Pyrrolopyridine derivatives for curing HIV infection

Abstract

The present invention relates to Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, which is for use in a method of curing HIV-1 infection, wherein the method comprises administering Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, to a subject and determining the efficacy of the curing by determining the reactivation potential of a retroviral reservoir of the subject, so as to determine whether or not treatment can be stopped.

Description

PYRROLOPYRIDINE DERIVATIVES FOR CURING HIV INFECTION

[0001] The present invention relates to pyrrolopyridine derivatives for use in methods of curing HIV infections. The present invention further relates to compositions suitable for use in these methods. The present invention further relates to the use of allosteric integrase inhibitors (ALLINIs) in retroviral gene therapy

[0002] Acquired immune deficiency syndrome (AIDS) is caused by human immunodeficiency virus (HIV) infection. There are two types of HIVs, HIV-1 and HIV-2, and the type most prevalent globally is HIV-1. For the treatment of AIDS, enzyme inhibitors have been developed in accordance with action mechanisms of HIV. Depending on the point of action, those inhibitors are classified into Nucleoside Reverse Transcriptase Inhibitor (NRTI), Protease Inhibitor (PI), Fusion Inhibitor, and Integrase Inhibitor. These four different classes are combined in different ways, which are referred to as anti-retroviral therapy (ART), combination anti-retroviral therapy (cART) or highly active anti-retroviral therapy (HAART).

[0003] The allosteric integrase inhibitor (ALLINI) binds to the LEDGF / p75 binding pocket of HIV-1 integrase (Frauke Christ, Zeger Debyser et al., Nature Chemical Biology, 2010, Vol. 6, 442), and thus, the development of inhibitors for affecting late-stage HIV replication has been conducted. It has been suggested that their potency is mainly determined by their effect on HIV particle maturation (Jurado et al., 2013). Based thereon, their therapeutic value would primarily be in combination with other antiretroviral drugs to suppress HIV replication in a chronic therapy.

[0004] It has been known that HIV infection is currently incurable due to latently infected cells that induce viral reinfection upon drug discontinuation, which is considered as a major limitation on HIV cure. Attempts to cure HIV through shock-and-kill (Cell Host Microbe. 2018 Jan 10; 23(1): 14-26.) and block-and-lock (Viruses 2020, 12, 84) methods have been studied at the laboratory level, but no success has been reported so far. Despite the success of antiretroviral therapy (ART), there is still a need for a cure, a drug that can completely eradicate the disease from HIV patients.

[0005] In an effort to meet the need for curative treatments of retroviral infections, the inventors have conducted intensive research to find a substance capable of curing HIV, and found that the pyrrolopyridine derivatives inhibit the HIV-1 reactivation from the HIV-1 T cell reservoirs.

[0006] Therefore, an object of the present invention is to provide a compound for use in methods of curing HIV infections.

[0007] Another object of the present invention is to provide a composition comprising the compound suitable for use in these methods.

[0008] A further object of the present invention is to provide a use of ALLINIs in retroviral gene therapy.

[0009] In order to achieve the above-described technical problems, the present invention provides Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, which is for use in a method of curing HIV-1 infection, wherein the method comprises administering Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, to a subject; and determining the efficacy of the curing by determining the reactivation potential of a retroviral reservoir of the subject, so as to determine whether or not treatment can be stopped.

[0010] Compound I

[0011]

[0012] According to the present invention, it has been provided a compound, which can restrict the HIV-1 reactivation from the HIV-1 T cell reservoirs, and thus, can be used in a method of curing HIV-1 infection, wherein the method comprises administering the compound to a subject and determining the efficacy of the curing by determining the reactivation potential of a retroviral reservoir of the subject, so as to determine whether or not treatment can be interrupted.

[0013] Figure 1 illustrates the suppression of HIV-1 reactivation in latently infected T cells by Compound I, in case that Compound I was removed from the culture medium after the 24-hour HIV-1 infection period, and the reactivation of the infected T cells was induced using IL-15.

[0014] Figure 2 illustrates the suppression of HIV-1 reactivation in latently infected T cells by Compound I, in case that Compound I was removed from the culture medium after the 24-hour HIV-1 infection period, and the reactivation of the infected T cells was induced using PMA / Ionomycin.

[0015] Figure 3 illustrates the suppression of HIV-1 reactivation in latently infected T cells by Compound I, in case that Compound I was removed from the culture medium after the 24-hour HIV-1 infection period, and the reactivation of the infected T cells was induced using LRA (Bryostation+Romidepsin).

[0016] Figure 4 illustrates the suppression of HIV-1 reactivation in latently infected T cells by Compound I, in case that Compound I was maintained in the culture medium until just before reactivation, and the reactivation of the infected T cells was induced using IL-15.

[0017] Figure 5 illustrates the suppression of HIV-1 reactivation in latently infected T cells by Compound I, in case that Compound I was maintained in the culture medium until just before reactivation, and the reactivation of the infected T cells was induced using PMA / Ionomycin.

[0018] Figure 6 illustrates the suppression of HIV-1 reactivation in latently infected T cells by Compound I, in case that Compound I was maintained in the culture medium until just before reactivation, and the reactivation of the infected T cells was induced using LRA (Bryostation+Romidepsin).

[0019] A first aspect of the present invention relates to Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, which is for use in a method of curing HIV-1 infection, wherein the method comprises administering the Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, to a subject; and determining the efficacy of the curing by determining the reactivation potential of a retroviral reservoir of the subject, so as to determine whether or not treatment can be stopped.

[0020] Compound I

[0021]

[0022] According to the present invention, it was identified that Compound I inhibit the enzymatic activity of retroviral integrase (IN) by disrupting the interaction between IN and LEDGF / p75. As a result, Compound I can reduce the number of integrated proviruses and retargets integration to safe genomic sites, thereby rendering the residual retroviral reservoirs transcriptionally inactive and preventing the reactivation of the retroviral reservoirs. Accordingly, appropriate dosing of Compound I can not only suppress viral replication but also establish non-reactivatable viral reservoirs, so that can offer the potential for functional cure or remission of HIV-1 infection.

[0023] The duration of administration and dosages of Compound I, eligibility criteria for treatment, determining the discontinuation of treatment, measuring reactivation potential of a retroviral reservoir, etc. can be determined in a manner similar to the conventional methods, for example, as disclosed in WO 2016 / 180770.

[0024] Therefore, in an embodiment of the invention, the Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, is administered until the HIV-1 reservoir is resistant to reactivation.

[0025] In another embodiment of the invention, the daily dosage of the Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, is equal to or higher than the dosage envisaged for use in standard therapy.

[0026] In another embodiment of the invention, the daily dosage of the Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, is a dosage which ensures a trough concentration in the blood which is at least times 5 times the antiviral EC50of Compound I.

[0027] In another embodiment of the invention, the subject is one who has received standard anti-retroviral combination therapy and the antiretroviral combination therapy is interrupted prior to the administration of Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof.

[0028] In another embodiment of the invention, the Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, is administered daily during a discrete period of 1 to 24 weeks, and the efficacy of cure is determined after said discrete period of 1 to 24 weeks.

[0029] In another embodiment of the invention, the method comprises administering the Compound I, or a racemate, stereoisomer or a pharmaceutically acceptable salt thereof, as a monotherapy.

[0030] Examples

[0031] In the following, the invention will be described in more detail by way of examples. However, the following embodiments are intended to illustrate the present invention and are not intended to limit the scope of the present invention.

[0032] Example 1: Test for the Compound I effect on HIV-1 reactivation in latently infected T cells

[0033] Activated CD4+T cells were isolated from at least 4 health donors (NY Blood Center). For Control Group, Compound I was not treated. For Experimental Group 1, activated CD4+T cells (2x106cells / ml, 6μg / ml PHA) were treated with Compound I (0.01μM, 0.1μM, 1.0μM and 10μM) for 2 hours, and then infected with dual-tropic HIV-1 89.1 strain for 24 hours. The media containing infected CD4+T cells were washed 3 times to remove Compound I and cultured in a media containing HAART (5nM Raltegravir, Efavirenz, and 3TC) without IL-2 for 6 days to establish the latently infected T cells.

[0034] The reactivation of the infected CD4+T cells was conducted for 48 hours with three different conditions, i.e., with IL-15 at 10ng / ml, PMA / Ionomycin at 100nM / 2μM, and with latency reversing agents (LRA, Bryostation+Romidepsin) at 10μM, respectively. The produced viruses in the supernatants were determined by p24 ELISA assay.

[0035] For Experimental Group 2, the same procedure as for Experimental Group 1 was followed except for removing Compound 1 just before the reactivation.

[0036] Figures 1 to 6 show the suppression of HIV-1 reactivation by Compound I in latently infected T cells. Figures 1 to 3 show the results when Compound I was removed from the media after the 24-hour HIV-1 infection period (Experimental Group 1), whereas Figures 4 to 6 show the results when Compound I was removed from the media just before the reactivation (Experimental Group 2).

[0037] HIV-1 productions were elevated by all three reactivation conditions, compared to the un-reactivated control conditions, indicating that thein vitroHIV-1 T cell latency model functioned as expected. However, the 2-hour pre-treatment with Compound I before HIV-1 infection and maintaining Compound I throughout the 24-hour HIV-1 infection period greatly suppressed the HIV-1 production even after the reactivations were induced with three different agents (Figures 1 to 3). More effective suppressions of HIV-1 reactivation were observed when Compound I was maintained throughout the latency until before the reactivation (Figures 4 to 6). Overall, these data confirm that allosteric integrase inhibitor, Compound I, restricts the HIV-1 reactivation from the HIV-1 T cell reservoirs by altering the HIV-1 proviral DNA integration site profiling. For Figure 2, although Compound I at 1μM was not tested, it is expected that it can also inhibit at 1μM the HIV-1 reactivation efficiently in view that Compound I at 0.01μM and 10μM inhibited more than 85% upon the PMA / Ionomycon treatment.

Claims

1.Compound I, or a racemate, stereoisomer or a pharmaceutically acceptable salt thereof, for use in a method of curing HIV-1 infection, wherein the method comprises administering Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, to a subject; and determining the efficacy of curing by determining the reactivation potential of a retroviral reservoir of the subject, so as to determine whether or not treatment can be stopped.Compound I2.The Compound I, or a racemate, stereoisomer or a pharmaceutically acceptable salt thereof, according to claim 1, wherein Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, is administered until the HIV-1 reservoir is resistant to reactivation.3.The Compound I, or a racemate, stereoisomer or a pharmaceutically acceptable salt thereof, according to claim 1, wherein daily dosage of Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, is equal to or higher than the dosage envisaged for use in standard therapy.4.The Compound I, or a racemate, stereoisomer or a pharmaceutically acceptable salt thereof, according to claim 1, wherein daily dosage of Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, is a dosage which ensures a trough concentration in the blood which is at least times 5 times the antiviral EC50of Compound I.5.The Compound I, or a racemate, stereoisomer or a pharmaceutically acceptable salt thereof, according to claim 1, wherein the subject has received standard anti-retroviral combination therapy and the antiretroviral combination therapy is interrupted prior to the administration of Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof.6.The Compound I, or a racemate, stereoisomer or a pharmaceutically acceptable salt thereof, according to claim 1, wherein Compound I, or a racemate, a stereoisomer or a pharmaceutically acceptable salt thereof, is administered daily during a discrete period of 1 to 24 weeks, and the efficacy of cure is determined after said discrete period of 1 to 24 weeks.7.The Compound I, or a racemate, stereoisomer or a pharmaceutically acceptable salt thereof, according to claim 1, wherein the method comprises administering Compound I, or a racemate, stereoisomer or a pharmaceutically acceptable salt thereof, as a monotherapy.

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