

[1.2 Condensation of monocyclohexyl 4-aminopiperidine with aromatic ring unit]
Scheme 4 below shows a method for synthesizing a monocyclohexyl-type CD4 mimic derivative.

A variety of desired monocyclohexyl-type CD4 mimic derivatives were synthesized by condensing 2-cyclohexyl-4-aminopiperidine (Compound 10) with a carboxylic acid derivative of each aromatic ring unit using a condensing agent (Scheme 4). ..

[2. Synthesis of monocyclohexyl-type CD4 mimic derivative with functional group introduced on nitrogen atom of piperidine]
By making the piperidine unit a monocyclohexyl type, steric hindrance around the piperidine nitrogen atom is eliminated compared to NBD-556 and HAR-171, so it was difficult for NBD-556 and HAR-171 Modification is possible. Therefore, various monocyclohexyl-type CD4 mimic derivatives in which a functional group was introduced on the piperidine nitrogen atom of YIR-329 (Compound 11) were designed and synthesized (Scheme 5-12).

YIR-329 (Compound 11) was reacted with 1H-pyrazole-1-carboxyamidine hydrochloride to obtain guanidinoylated YIR-720 (Compound 15). The obtained compound was purified by HPLC and isolated as a trifluoroacetic acid salt (Scheme 5).

By reacting YIR-329 (Compound 11) with benzyl bromide, benzylated YIR-327 (Compound 16) was obtained (Scheme 6).

YIR-703 (Compound 17) was obtained by allowing 2-(N-(tert-butoxycarbonyl)amino)-O-(4-methylphenylsulfonyl)ethanol to act on YIR-329 (Compound 11). (Scheme 7).

YIR-329 (Compound 11) was reacted with bromoacetonilolyl to induce YIR-631 (Compound 18), and then the nitrile group was reduced with LiAlH ₄ to give YIR-723 (Compound 19). ) Got. The obtained compound was purified by HPLC and isolated as a trifluoroacetic acid salt (Scheme 8).

YIR-723 (Compound 19) was reacted with 1H-pyrazole-1-carboxyamidine hydrochloride to obtain guanidinoylated YIR-737 (Compound 20). The obtained compound was purified by HPLC and isolated as a trifluoroacetic acid salt (Scheme 9).

YIR-723 (Compound 19) was condensed with N-(tert-butoxycarbonyl)glycine using a condensing agent, and then the Boc group was deprotected with trifluoroacetic acid to obtain YIR-818 (Compound 22). .. Then, by reacting with 1H-pyrazole-1-carboxyamidine hydrochloride, guanidinoylated YIR-819 (Compound 23) was obtained. The obtained YIR-818 (Compound 22) and YIR-819 (Compound 23) were purified by HPLC and isolated as a trifluoroacetate salt (Scheme 10).

YIR-723 (compound 19) was condensed with N-(tert-butoxycarbonyl)-5-aminovaleric acid using a condensing agent, and then the Boc group was deprotected with trifluoroacetic acid to remove YIR-820 (compound 19). 25) got Then, by reacting with 1H-pyrazole-1-carboxyamidine hydrochloride, guanidinoylated YIR-821 (Compound 26) was obtained. The obtained YIR-820 (Compound 25) and YIR-821 (Compound 26) were purified by HPLC and isolated as a trifluoroacetate salt (Scheme 11).

YIR-723 (Compound 28) was treated with bromoacetonitrile to induce YIR-738 (Compound 27) and then reducing the nitrile group with LiAlH _4. Obtained. Then, by reacting with 1H-pyrazole-1-carboxyamidine hydrochloride, guanidinoylated YIR-802 (Compound 29) was obtained. The obtained YIR-801 (Compound 28) and YIR-802 (Compound 29) were purified by HPLC and isolated as a trifluoroacetate salt (Scheme 12).

As a result of evaluating anti-HIV activity using the various compounds obtained above, first, a monocyclohexyl compound YIR-329 (compound 11) having a chloro group at the para position was found by the present inventors previously. It was found to have better anti-HIV activity than the compound HAR-171. Moreover, the molecular modeling analysis of YIR-329 (Compound 11) suggested that the cyclohexyl group had a hydrophobic interaction with Val430. Next, in the derivative synthesized based on YIR-329 (Compound 11) with the intention of improving the electrostatic interaction with Asp368, the compound in which a guanidino group was introduced on the piperidine nitrogen atom had further anti-HIV activity. It was found to improve. Molecular modeling analysis of the obtained compound suggested that the introduced guanidino group formed an electrostatic interaction with the carboxy group of the Asp368 side chain.

Even compounds with high anti-HIV activity are not suitable as pharmaceuticals if they are highly toxic. Therefore, the obtained compound was evaluated for cytotoxicity in addition to anti-HIV activity. As a result, the compound represented by the above general formula (I) or a salt thereof has low anti-HIV activity and low cytotoxicity, and, surprisingly, may act synergistically in combination with an anti-HIV antibody. It was found. The compounds of the present invention are predicted to be capable of hydrophobic interactions with Val430 and electrostatic interactions with Asp368 or Asp474 and also to achieve adequate water solubility for application in pharmaceutical formulations. , Has a structure in which one cyclohexyl group is bonded to the piperidine ring and a guanidino group is bonded via a linker (spacer) of a certain length.

The water solubility of the compound of the present invention can be described, for example, by the retention time in high performance liquid chromatography (HPLC). The present inventors have carried out HPLC measurement on the compound of the present invention under the following conditions, and obtained the following results.
Eluent: 2-component gradient mode column using water and acetonitrile (both containing 0.1% TFA): 5C18-AR-II (4.6 x 250 mm) (Nacalai Tesque)
Flow rate: 1 mL/min [Pump: JASCO JV-2075 plus (manufactured by JASCO Corporation)]
Detection wavelength: 254 nm [Detector: JASCO PU-2089 plus (manufactured by JASCO Corporation)]

For example, when the gradient is [0 to 5 minutes: water/acetonitrile (100/0), 6 to 36 minutes: water/acetonitrile (80/20 → 50/50)], the retention time of YIR-737 (compound 20) Was 25.0 minutes and the retention time of YIR-802 (Compound 29) was 25.6 minutes.

When the gradient is [0 to 5 minutes: water/acetonitrile (100/0), 5 to 65 minutes: water/acetonitrile (0/100 → 100/0)], retention of YIR-819 (compound 23) The time was 34.5 minutes, and the retention time of YIR-821 (Compound 26) was 34.8 minutes.

Further, the water solubility of the compound of the present invention can be represented by using a partition coefficient representing the lipophilicity of the compound, for example, a clogP value which is a LogP value calculated by calculation. The present inventors, for the compound of the present invention, by the program Molinspiration property engine v2014.11 provided by Molinspiration property calculation service (http://www.molinspiration.com/), miLogP (Molinspiration calculated logP) value I calculated. The miLogP value is a numerical value corresponding to the clogP value.

As a result, the YIR-737 (compound 20) had a miLogP value of 2.16, the YIR-819 (compound 23) had a miLogP value of 1.76, the YIR-821 (compound 26) had a miLogP value of 2.31, and the YIR-802 (compound 29) had a miLogP value of 2.31. The miLogP value was 1.05. As a result of calculating the miLogP value for various compounds, it was found that the miLogP value of the compound of the present invention suitable as an HIV infection inhibitor falls within the range of about 1 to 2.5.

The compound of the present invention or a salt thereof can competitively inhibit the binding between Phe43-cavity of HIV gp120 and CD4. Also, without wishing to be bound by any theory, the conformational changes in gp120 caused by the compounds of the invention or salts thereof are not the same as those caused by the interaction with CD4 and expose the V3 loop, although Since it does not cause subsequent binding with coreceptors, it can inhibit the entry of HIV into host cells. Therefore, the present invention provides an HIV infection inhibitor containing the above-mentioned compound of the present invention or a salt thereof as an active ingredient.

The compound of the present invention or a salt thereof can be directly administered as an HIV infection inhibitor, but in addition to this active ingredient, carriers, excipients, preservatives, and oxidative stabilizers that are usually used in pharmaceutical compositions are provided. It is also possible to add a drug or the like as appropriate and administer it as a pharmaceutical composition. Therefore, the present invention also provides a pharmaceutical composition for treating or preventing HIV infection, which comprises the HIV infection inhibitor described above.

The HIV infection inhibitor and the pharmaceutical composition of the present invention can be orally and parenterally administered, for example, orally; by intravenous, intramuscular, transdermal, subcutaneous, intradermal, or intraperitoneal injection or infusion. However, it is not particularly limited. Those skilled in the art can appropriately determine a suitable administration route for administration of the HIV infection inhibitor and the pharmaceutical composition of the present invention.

The human dose of the HIV infection inhibitor of the present invention depends on the age, body weight, symptoms, etc. of the patient to be administered and is not particularly limited, but for example, 100 μg/kg body weight to 100 mg/kg body weight per day, and preferably Can be in the range of 500 μg/kg body weight to 50 mg/kg body weight, more preferably 1 mg/kg to 30 mg/kg body weight.

Furthermore, the HIV-infection inhibitor of the present invention can be used alone, but is also intended to be used in combination with other anti-HIV agents having inhibitory effects by different mechanisms. Other anti-HIV agents include, but are not limited to, reverse transcriptase inhibitors, protease inhibitors, integrase inhibitors and the like. Although not intended to be limited, for example, as a reverse transcriptase inhibitor, zidovudine, lamivudine, abacavir, tenofovir, emtricitabine, efavirenz, etc. as a protease inhibitor, atazanavir, darunavir, ritonavir, etc., as an integrase inhibitor, Raltegravir and the like can be mentioned. The anti-HIV infection inhibitor and the other anti-HIV drug can be included in the same or different pharmaceutical composition. Administration of the HIV infection inhibitor of the present invention and the other anti-HIV drug may be simultaneous, continuous, or completely different. The administration route of the HIV infection inhibitor of the present invention and the other anti-HIV drug may be the same or different.

The HIV-infection inhibitor of the present invention is also intended to be used in combination with an antibody specific to HIV. The antibody is not particularly limited, but it was revealed that the HIV-infection inhibitor of the present invention causes a structural change in gp120 by binding to HIV and exposes the V3 loop, and thus neutralization It is preferred to use antibodies, particularly neutralizing monoclonal antibodies and functional fragments thereof specific for the V3 loop of HIV.

Regarding anti-HIV monoclonal antibody, various researches and developments have been made in this field, and KD-247 (generic name: suvizumab), which is currently undergoing clinical trials, is being represented as a representative monoclonal antibody against the V3 loop of HIV. Can be mentioned. Details of the anti-HIV monoclonal antibody are described in, for example, Journal of Virology, June 2006, p.5552-5562; Journal of Virology, June 2006, p.5563-5570; Kaketsu Kenkyusho, Dawn, 23: 42-54 (2014). Patent Nos. 2989862 and 5526386 are listed, and the amino acid sequences of the VH and VL regions of KD-247 are also disclosed in the above Journal of Virology, June 2006, p.5552-5562. Those skilled in the art can obtain the KD-247 antibody and other anti-HIV antibodies based on these information.

Accordingly, the present invention provides the above HIV infection inhibitor or the above pharmaceutical composition, which is administered in combination with an anti-HIV antibody. In a preferred but non-limiting embodiment, the anti-HIV antibody is a neutralizing antibody specific for the V3 loop on the HIV-1 surface.

Administration of the HIV infection inhibitor of the present invention and the anti-HIV antibody may be simultaneous, sequential, or completely different. The HIV infection inhibitor of the present invention and the anti-HIV antibody may have the same administration route or different administration routes.

The present invention will be specifically described with reference to the following examples, but the present invention is not limited to these examples.

<Example 1>
[Synthesis of Compound of the Present Invention]
¹ H-NMR and ¹³ C-NMR spectra for compound analysis were recorded on a Bruker Avance III spectrometer. Chemical shifts are shown in values (ppm) relative to Me ₄ Si (in CDCl ₃ ) as an internal standard. Mass spectrometry was recorded on Bruker Daltonics microTOF focus in positive and negative modes. Wakogel C-200 (Wako Pure Chemical Industries, Ltd.) and silica gel 60 N (Kanto Chemical Co., Inc.) were used for flash chromatography. A Cosmosil 5C ₁₈ -ARII column (4.6 × 250 mm, Nacalai Tesque, Inc., Kyoto, Japan) was used for HPLC analysis, and a linear gradient of CH ₃ CN containing 0.1% (v/v) trifluoroacetic acid was used. Was used with JASCO PU-2089 plus (JASCO Corporation, Ltd., Tokyo, Japan) at a flow rate of 1 cm ³ min ^-1 , and elution of the product was detected by UV at 220 nm. Preparative HPLC was performed using a Cosmosil 5C ₁₈ -ARII column (20 × 250 mm, Nacalai Tesque, Inc.) with a JASCO PU-2087 plus (JASCO Corporation, Ltd., Tokyo, Japan) at 0.1% (v/v). ) Performed at a flow rate of 7 cm ³ min ⁻¹ with an appropriate gradient of CH ₃ CN solution containing trifluoroacetic acid. Microwave reactions were performed in the Initiator™ (Biotage) Biotage Microwave Reaction Kit (sealed vials).

[Synthesis of 2-cyclohexyl-4-oxopiperidine (Compound 7):]

The synthesis of compound 1 to compound 4 was performed using an existing protocol (Rasapalli, S. et al., Org. Biomol. Chem. 11, 4133-4137 (2013); Bosch, J. et al., J. Chem. Soc. , Perkin Trans. 1. 1533-1539 (1986)).

Cyclohexanone (1.71 mL, 16.5 mmol), paratoluenesulfonic acid monohydrate (143 mg, 0.75 mmol) and compound 4 in a stirred mixture of dry MgSO ₄ (7.5 g) in CH ₂ Cl ₂ (12 mL). (1.97 g, 15 mmol) was added at room temperature. The mixture was stirred at room temperature for 1.5 hours, and BF ₃ ·OEt ₂ (2.83 mL, 22.5 mmol) was added at room temperature. The mixture was stirred by microwave irradiation at 90° C. for 3 hours, quenched with a saturated NaHCO ₃ aqueous solution at 0° C., and extracted with CH ₂ Cl ₂ . The organic phase was dried over Na ₂ SO ₄ and then concentrated under reduced pressure to give a crude mixture as a dark brown oil. The crude mixture was dissolved in acetone (150 mL), and 15% aqueous HCl solution (8.75 mL, 45 mmol) was added. The mixture was stirred at room temperature for 93 hours and then concentrated under reduced pressure. The residue was diluted with an excess of 0° C. aqueous 3M NaOH solution (20 mL, 60 mmol) (pH=13-14) and extracted with CH ₂ Cl ₂ . The organic phase was dried over Na ₂ SO ₄ , then concentrated under reduced pressure and purified by column chromatography (CHCl ₃ /MeOH = 12/1-10/1) to give the title compound 7 as a dark brown oil. Obtained (817 mg, yield 33%). The synthetic route of compound 7 is also described in Ciblat, S. et al., Tetrahedron Lett. 42, 4815-4817 (2001).

[Synthesis of N-benzyl-2-cyclohexyl-4-oxopiperidine (Compound 8)]

Benzyl bromide (1.72 mL, 14.4 mmol) and i-Pr ₂ NEt (3.28 mL, 19.2 mmol) were added to a THF solution (23.9 mL) of compound 7 (800 mg, 4.79 mmol) at room temperature. The reaction mixture was stirred at 60° C. for 56 hours, quenched with a saturated NaHCO ₃ aqueous solution at 0° C., and extracted with CHCl ₃ . The organic phase was dried over MgSO ₄ and then concentrated under reduced pressure, followed by purification by column chromatography (hexane/AcOEt=5/1) to give the title compound 8 as a brown oil (950 mg, Yield 82%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.4-1.6 (m, 6H), 1.7-1.9 (m, 4H), 2.28-2.31 (t, J = 12.4 Hz, 2H), 2.39 (s, 2H) , 2.94-2.97 (t, J = 12.4 Hz, 2H), 3.84 (s, 2H), 7.24-7.28 (m, 1H), 7.32-7.36 (m, 2H), 7.40-7.42 (m, 2H); ¹³ C-NMR (125 MHz, CDCl ₃ ) δ 21.4, 25.9, 34.5, 37.5, 43.9, 49.3, 49.5, 60.5, 126.8, 128.1, 128.3, 140.6, 209.8; HRMS (ESI), C ₁₇ H ₂₄ NO [M+ H] ⁺ m/z: calculated 258.1852, found 258.1850.

[Synthesis of N-benzyl-2-cyclohexyl-4-((4-methoxyphenyl)methyl)aminopiperidine (Compound 9)]

4-Methoxybenzylamine (1.43 mL, 11.1 mmol) was added to a solution of compound 8 (950 mg, 3.69 mmol) in MeOH (36.9 mL). The reaction mixture was stirred at room temperature for 3 hours, then NaBH(OAc) ₃ (2.82 g, 13.3 mmol) was added at room temperature. The reaction mixture was stirred at room temperature for 15.5 hours, quenched with a saturated aqueous NaHCO ₃ solution at 0° C., and extracted with ethyl acetate. The organic phase was dried over MgSO ₄ , concentrated under reduced pressure and subsequently purified by column chromatography (CHCl ₃ /MeOH = 15/1-10/1) to give the title compound 9 as a yellow oil. (1.21 g, yield 87%).
¹ H-NMR (500 MHz, CDCl ₃ ) δ 1.02-1.07 (m, 1H), 1.2-1.75 (m, 6H), 1.77-1.83 (m, 1H), 2.18-2.22 (m, 1H), 2.56- 2.58 (m, 2H), 2.71-2.77 (m, 1H), 3.26-3.28 (m, 1H), 3.76-3.76 (m, 2H), 3.80 (s, 3H), 4.00-4.03 (m, 1H), 6.85-6.87 (m, 2H), 7.18-7.21 (m, 1H), 7.23-7.25 (m, 2H), 7.27-7.30 (m, 2H), 7.35-7.36 (m, 2H); ¹³ C-NMR ( 125 MHz, CDCl ₃ ) δ 22.2, 26.5, 31.6, 37.7, 39.4, 44.7, 50.3, 50.8, 51.2, 55.2, 56.1, 113.8, 126.3, 128.1, 128.1, 129.2, 133.0, 142.0, 158.5; HRMS (ESI), C ₂₅ H ₃₅ N ₂ O [M+H] ⁺ m/z: calculated 379..2744, found 379.2742.

[Synthesis of 2-cyclohexyl-4-aminopiperidine (Compound 10)]

Dicerium ammonium nitrate (5.22 g, 9.51 mmol) was added to a CH ₃ CN/H ₂ O (23.8/7.92 mL) solution of compound 9 (1.2 g, 3.17 mmol). The reaction mixture was stirred at room temperature for 14 hours, then ceric ammonium nitrate (3.48 g, 6.34 mmol) was added at room temperature. The reaction mixture was stirred at 1 room temperature for 7.5 hr, quenched with 3 M HCl aqueous solution (10.6 mL, 10 eq) at 0° C., and extracted with CH ₂ Cl ₂ . The aqueous phase was made basic with 5 M aqueous NaOH (12.7 mL, 20 eq) then filtered through Celite and washed with AcOEt. Extraction with AcOEt, followed by dehydration of the organic phase with Na ₂ SO ₄ and concentration under reduced pressure gave the title compound 10 (407 mg, yield 76%) as a yellow oil.
¹ H-NMR (400 MHz, CDCl ₃ ) δ0.81-0.87 (m, 1H), 1.03-1.14 (m, 1H), 1.3-1.7 (m, 13H), 1.81-1.88 (m, 2H), 2.77 -2.92 (m, 3H); ¹³ C-NMR (125 MHz, CDCl ₃ ) δ 21.2, 25.9, 31.0, 37.7, 39.2, 41.1, 44.5, 51.4; HRMS (ESI), C ₁₀ H ₂₁ N ₂ [M+ H] ⁺ m/z: calculated 169.1699, found 169.1695.

[Synthesis of N ¹ -(4-chlorophenyl)-N ² -(2-cyclohexylpiperidin-4-yl)oxalamide (Compound 11, YIR-329)]

N-(4-chlorophenyl)oxamic acid (473 mg, 2.38 mmol) in THF (10 mL) was added to HOBt・H ₂ O (729 mg, 4.76 mmol), EDCI・HCl (912 mg, 4.76 mmol). ), compound 10 (200 mg, 1.19 mmol) and NEt ₃ (0.824 mL, 5.95 mmol) were added at 0°C. The reaction mixture was stirred at room temperature for 13 hours, quenched with saturated NaHCO ₃ aqueous solution at 0° C., and extracted with CHCl ₃ . The organic phase was dried over Na ₂ SO ₄ , then concentrated under reduced pressure and purified by column chromatography (CHCl ₃ /MeOH = 7/1 to 4/1) to give the title compound 11 (270 mg, yield 65%). %) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.10-1.16 (m, 1H), 1.3-1.9 (m, 12H), 1.97-2.00 (m, 2H), 2.91-2.93 (m, 2H), 4.00- 4.11 (m, 1H), 7.33-7.35 (d, J = 8.8 Hz, 2H), 7.43-7.45 (m, 1H), 7.58-7.60 (d, J = 2.0 Hz, 2H), 9.35 (br, 1H) ¹³ C-NMR (125 MHz, CDCl ₃ ) δ 21.5, 26.1, 31.2, 33.4, 39.1, 41.2, 44.6, 51.9, 120.9, 129.2, 130.4, 134.9, 157.5, 158.7; HRMS (ESI), C ₁₈ H ₂₅ ClN ₃ O ₂ [M+H] ⁺ m/z: calculated value 350.1630, found value 350.1632.

[Synthesis of N ¹ -(3,4-methylenedioxyphenyl)-N ² -(2-cyclohexylpiperidin-4-yl)oxalamide (Compound 12, YIR-438)]

N-(3,4-methylenedioxyphenyl) oxamic acid (230 mg, 1.1 mmol) THF (7.5 mL) was added HOBt · H ₂ O in (337 mg, 2.2 mmol), EDCI · HCl (422 mg, 2.2 mmol), compound 10 (150 mg, 0.892 mmol) and NEt ₃ (0.381 mL, 2.75 mmol) were added at 0°C. The reaction mixture was stirred at room temperature for 50 hours, quenched with a saturated NaHCO ₃ aqueous solution at 0° C., and extracted with CHCl ₃ . The organic phase was dried over Na ₂ SO ₄ , then concentrated under reduced pressure and purified by column chromatography (CHCl ₃ /MeOH = 4/1) to give the title compound 12 as a solid (161 mg, yield 50%).
¹ H-NMR (500 MHz, CDCl ₃ ) δ 1.09-1.14 (m, 1H), 1.2-1.8 (m, 12H), 1.96-1.98 (m, 2H), 2.91-2.93 (m, 2H), 4.02- 4.10 (m, 1H), 5.97 (s, 2H), 6.77-6.79 (m, 1H), 6.99-7.01 (m, 1H), 7.34-7.34 (m, 1H), 7.48-7.50 (m, 1H), 9.38 (br, 1H); ¹³ C-NMR (125 MHz, CDCl ₃ ) δ 21.6, 26.1, 31.2, 33.4, 39.2, 41.2, 44.6, 52.0, 101.4, 102.3, 108.2, 113.4, 130.9, 144.9, 147.9, 157.4 , 159.1; HRMS (ESI), C ₁₉ H ₂₆ N ₃ O ₄ [M+H] ⁺ m/z: Calculated value 360.1918, Found value 360.1919.

[Synthesis of N ¹ -(4-chloro-3-fluorophenyl)-N ² -(2-cyclohexylpiperidin-4-yl)oxalamide (Compound 13, YIR-501)]

To a solution of N-(3,4-methylenedioxyphenyl)oxamic acid (239 mg, 1.1 mmol) in THF (7.5 mL), HOBt・H ₂ O (337 mg, 2.2 mmol), EDCI・HCl (422 mg, 2.2 mmol), compound 10 (150 mg, 0.892 mmol) and NEt ₃ (0.381 mL, 2.75 mmol) were added at 0°C. The reaction mixture was stirred at room temperature for 73.5 hours, then quenched with a saturated NaHCO ₃ aqueous solution at 0° C., and extracted with CHCl ₃ . The organic phase was dried over Na ₂ SO ₄ , then concentrated under reduced pressure and purified by column chromatography (CHCl ₃ /MeOH = 4/1) to give the title compound 13 as a solid (64.9 mg, yield 20%).
¹ H-NMR (500 MHz, CDCl ₃ ) δ 1.08-1.13 (m, 1H), 1.2-1.8 (m, 12H), 1.96-1.99 (m, 2H), 2.92-2.94 (m, 2H), 4.02- 4.10 (m, 1H), 7.24-7.26 (m, 2H), 7.30-7.32 (m, 1H), 7.36-7.39 (m, 1H), 7.70-7.72 (m, 1H), 9.36 (br, 1H); ¹³ C-NMR (125 MHz, CDCl ₃ ) δ 21.6, 26.1, 31.3, 33.5, 39.2, 41.3, 44.8, 51.9, 108.5, 116.0, 117.1, 130.8, 136.3, 157.7, 158.5, 159.1; HRMS (ESI), C ₁₈ H ₂₄ ClFN ₃ O ₂ [M+H] ⁺ m/z: Calculated 368.1536, Found 368.1533.

[Synthesis of N ¹ -(4-methylphenyl)-N ² -(2-cyclohexylpiperidin-4-yl)oxalamide (Compound 14, YIR-504)]

Compound 10 (150 mg, 0.892 mmol) and NEt ₃ (0.381 mL, 2.75 mmol) were added to a solution of N-(4-methylphenyl)ethoxalamide (ethoxalamide) (228 mg, 1.1 mmol) in EtOH (17.8 mL) at room temperature. did. The reaction mixture was stirred by microwave irradiation at 150° C. for 3 hours, then N-(4-methylphenyl)ethoxalamide (148 mg, 0.714 mmol) and NEt ₃ (0.247 mL, 1.78 mmol) were added at room temperature. The reaction mixture was stirred at 150° C. for 3 hours and then concentrated under reduced pressure. Extracted with CHCl ₃ and saturated aqueous NaHCO ₃ , the organic phase was dried over Na ₂ SO ₄ and concentrated under reduced pressure. The crude mixture was purified by column chromatography (CHCl ₃ /MeOH= ₄ /1) to give the title compound 14 as a solid (213 mg, yield 73%).
¹ H-NMR (500 MHz, CDCl ₃ ) δ 1.07-1.12 (m, 1H), 1.2-1.8 (m, 12H), 1.96-1.99 (m, 2H), 2.91-2.92 (m, 2H), 4.02- 4.10 (m, 1H), 7.16-7.18 (m, 2H), 7.41 (br, 1H), 7.51-7.52 (m, 2H), 9.28 (br, 1H); ¹³ C-NMR (125 MHz, CDCl ₃ ). δ 20.9, 21.5, 26.1, 31.1, 33.4, 39.2, 41.2, 44.5, 51.8, 119.9, 129.5, 133.9, 134.9, 157.5, 159.1; HRMS (ESI), C ₁₉ H ₂₈ N ₃ O ₂ [M+H] ⁺ m/z: calculated value 330.2176, measured value 330.2175.

[Synthesis of N ¹ -(4-chlorophenyl)-N ² -(1-amidino-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 15, YIR-720)]

Compound 11 (50 mg, 0.143 mmol) in DMF (1.43 mL) was added with i-Pr ₂ NEt (0.147 mL, 0.859 mmol) and 1H-pyrazole-1-carboxyamidine hydrochloride (84 mg, 0.573 mmol) at room temperature. Added in. The reaction mixture was stirred at 65° C. for 24 hours, concentrated under reduced pressure, and purified by HPLC to give the trifluoroacetic acid salt of title compound 15 as a white powder (9.39 mg, yield 13%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.4-1.7 (m, 10H), 2.15 (s, 3H), 2.20-2.31 (m, 2H), 2.58-2.63 (m, 1H), 3.07-3.11 ( m, 1H), 3.40-3.46 (m, 1H), 3.59-3.66 (m, 1H), 4.14-4.25 (m, 1H), 7.34-7.36 (m, 2H), 7.42-7.44 (m, 1H), 7.57-7.59 (m, 2H), 9.22 (br, 1H); ¹³ C-NMR (125 MHz, CDCl ₃ ) δ 22.4, 22.8, 25.3, 25.9, 30.9, 31.6, 35.5, 37.8, 41.6, 42.9, 60.2, 121.0, 129.4, 130.6, 134.8, 157.3, 159.0, 171.5; HRMS (ESI), C ₁₉ H ₂₆ ClKN ₅ O ₂ [M+K] ⁺ m/z: Calculated 430.1407, Found 430.1406.

[Synthesis of N ¹ -(4-chlorophenyl)-N ² -(1-benzyl-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 16, YIR-327)]

To a DMF (2.86 mL) solution of compound 11 (100 mg, 0.286 mmol), benzyl bromide (0.103 mL, 0.859 mmol) and i-Pr ₂ NEt (0.196 mL, 1.15 mmol) were added at room temperature. The reaction mixture was stirred at 65° C. for 19 hr, quenched with a saturated NaHCO ₃ aqueous solution at 0° C., and extracted with CHCl ₃ . The organic phase was dried over Na ₂ SO ₄ , then concentrated under reduced pressure and purified by column chromatography (hexane/AcOEt = 3/1 to 1/2) to give the title compound 16 (94.2 mg, yield 75%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.3-1.7 (m, 9H), 1.7-1.93 (m, 4H), 2.27-2.32 (m, 1H), 2.60-2.72 (m, 2H), 3.24- 3.28 (m, 1H), 3.97-4.07 (m, 1H), 4.04-4.08 (m, 1H), 7.20-7.24 (m, 1H), 7.28-7.32 (m, 2H), 7.33-7.36 (m, 5H ), 7.57-7.60 (m, 2H), 9.28 (br, 1H); ¹³ C-NMR (125 MHz, CDCl ₃ )δ 22.2, 22.3, 26.3, 26.4, 29.7, 30.8, 31.9, 37.4, 38.1, 44.4, 45.0, 51.4, 56.4, 121.0, 126.6, 128.2, 129.3, 129.4, 130.4, 135.0, 157.6, 158.9; HRMS (ESI), C ₂₅ H ₃₁ ClN ₃ O ₂ [M+K] ⁺ m/z: Calculated value 440.2099 , Found 440.2096.

[N ¹ -(4-chlorophenyl)-N ² -(1-(2-(N-(tert-butoxycarbonyl)amino)ethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 17, YIR-703 ) Synthesis]

Compound 11 (100 mg, 0.286 mmol) in CHCl ₃ (2.86 mL) was added with 2-(N-(tert-butoxycarbonyl)amino)-O-(4-methylphenylsulfonyl)ethanol (271 mg, 0.858 mmol). And i-Pr ₂ NEt (0.196 mL, 1.14 mmol) were added at room temperature. The reaction mixture was stirred at 65°C for 47 hours, quenched with a saturated NaHCO ₃ aqueous solution at 0°C, and extracted with CHCl ₃ . The organic phase was dried over Na ₂ SO ₄ , concentrated under reduced pressure, and purified by column chromatography (acetone only) to give the title compound 17 (14.3 mg, yield 10%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ 0.8-0.95 (m, 3H), 1.0-1.4 (m, 4H), 1.45 (s, 9H), 1.5-1.8 (m, 4H), 1.86-1.93 ( m, 2H), 2.15-2.18 (m, 1H), 2.32-2.35 (m, 1H), 2.66-2.75 (m, 2H), 2.95-3.03 (m, 2H), 3.15-3.3 (m, 1H), 3.91-4.01 (m, 1H), 4.92 (br, 1H), 7.33-7.35 (m, 3H), 7.57-7.60 (m, 2H), 9.26 (br, 1H); ¹³ C-NMR (125 MHz, CDCl ₃ ) δ 22.2, 25.3, 26.1, 28.3, 29.6, 31.5, 37.2, 38.0, 39.0, 44.7, 44.9, 46.9, 53.4, 56.3, 120.9, 129.2, 130.4, 134.9, 156.1, 157.4, 158.8; HRMS (ESI), C ₂₅ H ₃₈ ClN ₄ O ₄ [M+H] ⁺ m/z: calculated 493.2576, found 493.2574.

[Synthesis of N ¹ -(4-chlorophenyl)-N ² -(1-cyanomethyl-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 18, YIR-631)]

To a DMF (10.1 mL) solution of compound 11 (531 mg, 1.52 mmol), bromoacetonitrile (0.303 mL, 4.56 mmol) and i-Pr ₂ NEt (1.04 mL, 6.08 mmol) were added at room temperature. The reaction mixture was stirred at 65° C. for 19 hr, quenched with a saturated NaHCO ₃ aqueous solution at 0° C., and extracted with CHCl ₃ . The organic phase was dried over Na ₂ SO ₄ , then concentrated under reduced pressure and purified by column chromatography (hexane/AcOEt = 3/1 to 1/2) to give the title compound 18 (483 mg, yield 82%). ) Got.
¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.1-1.7 (m, 11H), 1.88-2.01 (m, 2H), 2.28-2.33 (m, 1H), 2.83-2.88 (m, 1H), 3.07- 3.14 (m, 1H), 3.63-3.64 (m, 2H), 3.97-4.07 (m, 1H), 7.33-7.37 (m, 3H), 7.57-7.59 (m, 2H), 9.24 (br, 1H); ¹³ C-NMR (125 MHz, CDCl ₃ ) δ 21.9, 22.0, 25.8, 26.4, 30.8, 37.1, 37.4, 37.5, 44.2, 46.2, 57.0, 118.3, 120.9, 129.3, 130.5, 134.8, 157.3, 158.9; HRMS ( ESI), C ₂₀ H ₂₆ ClN ₄ O ₂ [M+H] ⁺ m/z: calculated 389.1739, found 389.1740.

[Synthesis of N ¹ -(4-chlorophenyl)-N ² -(1-(2-aminoethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 19, YIR-723)]

LiAlH ₄ (156 mg, 4.11 mmol) was slowly added to a solution of Compound 18 (483 mg, 1.24 mmol) in THF (24.8 mL) at 0°C. The reaction mixture was stirred at room temperature for 1.5 hours, then quenched with saturated aqueous sodium potassium tartrate solution at 0° C., and made basic with 1N aqueous NaOH solution (5 mL, pH=13-14). Extracted with CHCl ₃ , then the organic phase was dried over Na ₂ SO ₄ . After concentration under reduced pressure, purification by HPLC gave the trifluoroacetate salt of the title compound 19 as a white powder (393 mg, yield 81%).
¹ H-NMR (500 MHz, CDCl ₃ ) δ 1.1-1.25 (m, 2H), 1.25-1.55 (m, 5H), 1.6-1.95 (m, 4H), 1.95-2.15 (m, 3H), 2.9- 3.3 (m, 2H), 3.3-3.65 (m, 4H), 3.65-3.8 (m, 1H), 3.8-4.02 (m, 1H), 4.02-4.2 (m, 1H), 7.29-7.30 (m, 3H ), 7.54-7.55 (m, 2H), 9.36 (br, 1H); ¹³ C-NMR (125 MHz, CDCl ₃ )δ 18.1, 21.4, 21.5, 24.3, 29.6, 31.4, 35.4, 36.5, 41.5, 58.1, 121.3, 129.1, 130.5, 134.9, 157.0, 159.7; HRMS (ESI), C ₂₀ H ₃₀ ClN ₄ O ₂ [M+H] ⁺ m/z: Calculated 393.2052, Found 393.2051.

[Synthesis of N ¹ -(4-chlorophenyl)-N ² -(1-(2-guanidinoethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 20, YIR-737)]

Compound 19 (20 mg, 0.051 mmol) in DMF (1.02 mL) was added with i-Pr ₂ NEt (0.035 mL, 0.204 mmol) and 1H-pyrazole-1-carboxyamidine hydrochloride (22.4 mg, 0.153 mmol) at room temperature. Added in. The reaction mixture was stirred at 65° C. for 44 hours, then concentrated under reduced pressure and purified by HPLC to give the trifluoroacetate salt of the title compound 20 as a white powder (9.50 mg, yield 34%).
¹ H-NMR (500 MHz, CDCl ₃ ) δ 1.1-1.3 (m, 1H), 1.3-1.55 (m, 4H), 1.55-1.9 (m, 5H), 1.9-2.2 (m, 3H), 2.66- 2.68 (m, 1H), 2.89-2.91 (m, 1H), 3.17 (m, 1H), 3.49-3.79 (m, 5H), 4.08 (m, 1H), 7.30-7.32 (m, 3H), 7.54- 7.56 (m, 2H), 7.81-7.82 (m, 1H), 8.51 (br, 1H), 9.33 (br, 1H), 10.10 (br, 1H); ¹³ C-NMR (125 MHz, CDCl ₃ ) δ 21.2 , 21.4, 24.0, 24.4, 28.2, 33.2, 34.6, 36.9, 41.6, 47.7, 48.2, 67.8, 121.3, 129.2, 130.7, 134.7, 156.9, 157.5, 159.5; HRMS (ESI), C ₂₁ H ₃₂ ClN ₆ O ₂ [M+H] ⁺ m/z: calculated 435.2270, found 435.2269.

[N ¹ -(4-chlorophenyl)-N ² -(1-(2-(N-(tert-butoxycarbonyl)glycinamido)ethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 21, YIR- 816) Synthesis]

N- (tert- butoxycarbonyl) glycine (134 mg, 0.765 mmol) in DMF (5.1 mL) solution _{of, HOBt · H 2 O (195} mg, 1.28 mmol), EDCI · HCl (244 mg, 1.28 mmol), compound 19 (100 mg, 0.255 mmol) and NEt ₃ (0.247 mL, 1.79 mmol) were added at 0°C. The reaction mixture was stirred at room temperature for 25 hours, quenched with saturated NaHCO ₃ aqueous solution at 0° C., and extracted with CHCl ₃ . The organic phase was dried over Na ₂ SO ₄ , then concentrated under reduced pressure and purified by column chromatography (CHCl ₃ /MeOH = 10/1 to 6/1) to give the title compound 21 (96.1 mg, yield 69 %) as a solid.
¹ H-NMR (500 MHz, CDCl ₃ ) δ 0.95-1.35 (m, 8H), 1.38 (s, 9H), 1.4-1.65 (m, 3H), 1.78-1.85 (m, 2H), 2.15-2.26 ( m, 2H), 2.60-2.69 (m, 2H), 2.90 (m, 1H), 3.07 (m, 1H), 3.35 (m, 1H), 3.73 (m, 2H), 3.89 (m, 1H), 5.17 (br, 1H), 6.45-6.60 (m, 1H), 7.25-7.27 (m, 3H), 7.35-7.45 (m, 1H), 7.53-7.55 (m, 2H), 9.35 (br, 1H); ¹³ C-NMR (125 MHz, CDCl ₃ ) δ 22.1, 25.5, 26.0, 28.3, 31.3, 37.2, 37.5, 37.9, 44.3, 44.5, 44.8, 46.2, 55.5, 80.0, 121.0, 129.2, 130.4, 134.9, 155.9, 157.5 , 158.9, 169.2; HRMS (ESI), C ₂₇ H ₄₁ ClN ₅ O ₅ [M+H] ⁺ m/z: calculated 550.2791, found 550.2792.

[Synthesis of N ¹ -(4-chlorophenyl)-N ² -(1-(2-(glycinamido)ethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 22, YIR-818)]

To a solution of compound 21 (50 mg, 0.091 mmol) in CH ₂ Cl ₂ (0.91 mL), trifluoroacetic acid (0.035 mL, 0.455 mmol) was added at 0°C. The reaction mixture was stirred at room temperature for 1.5 hours, then concentrated under reduced pressure and purified by HPLC to give the trifluoroacetate salt of the title compound 22 as a white powder (39.4 mg, yield 77%).
¹ H-NMR (400 MHz, MeOH) δ 1.2-1.35 (m, 1H), 1.5-1.9 (m, 8H), 1.95-2.3 (m, 4H), 2.79-2.93 (m, 1H), 3.41 (m , 2H), 3.67-3.76 (m, 6H), 4.17 (m, 1H), 7.34-7.37 (d, J = 8.8 Hz, 2H), 7.72-7.74 (d, J = 8.8 Hz, 2H); ¹³ C -NMR (125 MHz, MeOH) δ 22.4, 22.6, 25.1, 25.7, 29.9, 34.6, 36.4, 37.1, 41.4, 42.8, 49.8, 68.8, 122.9, 129.9, 131.0, 137.3, 159.4, 161.4, 169.2; HRMS (ESI ), C ₂₂ H ₃₃ ClN ₅ O ₃ [M+H] ⁺ m/z: calculated 450.2266, found 450.2264.

Of (N ¹ -(4-chlorophenyl)-N ² -(1-(2-(N-(amidino)glycinamide)ethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 23, YIR-819) Synthesis]

Compound 22 (40 mg, 0.089 mmol) in DMF (1.78 mL) was added with i-Pr ₂ NEt (0.092 mL, 0.534 mmol) and 1H-pyrazole-1-carboxyamidine hydrochloride (52.2 mg, 0.356 mmol) at room temperature. Added in. The reaction mixture was stirred at 65° C. for 13 hours, concentrated under reduced pressure, and purified by HPLC to give the trifluoroacetic acid salt of title compound 23 as a white powder (24.1 mg, yield 45%).
¹ H-NMR (400 MHz, MeOH) δ 1.2-1.4 (m, 1H), 1.5-1.95 (m, 8H), 1.95-2.2 (m, 2H), 2.21-2.27 (m, 2H), 2.82-2.95 (m, 1H), 3.3-3.50 (m, 2H), 3.59-3.82 (m, 4H), 4.03 (s, 2H), 4.19-4.22 (m, 1H), 7.37-7.39 (d, J = 8.8 Hz , 2H), 7.74-7.76 (d, J = 8.8 Hz, 2H); ¹³ C-NMR (125 MHz, MeOH) δ 22.4, 22.7, 25.1, 25.8, 30.1, 34.8, 36.7, 37.5, 42.9, 44.6, 51.0 , 68.7, 123.0, 129.9, 131.1, 137.4, 159.4, 161.4, 172.3; HRMS (ESI), C ₂₃ H ₃₅ ClN ₇ O ₃ [M+H] ⁺ m/z: Calculated value 492.2484, Found value 492.2484.

[N ^1- (4-chlorophenyl)-N ² -(1-(2-(N-(tert-butoxycarbonyl)-5-aminovaleramido)ethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (compound 24, YIR-817) synthesis]

N- (tert- butoxycarbonyl) -5-amino valeric acid (216 mg, 0.994 mmol) in DMF (3.32 mL) solution _{of, HOBt · H 2 O (254} mg, 1.66 mmol), EDCI · HCl (318 mg, 1.66 mmol), compound 19 (130 mg, 0.332 mmol) and NEt ₃ (0.397 mL, 2.32 mmol) were added at 0°C. The reaction mixture was stirred at room temperature for 36 hours, then quenched with saturated NaHCO ₃ aqueous solution at 0° C., and extracted with CHCl ₃ . The organic phase was dried over Na ₂ SO _4, then concentrated under reduced pressure to give purified by column chromatography _{(CHCl 3 / MeOH = 10 /} 1~6 / 1) Compound 24 of the title as a solid (119 mg, yield 61%).
¹ H-NMR (500 MHz, CDCl ₃ ) δ 1.1-1.4 (m, 4H), 1.43 (s, 9H), 1.45-1.8 (m, 4H), 1.90-1.97 (m, 2H), 2.19-2.23 ( m, 4H), 2.29-2.39 (m, 3H), 2.73-2.79 (m, 1H), 2.92-2.95 (m, 1H), 3.07-3.19 (m, 8H), 3.46-3.52 (m, 1H), 3.95-4.03 (m, 1H), 4.75 (br, 1H), 5.19 (br, 1H), 6.78 (br, 1H), 7.33-7.35 (m, 2H), 7.59-7.62 (m, 3H), 9.37 ( br, 1H); ¹³ C-NMR (125 MHz, CDCl ₃ )δ 22.1, 22.8, 25.4, 25.9, 28.5, 29.6, 29.8, 30.7, 36.1, 36.5, 37.1, 40.1, 40.4, 44.1, 45.6, 47.3, 58.6 , 79.0, 121.1, 129.3, 130.5, 135.1, 156.2, 157.5, 159.2, 173.1; HRMS (ESI), C ₃₀ H ₄₇ ClN ₅ O ₅ [M+H] ⁺ m/z: Calculated 592.3260, Found 592.3259.

[Synthesis of N ¹ -(4-chlorophenyl)-N ² -(1-(2-(5-aminovaleramido)ethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 25, YIR-820)]

To a solution of compound 24 (60 mg, 0.102 mmol) in CH ₂ Cl ₂ (1.02 mL), trifluoroacetic acid (0.039 mL, 0.508 mmol) was added at 0°C. The reaction mixture was stirred at room temperature for 2.5 hours, then concentrated under reduced pressure and purified by HPLC to give the trifluoroacetate salt of the title compound 25 as a white powder (45.3 mg, yield 74%).
¹ H-NMR (500 MHz, MeOH) δ 1.25-1.35 (m, 1H), 1.5-1.9 (m, 12H), 1.93-2.05 (m, 2H), 2.19-2.24 (m, 2H), 2.33-2.40 (m, 2H), 2.79-2.82 (m, 1H), 2.9-3.0 (m, 2H), 3.36-3.51 (m, 2H), 3.57-3.78 (m, 4H), 4.16 (m, 1H), 7.34 -7.36 (d, J = 8.8 Hz, 2H), 7.71-7.73 (d, J = 8.8 Hz, 2H); ¹³ C-NMR (125 MHz, MeOH) δ 21.2, 21.8, 23.6, 24.2, 26.5, 28.6, 32.5, 33.2, 34.2, 35.3, 36.1, 38.9, 41.5, 50.2, 50.7, 66.7, 121.5, 128.4, 129.6, 135.9, 157.9, 159.9, 175.4; HRMS (ESI), C ₂₅ H ₃₉ ClN ₅ O ₃ [M+ H] ⁺ m/z: calculated 492.2736, found 492.2737.

[Synthesis of N ¹ -(4-chlorophenyl)-N ² -(1-(2-(5-guanidinovaleramido)ethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 26, YIR-821)]

Compound 25 (50 mg, 0.102 mmol) in DMF (2.04 mL) was added with i-Pr ₂ NEt (0.105 mL, 0.611 mmol) and 1H-pyrazole-1-carboxyamidine hydrochloride (59.7 mg, 0.407 mmol) at room temperature. Added in. The reaction mixture was stirred at 65° C. for 38.5 hours, then concentrated under reduced pressure and purified by HPLC to give the trifluoroacetate salt of the title compound 26 as a white powder (33.6 mg, yield 51%).
¹ H-NMR (400 MHz, MeOH) δ 1.2-1.4 (m, 1H), 1.5-1.95 (m, 12H), 1.98-2.08 (m, 2H), 2.22-2.27 (m, 2H), 2.34-2.38 (m, 2H), 2.82-2.93 (m, 1H), 3.20-3.24 (m, 2H), 3.38-3.53 (m, 2H), 3.60-3.82 (m, 4H), 4.15-4.3 (m, 1H) , 7.37-7.39 (m, 2H), 7.73-7.76 (m, 2H); ¹³ C-NMR (125 MHz, MeOH) δ 20.9, 21.2, 22.1, 23.6, 24.3, 27.9, 28.8, 33.3, 34.4, 35.5, 36.2, 40.6, 41.4, 50.5, 66.6, 121.5, 128.4, 129.6, 135.9, 157.2, 157.9, 159.9, 177.2; HRMS (ESI), C ₂₆ H ₄₁ ClN ₇ O ₃ [M+H] ⁺ m/z: Calculation Value 534.2954, Found 534.2954.

[N ¹ -(4-chlorophenyl)-N ² -(1-(2-(N,N-bis(cyanomethyl))aminoethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 27, YIR-738 ) Synthesis]

To a DMF (3.83 mL) solution of compound 19 (300 mg, 0.765 mmol), i-Pr ₂ NEt (1.05 mL, 6.12 mmol) and bromoacetonitrile (0.256 mL, 3.83 mmol) were added at room temperature. The reaction mixture was stirred at 65° C. for 63.5 hours, quenched with 0° C. saturated NaHCO ₃ aqueous solution, and extracted with CHCl ₃ . The organic phase was dried over Na ₂ SO ₄ , then concentrated under reduced pressure and purified by column chromatography (hexane/AcOEt = 1/1 to 1/2) to give the title compound 27 (224 mg, yield. 62%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.0-1.8 (m, 11H), 1.86-1.97 (m, 2H), 2.25-2.38 (m, 2H), 2.60-2.84 (m, 4H), 3.08- 3.14 (m, 1H), 3.73-3.85 (m, 4H), 3.92-4.00 (m, 1H), 7.33-7.35 (m, 3H), 7.58-7.60 (m, 2H), 9.27 (br, 1H); ¹³ C-NMR (125 MHz, MeOH) δ 23.3, 23.4, 23.8, 26.0, 27.2, 31.8, 36.7, 38.2, 38.3, 43.6, 45.4, 47.1, 47.6, 53.6, 58.2, 65.7, 116.3, 122.9, 129.9, 131.0 , 137.4, 159.8, 160.9; HRMS (ESI), C ₂₄ H ₃₂ ClN ₆ O ₂ [M+H] ⁺ m/z: calculated 471.2270, found 471.2270.

[N ¹ -(4-chlorophenyl)-N ² -(1-(2-(N,N-bis(2-aminoethyl))aminoethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 28, Synthesis of YIR-801)]

LiAlH ₄ (17.8 mg, 0.468 mmol) was slowly added to a solution of compound 27 (50 mg, 0.106 mmol) in THF (2.13 mL) at 0°C. The reaction mixture was stirred at room temperature for 2 hours, then quenched with a saturated aqueous potassium sodium tartrate solution at 0°C, and made basic with a 1N aqueous NaOH solution (5 mL, pH = 13 to 14). It was extracted with CHCl ₃ and the organic phase was dried over Na ₂ SO ₄ . Concentration under reduced pressure and subsequent purification by HPLC gave the trifluoroacetate salt of the title compound 28 as a white powder (41.3 mg, yield 55%).
¹ H-NMR (500 MHz, MeOH) δ 1.1-1.8 (m, 11H), 1.85-2.2 (m, 5H), 2.65-2.68 (m, 1H), 2.75 (m, 4H), 2.86-2.91 (m , 3H), 3.01 (m, 4H), 4.06 (m, 1H), 7.25-7.27 (m, 2H), 7.63-7.65 (m, 2H); ¹³ C-NMR (125 MHz, MeOH) δ 22.4, 22.7 , 25.2, 25.6, 29.7, 29.7, 30.8, 34.6, 36.1, 38.0, 42.8, 47.4, 52.3, 69.1, 122.9, 129.9, 131.0, 137.4, 159.4, 161.4; HRMS (ESI), C ₂₄ H ₄₀ ClN ₆ O ₂ [M+H] ⁺ m/z: calculated 479.2896, found 479.2896.

[N ¹ -(4-chlorophenyl)-N ² -(1-(2-(N,N-bis(2-guanidinoethyl))aminoethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 29, Synthesis of YIR-802)]

Compound 28 (40 mg, 0.084 mmol) in DMF (1.68 mL) was charged with i-Pr ₂ NEt (0.086 mL, 0.502 mmol) and 1H-pyrazole-1-carboxyamidine hydrochloride (49 mg, 0.334 mmol) at room temperature. Added in. The reaction mixture was stirred at 65° C. for 17.5 hours, then concentrated under reduced pressure and purified by HPLC to give the trifluoroacetic acid salt of title compound 29 as a white powder (15.3 mg, yield 23%).
¹ H-NMR (500 MHz, MeOH) δ 1.3-1.9 (m, 13H), 2.0-2.2 (m, 3H), 2.67-2.72 (m, 1H), 2.81-2.82 (m, 3H), 2.9-3.05 (m, 2H), 3.4-3.9 (m, 6H), 4.17-4.21 (m, 1H), 7.37-7.39 (m, 2H), 7.74-7.76 (m, 2H); ¹³ C-NMR (125 MHz, MeOH) δ 22.1, 22.7, 26.5, 26.6, 29.8, 30.0, 31.3, 33.6, 36.6, 37.5, 41.4, 49.4, 50.3., 68.9, 121.9, 129.1, 131.4, 138.2, 157.2, 159.7, 161.8; HRMS (ESI) , C ₂₆ H ₄₄ ClN ₁₀ O ₂ [M+H] ⁺ m/z: calculated value 563.3332, measured value 563.3331.

[N ¹ -(4-chlorophenyl)-N ² -(1-(2-(N-(2-guanidinoethyl))aminoethyl)-2-cyclohexylpiperidin-4-yl)oxalamide (Compound 30, YIR-913 ) Synthesis]

LiAlH ₄ (3.57 mg, 0.0941 mmol) was slowly added to a solution of compound 23 (14 mg, 0.0285 mmol) in THF (0.57 mL) at 0°C. The reaction mixture was stirred at room temperature for 4 hours, and then the reaction was stopped with a saturated aqueous solution of potassium sodium tartrate at 0°C. The mixture was filtered, then the filtrate was concentrated under reduced pressure, and purified by HPLC to give the trifluoroacetate salt of the title compound YIR-913 as a white powder (2 mg, yield 10%).

<Example 2>
[Activity evaluation]
The various novel compounds synthesized in Example 1 were evaluated for anti-HIV activity, cytotoxicity, and ability to induce structural changes using the following methods. The cells and antibodies used were obtained as described in K. Yoshimura et al., J. Virology, Aug. 2010, p.7558-7568.

[Anti-HIV activity]
HIV-1 cYTA48P strain (infectious clone virus produced by the present inventors by replacing the virus env from HIV patients with env of NL43) and TZM-bl cells (NIH AIDS Reagent Program (https://www.aidsreagent .org/Index.cfm))). TZM-bl cells have CD4/CCR5/CXCR4 on the surface and are indicator cells expressing luciferase upon HIV infection. In this cell, the β-galactosidase gene linked to the HIV-1 LTR sequence was integrated, and when infected with HIV-1, the transcriptional activator Tat is expressed from the infected HIV-1 tat gene. Tat acts on the promoter region of LTR so that β-galactosidase is expressed. By adding a substrate thereto, enzymatic cleavage occurs and galactose and luciferin are produced.Therefore, the number of HIV-1-infected cells is measured by detecting the chemiluminescence amount emitted by luciferin oxidized by luciferase with a luminometer. be able to.

Specifically, TZM-bl cells (1×10 ⁴ cells) were cultured with 100 TCID ₅₀ of the cYTA48P strain and a dilution series inhibitor (the compound of the present invention or a control compound). After 48 hours of culture, β-galactosidase activity was measured with a luminometer ARVO (PerkinElmer) according to the instructions attached to Beta-Glo(TM) Assay Reagent (Promega) to compare the sensitivity of anti-HIV entry inhibitors. did. The IC ₅₀ value is the concentration at which HIV-1 inhibits 50% of the cytopathogenicity of HIV-1 on PM1/CCR5 cells (CCR5-highly expressing T cells donated by Dr. Yosuke Maeda of Kumamoto University).

[Cytotoxicity]
It was evaluated by the WST-8 assay using PM1/CCR5 cells. Intracellular dehydrogenase (NADH) present in living cells is directly or indirectly involved in various reduction reactions. When tetrazolium salt (WST-8) is added to a sample containing living cells, it is reduced to produce orange water-soluble formazan. Since the reduction reaction of this tetrazolium salt occurs only when the cells are alive, there is a linear proportional relationship between the amount of formazan produced and the number of living cells. Using this, the amount of formazan produced was measured by the absorbance (450 nm) to quantify the cell proliferation rate and cell survival rate. The CC ₅₀ value is the concentration when PM1/CCR5 cells were mock-infected and the survival rate reached 50%.

[Structural change inducing ability]
HIV-1 JR-FL strain persistently infected with CD4 mimic derivative PM1 cells (PM1 cells obtained from the NIH AIDS Reagent Program were added to the HIV-1 JR-FL strain by Professor Yoshio Koyanagi, Institute for Virus Research, Kyoto University) Of the persistently infected cell line established by the present inventors by infecting E.coli. Using a CD4i antibody (4C11, a monoclonal antibody established by the group of the present inventors) known to specifically bind to a site exposed when gp120 undergoes a structural change (CD4-inducing site), A structural change of gp120 can be detected using a goat-derived anti-human IgG antibody (secondary antibody) labeled with fluorescein isothiocyanate (FITC). The fluorescence intensity (mean fluorescence intensity: MFI) was measured by fluorescence activated cell sorting (FACS) analysis, and the extent of structural change of gp120 was quantified from the binding amount of the CD4i antibody (4C11).

For comparison of data, from the MFI of the CD4 mimic derivative such as the compound of the present invention and the MFI of NBD-556, the relative FACS (rel.FACS) was calculated using the following formula, and the CD4 mimic derivative was NBD-556. It is used as an index as to how much the structural change of gp120 is induced as compared with.
rel. FACS = ([MFI of CD4 mimic derivative]-[MFI of 4C11])
/ ([NBD-556 MFI]-[4C11 MFI])

Since the CD4i antibody alone induces a slight structural change in gp120, subtracting the MFI of 4C11 alone from each MFI eliminates the background noise.

[Example 2-1. Evaluation of activity of monocyclohexyl-type CD4 mimic derivative]
Regarding the monocyclohexyl-type CD4 mimic derivative obtained in the above-mentioned synthesis example, first, compounds having different aromatic ring structures were compared and evaluated for anti-HIV activity, cytotoxicity, and ability to induce structural change (Table 1).

As shown in Table 1, by using a monocyclohexyl-type piperidine structure (R ^c ), reduction in cytotoxicity was generally observed. Among them, YIR-329 (compound 11) having a chloro group in the para position was not as potent in anti-HIV activity as NBD-556, but its cytotoxicity was reduced more than 3-fold compared to NBD-556, and gp120 It was found that the compound has the same effect as NBD-556 on the structure change inducing ability.

From this result, it is considered that the aromatic ring unit having a chloro group at the para position is the most suitable as the aromatic ring unit in the CD4 mimic derivative having a monocyclohexyl piperidine structure. -I decided to fix it on the chloro form. Similar to the p-chloro compound, a compound having a bromo group or a fluoro group at the para position is expected to have similar activity.

[Example 2-2. Activity evaluation of amino-type monocyclohexyl CD4 mimic derivative]
The monocyclohexyl-type CD4 mimic derivative (YIR-329, Compound 11) had lower cytotoxicity but lower anti-HIV activity than NBD-556 (Table 1). In addition, docking simulations of YIR-329 (Compound 11) and gp120 showed a hydrophobic interaction with Val430 but no significant interaction with Asp368 (data not shown). Therefore, it was investigated whether modification of the piperidine nitrogen atom of YIR-329 (Compound 11) could form an interaction with Asp368.

Specifically, in order to form an electrostatic interaction with the side chain carboxy group of Asp368, a monocyclohexyl CD4 mimic derivative in which an amino group was introduced on the piperidine nitrogen atom of YIR-329 (Compound 11). Was evaluated for anti-HIV activity and cytotoxicity (Table 2).

As shown in Table 2, the introduction of the amino group generally improved the anti-HIV activity as compared with YIR-329 (Compound 11). Among them, YIR-801 (compound 28) having two amino groups showed higher anti-HIV activity than NBD-556, but it was also found that cytotoxicity was increased.

[Example 2-3. Activity evaluation of guanidino type monocyclohexyl CD4 mimic derivative]
The anti-HIV activity and cytotoxicity of the monocyclohexyl CD4 mimic derivative in which a guanidino group was introduced on the piperidine nitrogen atom of YIR-329 (Compound 11) were evaluated in the same manner as in Example 2-2 (Table 3).

As shown in Table 3, the introduction of the guanidino group showed a significant improvement in anti-HIV activity, and higher anti-HIV activity than NBD-556 was observed except for YIR-720 (Compound 15). Among them, YIR-821 (Compound 26) showed a remarkable increase in anti-HIV activity and significantly decreased cytotoxicity.

These results suggest that the presence of guanidino group greatly contributes to the anti-HIV activity. It was also suggested that the presence of a guanidino group via a linker of a certain length is important.

[Example 2-4. Activity evaluation of other types of monocyclohexyl CD4 mimic derivatives]
The derivatives having other types of functional groups obtained in Example 1 were evaluated for anti-HIV activity and cytotoxicity in the same manner as described above (Table 4).

As shown in Table 4, the cyano group-introduced compounds (YIR-631 (Compound 18) and YIR-738 (Compound 27)) exhibited higher anti-HIV activity than NBD-556. In the case where the Boc group was introduced, anti-HIV activity was observed in the case where the linker had a certain length, and YIR-816 (Compound 21) showed higher anti-HIV activity than NBD-556. Those introduced with a phenyl group did not show anti-HIV activity. On the other hand, it was found that all of the active compounds were highly cytotoxic.

<Example 3>
[Combination test with neutralizing antibody KD-247]
Neutralizing antibody KD-247 that recognizes the V3 loop against YIR-802 (Compound 29), YIR-819 (Compound 23), and YIR-821 (Compound 26), which showed high anti-HIV activity. (Provided by the Serum Therapy Research Institute) was performed. It was evaluated whether the region recognized by the neutralizing antibody by CD4 mimic is exposed on the surface to enhance the neutralizing activity of the antibody. The assay method was the β-galactosidase reporter assay used in Example 2. KD-247 was also examined at several concentrations simultaneously with the compound. In addition, NBD-556 and YIR-329 (Compound 11) were used as comparative compounds. Results are shown in FIG.

The results shown in FIG. 3 indicate that the compounds of the present invention, YIR-802 (Compound 29), YIR-819 (Compound 23), and YIR-821 (Compound 26), have almost no activity when KD-247 alone is used. ), the inhibitory activity was enhanced, and a synergistic inhibitory action between the compound of the present invention and the anti-HIV antibody was shown. This means that when the compound of the present invention is used in combination with an anti-HIV antibody, a high amount of antibody can bring about a high inhibitory activity.

<Example 4>
[Comparison of interaction modes by docking simulation]
Based on the co-crystal structure of NBD-556 and Phe43-cavity (PDB: 3TGS), MOE was used to perform docking simulation of the compound of the present invention and Phe43-cavity, focusing on the piperidine ring site to study the binding mode. did. The interaction modes were compared between YIR-821 (compound 26), which showed the best anti-HIV activity, and YIR-329 (compound 11) and NBD-556 as comparative compounds (FIG. 4).

As a result, in NBD-556, the dimethyl group on the piperidine ring and Val430 had a hydrophobic interaction, but in YIR-329 (Compound 11), the cyclohexyl group had a more marked hydrophobic interaction with Val430. It was suggested that Furthermore, it was confirmed that YIR-821 (Compound 26) has a significant electrostatic interaction with Asp368 in addition to the hydrophobic interaction with Val430.

From these results, the compound of the present invention, YIR-821 (Compound 26), is a novel CD4 mimic compound capable of effectively interacting with both Val430 and Asp368, showing a significant improvement in the interaction with Asp368. It is considered to contribute to the improvement of HIV activity.

In the same manner, the interaction mode of YIR-819 (Compound 23), which showed the same high anti-HIV activity as YIR-821 (Compound 26), was examined (Fig. 5).

In YIR-819 (Compound 23), the cyclohexyl group is a little further away from Val430 than in the above YIR-329 (Compound 11) and YIR-821 (Compound 26) (Fig. 4), and the hydrophobic interaction is a little. It is suggested that it may be decreasing. In YIR-821 (Compound 26), the guanidino group introduced on the piperidine nitrogen atom interacted with Asp368, whereas in YIR-819 (Compound 23), the guanidino group was opposite to Asp368. It was confirmed that they had an electrostatic interaction with Asp474.

From the above, we have focused so far on the two amino acids Val430 and Asp368 located near the entrance of the Phe43-cavity.In addition to these two amino acids, Asp474 may also be an important amino acid residue for anti-HIV activity. Was suggested.

The HIV infection inhibitor of the present invention targets a non-traditional mechanism of HIV invasion into host cells, and is used in a drug repertoire for resisting the emergence of drug resistant virus in multidrug combination therapy. Expansion can be achieved. In particular, the HIV infection inhibitor of the present invention exerts a synergistic effect with an anti-HIV monoclonal antibody, so that the amount of antibody required for treatment can be reduced and the cost of treatment can be reduced.

All publications, patents and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims (8)

General formula (I):

[In the formula,
X is a halogen atom selected from Cl, Br, and F,
A is an alkylene group having 1 to 5 carbon atoms,
B is represented by the following formulas (II) to (IV):

(In the formula, R ¹ and R ² are each independently an alkylene group having 1 to 5 carbon atoms which may include one carbonyl group),

(In the formula, R ³ is an alkylene group having 1 to 5 carbon atoms which may contain one carbonyl group), and

Is a group selected from. ]
Or a salt thereof. The compound or salt thereof according to claim 1, wherein X is Cl and A is ethylene. The compound or salt thereof according to claim 1 or 2, wherein both R ¹ and R ² are ethylene. The compound according to claim 1 or 2, or a salt thereof, wherein R ³ is —CO—(CH ₂ ) _n — (where n is 1 to 4). An HIV infection inhibitor comprising the compound according to any one of claims 1 to 4 or a salt thereof as an active ingredient. The HIV infection inhibitor according to claim 5, which is administered in combination with an anti-HIV antibody . The HIV infection inhibitor according to claim 6 , wherein the anti-HIV antibody is a neutralizing antibody specific for the V3 loop on the HIV-1 surface . A pharmaceutical composition for treating or preventing HIV infection, which comprises the HIV infection inhibitor according to any one of claims 5 to 7 .

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