Combination of TLR7 modulating compound and HIV vaccine

Abstract

The present disclosure describes methods, compositions, and kits related to a combination of a TLR7 modulating compound and an HIV vaccine. The combination may be used in a method of treating or preventing human HIV infection.

Description

[0001] This application is a divisional application of a patent application with an application date of May 21, 2020, application number 202080037405.6, and name “Combination of TLR7 regulatory compounds and HIV vaccines”.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Application No. 62 / 851,363, filed May 22, 2019, which is incorporated herein by reference in its entirety for all purposes.

[0004] Sequence Listing

[0005] This application contains a sequence listing, which is submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. The ASCII copy was created on April 29, 2020, is named 1300PF_ST25.txt, and is 14,473 bytes in size. Background Art

[0006] The innate immune system provides the body with the first line of defense against invading pathogens. In the innate immune response, invading pathogens are recognized by germline-encoded receptors, whose activation initiates a signaling cascade that leads to the induction of cytokine expression. Innate immune system receptors have a broad spectrum of specificity, recognizing molecular structures that are highly conserved among different pathogens. One family of these receptors is known as Toll-like receptors (TLRs) due to their homology to the receptors originally identified and named in the genus Drosophila. TLRs are found in cells such as macrophages, dendritic cells, and epithelial cells.

[0007] There are at least ten different TLRs in mammals. Ligands and corresponding signal cascades for some of these receptors have been identified. For example, TLR2 is activated by lipoproteins of bacteria (e.g., E. coli), TLR3 is activated by double-stranded RNA, TLR4 is activated by lipopolysaccharides (i.e., LPS or endotoxin) of Gram-negative bacteria (e.g., Salmonella and E. coli O157:H7), TLR5 is activated by flagellin of motile bacteria (e.g., Listeria), TLR7 recognizes and responds to imiquimod, and TLR9 is activated by non-methylated CpG sequences of pathogen DNA. Stimulation of each of these receptors leads to the activation of transcription factors NF-κB and other signaling molecules involved in regulating the expression of cytokine genes (including those encoding tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), and certain chemokines). Agonists of TLR7 are immunostimulants and can induce endogenous interferon-α production in vivo.

[0008] There are many diseases, disorders and conditions associated with TLRs such that therapy using TLR agonists is considered promising, including but not limited to melanoma, non-small cell lung cancer, hepatocellular carcinoma, basal cell carcinoma, renal cell carcinoma, myeloma, allergic rhinitis, asthma, COPD, ulcerative colitis, liver fibrosis and viral infections.

[0009] TLR7 modulating compounds include TLR7 agonist compounds of US Pat. Nos. 8,367,670, 8,629,142, and 8,809,527, as demonstrated by the IFN-α minimum effective concentration (MEC). The activity of the TLR7 agonist GS-9620 has been discussed in the following articles: Lanford et al., Gastroenterology 2013, 144(7), 1508-17 (Gastroenterology, 2013, Vol. 144, No. 7, pp. 1508-1517), and Roethle, P. et al., J. Med. Chem. 2013, 56(18), 7324-7333 (Journal of Medicinal Chemistry, 2013, Vol. 56, No. 18, pp. 7324-7333), which discuss the TLR7 agonist activity of compounds in U.S. Pat. Nos. 8,367,670, 8,629,142, and 8,809,527, including those in Examples 4, 49, 89, 99, and 105.

[0010] Around the world, more than 36 million people are infected with HIV virus. Many drugs and combination therapies have been developed to treat human HIV infection. Although combined antiretroviral therapy (cART) and highly active antiretroviral therapy (HAART) have been able to reduce HIV virus activation, usually below 50 copies of HIV RNA / ml of plasma, the therapy of clearing HIV-infected cells is not yet provided, and the cells infected by this HIV will not actively replicate HIV, commonly referred to as the latent HIV reservoir of the patient. The strategy of the "attack and kill" method for treating HIV has been sought, wherein the cells of the latent reservoir "attack" HIV-infected cells to induce transcription of static, replication-competent HIV proviruses, thereby producing a state of transient viremia and making activated cells sensitive to "killing" from antiretroviral therapy. The “attack” procedure has tested a variety of agents, including histone deacetylase inhibitors, disulfiram, PD-1 antibodies, and HIV vaccines, as described in the following references: Barton, KM et al., Clin. Pharmacol. Ther. 2013, 93(1), 46-56; Marsden, MD et al., Cell 2014, 158(5), 971-972; Battistini, A. et al., Viruses 2014, 6(4), 1715-1758 (Virus, 2014, Vol. 6, No. 4, pp. 1715-1758); and Cillo, AR et al., Proc. Natl. Acad. Sci. 2014, 111(19), 7078-7083 (Proceedings of the National Academy of Sciences of the United States of America, 2014, Vol. 111, No. 19, pp. 7078-7083).

[0011] The increased use of highly effective combined antiretroviral therapy (cART) has led to a significant reduction in the morbidity and mortality associated with human immunodeficiency virus (HIV) infection. However, despite the availability of new classes of antiretroviral drugs, currently available cART regimens cannot eradicate HIV from the body. Therefore, cART cessation in participants who maintain undetectable viral loads is followed by a recurrence of viremia. This reflects that standard cART cannot clear the viral reservoir formed by latently infected cells, in which the integrated provirus remains static and stable from the early stages of infection, and the immune response cannot effectively contain viral rebound after treatment interruption.

[0012] Even though cART results in control of viral load (thus preventing the development of AIDS and viral transmission), it has several problems:

[0013] (a) Non-curative: cART is a lifelong treatment. If a person stops treatment, the viral load generally rebounds to the original level within 2 to 4 weeks, making the person reinfected.

[0014] (b) Adherence issues: 30% to 50% of patients cannot control their viral load because they do not strictly follow their treatment regimen. This is largely related to psychological stress (being infected with HIV and having no hope of being cured affects the patient's quality of life), and even without psychological stress, patients also feel inconvenienced to varying degrees by their treatment routine (also known as "pill fatigue").

[0015] (c) Resistance: HIV can develop resistance to cART.

[0016] (d) Side effects: Due to the long-term toxicity of cART, patients may suffer from cardiovascular disease, dyslipidemia, hypertension, diabetes, osteoporosis, or kidney disease.

[0017] (e) High cost: Treating a patient with cART costs approximately $20,000 per year, and the total cost to the health system over the patient's lifetime is calculated to be over $400,000.

[0018] (f) Social stigma: The stigma surrounding HIV makes people reluctant to get tested and disclose their HIV status; it also limits their access to HIV treatment.

[0019] There remains a need for new agents and therapies that can help activate latent HIV-infected cells to enhance the activity of antiretroviral therapy and immune responses. Summary of the invention

[0020] In one embodiment, the present disclosure provides a method of treating or preventing HIV infection in a human in need thereof who has HIV infection or is at risk of developing HIV infection, the method comprising administering to the human a therapeutically effective amount of a compound of formula (I):

[0021]

[0022] or a pharmaceutically acceptable salt thereof,

[0023] and a first virus comprising a nucleic acid encoding an immunogenic polypeptide comprising:

[0024] (i) a sequence having at least 90% sequence identity to SEQ ID NO: 1;

[0025] (ii) a sequence having at least 90% sequence identity to SEQ ID NO: 2;

[0026] (iii) a sequence having at least 90% sequence identity to SEQ ID NO: 3;

[0027] (iv) a sequence having at least 90% sequence identity to SEQ ID NO:4;

[0028] (v) a sequence having at least 90% sequence identity to SEQ ID NO:5;

[0029] (vi) a sequence having at least 90% sequence identity to SEQ ID NO:6;

[0030] (vii) a sequence having at least 90% sequence identity to SEQ ID NO:7;

[0031] (viii) a sequence having at least 90% sequence identity to SEQ ID NO: 8;

[0032] (ix) a sequence having at least 90% sequence identity to SEQ ID NO:9;

[0033] (x) a sequence having at least 90% sequence identity to SEQ ID NO: 10;

[0034] (xi) a sequence having at least 90% sequence identity to SEQ ID NO: 11;

[0035] (xii) a sequence having at least 90% sequence identity to SEQ ID NO: 12;

[0036] (xiii) a sequence having at least 90% sequence identity to SEQ ID NO: 13;

[0037] (xiv) a sequence having at least 90% sequence identity to SEQ ID NO: 14;

[0038] (xv) a sequence having at least 90% sequence identity to SEQ ID NO: 15; and

[0039] (xvi) a sequence having at least 90% sequence identity to SEQ ID NO: 16;

[0040] wherein at least two of (i) to (xvi) are linked by a single, double or triple alanine amino acid linker, wherein the linker results in the formation of an AAA sequence in the connecting region between the adjacent sequences, and wherein the sequence of each of (i) to (xvi) is 11 to 85 amino acids in length.

[0041] Also provided is a method comprising administering to a human a TLR7 modulating compound as described herein and an HIV vaccine. DETAILED DESCRIPTION

[0042] I. Overview

[0043] The present disclosure provides methods, compositions and kits for treating or preventing HIV infection in humans, which include a combination of a TLR7 modulating compound and an HIV vaccine.

[0044] II. Definitions

[0045] "Compounds of the present disclosure" include compounds disclosed herein, for example, compounds of the present disclosure include compounds of formula (I) and pharmaceutically acceptable salts thereof.

[0046] "Tris" or tris(hydroxymethyl)aminomethane, or tromethamine or THAM as it is known during medical usage, is a compound having the formula (HOCH2)3CNH2.

[0047] "Modulation", "modulating" and "modulator" refer to the effect of an agent on the agonism (activation or enhancement) or antagonism (inhibition or attenuation) of a biological target function. Agonists or enhancers include those modulators that increase the activity of a TLR7 receptor. In each method, combination, kit, use, composition and regimen described herein using or comprising a TLR7 modulator or a TLR7 modulating compound, there is a separate embodiment in which the TLR7 modulator or TLR7 modulating compound is an agonist of TLR7. TLR7 agonism can be determined by the PBMC assay protocol in U.S. Patent No. 8,367,670 (the contents of which are incorporated herein by reference), and Bioorg Med. Chem. Lett. 16, 4559 (2006) ("Bioorganic Chemistry and Medicinal Chemistry", Vol. 16, p. 4559, 2006).

[0048] As used herein, "pharmaceutically acceptable" with respect to a substance refers to a substance that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit / risk ratio, and effective for its intended purpose when used in a pharmaceutical composition.

[0049] As used herein, "pharmaceutically acceptable salts" are intended to mean salts of the compounds of the present disclosure that are, within the scope of sound medical judgment, suitable for contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., commensurate with a reasonable benefit / risk ratio, generally water-soluble or oil-soluble or dispersible, and effective for their intended use. The term includes, but is not limited to, pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. A list of suitable salts can be found, for example, in Berge, SM et al., J. Pharm. Sci., 1977, 66, 1-19 (Journal of Pharmaceutical Science, 1977, Vol. 66, pp. 1-19).

[0050] In the context of proteins, a functional equivalent or a fragment of a functional equivalent may have one or more conservative amino acid substitutions. The term "conservative amino acid substitution" refers to replacing an amino acid with another amino acid that has similar properties to the original amino acid. The groups of conservative amino acids are as follows:

[0051] Group Amino acid name Aliphatic Gly, Ala, Val, Leu, Ile Containing hydroxyl or thiol / selenium Ser, Cys, Thr, Met ring Pro Aromatic Phe, Tyr, Trp Alkaline His, Lys, Arg Acid and its amide Asp, Glu, Asn, Gln

[0052] Conservative substitutions may be introduced at any position of a predetermined peptide or fragment thereof. However, it may also be desirable to introduce non-conservative substitutions, particularly but not limited to non-conservative substitutions, at any one or more positions. Non-conservative substitutions resulting in functionally equivalent fragments of a peptide will differ significantly in polarity, charge and / or steric bulk, for example, while maintaining the function of the derivative or variant fragment.

[0053] "Percentage of sequence identity" is determined by comparing two optimally aligned sequences over a comparison window, wherein the polynucleotide or polypeptide sequence in the comparison window may have additions or deletions (i.e., gaps) compared to a reference sequence (which does not have additions or deletions) for optimal alignment of the two sequences. In some cases, the percentage can be calculated by determining the number of positions at which the same nucleic acid base or amino acid residue appears in the two sequences to obtain the number of matching positions, dividing the number of matching positions by the total number of positions in the comparison window, and multiplying the result by 100 to obtain the percentage of sequence identity.

[0054] In the context of two or more nucleic acid or polypeptide sequences, "identical" or percentage "identity" refers to two or more sequences or subsequences that are identical or have a specified percentage of identical amino acid residues or nucleotides when compared and aligned for maximum correspondence over a comparison window or specified region (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity over a specified region, such as an entire polypeptide sequence or a separate domain of a polypeptide). Such sequences are then referred to as "substantially identical". This definition also refers to the complementary sequence of a test sequence.

[0055] A DNA sequence that "encodes" a specific RNA is a DNA nucleic acid sequence that can be transcribed into RNA. A DNA polynucleotide can encode an RNA (mRNA) that is translated into a protein, or a DNA polynucleotide can encode an RNA that is not translated into a protein (e.g., tRNA, rRNA, or guide RNA; also referred to herein as "non-coding" RNA or "ncRNA"). A "protein coding sequence or a sequence encoding a specific protein or polypeptide" is a nucleic acid sequence that is transcribed into mRNA (in terms of DNA) and translated into a polypeptide (in terms of mRNA) in vitro or in vivo when placed under the control of an appropriate regulatory sequence.

[0056] A "vector," "expression vector," or "construct" is a nucleic acid used to introduce a heterologous nucleic acid into a cell, which has regulatory elements to provide for expression of the heterologous nucleic acid in the cell. Vectors include, but are not limited to, plasmids, minicircles, yeast, and viral genomes. In some embodiments, the vector is a plasmid, minicircle, yeast, or viral genome. In some embodiments, the vector is a viral vector.

[0057] As used herein, "treatment" or "treat" or "treating" refers to a method for obtaining a beneficial or desired result. For the purposes of this disclosure, a beneficial or desired result includes, but is not limited to, alleviation of symptoms and / or reduction of the extent of symptoms and / or prevention of worsening of symptoms associated with a disease or disorder. In one embodiment, "treatment" or "treating" includes one or more of the following: a) inhibiting a disease or disorder (e.g., reducing one or more symptoms caused by a disease or disorder, and / or reducing the extent of a disease or disorder); b) slowing down or preventing the development of one or more symptoms associated with a disease or disorder (e.g., stabilizing a disease or disorder, delaying worsening or progression of a disease or disorder); and c) alleviating a disease or disorder, such as causing regression of clinical symptoms, improving the disease state, delaying progression of the disease, improving quality of life, and / or prolonging survival.

[0058] As used herein, "therapeutically effective amount" or "effective amount" refers to an amount that effectively causes a desired biological or medical response, including an amount of an agent, such as a compound of formula (I) or an HIV vaccine, sufficient to achieve such treatment of the disease when administered to a patient to treat the disease. The effective amount will vary depending on the agent, the disease and its severity, and the age, weight, etc. of the patient to be treated. The effective amount may include a range of amounts. As understood in the art, an effective amount may be one or more doses, i.e., a single dose or multiple doses may be required to achieve the desired treatment endpoint. An effective amount may be considered in the context of administering one or more therapeutic agents, and if combined with one or more other agents, a desired or beneficial result can be achieved or realized, then a single agent may be considered to be administered in an effective amount. The appropriate dose of any co-administered agent may optionally be reduced due to the combined action (e.g., additive or synergistic effect) of the agents.

[0059] As used herein, "delay" refers to the development of a disease or condition, which means to slow down, hinder, slow down, retard, stabilize and / or postpone the development of the disease or condition. The delay can be of varying lengths of time, depending on the history of the disease and / or the individual to be treated. It will be apparent to one skilled in the art that a sufficient or significant delay may actually encompass prevention, in that the individual does not develop the disease or condition.

[0060] As used herein, "prevention" or "prevention" or "preventing" refers to a scheme that prevents the onset of a disease or disorder so that the clinical symptoms of the disease do not develop. Therefore, "prevention" involves administering therapy (e.g., administering a therapeutic substance) to a patient before signs of disease can be detected in the patient (e.g., administering a therapeutic substance to the patient in the absence of a detectable infectious agent (e.g., virus) in the patient). The patient may be an individual with a risk of developing a disease or disorder, such as an individual with one or more risk factors known to be associated with the development or onset of a disease or disorder. Therefore, in certain embodiments, the term "prevention of HIV infection" refers to administering anti-HIV therapeutic substances to a patient who does not have a detectable HIV infection. It should be understood that the patient used for anti-HIV preventive therapy can be an individual with a risk of being infected with HIV virus. It should also be understood that prevention does not require a 100% success rate. In some cases, prevention can be understood as reducing the risk of infection, rather than completely eliminating the occurrence of infection.

[0061] As used herein, "human at risk" refers to a person who is at risk of developing a condition to be treated. A person "at risk" may or may not have a detectable disease or condition, and may or may not show detectable disease prior to treatment with the methods described herein. "At risk" means that a person has one or more risk factors that are measurable parameters known in the art that are associated with the development of a disease or condition. A person who has one or more of these risk factors has a higher probability of developing a disease or condition than a person who does not have these risk factors.

[0062] "Viral infection" describes a disease state in which a virus invades a healthy cell, uses the cell's reproductive machinery to multiply or replicate, and ultimately lyses the cell, resulting in cell death, release of viral particles, and infection of other cells by the newly produced progeny viruses. Latent infection with certain viruses (e.g., HIV) is also a possible outcome of viral infection.

[0063] As used herein, "ART" refers to antiretroviral therapy. Generally speaking, the term refers to a combination of antiretroviral drugs used to treat human viral infections (including HIV infection). Combinations and regimens may include multiple (usually three or more) drugs, such as nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), protease inhibitors (PIs), fusion inhibitors, CCR5 agonists and / or integrase inhibitors.

[0064] "Viral load" and "HIV viral load" refer to the level of HIV detectable in the blood of HIV-infected humans. It can be calculated by estimating the amount of virus in the body fluid in question. For example, it can be given as HIV RNA copies per milliliter of blood or plasma. "Undetectable" HIV viral load includes conditions in which HIV RNA copies cannot be routinely detected by standard viral load tests, such as less than 50 copies of HIV RNA per milliliter of blood or plasma.

[0065] "Viraemia" refers to the presence of measurable viruses or viral particles in the circulation of a virus-infected human. Transient viremia refers to a brief, transient or temporary increase in the presence of measurable viruses or viral particles in the circulation of a virus-infected human. Examples of transient HIV viremia include periods in which HIV-1 RNA levels in the blood or plasma of an HIV-infected human that have been maintained at a concentration of less than 50 copies HIV-1 RNA / mL for a period of time are transiently, transiently or temporarily increased to a concentration greater than 50 copies / mL, such as from 50 copies / mL to 2,000 copies / mL.

[0066] The terms "chronic set point", "set point for chronic HIV infection", "viral load set point" and "viral set point for chronic HIV infection" refer to the steady-state HIV viral load established in the blood of an HIV-infected human. The chronic set point can refer to the value of the steady-state HIV viral load after infection, after the introduction of antiretroviral therapy or treatment (including the administration of ART, TLR7 modulating compounds and / or HIV vaccines described herein), or after cessation of antiretroviral therapy or treatment. The chronic set point can be determined in a single HIV-infected human, or as a median chronic set point in a group of HIV-infected humans. When comparing two chronic set points, the first chronic set point can be a percentage of the second chronic set point, or the second chronic set point can be a multiple of the first chronic set point. For example, a first chronic set point of 100 copies of HIV-1 RNA / mL is 10% of a second chronic set point of 1000 copies of HIV-1 RNA / mL, and can alternatively be described as a second chronic set point that is 10 times higher than the first chronic set point.

[0067] "Viral rebound" refers to the following observations: the undetectable HIV viral load in virologically suppressed HIV-infected humans after treatment with ART is usually restored to a detectable pre-treatment HIV viral load after ART is stopped. Viral rebound can occur within a few days or weeks after ART is stopped, for example, 4 weeks. "Viral rebound delay" refers to the expected observation (e.g., 4 weeks) of viral rebound after stopping ART compared to the viral rebound (e.g., 12 weeks) actually observed after stopping another therapy (e.g., administering ART, TLR7 modulating compounds and HIV vaccines) according to the methods described herein. In the above hypothetical example, the delay of viral rebound is 8 weeks after the treatment of ART, TLR7 modulating compounds and HIV vaccines. The delay of viral rebound can be determined in a single HIV-infected human, or in a group of HIV-infected humans as the median delay of viral rebound.

[0068] III. Compounds

[0069] TLR7 modulating compounds useful in the methods, compositions and / or kits of the present disclosure are described herein. In some embodiments, the TLR7 modulating compound is a compound of formula (I):

[0070]

[0071] or a pharmaceutically acceptable salt thereof.

[0072] Pharmaceutically acceptable salts of the TLR7 modulating compounds described herein include, but are not limited to, those salts formed with inorganic and organic acids that retain the biological effectiveness and properties of the free base and are not biologically or otherwise undesirable, including, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, nitric acid, phosphoric acid, and the like, and organic acids including, but are not limited to, acetic acid, trifluoroacetic acid, adipic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, butyric acid, camphoric acid, camphorsulfonic acid, cinnamic acid, Citric acid, digluconic acid, ethanesulfonic acid, glutamic acid, glycolic acid, glycerophosphoric acid, hemisulfonic acid, caproic acid, formic acid, fumaric acid, 2-hydroxyethanesulfonic acid (hydroxyethylsulfonic acid), lactic acid, hydroxymaleic acid, malic acid, malonic acid, mandelic acid, mesitylenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, nicotinic acid, 2-naphthalenesulfonic acid, oxalic acid, pamonic acid, pectinic acid, phenylacetic acid, 3-phenylpropionic acid, pivalic acid, propionic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, tartaric acid, p-toluenesulfonic acid, undecanoic acid, etc.

[0073] Solid forms of the compounds of the present disclosure are also included. Crystalline forms of TLR7 modulating compounds of formula (I) are described in U.S. Patent Nos. 9,738,646 and 10,202,384, each of which is incorporated herein by reference in its entirety. Exemplary crystalline forms of compounds of formula (I) may be characterized by having an X-ray powder diffraction (XRPD) pattern with peaks at 5.8, 11.4, 11.6, 17.7, 20.1, 20.9, 22.3, 23.9, 26.0, and 26.8 degrees 2θ (± 0.2 degrees 2θ), wherein XRPD is characterized by using CuK α1 irradiation, and a differential scanning calorimetry (DSC) pattern with endotherms at about 133° C., 170° C., and 273° C. Another exemplary crystalline form of the compound of Formula (I) can be characterized by an XRPD pattern having peaks at 4.6, 9.2, 15.8, 17.8, 18.3, 19.2, 19.9, 22.4, 25.5, and 29.1 degrees 2θ (±0.2 degrees 2θ), wherein the XRPD is performed using a CuK α1 Irradiation, and DSC endotherms at about 98°C and 253°C were performed.

[0074] IV. Vaccines

[0075] HIV vaccines in the region on Gag, Pol, Vif and Nef proteins of specific targeting HIV virus are described herein. Such HIV vaccines can induce immune responses for one or more HIV proteins, and can protect humans without HIV infection from being infected with viruses, or can have therapeutic effects on people who infect HIV or infect HIV later. Vaccines generally include delivery mechanisms (e.g., viral vectors) and packaging, such as immunogenic compositions or nucleic acids encoding immunogenic compositions, which are designed to produce desired immune responses. In some embodiments, immunogenic compositions include immunogenic polypeptides, which are antigens that can induce adaptive immune responses (i.e., humoral or cell-mediated immune responses) when introduced into the body.

[0076] Any viral vector capable of introducing the desired package into the body to promote an adaptive response can be used in the methods, compositions and / or kits described herein. In some embodiments, the viral vector comprises a live vector vaccine, an inactivated vaccine or a modified envelope vaccine. In some embodiments, the viral vector comprises an adenoviridae, a poxviridae, a herpesviridae, an adeno-associated virus, a cytomegalovirus, a carynpox, a rubella polio virus, a Venezuelan equine encephalitis virus, a lentivirus or a Sendai virus vector. In some embodiments, the viral vector comprises an adenoviridae or a poxviridae viral vector. In some embodiments, the viral vector comprises a poxvirus vector, such as a modified vaccinia virus Ankara (MVA) vector. Exemplary MVA vectors are described in the following literature: Barouch, DH et al., Cell 2013, 155 (3), 531-539 ("Cell", 2013, Vol. 155, No. 3, pp. 531-539) (the entire text is incorporated herein by reference). In some embodiments, the viral vector comprises an adenoviral viral vector, such as a chimpanzee adenovirus, e.g., a replication-defective chimpanzee adenovirus. Exemplary chimpanzee adenovirus vectors have been described, e.g., in U.S. Pat. No. 9,714,435 (incorporated herein by reference in its entirety).

[0077] International publication WO 2013 / 110818 and U.S. Pat. No. 9,988,425 (each of which is incorporated herein by reference in its entirety) describe immunogens for HIV vaccination. Sixteen regions in the Gag, Pol, Vif and Nef proteins of the HIV-1 virus are relatively conserved and are targeted by HIV patients whose viral load is reduced to <5000 copies of HIV-1 RNA / mL. Hancock, G. et al., PLOS Pathogens 2015, 11 (2) ("Public Library of Science Pathogens", 2015, Vol. 11, No. 2), e1004658; Mothe, B. et al., J. Translational Med. 2015, 13, 60 ("Journal of Translational Medicine", 2015, Vol. 13, No. 60). These regions of HIV proteins form the basis of immunogens for therapeutic vaccination of HIV. The following table summarizes the regions of HIV-1 targeted by immunogens:

[0078] HIV-1 Protein Location (HXB2) SEQ ID NO p17 17-94 1 p24 30-43 2 p24 61-71 3 p24 91-150 4 p24 164-177 5 p24 217-231 6 p2p7p1p6 63-89 7 Protease 45-99 8 Reverse transcriptase 34-50 9 Reverse transcriptase 210-264 10 Reverse transcriptase 309-342 11 Integrase 210-243 12 Integrase 266-282 13 Vif 25-50 14 Vif 166-184 15 Nef 56-68 16

[0079] HIV numbering is as described in Korber, BT et al., (1998) Numbering positions in HIV relative to HXB2CG. Reference is made to Korber, CK, Foley, B., Hahn, B., McCutchan, F., Mellors, J. and Sodroski, J (eds.), Human Retroviruses and AIDS 1998, Theoretical Biology and Biophysics Group, Los Alamos National Laboratory, Los Alamos, NM, pp. III-102-111.

[0080] In some embodiments, an HIV vaccine comprises a virus comprising an immunogenic polypeptide or a nucleic acid encoding an immunogenic polypeptide, wherein the immunogenic polypeptide comprises:

[0081] (i) having at least 90%, 91%, 92%, 93%, 94%,

[0082] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0083] (ii) having at least 90%, 91%, 92%, 93%, 94%,

[0084] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0085] (iii) having at least 90%, 91%, 92%, 93%, 94%,

[0086] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0087] (iv) having at least 90%, 91%, 92%, 93%, 94%,

[0088] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0089] (v) having at least 90%, 91%, 92%, 93%, 94%,

[0090] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0091] (vi) having at least 90%, 91%, 92%, 93%, 94%,

[0092] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0093] (vii) having at least 90%, 91%, 92%, 93%, 94%,

[0094] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0095] (viii) having at least 90%, 91%, 92%, 93%, 94%,

[0096] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0097] (ix) having at least 90%, 91%, 92%, 93%, 94%,

[0098] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0099] (x) has at least 90%, 91%, 92%, 93%, 94%,

[0100] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0101] (xi) having at least 90%, 91%, 92%, 93%, 94%,

[0102] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0103] (xii) has at least 90%, 91%, 92%, 93%, 94%,

[0104] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0105] (xiii) having at least 90%, 91%, 92%, 93%, 94%,

[0106] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0107] (xiv) having at least 90%, 91%, 92%, 93%, 94%,

[0108] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0109] (xv) having at least 90%, 91%, 92%, 93%, 94%,

[0110] sequences with 95%, 96%, 97%, 98% or 99% sequence identity; and

[0111] (xvi) has at least 90%, 91%, 92%, 93%, 94%,

[0112] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0113] wherein at least two of (i) to (xvi) are linked by a single, double or triple alanine amino acid linker, wherein the linker results in the formation of an AAA sequence in the connecting region between the adjacent sequences, and wherein the sequence of each of (i) to (xvi) is 11 to 85 (e.g., 11 to 82, 11 to 80, or 11 to 78) amino acids in length. In some embodiments, the immunogenic polypeptide comprises a sequence having no more than 1, 2 or 3 substitutions of an amino acid sequence in any one of SEQ ID NO: 1-SEQ ID NO: 16. In some embodiments, the immunogenic polypeptide comprises a sequence having an amino acid sequence according to SEQ ID NO: 1-SEQ ID NO: 16.

[0114] In some embodiments, the immunogenic polypeptide comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 17. In some embodiments, the immunogenic polypeptide comprises an amino acid sequence according to SEQ ID NO: 17.

[0115] The immunogenic polypeptide can be encoded by any suitable nucleic acid sequence. In some embodiments, the nucleic acid encoding the immunogenic polypeptide comprises a nucleic acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 18. In some embodiments, the nucleic acid encoding the immunogenic polypeptide comprises a nucleic acid sequence according to SEQ ID NO: 18.

[0116] In some embodiments, the HIV vaccine comprises a modified vaccinia virus Ankara (MVA) comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence according to SEQ ID NO: 17. In some embodiments, the HIV vaccine comprises a replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence according to SEQ ID NO: 17.

[0117] V. Composition

[0118] In some embodiments, the present disclosure provides a pharmaceutical composition comprising a TLR7 modulating compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0119] In some embodiments, a pharmaceutical composition comprises an HIV vaccine as described herein and a pharmaceutically acceptable excipient.

[0120] In some embodiments, the pharmaceutical compositions comprise one or more additional therapeutic agents, as described more fully below.

[0121] Pharmaceutical compositions comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof can be prepared with one or more pharmaceutically acceptable excipients, which can be selected according to conventional practice. Tablets can contain excipients, including glidants, fillers, binders, etc. In certain embodiments, compositions comprising TLR7 modulating compounds are provided as solid dosage forms, including solid oral dosage forms.

[0122] Compositions suitable for oral administration described herein may be presented as discrete units (unit dosage forms) including, but not limited to, capsules, sachets or tablets, each containing a predetermined amount of the active ingredient. In one embodiment, the pharmaceutical composition is a tablet.

[0123] The pharmaceutical compositions disclosed herein comprise one or more therapeutic agents disclosed herein, such as compounds of the present disclosure or HIV vaccines, together with pharmaceutically acceptable excipients and optionally other therapeutic agents. The pharmaceutical composition containing the active ingredient may be in any form suitable for the intended method of administration. A pharmaceutically acceptable excipient may be any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the U.S. Food and Drug Administration as acceptable for human use.

[0124] Compositions intended for oral use can be prepared according to any method known in the art for making pharmaceutical compositions, and such compositions may contain one or more excipients, including sweeteners, flavoring agents, colorants, and preservatives, in order to provide a palatable preparation. Tablets containing active ingredients mixed with non-toxic pharmaceutically acceptable excipients suitable for making tablets are acceptable. These excipients may be, for example, inert diluents such as calcium carbonate or sodium carbonate, lactose, lactose monohydrate, cross-linked sodium carboxymethylcellulose, polyvinylpyrrolidone, calcium phosphate or sodium phosphate; granulating agents and disintegrants such as corn starch or alginic acid; binders such as cellulose, microcrystalline cellulose, starch, gelatin, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. Tablets may be uncoated or may be coated by known techniques including microencapsulation to delay disintegration and adsorption in the gastrointestinal tract, thereby providing a sustained effect over a longer period. For example, a time-delay material such as glyceryl monostearate or glyceryl distearate may be used alone or with wax.

[0125] The amount of active ingredients that can be combined with inactive ingredients to produce a dosage form can vary depending on the intended treatment of the patient and the specific mode of administration. For example, in some embodiments, a dosage form of a compound of formula (I) for oral administration to humans may contain about 1 mg to 10 mg of active material, such as about 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, or about 8 mg, formulated with an appropriate and convenient amount of a pharmaceutically acceptable excipient. In certain embodiments, a pharmaceutically acceptable excipient varies between about 5% to about 95% of the total composition (weight: weight).

[0126] In certain embodiments, the composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof in one variation does not include an agent that affects the metabolic rate of the active ingredient. Therefore, it should be understood that in one aspect, the composition comprising a compound of the present disclosure does not include an agent that will affect (e.g., slow down, hinder or retard) the metabolism of the compound of the present disclosure or any other active ingredient that is administered separately, sequentially or simultaneously with the compound of the present disclosure. It should also be understood that any of the methods, kits, products, etc. described in detail herein in one aspect does not include an agent that will affect (e.g., slow down, hinder or retard) the metabolism of the compound of the present disclosure or any other active ingredient that is administered separately, sequentially or simultaneously with the compound of the present disclosure.

[0127] Aqueous compositions, such as those used to prepare HIV vaccine formulations, can be prepared in sterile form and, when intended for delivery by non-oral administration, can generally be isotonic. All compositions may optionally contain excipients such as those described in Rowe et al., Handbook of Pharmaceutical Excipients, 6 th The excipients described in the 6th edition, American Pharmacists Association, 2009. Excipients may include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextrin, hydroxyalkyl cellulose, hydroxyalkyl methyl cellulose, stearic acid, etc.

[0128] The amount of virus in HIV vaccine preparation can be measured by any method known in the art. The amount can be determined by the overall measurement of the number of viral particles (vp) in a certain amount of aqueous composition, for example, by flow cytometry. Alternatively, the amount can be determined by the activity of the virus in the composition, for example, by average plaque determination. Plaque-based determination can be used to determine the concentration of the virus with an infectious dose. The virus plaque assay determines the number of plaque forming units (pfu) in a virus sample, which can be used as a measure of the amount of virus. See, for example, Kaufmann, SH, Kabelitz, D. (2002), Methods in Microbiology Vol. 32: Immunology of Infection. Academic Press. ("Microbiology Methods", Vol. 32, Academic Press), ISBN 0-12-521532-0.

[0129] The compositions include those suitable for various routes of administration, including oral and intramuscular administration. The compositions may be in unit dosage form and may be prepared by any of the methods well known in the pharmaceutical art. Such methods include the steps of associating the active ingredient (e.g., a compound of the present disclosure or a pharmaceutically acceptable salt thereof) with one or more pharmaceutically acceptable excipients. The composition may be prepared by uniformly and intimately associating the active ingredient with a liquid excipient or a finely dispersed solid excipient or both, and then, if necessary, shaping the product. Techniques and formulations generally can be found in Remington: The Science and Practice of Pharmacy, 21 st Edition, Lippincott Williams and Wilkins, Philadelphia, Pa., 2006 (Remington: The Science and Practice of Pharmacy 21st Edition, Lippincott Williams and Wilkins Publishing Company, Philadelphia, Pa., 2006).

[0130] In some embodiments, the composition comprises about 2 mg to about 6 mg, such as about 2 mg, 3 mg, 4 mg, 5 mg or about 6 mg, for example 2 mg or 4 mg, of a compound of formula (I), lactose, microcrystalline cellulose, croscarmellose sodium, magnesium stearate, polyethylene glycol, polyvinyl alcohol, talc and titanium dioxide.

[0131] In some embodiments, the composition comprises about 1×10 50 μL of a replication-defective chimpanzee adenovirus in about 0.5 mL of formulation buffer. 10 About 1×10 11 , for example about 1×10 10 , 2×10 10, 3×10 10 , 4×10 10 , 5×10 10 , 6×10 10 ,7×10 10 , 8×10 10 ,9×10 10 , or about 1×10 11 Virus particles (vp), the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17. In some embodiments, the formulation buffer comprises about 10 mM L-histidine. In some embodiments, the formulation buffer comprises about 35 mM NaCl. In some embodiments, the formulation buffer comprises about 7.5% (w / v) sucrose. In some embodiments, the formulation buffer comprises about 1 mM MgCl2. In some embodiments, the formulation buffer comprises about 0.1 mM EDTA disodium. In some embodiments, the formulation buffer comprises about 0.1% (w / v) polysorbate-80. In some embodiments, the formulation buffer comprises about 0.5% (v / v) ethanol. In some embodiments, the formulation buffer has a pH of about 6.6. In some embodiments, the composition comprises 5×10 10 The invention relates to a method for preparing a novel virus particle (vp), wherein the replication-deficient chimpanzee adenovirus comprises a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, wherein the formulation buffer comprises 10 mM L-histidine, 35 mM NaCl, 7.5% (w / v) sucrose, 1 mM MgCl2, 0.1 mM disodium EDTA, 0.1% (w / v) polysorbate-80, 0.5% (v / v) ethanol, and has a pH of 6.6.

[0132] In some embodiments, the composition comprises about 0.5×10 50 μL of modified vaccinia virus Ankara (MVA) in about 0.5 mL of Tris buffer. 8 About 5×10 8 For example, about 1×10 8 , 2×10 8 , 3×10 8 , 4×10 8 or about 5×10 8 Plaque forming units (pfu) of modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17. In some embodiments, the composition comprises 2×10 pfu of modified vaccinia virus Ankara (MVA) in 0.5 mL of Tris buffer. 8The modified vaccinia virus Ankara comprises a nucleic acid encoding an immunogenic polypeptide having the amino acid sequence of SEQ ID NO: 17.

[0133] VI. Methods

[0134] As will be appreciated by those skilled in the art, when treating a viral infection such as HIV, such treatment can be characterized in a variety of ways and measured by a variety of endpoints. The scope of the present disclosure is intended to encompass all such characterizations.

[0135] In some embodiments, a method for treating or preventing HIV infection in a human being who is in need of HIV infection or who is at risk of developing HIV infection comprises administering a TLR7 modulating compound of the present disclosure (e.g., a compound of formula (I) or a pharmaceutically acceptable salt thereof) and an HIV vaccine as described herein to a human being. In some embodiments, the method can effectively induce an immune response to one or more clades of HIV. In some embodiments, the method can be used to induce an immune response to multiple epitopes of human viral infection. The induction of an immune response to viral infection can be assessed using any technique known to those skilled in the art for determining whether an immune response has occurred. Among other things, suitable methods for detecting an immune response of the present disclosure include detecting a decrease in viral load or antigen in patient serum, detecting antigen-specific T cells that secrete interferon (IFN)-γ, and detecting elevated levels of one or more liver enzymes, such as alanine transferase (ALT) and aspartate transferase (AST). In one embodiment, the detection of antigen-specific T cells that secrete IFN-γ is accomplished using ELISPOT assays or FACS analysis. Another embodiment includes reducing viral load associated with HIV infection, including reduction as measured by PCR testing.

[0136] TLR7 modulating compounds can induce transient viremia from latent HIV reservoirs. See, for example, U.S. Patent Publication 20160008374 (incorporated herein by reference in its entirety). Latent HIV reservoirs and latent HIV infection refer to conditions in which resting CD4+T lymphocytes or other cells are infected with HIV but do not actively produce HIV. Inactive HIV-infected cells are commonly referred to as latently infected cells. Antiretroviral therapy (ART) can reduce the level of HIV in the blood to undetectable levels, while latent HIV reservoirs continue to survive. When latently infected cells are reactivated, the cells begin to produce HIV (HIV replication).

[0137] Methods for treating or preventing HIV infection comprising the "attack and kill" combination of the TLR7 modulating compounds of the present disclosure and HIV vaccines as described herein can target and remove active HIV viruses and activate latent HIV viruses so as to target latent reservoirs and remove HIV viruses from latent reservoirs. Methods for determining the level of HIV in latent reservoirs are known in the art and include, for example, direct measurement of the level of HIV DNA in CD4+T cells and indirect measurement of the time of viral rebound after cessation of anti-HIV therapy. Compared to the viral rebound observed with standard HIV therapy, the control of HIV viremia improved by the treatment or prevention methods described herein can be reflected as a delay in viral rebound.

[0138] In some embodiments, the method for treating or preventing human HIV infection comprising administering a TLR7 modulating compound of the present disclosure and an HIV vaccine as described herein to a human also includes maintaining a low viral load (e.g., less than about 200 copies, 100 copies, 50 copies, or about 20 copies of HIV-1 RNA per milliliter of blood or plasma) for a period of time, such as about one week to about 9 months, after stopping anti-HIV viral therapy (including ART, TLR7 modulating compounds, and HIV vaccines). In some embodiments, the time period after administering a TLR7 modulating compound and an HIV vaccine is longer than administering only a TLR7 modulating compound or an HIV vaccine. In some embodiments, the time period is one week, two weeks, three weeks, one month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, or longer.

[0139] In some embodiments, a method for treating or preventing HIV infection in a human with HIV infection or at risk of developing HIV infection in need thereof comprises administering to the human a TLR7 modulatory compound of formula (I) or a pharmaceutically acceptable salt thereof and an HIV vaccine. In some embodiments, the TLR7 modulatory compound is a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the HIV vaccine comprises a virus comprising a nucleic acid encoding an immunogenic polypeptide comprising a sequence having an amino acid sequence according to SEQ ID NO: 1-EQ ID NO: 16, for example, an immunogenic polypeptide having a sequence of SEQ ID NO: 17.

[0140] In some embodiments, the human with HIV infection in need is a human being that is virologically suppressed, that is, a human being infected with the virus, which is maintained at or below the desired viremia level for a specified human or antiviral treatment or regimen. An example of HIV viral suppression in HIV infected humans can be a measurable HIV viral load of less than 200 copies of HIV-1 RNA per milliliter of blood or plasma maintained in humans. Other examples of virological suppression would be maintaining less than 100 copies / mL, less than 50 copies / mL, less than 40 copies / mL, less than 30 copies / mL, and less than 20 copies / mL of viral load in humans.

[0141] In some embodiments, the method of treating or preventing HIV infection comprises achieving virological suppression in a human. In some embodiments, the method of treating or preventing HIV infection comprises maintaining virological suppression in a human.

[0142] In some embodiments, virological suppression can be achieved by other anti-HIV treatments, such as antiretroviral therapy (ART). In some embodiments, antiretroviral therapy includes HIV reverse transcriptase inhibitors (e.g., nucleoside or non-nucleoside reverse transcriptase inhibitors), HIV integrase inhibitors, HIV non-catalytic site (or allosteric) integrase inhibitors, HIV entry (fusion) inhibitors, HIV maturation inhibitors, or combinations thereof. Exemplary antiretroviral agents include HIV integrase catalytic site inhibitors Raltegravir ( Merck, Inc. (Merck), bictegravir (Gilead), elvitegravir (Gilead), dolutegravir (GSK, ViiV), cabotegravir (GSK 1265744, GSK744, GSK ViiV), and dolutegravir; the HIV nucleoside reverse transcriptase inhibitor abacavir ( GlaxoSmithKline (GSK), didanosine ( Bristol-Myers Squibb Company (BMS)), tenofovir disoproxil fumarate ( Gilead), tenofovir alafenamide (TAF), emtricitabine ( Gilead), lamivudine ( GlaxoSmithKline / Shire), Stavudine ( BMS), Zidovudine ( GlaxoSmithKline), abacavir, efavirenz (Achillion), ethenofovir (CMX-157, Chimerix), and fetofavir (Oncolys); the HIV non-nucleoside reverse transcriptase inhibitor nevirapine ( BI (BI)), Efavirenz ( BMS), etravirine ( Johnson & Johnson (J&J), rilpivirine (TMC278, R278474, Johnson & Johnson), fordavidin (GSK ViiV), doravirine (MK-1439, Merck), and risvirine (Pfizer / ViiV); HIV protease inhibitor atazanavir ( BMS), darunavir ( Johnson & Johnson), indinavir ( Merck), Lopinavir ( Abbvie, Inc. (Abbvie), nelfinavir ( Pfizer), saquilavir ( Roche (Hoffmann-LaRoche), Tipranavir ( BI Company), ritonavir ( AbbVie) and fosamprenavir ( GlaxoSmithKline / Vertex); HIV entry inhibitor maraviroc ( Pfizer), Enfuvirtide ( Trimeris) and forsteravir (BMS-663068, BMS); and the HIV maturation inhibitor bevirimab (Myriad Genetics).

[0143] In some embodiments, the antiretroviral therapy includes one or more agents selected from the group consisting of raltegravir, elvitegravir, dolutegravir, cabotegravir, derogavir, abacavir, didanosine, tenofovir disoproxil fumarate, tenofovir alafenamide, emtricitabine, lamivudine, stavudine, zidovudine, abacavir, efcitabine, ethanofovir, fetenavir, nevirapine, efavirenz, etravirine, rilpivirine, fodavirine, doravirine, risvirine, atazanavir, darunavir, indinavir, lopinavir, nelfinavir, saquelavir, tipranavir, ritonavir, fosamprenavir, maraviroc, enfuvirtide, forsteraxvir, bevirimab, cobicistat, and bictegravir; or a pharmaceutically acceptable salt thereof. In some embodiments, the antiretroviral therapy comprises one or more agents selected from the group consisting of raltegravir, dolutegravir, cabotegravir, derogavir, abacavir, didanosine, tenofovir disoproxil fumarate, tenofovir alafenamide, emtricitabine, lamivudine, stavudine, zidovudine, abacavir, efcitabine, eptinofovir, fetenavir, rilpivirine, fodovirine, doravirine, risvirine, maraviroc, enfuvirtide, forsteravir, bevirimab, and bictegravir; or a pharmaceutically acceptable salt thereof. In some embodiments, the antiretroviral therapy comprises three or more agents, for example, two nucleoside reverse transcriptase inhibitors and a non-nucleoside reverse transcriptase inhibitor or an integrase inhibitor.

[0144] In some embodiments, the method for treating or preventing HIV infection comprises administering a TLR7-regulated compound of formula (I) and an HIV vaccine after administering ART. In some embodiments, the method for treating or preventing HIV infection comprises administering a TLR7-regulated compound of the present disclosure and an HIV vaccine simultaneously with ART. In some embodiments, before and during the administration of the TLR7-regulated compound and the HIV vaccine, the therapeutic agent of ART is the same. In some embodiments, before and during the administration of the TLR7-regulated compound and the HIV vaccine, the therapeutic agent of ART is different.

[0145] HIV vaccination schemes have been developed in non-human primates, which include two different viruses, such as an adenovirus primary immunization vector and a modified vaccinia virus Ankara (MVA) booster vector. See, for example, Barouch, DH et al., Cell 2013, 155 (3), 531-539 ("Cell", 2013, Vol. 155, No. 3, pp. 531-539). This heterologous primary booster vaccination method can provide a more effective HIV vaccine than using a single viral vector. Therefore, in some embodiments, the HIV vaccine includes a first virus and a second virus. In some embodiments, the first virus includes an adenoviridae or poxviridae virus vector, such as an adenovirus virus vector, such as a chimpanzee adenovirus, such as a replication-deficient chimpanzee adenovirus. In some embodiments, the second virus includes a poxviridae virus vector, such as a modified vaccinia virus Ankara (MVA).

[0146] In some embodiments, a method of treating or preventing HIV infection in a human having HIV infection or at risk of developing HIV infection in need thereof comprises administering to the human a therapeutically effective amount of a compound of formula (I):

[0147]

[0148] or a pharmaceutically acceptable salt thereof,

[0149] and a first virus comprising an immunogenic polypeptide or a nucleic acid encoding an immunogenic polypeptide, wherein the immunogenic polypeptide comprises:

[0150] (i) having at least 90%, 91%, 92%, 93%, 94%,

[0151] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0152] (ii) having at least 90%, 91%, 92%, 93%, 94%,

[0153] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0154] (iii) having at least 90%, 91%, 92%, 93%, 94%,

[0155] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0156] (iv) having at least 90%, 91%, 92%, 93%, 94%,

[0157] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0158] (v) having at least 90%, 91%, 92%, 93%, 94%,

[0159] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0160] (vi) having at least 90%, 91%, 92%, 93%, 94%,

[0161] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0162] (vii) having at least 90%, 91%, 92%, 93%, 94%,

[0163] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0164] (viii) having at least 90%, 91%, 92%, 93%, 94%,

[0165] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0166] (ix) having at least 90%, 91%, 92%, 93%, 94%,

[0167] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0168] (x) has at least 90%, 91%, 92%, 93%, 94%,

[0169] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0170] (xi) having at least 90%, 91%, 92%, 93%, 94%,

[0171] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0172] (xii) has at least 90%, 91%, 92%, 93%, 94%,

[0173] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0174] (xiii) having at least 90%, 91%, 92%, 93%, 94%,

[0175] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0176] (xiv) having at least 90%, 91%, 92%, 93%, 94%,

[0177] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0178] (xv) having at least 90%, 91%, 92%, 93%, 94%,

[0179] sequences with 95%, 96%, 97%, 98% or 99% sequence identity; and

[0180] (xvi) has at least 90%, 91%, 92%, 93%, 94%,

[0181] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0182] wherein at least two of (i) to (xvi) are linked by a single, double or triple alanine amino acid linker, wherein the linker results in the formation of an AAA sequence in the connecting region between the adjacent sequences, and wherein the sequence of each of (i) to (xvi) is 11 to 85 (e.g., 11 to 82, 11 to 80, or 11 to 78) amino acids in length. In some embodiments, the immunogenic polypeptide comprises a sequence having no more than 1, 2 or 3 substitutions of an amino acid sequence in any one of SEQ ID NO: 1-SEQ ID NO: 16. In some embodiments, the immunogenic polypeptide comprises a sequence having an amino acid sequence according to SEQ ID NO: 1-SEQ ID NO: 16.

[0183] In some embodiments, the method comprises an immunogenic polypeptide comprising an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 17. In some embodiments, the immunogenic polypeptide comprises an amino acid sequence according to SEQ ID NO: 17.

[0184] In some embodiments, the method comprises a nucleic acid encoding an immunogenic polypeptide comprising a nucleic acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 18. In some embodiments, the nucleic acid encoding the immunogenic polypeptide comprises a nucleic acid sequence according to SEQ ID NO: 18.

[0185] In some embodiments, a method of treating or preventing HIV infection in a human having HIV infection or at risk of developing HIV infection in need thereof comprises administering to the human a therapeutically effective amount of a compound of formula (I):

[0186]

[0187] or a pharmaceutically acceptable salt thereof,

[0188] and a first virus comprising a nucleic acid encoding an immunogenic polypeptide comprising:

[0189] (i) a sequence having at least 90% sequence identity to SEQ ID NO: 1;

[0190] (ii) a sequence having at least 90% sequence identity to SEQ ID NO: 2;

[0191] (iii) a sequence having at least 90% sequence identity to SEQ ID NO: 3;

[0192] (iv) a sequence having at least 90% sequence identity to SEQ ID NO:4;

[0193] (v) a sequence having at least 90% sequence identity to SEQ ID NO:5;

[0194] (vi) a sequence having at least 90% sequence identity to SEQ ID NO:6;

[0195] (vii) a sequence having at least 90% sequence identity to SEQ ID NO:7;

[0196] (viii) a sequence having at least 90% sequence identity to SEQ ID NO: 8;

[0197] (ix) a sequence having at least 90% sequence identity to SEQ ID NO:9;

[0198] (x) a sequence having at least 90% sequence identity to SEQ ID NO: 10;

[0199] (xi) a sequence having at least 90% sequence identity to SEQ ID NO: 11;

[0200] (xii) a sequence having at least 90% sequence identity to SEQ ID NO: 12;

[0201] (xiii) a sequence having at least 90% sequence identity to SEQ ID NO: 13;

[0202] (xiv) a sequence having at least 90% sequence identity to SEQ ID NO: 14;

[0203] (xv) a sequence having at least 90% sequence identity to SEQ ID NO: 15; and

[0204] (xvi) a sequence having at least 90% sequence identity to SEQ ID NO: 16;

[0205] wherein at least two of (i) to (xvi) are linked by a single, double or triple alanine amino acid linker, wherein the linker results in the formation of an AAA sequence in the connecting region between the adjacent sequences, and

[0206] wherein the sequence of each of (i) to (xvi) is 11 to 85 amino acids in length.

[0207] In some embodiments of the method, the immunogenic polypeptide comprises:

[0208] (i) a sequence having at least 95% sequence identity to SEQ ID NO: 1;

[0209] (ii) a sequence having at least 95% sequence identity to SEQ ID NO: 2;

[0210] (iii) a sequence having at least 95% sequence identity to SEQ ID NO: 3;

[0211] (iv) a sequence having at least 95% sequence identity to SEQ ID NO:4;

[0212] (v) a sequence having at least 95% sequence identity to SEQ ID NO:5;

[0213] (vi) a sequence having at least 95% sequence identity to SEQ ID NO:6;

[0214] (vii) a sequence having at least 95% sequence identity to SEQ ID NO:7;

[0215] (viii) a sequence having at least 95% sequence identity to SEQ ID NO: 8;

[0216] (ix) a sequence having at least 95% sequence identity to SEQ ID NO:9;

[0217] (x) a sequence having at least 95% sequence identity to SEQ ID NO: 10;

[0218] (xi) a sequence having at least 95% sequence identity to SEQ ID NO: 11;

[0219] (xii) a sequence having at least 95% sequence identity to SEQ ID NO: 12;

[0220] (xiii) a sequence having at least 95% sequence identity to SEQ ID NO: 13;

[0221] (xiv) a sequence having at least 95% sequence identity to SEQ ID NO: 14;

[0222] (xv) a sequence having at least 95% sequence identity to SEQ ID NO: 15; and

[0223] (xvi) a sequence having at least 95% sequence identity to SEQ ID NO:16.

[0224] In some embodiments of the method, the immunogenic polypeptide comprises a sequence of SEQ ID NO: 1-SEQ ID NO: 16, wherein at least two of SEQ ID NO: 1-SEQ ID NO: 16 are linked by a single, double or triple alanine amino acid linker, and wherein the linker results in the formation of an AAA sequence in the connecting region between the adjacent sequences.

[0225] In some embodiments of the method, the immunogenic polypeptide has an amino acid sequence according to SEQ ID NO:17.

[0226] In some embodiments of the method, the nucleic acid has a nucleic acid sequence according to SEQ ID NO:18.

[0227] In some embodiments of the method, the first virus comprises an adenoviridae or poxviridae viral vector. In some embodiments, the first virus comprises an adenoviral vector. In some embodiments, the first virus comprises a chimpanzee adenoviral vector. In some embodiments, the first virus comprises a replication-defective chimpanzee viral vector. In some embodiments, about 5×10 of the first virus is administered. 10 In some embodiments, the first virus is administered once every 12 weeks. In some embodiments, the first virus is administered twice.

[0228] In some embodiments, the method further comprises administering a second virus comprising a nucleic acid encoding an immunogenic polypeptide. In some embodiments, the second virus comprises a modified vaccinia virus Ankara (MVA) vector. In some embodiments, administering about 2×10 8 In some embodiments, the second virus is administered once every 12 weeks. In some embodiments, the second virus is administered twice.

[0229] In some embodiments of the method, the first virus and the second virus are administered intramuscularly.

[0230] In some embodiments of the method, the first virus is administered at weeks 0 and 12, and the second virus is administered at weeks 24 and 36.

[0231] In some embodiments of the method, about 6 mg to about 8 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof is administered. In some embodiments, a compound of formula (I) or a pharmaceutically acceptable salt thereof is administered every two weeks after the third administration of the virus. In some embodiments, a compound of formula (I) is administered at weeks 26, 28, 30, 32, 34, 38, 40, 42, 44, and 46.

[0232] In some embodiments of the method, the human is virologically suppressed. In some embodiments, a virologically suppressed human has a viral load of less than about 200 copies, 100 copies, 50 copies, or about 20 copies of HIV-1 RNA per milliliter of plasma or blood. In some embodiments, virological suppression is caused by the administration of antiretroviral therapy. In some embodiments, the antiretroviral therapy includes one or more agents selected from the group consisting of raltegravir, elvitegravir, dolutegravir, cabotegravir, derogavir, abacavir, didanosine, tenofovir disoproxil fumarate, tenofovir alafenamide, emtricitabine, lamivudine, stavudine, zidovudine, abacavir, efcitabine, ethanofovir, fetenavir, nevirapine, efavirenz, etravirine, rilpivirine, fodavirine, doravirine, risvirine, atazanavir, darunavir, indinavir, lopinavir, nelfinavir, saquelavir, tipranavir, ritonavir, fosamprenavir, maraviroc, enfuvirtide, forsteraxvir, bevirimab, cobicistat, and bictegravir; or a pharmaceutically acceptable salt thereof.

[0233] In some embodiments, a method of treating or preventing HIV infection in a human having HIV infection or at risk of developing HIV infection in need thereof comprises administering to the human a compound of formula (I):

[0234]

[0235] The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus. 10 A virus particle, the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and

[0236] A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8 plaque forming units, the modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17;

[0237] wherein the first virus is administered at weeks 0 and 12, the second virus is administered at weeks 24 and 36, 4 mg of the compound of formula (I) is administered at weeks 26 and 28, and 6 mg of the compound of formula (I) is administered at weeks 30, 32, 34, 38, 40, 42, 44 and 46.

[0238] In some embodiments, a method of treating or preventing HIV infection in a human having HIV infection or at risk of developing HIV infection in need thereof comprises administering to the human a compound of formula (I):

[0239]

[0240] The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus. 10 A virus particle, the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and

[0241] A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8 plaque forming units, the modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17;

[0242] wherein the first virus is administered at weeks 0 and 12, the second virus is administered at weeks 24 and 36, 4 mg of the compound of formula (I) is administered at weeks 26, 28 and 30, and 6 mg of the compound of formula (I) is administered at weeks 32, 34, 38, 40, 42, 44 and 46.

[0243] In some embodiments, a method of treating or preventing HIV infection in a human having HIV infection or at risk of developing HIV infection in need thereof comprises administering to the human a compound of formula (I):

[0244]

[0245] The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus. 10 A virus particle, the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and

[0246] A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8 plaque forming units, the modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17;

[0247] wherein the first virus is administered at weeks 0 and 12, the second virus is administered at weeks 24 and 36, 4 mg of the compound of formula (I) is administered at weeks 26, 28, 30 and 32, and 6 mg of the compound of formula (I) is administered at weeks 34, 38, 40, 42, 44 and 46.

[0248] In some embodiments, a method of treating or preventing HIV infection in a human having HIV infection or at risk of developing HIV infection in need thereof comprises administering to the human a compound of formula (I):

[0249]

[0250] The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus. 10 A virus particle, the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and

[0251] A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8 plaque forming units, the modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17;

[0252] wherein the first virus is administered at weeks 0 and 12, the second virus is administered at weeks 24 and 36, and 6 mg of the compound of formula (I) is administered at weeks 26, 28, 30, 32, 34, 38, 40, 42, 44 and 46.

[0253] In some embodiments, a method of treating or preventing HIV infection in a human having HIV infection or at risk of developing HIV infection in need thereof comprises administering to the human a compound of formula (I):

[0254]

[0255] The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus. 10 A virus particle, the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and

[0256] A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8 plaque forming units, the modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17;

[0257] wherein the first virus is administered at weeks 0 and 12, the second virus is administered at weeks 24 and 36, 6 mg of the compound of formula (I) is administered at weeks 26 and 28, and 8 mg of the compound of formula (I) is administered at weeks 30, 32, 34, 38, 40, 42, 44 and 46.

[0258] In some embodiments, a method of treating HIV infection in a human suffering from HIV infection in need thereof comprises administering to the human a compound of formula (I):

[0259]

[0260] The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus. 10 A virus particle, the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and

[0261] A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8 plaque forming units, the modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17;

[0262] wherein the first virus is administered at weeks 0 and 12, the second virus is administered at weeks 24 and 36, 6 mg of the compound of formula (I) is administered at weeks 26 and 28, and 8 mg of the compound of formula (I) is administered at weeks 30, 32, 34, 38, 40, 42, 44 and 46.

[0263] The method comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof and an HIV vaccine as described herein to a human being is expected to produce cellular and humoral responses against HIV in humans. In some embodiments, the vaccine produces an effective cytotoxic T cell response. Cytotoxic T cells or cytotoxic T lymphocytes (CTL) assays can be used to monitor cellular immune responses after subgenomic immunization with viral sequences against homologous and heterologous HIV strains. See Burke, S. et al., J. Inf. Dis. 1994; 170: 1110-1119 (Journal of Infectious Diseases, 1994, Vol. 170, pp. 1110-1119) and Tigges, M. et al., J. Immunol, 1996; 156: 3901-3910 (Journal of Immunology, 1996, Vol. 156, pp. 3901-3910). Conventional assays for detecting T cell responses include, for example, proliferation assays, lymphokine secretion assays, direct cytotoxicity assays, and limiting dilution assays. For example, the ability of antigen presenting cells incubated with peptides to induce CTL responses in a responding cell population can be determined. Antigen presenting cells can be cells such as peripheral blood mononuclear cells (PBMCs) or dendritic cells (DCs). Alternatively, a mutant non-human mammalian cell line can be used to test the ability of the target peptide to induce primary CTL responses in vitro, which is defective in the ability to load MHC I class molecules with internally processed peptides and has been transfected with suitable human MHC I class genes. PBMC can be used as a responding cell source for CTL precursors. Appropriate antigen presenting cells are incubated with peptides, and then the antigen presenting cells loaded with proteins are incubated with the responding cell population under optimized culture conditions. Positive CTL activation can be determined by measuring the presence of CTL in the culture, which kills both radiolabeled target cells, specific peptide pulsed targets, and target cells expressing endogenous processed forms of antigens, from which the peptide sequence is derived. For example, target cells can be used 51Cr radiolabeling, and cytotoxic activity can be calculated by the radioactivity released by the target cells. Another suitable method allows direct quantification of antigen-specific T cells by staining with HLA tetramer complexes labeled with fluorescein. See Altman J, et al., Proc. Natl. Acad. Sci. USA 1993; 90: 10330-10334 ("Proceedings of the National Academy of Sciences of the United States of America", 1993, Vol. 90, pp. 10330-10334) and Altman J et al., Science 1996; 274: 94-96 ("Science", 1996, Vol. 274, pp. 94-96). Other relatively recent technical developments include staining of intracellular lymphokines and interferon release assays or ELISPOT assays. In some embodiments, a method of generating effective CTLs in a human in need thereof comprises administering to the human a therapeutically effective amount of a compound of the present disclosure, and a virus encoding an immunogenic polypeptide comprises a sequence according to SEQ ID NO: 1-SEQ ID NO: 16, wherein the CTLs are directed against one or more of the following regions of the HIV virus: p17 17-94, p24 30-43, p24 61-71, p2491-150, p24 164-177, p24 217-231, p2p7p1p6 63-89, protease 45-99, reverse transcriptase 34-50, reverse transcriptase 210-264, reverse transcriptase 309-342, integrase 210-243, integrase 266-282, Vif 25-50, Vif 166-184, and Nef 56-68, wherein the amino acid numbering is according to HIV-1 HXB2.

[0264] Also provided is a method of enhancing the efficacy of an HIV vaccine, the method comprising administering a pharmaceutically effective amount of a TLR7 modulating compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein and an HIV vaccine to a human in need thereof.

[0265] Clinical improvement in treated HIV-infected humans is expected to be greater than in control HIV-infected humans treated with standard of care. Clinical improvement may include one or more of a lower peak viral load, a lower chronic set point, or an increase in delayed viral rebound.

[0266] In some embodiments, the methods described herein have an impact on the treatment of HIV infection, for example, as measured by a lower peak viral load compared to standard therapy, such as ART alone. As generally understood in the art, a comparison of the first peak viral load in a first HIV-infected human and the second peak viral load in a second HIV-infected human is measured during the same time period. In some embodiments, the measurement is performed after all antiviral therapies have stopped. In some embodiments, after treatment with ART, TLR7 modulating compounds, and HIV vaccines, the viral load is maintained at an undetectable level in the first HIV-infected human.

[0267] In some embodiments, the first peak viral load in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is lower than the second peak viral load in a second HIV-infected human after treatment with ART alone. In some embodiments, the second peak viral load in a second HIV-infected human after treatment with ART alone is higher, for example, about 1.2 times to about 10,000 times, about 2 times to about 10,000 times, about 5 times to about 10,000 times, about 10 times to about 10,000 times higher than the first peak viral load in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine. In some embodiments, the second peak viral load in a second HIV-infected human after treatment with ART alone is about 1.2 times, about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, about 10 times, about 20 times, about 50 times, about 100 times, about 200 times, about 500 times, about 1000 times, about 2000 times, about 5000 times, about 1000 times, about 2000 times, about 5000 times, or about 10000 times higher than the first peak viral load in a first HIV-infected human after treatment with ART alone. In some embodiments, the second peak viral load in a second HIV-infected human after treatment with ART alone is about 1000 times higher than the first peak viral load in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine.

[0268] In some embodiments, the first peak viral load in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is lower than the second peak viral load in a second HIV-infected human after treatment with ART and a TLR7 modulating compound. In some embodiments, the second peak viral load in a second HIV-infected human after treatment with ART and a TLR7 modulating compound is higher, for example, about 1.2 times to about 10,000 times, about 2 times to about 10,000 times, about 5 times to about 10,000 times, about 10 times to about 10,000 times higher than the first peak viral load in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine. In some embodiments, the second peak viral load in a second HIV-infected human after treatment with ART and a TLR7 modulating compound is about 1.2 times, about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, about 10 times, about 20 times, about 50 times, about 100 times, about 200 times, about 500 times, about 1000 times, about 2000 times, about 5000 times, about 1000 times, about 2000 times, about 5000 times, or about 10000 times higher than the first peak viral load in a first HIV-infected human after treatment with ART and a TLR7 modulating compound. In some embodiments, the second peak viral load in a second HIV-infected human after treatment with ART and a TLR7 modulating compound is about 20 times higher than the first peak viral load in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine.

[0269] In some embodiments, the first peak viral load in the first HIV-infected human after treatment with ART, TLR7 modulating compound and HIV vaccine is lower than the second peak viral load in the second HIV-infected human after treatment with ART and HIV vaccine. In some embodiments, the second peak viral load in the second HIV-infected human after treatment with ART and HIV vaccine is higher, for example, about 1.2 times to about 10,000 times, about 2 times to about 10,000 times, about 5 times to about 10,000 times, about 10 times to about 10,000 times higher than the first peak viral load in the first HIV-infected human after treatment with ART, TLR7 modulating compound and HIV vaccine. In some embodiments, the second peak viral load in a second HIV-infected human after treatment with ART and an HIV vaccine is about 1.2 times, about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, about 10 times, about 20 times, about 50 times, about 100 times, about 200 times, about 500 times, about 1000 times, about 2000 times, about 5000 times, about 1000 times, about 2000 times, about 5000 times, or about 10000 times higher than the first peak viral load in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine. In some embodiments, the second peak viral load in a second HIV-infected human after treatment with ART and an HIV vaccine is about 100 times higher than the first peak viral load in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine.

[0270] In some embodiments, the methods described herein have an impact on the treatment of HIV infection, for example, as measured by a lower chronic set point compared to standard therapy such as ART. As is generally understood in the art, a comparison of a first chronic set point in a first HIV-infected human to a second chronic set point in a second HIV-infected human is measured at the same time point. In some embodiments, the measurement is made after all antiviral therapies have been stopped.

[0271] In some embodiments, the first chronic set point in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is lower than the second chronic set point in a second HIV-infected human after treatment with ART alone. In some embodiments, the second chronic set point in a second HIV-infected human after treatment with ART alone is higher, for example, about 1.2 times to about 10,000 times, about 2 times to about 10,000 times, about 5 times to about 10,000 times, about 10 times to about 10,000 times higher than the first chronic set point in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine. In some embodiments, the second chronic set point in a second HIV-infected human following treatment with ART alone is about 1.2-fold, about 1.5-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 10-fold, about 20-fold, about 50-fold, about 100-fold, about 200-fold, about 500-fold, about 1000-fold, about 2000-fold, about 5000-fold, about 1000-fold, about 2000-fold, about 5000-fold, or about 10000-fold higher than the first chronic set point in a first HIV-infected human following treatment with ART alone. In some embodiments, the second chronic set point in a second HIV-infected human following treatment with ART alone is about 10-fold higher than the first chronic set point in a first HIV-infected human following treatment with ART, a TLR7 modulating compound, and an HIV vaccine.

[0272] In some embodiments, the first chronic set point in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is lower than the second chronic set point in a second HIV-infected human after treatment with ART and a TLR7 modulating compound. In some embodiments, the second chronic set point in a second HIV-infected human after treatment with ART and a TLR7 modulating compound is higher, for example, about 1.2 times to about 10,000 times, about 2 times to about 10,000 times, about 5 times to about 10,000 times, about 10 times to about 10,000 times higher than the first chronic set point in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine. In some embodiments, the second chronic set point in a second HIV-infected human following treatment with ART and a TLR7 modulating compound is about 1.2-fold, about 1.5-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 10-fold, about 20-fold, about 50-fold, about 100-fold, about 200-fold, about 500-fold, about 1000-fold, about 2000-fold, about 5000-fold, about 1000-fold, about 2000-fold, about 5000-fold, or about 10000-fold higher than the first chronic set point in a first HIV-infected human following treatment with ART and a TLR7 modulating compound. In some embodiments, the second chronic set point in a second HIV-infected human following treatment with ART and a TLR7 modulating compound is about 2-fold higher than the first chronic set point in a first HIV-infected human following treatment with ART, a TLR7 modulating compound, and an HIV vaccine.

[0273] In some embodiments, the first chronic set point in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is lower than the second chronic set point in a second HIV-infected human after treatment with ART and an HIV vaccine. In some embodiments, the second chronic set point in a second HIV-infected human after treatment with ART and an HIV vaccine is higher, for example, about 1.2 times to about 10,000 times, about 2 times to about 10,000 times, about 5 times to about 10,000 times, about 10 times to about 10,000 times higher than the first chronic set point in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine. In some embodiments, the second chronic set point in a second HIV-infected human following treatment with ART and an HIV vaccine is about 1.2-fold, about 1.5-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 10-fold, about 20-fold, about 50-fold, about 100-fold, about 200-fold, about 500-fold, about 1000-fold, about 2000-fold, about 5000-fold, about 1000-fold, about 2000-fold, about 5000-fold, or about 10000-fold higher than the first chronic set point in a first HIV-infected human following treatment with ART, a TLR7 modulating compound, and an HIV vaccine. In some embodiments, the second chronic set point in a second HIV-infected human following treatment with ART and an HIV vaccine is about 10-fold higher than the first chronic set point in a first HIV-infected human following treatment with ART, a TLR7 modulating compound, and an HIV vaccine.

[0274] After all antiviral therapies are stopped, the present method can increase the delay of viral rebound compared to standard therapy. In some embodiments, after treatment with ART, TLR7 modulating compounds and HIV vaccines, viral load does not rebound in HIV-infected humans. In the absence of rebound, previously HIV-infected humans maintain undetectable viral loads after antiviral therapy stops for at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 1.5 years, 2 years, 3 years, 5 years, or at least 10 years or longer after antiviral therapy stops.

[0275] In some embodiments, the first viral rebound delay in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is longer than the second viral rebound delay in a second HIV-infected human after treatment with ART alone. In some embodiments, the first viral rebound delay in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is longer than the second viral rebound delay in a second HIV-infected human after treatment with ART alone by about 1 day to about 10 years, e.g., about 1 week to about 1 year, about 2 weeks to about 1 year, about 3 weeks to about 1 year, about 1 month to about 1 year, about 2 months to about 1 year, about 3 months to about 1 year, about 3 months to about 2 years, etc. In some embodiments, the delay of a first viral rebound in a first HIV-infected human following treatment with ART, a TLR7 modulating compound, and an HIV vaccine is 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 1.5 years, 2 years, 3 years, 5 years, 10 years, or more longer than the delay of a second viral rebound in a second HIV-infected human following treatment with ART alone. In some embodiments, the first viral rebound delay in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is about 1 week, about 2 weeks, about 3 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 1 year, about 1.5 years, about 2 years, about 3 years, or longer. In some embodiments, the first viral rebound delay in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is about 3 months longer than the second viral rebound delay in a second HIV-infected human after treatment with ART alone.

[0276] In some embodiments, the first viral rebound delay in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is longer than the second viral rebound delay in a second HIV-infected human after treatment with ART and a TLR7 modulating compound. In some embodiments, the first viral rebound delay in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is longer than the second viral rebound delay in a second HIV-infected human after treatment with ART and a TLR7 modulating compound by about 1 day to about 10 years, e.g., about 1 week to about 1 year, about 2 weeks to about 1 year, about 3 weeks to about 1 year, about 1 month to about 1 year, about 2 months to about 1 year, about 3 months to about 1 year, about 3 months to about 2 years, etc. In some embodiments, the delay of a first viral rebound in a first HIV-infected human following treatment with ART, a TLR7 modulating compound, and an HIV vaccine is 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 1.5 years, 2 years, 3 years, or longer than the delay of a second viral rebound in a second HIV-infected human following treatment with ART and a TLR7 modulating compound. In some embodiments, the first viral rebound delay in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is about 1 week, about 2 weeks, about 3 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 1 year, about 1.5 years, about 2 years, about 3 years, or longer. In some embodiments, the first viral rebound delay in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is about 3 months longer than the second viral rebound delay in a second HIV-infected human after treatment with ART and a TLR7 modulating compound.

[0277] In some embodiments, the first viral rebound delay in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is longer than the second viral rebound delay in a second HIV-infected human after treatment with ART and an HIV vaccine. In some embodiments, the first viral rebound delay in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is longer than the second viral rebound delay in a second HIV-infected human after treatment with ART and an HIV vaccine by about 1 day to about 10 years, e.g., about 1 week to about 1 year, about 2 weeks to about 1 year, about 3 weeks to about 1 year, about 1 month to about 1 year, about 2 months to about 1 year, about 3 months to about 1 year, about 3 months to about 2 years, etc. In some embodiments, the delay of a first viral rebound in a first HIV-infected human following treatment with ART, a TLR7 modulating compound, and an HIV vaccine is 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 1.5 years, 2 years, 3 years, or longer than the delay of a second viral rebound in a second HIV-infected human following treatment with ART and an HIV vaccine. In some embodiments, the delay of a first viral rebound in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is about 1 week, about 2 weeks, about 3 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 1 year, about 1.5 years, about 2 years, about 3 years, or more longer than the delay of a second viral rebound in a second HIV-infected human after treatment with ART and an HIV vaccine. In some embodiments, the delay of a first viral rebound in a first HIV-infected human after treatment with ART, a TLR7 modulating compound, and an HIV vaccine is about 1 month longer than the delay of a second viral rebound in a second HIV-infected human after treatment with ART and an HIV vaccine.

[0278] VII. Administration

[0279] The combination of a TLR7 modulating compound of the present disclosure (e.g., a compound of formula (I) or a pharmaceutically acceptable salt thereof) and an HIV vaccine as described herein is administered to humans to increase the likelihood of achieving the desired biological effect and to minimize adverse effects. In some embodiments, the TLR7 modulating compound and the HIV vaccine are administered simultaneously. In some embodiments, the TLR7 modulating compound and the HIV vaccine are administered sequentially, for example, on different days or different weeks.

[0280] HIV vaccines as described herein can be administered by any means known in the art, including but not limited to intravenous administration, intramuscular administration, intrathecal administration, intraperitoneal administration, intranasal administration or oral administration. In some embodiments, HIV vaccines are administered intramuscularly. In some embodiments, HIV vaccines are administered once every 8 weeks, 10 weeks, 12 weeks, 14 weeks or 16 weeks. In some embodiments, HIV vaccines are administered once every 12 weeks. In some embodiments, HIV vaccines comprise a first virus and a second virus. In some embodiments, the first virus is administered once or more, and then the second virus is administered once or more. In some embodiments, the first virus is administered twice, and the second virus is administered twice. In some embodiments, the first virus is administered at week 0 and week 12, and the second virus is administered at week 24 and week 36.

[0281] The TLR7 modulating compounds of the present disclosure may be administered by any means known in the art, including but not limited to intravenous administration, intramuscular administration, intrathecal administration, intraperitoneal administration, or oral administration. In some embodiments, the TLR7 modulating compound is administered orally.

[0282] In some embodiments, the TLR7 regulating compound of formula (I) or its pharmaceutically acceptable salt is applied once every week, every two weeks or every three weeks. In some embodiments, the compound of formula (I) or its pharmaceutically acceptable salt is applied once every two weeks, for example, every 12 days-16 days, 13 days-15 days or 14 days. In some embodiments, the compound of formula (I) or its pharmaceutically acceptable salt is applied every two weeks after the third administration of the virus. In some embodiments, the compound of formula (I) or its pharmaceutically acceptable salt is applied in a continuous manner, for example, once every two weeks in eight weeks, for a total of 5 times, that is, in the 26th week, the 28th week, the 30th week, the 32nd week and the 34th week, wherein the 0th week is the first administration of the first virus. In some embodiments, the compound of formula (I) or its pharmaceutically acceptable salt is applied intermittently. In some embodiments, a compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered at week 26, week 28, week 30, week 32, week 34, week 38, week 40, week 42, week 44, and week 46, wherein week 0 is the initial administration of the first virus.

[0283] In some embodiments, the compound of formula (I) is administered in a single tablet. In some embodiments, the compound of formula (I) is administered in two or more (e.g., 3, 4, or 5) tablets. When administered in two or more tablets, the compound of formula (I) may be present in the same dose, such as three tablets of 2 mg of the compound of formula (I) (i.e., 3×2 mg), for a total of 6 mg; or two tablets of 4 mg of the compound of formula (I), for a total of 8 mg; or in different doses, such as one tablet of 4 mg of the compound of formula (I) and one tablet of 2 mg of the compound of formula (I), for a total of 6 mg.

[0284] In some embodiments, about 4 mg to about 12 mg, such as about 6 mg to about 8 mg, for example about 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg or about 12 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof is administered. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered 10 times. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in 10 doses, with one 4 mg tablet for the 1st-2nd dose and 3×2 mg tablets for the 3rd-10th dose. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in 10 doses, with one 4 mg tablet for the 1st-3rd dose and 3×2 mg tablets for the 4th-10th dose. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in 10 doses, with one 4 mg tablet for the 1st-4th dose and 3×2 mg tablets for the 5th-10th dose. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in 10 doses, wherein 3×2 mg tablets are used for doses 1-10. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in 10 doses, wherein 3×2 mg tablets are used for doses 1-2, and 2×4 mg tablets are used for doses 3-10. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in 10 doses, wherein 5×2 mg tablets are used for doses 1-3, and 3×4 mg tablets are used for doses 4-10.

[0285] In some embodiments, the TLR7 modulating compound of formula (I) or a pharmaceutically acceptable salt thereof and HIV vaccine are administered simultaneously with ART. In some embodiments, the therapeutic agent of ART is the same before and during the administration of the compound of formula (I) and HIV vaccine. In some embodiments, the therapeutic agent of ART is different before and during the administration of the compound of formula (I) and HIV vaccine.

[0286] In some embodiments, the method comprises administering to a human a therapeutically effective amount of a compound of formula (I):

[0287]

[0288] or a pharmaceutically acceptable salt thereof,

[0289] and a first virus comprising an immunogenic polypeptide or a nucleic acid encoding an immunogenic polypeptide, wherein the immunogenic polypeptide comprises:

[0290] (i) having at least 90%, 91%, 92%, 93%, 94%,

[0291] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0292] (ii) having at least 90%, 91%, 92%, 93%, 94%,

[0293] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0294] (iii) having at least 90%, 91%, 92%, 93%, 94%,

[0295] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0296] (iv) having at least 90%, 91%, 92%, 93%, 94%,

[0297] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0298] (v) having at least 90%, 91%, 92%, 93%, 94%,

[0299] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0300] (vi) having at least 90%, 91%, 92%, 93%, 94%,

[0301] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0302] (vii) having at least 90%, 91%, 92%, 93%, 94%,

[0303] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0304] (viii) having at least 90%, 91%, 92%, 93%, 94%,

[0305] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0306] (ix) having at least 90%, 91%, 92%, 93%, 94%,

[0307] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0308] (x) has at least 90%, 91%, 92%, 93%, 94%,

[0309] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0310] (xi) having at least 90%, 91%, 92%, 93%, 94%,

[0311] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0312] (xii) has at least 90%, 91%, 92%, 93%, 94%,

[0313] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0314] (xiii) having at least 90%, 91%, 92%, 93%, 94%,

[0315] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0316] (xiv) having at least 90%, 91%, 92%, 93%, 94%,

[0317] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0318] (xv) having at least 90%, 91%, 92%, 93%, 94%,

[0319] sequences with 95%, 96%, 97%, 98% or 99% sequence identity; and

[0320] (xvi) has at least 90%, 91%, 92%, 93%, 94%,

[0321] sequences with 95%, 96%, 97%, 98% or 99% sequence identity;

[0322] wherein at least two of (i) to (xvi) are linked by a single, double or triple alanine amino acid linker, wherein the linker results in the formation of an AAA sequence in the connecting region between the adjacent sequences, and wherein the sequence of each of (i) to (xvi) is 11 to 85 (e.g., 11 to 82, 11 to 80, or 11 to 78) amino acids in length. In some embodiments, the immunogenic polypeptide comprises a sequence having no more than 1, 2 or 3 substitutions of an amino acid sequence in any one of SEQ ID NO: 1-SEQ ID NO: 16. In some embodiments, the immunogenic polypeptide comprises a sequence having an amino acid sequence according to SEQ ID NO: 1-SEQ ID NO: 16.

[0323] In some embodiments, an immunogenic polypeptide is administered that comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 17. In some embodiments, the immunogenic polypeptide comprises an amino acid sequence according to SEQ ID NO: 17.

[0324] In some embodiments, a nucleic acid encoding an immunogenic polypeptide is administered that comprises a nucleic acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 18. In some embodiments, a nucleic acid encoding an immunogenic polypeptide comprises a nucleic acid sequence according to SEQ ID NO: 18.

[0325] In some embodiments, the method comprises administering to a human a compound of formula (I):

[0326]

[0327] The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus. 10 A virus particle, the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and

[0328] A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8plaque forming units, the modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17;

[0329] wherein the first virus is administered at weeks 0 and 12, the second virus is administered at weeks 24 and 36, 4 mg of the compound of formula (I) is administered at weeks 26 and 28, and 6 mg of the compound of formula (I) is administered at weeks 30, 32, 34, 38, 40, 42, 44 and 46.

[0330] In some embodiments, the method comprises administering to a human a compound of formula (I):

[0331]

[0332] The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus. 10 A virus particle, the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and

[0333] A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8 plaque forming units, the modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17;

[0334] wherein the first virus is administered at weeks 0 and 12, the second virus is administered at weeks 24 and 36, 4 mg of the compound of formula (I) is administered at weeks 26, 28 and 30, and 6 mg of the compound of formula (I) is administered at weeks 32, 34, 38, 40, 42, 44 and 46.

[0335] In some embodiments, the method comprises administering to a human a compound of formula (I):

[0336]

[0337] The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus. 10 A virus particle, the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and

[0338] A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8plaque forming units, the modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17;

[0339] wherein the first virus is administered at weeks 0 and 12, the second virus is administered at weeks 24 and 36, 4 mg of the compound of formula (I) is administered at weeks 26, 28, 30 and 32, and 6 mg of the compound of formula (I) is administered at weeks 34, 38, 40, 42, 44 and 46.

[0340] In some embodiments, the method comprises administering to a human a compound of formula (I):

[0341]

[0342] The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus. 10 A virus particle, the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and

[0343] A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8 plaque forming units, the modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17;

[0344] wherein the first virus is administered at weeks 0 and 12, the second virus is administered at weeks 24 and 36, and 6 mg of the compound of formula (I) is administered at weeks 26, 28, 30, 32, 34, 38, 40, 42, 44 and 46.

[0345] In some embodiments, the method comprises administering to a human a compound of formula (I):

[0346]

[0347] The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus. 10 A virus particle, the replication-deficient chimpanzee adenovirus comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and

[0348] A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8plaque forming units, the modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17;

[0349] wherein the first virus is administered at weeks 0 and 12, the second virus is administered at weeks 24 and 36, 6 mg of the compound of formula (I) is administered at weeks 26 and 28, and 8 mg of the compound of formula (I) is administered at weeks 30, 32, 34, 38, 40, 42, 44 and 46.

[0350] VIII. Kit

[0351] The present disclosure provides a kit comprising a TLR7 modulating compound of formula (I) as described herein or a pharmaceutically acceptable salt thereof and an HIV vaccine. The kit may also include instructions for use, for example, for treating viral infections. The instructions for use are typically written instructions, but electronic storage media (e.g., disks or optical disks) containing instructions are also acceptable.

[0352] The present disclosure also provides a drug kit including one or more containers, the container comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and an HIV vaccine. Optionally associated with such containers may be a notification in a form prescribed by a government agency that manages the manufacture, use or sale of the drug, which reflects the approval of the agency for manufacture, use or sale for human administration. Each component may be packaged in a separate container, or some components may be combined in one container when cross-reactivity and shelf life permit. The kit may be a unit dosage form, a batch package (e.g., a multi-dose package) or a subunit dose. The kit may also include a compound and an HIV vaccine in multiple unit doses, which have instructions for use, and are packaged in an amount sufficient to be stored and used in a pharmacy (e.g., a hospital pharmacy and a compounding pharmacy).

[0353] Also provided are articles of manufacture comprising a unit dose of a compound of the present disclosure or a pharmaceutically acceptable salt thereof and an HIV vaccine in suitable packaging for use in the methods described herein. Suitable packaging is known in the art and includes, for example, vials, containers, ampoules, bottles, jars, flexible packaging, etc. The articles of manufacture may be further sterilized and / or sealed.

[0354] IX. Examples

[0355] Example 1: HIV treatment regimen of TLR7 modulating compounds of formula (I) and HIV vaccines

[0356] Phase IIa, randomized, double-blind, double-dummy, placebo-controlled studies are described in this embodiment, which evaluate the safety and tolerability of the sequential regimen of HIV vaccines and compounds of formula (I) in HIV-1 infections of early diagnosis and early treatment. This study screened participants infected with HIV-1, who had started ART within 180 days (6 months) of the estimated date of HIV-1 acquisition, and had achieved virological suppression for at least 1 year. Participants who provided informed consent and met the study inclusion criteria were randomly divided into 1 of 4 parallel treatment groups. The study was conducted in 3 cycles: the first cycle lasted 48 weeks, during which participants received the administration regimen and continued the ART regimen; the second cycle lasted up to 24 weeks, during which participants discontinued the ART regimen (i.e., analytical treatment interruption cycle); and the third cycle lasted up to 12 weeks, during which participants were monitored after restarting their ART.

[0357] For the participants to be included in the study, the following criteria were included: Patients:

[0358] (1) 18 to 60 years old, diagnosed with HIV-1 infection;

[0359] (2) Currently receiving ART, i.e., three or more antiretroviral drugs, initiated within 6 months of the estimated date of HIV-1 infection. Documentation of early treatment initiation is required according to at least 1 of the following criteria (a)-(h):

[0360] (a) Third-generation or fourth-generation assay for HIV-1 / 2 negative and positive plasma HIV-1 RNA < 160 days before ART initiation date,

[0361] (b) Third-generation assay for HIV1 / 2 negative and positive plasma HIV-1p24Ag and positive plasma HIV-1 RNA <158 days before ART initiation date,

[0362] (c) HIV1 / 2 positive and negative HIV-1 and 2 antibody differentiation immunoassay and positive HIV-1 RNA by fourth-generation assay <158 days before ART initiation date,

[0363] (d) positive and negative western blot (WB) test (no band detected) for HIV-1 / 2 and positive HIV-1 RNA by third-generation or fourth-generation assay <157 days before ART initiation date,

[0364] (e) A third-generation or fourth-generation assay for HIV-1 / 2 positivity and indeterminate WB test (<2 envelope bands) and positive HIV-1 RNA <151 days before ART initiation date,

[0365] (f) third-generation or fourth-generation assays for HIV-1 / 2 positive and indeterminate HIV-1 and 2 antibody differentiation immunoassays and positive HIV-1 RNA,

[0366] (g) HIV seroconversion (negative HIV test before first positive HIV test)

[0367] <160 days), and <90 days since HIV-1 diagnosis, ART initiation, and

[0368] (h) third- or fourth-generation assay for HIV-1 / 2 positivity without p31 band and positive WB test <90 days before ART start date in the context of compatible medical history (clearly reported risk of transmission and / or documented acute retroviral syndrome <5 months before HIV diagnosis) <151 days before ART start date;

[0369] (3) Virologically suppressed for at least 1 year before screening, defined as pVL < 50 copies

[0370] / mL; markers that allow separation (<200 copies / mL, non-continuous, indicating less than or equal to twice-yearly total determinations or <10% incidence);

[0371] (4) Stable CD4 count ≥ 450 cells / mm in the 6 months before screening 3 ;as well as

[0372] (5) Since HIV diagnosis, the lowest CD4 count is ≥ 200 cells / mm 3 ; Lower counts isolated during acute HIV-1 infection are allowed only if adequate immune recovery has occurred after ART initiation (see inclusion criterion #4).

[0373] Participants were screened for participation in the study. After providing informed consent, participants were randomly assigned in a ratio of 5:1:1:2 using an interactive response technique (IRT) to receive HIV vaccine and a compound of formula (I), HIV vaccine alone, a compound of formula (I) alone, or placebo. Randomization was stratified by sentinel participants and non-sentinel participants. The sentinel population, consisting of the first 9 participants, was randomly assigned and dosed in a ratio of 5:1:1:2. A blinded, independent safety monitoring committee (SMC) reviewed sentinel population data collected within 1 week after the last sentinel participant received his / her first injection, and then enrolled the remaining 81 participants (non-sentinel population). The SMC reviewed safety data for all participants for the remainder of the study. Sentinel participants who discontinued the study within 1 week after the first dose of investigational product (IMP) were replaced for reasons unrelated to safety, while sentinel participants who discontinued the study after this time point and non-sentinel participants who discontinued the study after the first IMP dose were not replaced. The IMP administration schedule will be managed by the IRT.

[0374] Because the compound of formula (I) is metabolized primarily by cytochrome P450 (CYP) 3A4 enzyme (CYP3A) in vitro, with smaller contributions from CYP2C8 and CYP2D6, and because the compound of formula (I) is a substrate of P-glycoprotein and breast cancer resistance protein in vitro, compound plasma exposure may be increased or decreased when co-administered with CYP3A, P-glycoprotein or breast cancer resistance protein inhibitors or inducers. Any ART agent known to inhibit or induce CYP3A, P-glycoprotein or breast cancer resistance protein was not used in this study during treatment (cycle 1). For those participants who may be taking a regimen that includes one of these drugs, a regimen switch from one of the prohibited drugs to an allowed drug was allowed between screening and baseline visits. The following agents were excluded from the ART regimen during the study: HIV protease inhibitors (including low-dose ritonavir), regimens containing cobicistat, elvitegravir, efavirenz, etravirine and nevirapine.

[0375] Participants had a screening visit within 28 days before the first dose of HIV vaccine. In Cycle 1 (Weeks 0 to 48), participants were randomized to treatment at Week 0 (baseline) and received the first dose of HIV vaccine or matching vaccine placebo administration on the same day. Participants who needed to switch their ART from prohibited drugs to permitted drugs during the study did so between screening and baseline, and for these participants, the screening period was extended to 45 days before the first dose.

[0376] Participants continued to take ART during Cycle 1. Participants in the sentinel population had an additional Cycle 1 visit for additional monitoring and assessment compared to the non-sentinel population. Participants received an HIV vaccine during Cycle 1 comprising a first virus comprising 5×10 50 μL of a replication-defective chimpanzee adenovirus in 0.5 mL of formulation buffer and a second virus: up to two doses of the first virus, two doses of the second virus, and ten doses of a compound of Formula (I), or matching placebo. 10 The replication-deficient chimpanzee adenovirus comprises a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17; the second virus comprises 2×10 8 Plaque forming units of modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17. All participants had HIV-1 viral load monitored throughout Cycle 1. At the end of Cycle 1, participants met the analysis of treatment interruption (ATI) eligibility criteria to start ATI before entering Cycle 2. If the ATI eligibility criteria were not met, entry into Cycle 2 was postponed or the participant was discontinued from the study and the Cycle 1 early termination procedure was performed. Participants in Cycle 1 who prematurely discontinued the vaccine were withdrawn from the study and completed the Cycle 1 early termination assessment, while participants who prematurely discontinued compound administration had the option to continue the study and proceed to Cycle 2.

[0377] The first virus includes a replication-deficient recombinant chimpanzee adenovirus (ChAd) vector based on the chimpanzee adenovirus isolate ChAdY25 described in U.S. Pat. No. 9,714,435 (incorporated herein by reference in its entirety), wherein the vector encodes an immunogenic polypeptide according to SEQ ID NO: 17. The vector was obtained by subcloning the immunogenic polypeptide sequence into the universal ChAdOx1 bacterial artificial chromosome (BAC) system (Oxford University, Oxford, United Kingdom). The plasmid (pC255, 40,483 kbp) generated by the subclone was linearized and transfected into a commercial HEK293 Cells (ThermoFisher Scientific, Waltham, MA USA) were used to produce the first virus, which was formulated into a suspension for intramuscular injection. The buffer for injection contained 10 mM L-histidine, 35 mM NaCl, 7.5% (w / v) sucrose, 1 mM MgCl2, 0.1 mM disodium EDTA, 0.1% (w / v) polysorbate-80, and 0.5% (v / v) ethanol. The pH was adjusted to 6.6 with HCl. The vials were stored at -80°C.

[0378] The schedule for administration in cycle 1 was as follows: at weeks 0 and 12, administration of the first virus comprised 5 × 10 cells / mL of a replication-defective chimpanzee adenovirus in 0.5 mL of formulation buffer. 10 The replication-deficient chimpanzee adenovirus contained a nucleic acid encoding an immunogenic polypeptide having the amino acid sequence of SEQ ID NO: 17. At weeks 24 and 36, a second virus was administered, which contained 2×10 vaccinia virus Ankara (MVA) in 0.5 mL Tris buffer (10 mM Tris HCl, pH 7.7, 140 mM NaCl). 8 Plaque forming units of modified vaccinia virus Ankara comprising a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17. At Weeks 26 and 28, 6 mg of the compound of Formula (I) was administered. At Weeks 30, 32, 34, 38, 40, 42, 44, and 46, 8 mg of the compound of Formula (I) was administered, provided that no compound-related Grade 3 / 4 adverse events occurred when 6 mg of the compound was administered.

[0379] In cycle 2 (week 48 to week 72), participants were instructed to discontinue ART after the 48th week visit. Participants were monitored for rebound of HIV-1 plasma viremia at weekly visits. If specific criteria were met, participants restarted their ART during cycle 2. Participants with viral load <50 copies / mL at the end of cycle 2 and who did not restart ART during cycle 2 underwent additional assessments at the 72nd week visit. Participants who met the criteria for restarting ART during cycle 2 restarted ART, and participants entered cycle 3. If a participant discontinued the study prematurely during cycle 2, the participant's ART regimen should be restarted, and the participant should undergo a 2nd cycle early termination procedure.

[0380] The 24-week second cycle consisted of weekly visits. Participants discontinued ART at week 48. Once all participants met the criteria for doing so or at the latest visit in week 72, they restarted ART. All participants who restarted their ART before the end of cycle 2 continued to cycle 3 when restarting their ART medication. All participants who restarted their ART during cycle 2 (either during cycle 2 or at the end of cycle 2 (i.e., during week 72)) monitored their viral load after restarting ART.

[0381] During cycle 2, careful clinical monitoring for symptoms was performed by the investigator. Participants provided routine blood samples for determination of HIV-1 pVL (ie, plasma viral load), CD4 and CD8 counts, and for ART pharmacokinetics. Participants with a viral load <50 copies / mL after completing 12 weeks of ATI (ie, week 60) and who had not yet restarted ART underwent additional assessments at the week 72 visit; specifically, blood samples for immunological and virological assays were collected.

[0382] In Cycle 3 (Weeks 72 to 84), all participants who restarted their ART during Cycle 2 (either during Cycle 2 or after completion of Cycle 2 (i.e., at the Week 72 visit)) had viral load monitored at Weeks 4 and 12 after restarting ART (Weeks 76 and 84). Participants had an end-of-study visit at Week 84. The Week 84 visit also served as an early termination visit for participants who discontinued the study prematurely during Cycle 3.

[0383] Efficacy endpoints were assessed by changes in HIV-1 pVL over time and changes in viral reservoirs in the periphery and gut-associated lymphoid tissue (GALT). Participants were monitored for viremia throughout the study.

[0384] Blood samples for assessment of HIV-1 pVL were collected to determine pathological control of the virus and viral rebound, such as viral load <50 copies / mL or <2000 copies / mL. During the ATI period (cycle 2, from week 48 to week 72), sustained pathological control was usually less than 50 copies / mL.

[0385] Immunogenicity and pharmacodynamic endpoints were assessed based on the following laboratory tests: (1) interferon-γ ELISPOT assay to determine the breadth and magnitude of de novo T cell responses to HIV vaccine-targeted regions of HIV-1 proteins and the total vaccine-induced HIV-1-specific response, (2) changes from before administration of a compound of formula (I) to 24 hours after administration of the compound: (a) serum / plasma cytokines, (b) gene expression in whole blood (including interferon-stimulated genes), (c) immune cell phenotype / activation in peripheral blood, (3) microbiome based on stool sample collection, and (4) changes in baseline GALT immune cell phenotype / activation, gene expression (including interferon-stimulated genes), HIV-1-specific T cell responses, and HIV-1 reservoirs.

[0386] Fecal samples were collected for microbiome analysis at Weeks 0 and 26 and upon early termination of the protocol, if applicable.

[0387] Example 2: Administration to HIV patients

[0388] A 40-year-old male patient who had been diagnosed with HIV-1 infection and was receiving ART was administered the HIV vaccine according to Example 1 and the compound of formula (I) to treat HIV-1 infection.

[0389] X. Sequence

[0390] In addition to sequences disclosed elsewhere in this disclosure, the following sequences are provided because they are referred to or used in various exemplary embodiments of the disclosure, which are provided for illustrative purposes.

[0391]

[0392]

[0393]

[0394] Although the foregoing disclosure has been described in some detail by way of illustration and example for clarity of understanding and purpose, it will be appreciated by those skilled in the art that certain variations and modifications may be practiced within the scope of the appended claims. In addition, each reference provided herein is incorporated by reference in its entirety to the same extent as if each reference were individually incorporated by reference. In the event of a conflict between the present application and the references provided herein, the present application shall prevail.

Claims

1. A method of treating or preventing HIV infection in a human being in need thereof who has HIV infection or is at risk of developing HIV infection, the method comprising administering to the human being a therapeutically effective amount of a compound of formula (I): or a pharmaceutically acceptable salt thereof, and a first virus comprising a nucleic acid encoding an immunogenic polypeptide comprising: (i) a sequence having at least 90% sequence identity to SEQ ID NO: 1; (ii) a sequence having at least 90% sequence identity to SEQ ID NO: 2; (iii) a sequence having at least 90% sequence identity to SEQ ID NO: 3; (iv) a sequence having at least 90% sequence identity to SEQ ID NO:4; (v) a sequence having at least 90% sequence identity to SEQ ID NO:5; (vi) a sequence having at least 90% sequence identity to SEQ ID NO:6; (vii) a sequence having at least 90% sequence identity to SEQ ID NO:7; (viii) a sequence having at least 90% sequence identity to SEQ ID NO: 8; (ix) a sequence having at least 90% sequence identity to SEQ ID NO:9; (x) a sequence having at least 90% sequence identity to SEQ ID NO: 10; (xi) a sequence having at least 90% sequence identity to SEQ ID NO: 11; (xii) a sequence having at least 90% sequence identity to SEQ ID NO: 12; (xiii) a sequence having at least 90% sequence identity to SEQ ID NO: 13; (xiv) a sequence having at least 90% sequence identity to SEQ ID NO: 14; (xv) a sequence having at least 90% sequence identity to SEQ ID NO: 15; and (xvi) a sequence having at least 90% sequence identity to SEQ ID NO: 16; wherein at least two of (i) to (xvi) are linked by a single, double or triple alanine amino acid linker, wherein the linker results in the formation of an AAA sequence in the connecting region between the adjacent sequences, and wherein the sequence of each of (i) to (xvi) is 11 to 85 amino acids in length.

2. The method of claim 1, wherein the immunogenic polypeptide comprises: (i) a sequence having at least 95% sequence identity to SEQ ID NO: 1; (ii) a sequence having at least 95% sequence identity to SEQ ID NO: 2; (iii) a sequence having at least 95% sequence identity to SEQ ID NO: 3; (iv) a sequence having at least 95% sequence identity to SEQ ID NO:4; (v) a sequence having at least 95% sequence identity to SEQ ID NO:5; (vi) a sequence having at least 95% sequence identity to SEQ ID NO:6; (vii) a sequence having at least 95% sequence identity to SEQ ID NO:7; (viii) a sequence having at least 95% sequence identity to SEQ ID NO: 8; (ix) a sequence having at least 95% sequence identity to SEQ ID NO:9; (x) a sequence having at least 95% sequence identity to SEQ ID NO: 10; (xi) a sequence having at least 95% sequence identity to SEQ ID NO: 11; (xii) a sequence having at least 95% sequence identity to SEQ ID NO: 12; (xiii) a sequence having at least 95% sequence identity to SEQ ID NO: 13; (xiv) a sequence having at least 95% sequence identity to SEQ ID NO: 14; (xv) a sequence having at least 95% sequence identity to SEQ ID NO: 15; and (xvi) a sequence having at least 95% sequence identity to SEQ ID NO:

16.

3. The method of claim 1 or 2, wherein the immunogenic polypeptide comprises a sequence of SEQ ID NO: 1 to SEQ ID NO: 16, wherein at least two of SEQ ID NO: 1 to SEQ ID NO: 16 are linked by a single, double or triple alanine amino acid linker, and wherein the linker results in the formation of an AAA sequence in the connecting region between the adjacent sequences.

4. The method according to any one of claims 1 to 3, wherein the immunogenic polypeptide has an amino acid sequence according to SEQ ID NO:

17.

5. The method according to any one of claims 1 to 4, wherein the nucleic acid has a nucleic acid sequence according to SEQ ID NO:

18.

6. The method of any one of claims 1 to 5, wherein the first virus comprises an adenoviridae or poxviridae viral vector.

7. The method of any one of claims 1 to 6, wherein the first virus comprises an adenoviral viral vector.

8. The method of claim 7, wherein the first virus is a chimpanzee adenovirus.

9. The method of claim 7 or 8, wherein the first virus is a replication-defective chimpanzee adenovirus.

10. The method of any one of claims 7 to 9, wherein about 5×10 10 Virus particles.

11. The method of any one of claims 1 to 10, wherein the first virus is administered once every 12 weeks.

12. The method of claim 11, wherein the first virus is administered twice.

13. The method of any one of claims 1 to 12, wherein the method further comprises administering a second virus comprising the nucleic acid encoding the immunogenic polypeptide.

14. The method of claim 13, wherein the second virus comprises a modified vaccinia virus Ankara (MVA) vector.

15. The method of claim 14, wherein about 2×10 8 plaque forming units.

16. The method of any one of claims 13 to 15, wherein the second virus is administered once every 12 weeks.

17. The method of claim 16, wherein the second virus is administered twice.

18. The method of any one of claims 13 to 17, wherein the first virus and the second virus are administered intramuscularly.

19. The method of any one of claims 13 to 18, wherein the first virus is administered at week 0 and week 12, and the second virus is administered at week 24 and week 36.

20. The method of claim 19, wherein about 6 mg to about 8 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered.

21. The method of claim 20, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered every two weeks after the third viral administration.

22. The method of any one of claims 19 to 21, wherein the compound of formula (I) is administered at week 26, week 28, week 30, week 32, week 34, week 38, week 40, week 42, week 44 and week 46.

23. The method of any one of claims 1 to 22, wherein the human is virologically suppressed.

24. The method of claim 23, wherein the virologically suppressed human has a viral load of less than about 200 copies, 100 copies, 50 copies, or about 20 copies of HIV-1 RNA per milliliter of plasma or blood.

25. The method of claim 23 or 24, wherein the virological suppression is caused by the administration of antiretroviral therapy.

26. The method of claim 25, wherein the antiretroviral therapy comprises one or more agents selected from the group consisting of raltegravir, elvitegravir, dolutegravir, cabotegravir, derogavir, abacavir, didanosine, tenofovir disoproxil fumarate, tenofovir alafenamide, emtricitabine, lamivudine, stavudine, zidovudine, abacavir, efcitabine, ethanofovir, fetenavir, nevirapine, efavirenz, etravirine, rilpivirine, fodavirine, doravirine, risvirine, atazanavir, darunavir, indinavir, lopinavir, nelfinavir, saquelavir, tipranavir, ritonavir, fosamprenavir, maraviroc, enfuvirtide, forsenoside, bevirimab, cobicistat, and bictegravir; or a pharmaceutically acceptable salt thereof.

27. A method of treating HIV infection in a virologically suppressed human with HIV infection in need thereof, said method comprising administering to said human a compound of formula (I): The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus 10 A virus particle, wherein the replication-deficient chimpanzee adenovirus comprises a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8 plaque forming units, wherein the modified vaccinia virus Ankara comprises a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17; in The first virus is administered at week 0 and week 12, The second virus is administered at weeks 24 and 36, 6 mg of the compound of formula (I) is administered at week 26 and week 28, and 8 mg of the compound of formula (I) was administered at 30 weeks, 32 weeks, 34 weeks, 38 weeks, 40 weeks, 42 weeks, 44 weeks and 46 weeks.

28. A method comprising administering to a human a compound of formula (I): The first virus comprises 5×10 of a replication-defective chimpanzee adenovirus 10 A virus particle, wherein the replication-deficient chimpanzee adenovirus comprises a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17, and A second virus comprising 2×10 of modified vaccinia virus Ankara (MVA) 8 plaque forming units, wherein the modified vaccinia virus Ankara comprises a nucleic acid encoding an immunogenic polypeptide having an amino acid sequence of SEQ ID NO: 17; in The first virus is administered at week 0 and week 12, The second virus is administered at weeks 24 and 36, 6 mg of the compound of formula (I) is administered at week 26 and week 28, and 8 mg of the compound of formula (I) was administered at 30 weeks, 32 weeks, 34 weeks, 38 weeks, 40 weeks, 42 weeks, 44 weeks and 46 weeks.

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