TW202110476A - Methods for inducing an immune response against human immunodeficiency virus infection in subjects undergoing antiretroviral treatment - Google Patents

Abstract

Methods for inducing an immune response against Human Immunodeficiency Virus (HIV) in HIV-infected subjects undergoing antiretroviral therapy (ART) are described. The methods involve an adenovirus vector primer vaccine, booster vaccines of an adenovirus vector vaccine, a Modified Vaccinia Ankara virus (MVA) vector vaccine, optionally in combination with isolated HIV envelope polypeptides, and a TLR7 agonist such as vesatolimod or a pharmaceutically acceptable salt thereof.

Description

Method for inducing immune response against human immunodeficiency virus infection in individuals receiving antiretroviral therapy

The number of new HIV infections and the number of acquired immunodeficiency syndrome (AIDS) related deaths has declined. However, globally, in 2016, an estimated 36.7 million people were carrying the human immunodeficiency virus (HIV) (http://www.unaids.org/en/resources/fact-sheet), which is due to the wider use of Life-saving antiretroviral therapy (ART) has increased compared with previous years.

Although it has been proven to successfully inhibit viral replication and save lives, there are still significant challenges in initiating and maintaining ART for all HIV-infected patients in the world who need it. For example, ART does not eliminate virus reservoirs, and treatment is related to incomplete recovery of the host's immune system. In particular, although ART promotes CD4 ⁺ T cell remodeling in the blood, there is only a limited improvement in the function of anti-HIV-specific CD8 ^{+ T cell response.} In addition, to be effective, ART must be performed for life with near-perfect compliance. This puts tremendous pressure and costs on the overtaxed health systems of international donors and developing countries with the highest HIV incidence. In addition, ART has short-term and long-term side effects on users, and as more people undergo longer-term treatment, the drug resistance rate increases. Therefore, alternative or complementary therapies, including therapeutic vaccines that can induce a true or "functional" cure of HIV infection and reduce or eliminate the need for life-long ART for HIV-infected individuals, will therefore have great benefits. The concept of "functional cure" includes treatment strategies that achieve host control of the virus without the need for treatment.

Therefore, there is a need for improved methods for the treatment of individuals infected with HIV, especially novel therapies that seek to achieve a functional cure for HIV, including preferably improving the immune response to HIV and possibly allowing at least some treated individuals to discontinue ART while maintaining viremia Controlled therapeutic vaccine.

The present invention relates to a method for inducing an immune response against HIV in individuals receiving antiretroviral therapy (ART) infected with human immunodeficiency virus (HIV): comprising adenovirus 26 (Ad26) encoding mosaic HIV antigen The first vaccine of the vector, the second vaccine containing the modified Ankara acne (MVA) vector encoding the embedded HIV antigen, and optionally the third vaccine containing the isolated adjuvant branch C gp140 and the embedded gp140 protein, and Toll-like receptor 7 (TLR-7) agonist or a pharmaceutically acceptable salt thereof, preferably vesatolimod (VES) or a pharmaceutically acceptable salt thereof.

In a general aspect, the present invention relates to a method for inducing an immune response against HIV in human subjects infected with human immunodeficiency virus (HIV) receiving antiretroviral therapy (ART), the method comprising: (i) administer an immunogenically effective amount of Ad26 vaccine to the human individual, and the Ad26 vaccine includes an amine that encodes SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4. One or more adenovirus 26 (Ad26) vectors and pharmaceutically acceptable carriers of four HIV antigens based on the base acid sequence; (ii) administering an immunogenically effective amount of MVA vaccine to the human individual, the MVA vaccine comprising one of the four HIV antigens (ie, the HIV antigens of SEQ ID NO: 1, 2, 3, and 4) or Various modified Ankara acne (MVA) carriers and pharmaceutically acceptable carriers; and (iii) Administer an effective amount of Vixamod (VES) or a pharmaceutically acceptable salt thereof to the human individual.

In one embodiment, the method further comprises: re-administering an immunogenically effective amount of Ad26 vaccine to the human individual.

In certain embodiments, the method further comprises: re-administering to the individual an immunogenically effective amount of MVA vaccine.

In certain embodiments, Vixamod or a pharmaceutically acceptable salt thereof is administered at about 3 to 15 mg, such as about 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg per administration. The individual is administered orally in doses of mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, or 15 mg visamod.

In certain embodiments, Vixamod or a pharmaceutically acceptable salt thereof is repeatedly administered to the individual.

In certain embodiments, the Ad26 vaccine consists of the following: a first Ad26 vector encoding the HIV antigen of SEQ ID NO: 1, a second Ad26 vector encoding the HIV antigen of SEQ ID NO: 2, and a second Ad26 vector encoding the HIV antigen of SEQ ID NO: 3. The third Ad26 vector for HIV antigen and the fourth Ad26 vector for HIV antigen encoding SEQ ID NO: 4.

In some embodiments, the MVA vaccine system consists of a single MVA vector encoding the four HIV antigens of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4.

In other embodiments, the MVA vaccine consists of more than one MVA vector encoding four HIV antigens together.

In some embodiments, the first, second, third, and fourth Ad26 carrier systems together use about 5×10 ⁹ to about 1×10 ¹¹ virus particles (vp), preferably about 5×10 ¹⁰ vp. The total dose of Ad26 carrier is administered intramuscularly; and a single MVA carrier or more than one MVA carrier system is used together to provide about 1×10 ⁷ to about 5×10 ⁸ infectious units (IU), preferably about 2×10 ⁸ The total dose of IU of MVA carrier is administered intramuscularly. Preferably, the MVA carrier system MVA-BN carrier.

In some embodiments, the method further comprises administering to the human individual an immunogenically effective amount of a gp140 vaccine comprising one or more isolated HIV gp140 envelope polypeptides, and the gp140 vaccine is administered in combination with the Ad26 vaccine or the MVA vaccine Preferably, the gp140 vaccine is administered in combination with the MVA vaccine. Preferably, the one or more isolated HIV gp140 mantle polypeptides are selected from the group consisting of: two HIV gp140 mantle polypeptides having the amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 10. More preferably, the gp140 vaccine comprises two HIV gp140 polypeptides having the amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 10, respectively. Preferably, the gp140 vaccine further comprises an adjuvant or is co-administered with an adjuvant.

In certain embodiments, the gp140 vaccine is administered in a total dose of about 125 to 350 μg, preferably about 250 μg of glycoprotein of the HIV gp140 mantle polypeptide per administration.

In a specific embodiment, the Ad26 vaccine is administered about 10 to 14 weeks (e.g., 12 weeks) after the initial administration of the Ad26 vaccine.

In certain embodiments, the MVA vaccine is first administered about 22 to 26 weeks after the initial administration of the Ad26 vaccine, for example about 24 weeks.

In certain embodiments, the MVA vaccine is administered approximately 34 to 38 weeks after the initial administration of the Ad26 vaccine, such as 36 weeks.

In certain embodiments, 26 to 34 weeks after the initial administration of the Ad26 vaccine, Vixamod is administered once every two weeks.

In certain embodiments, the Ad26 vaccine is administered 38 to 46 weeks after the initial administration, and furthermore, Vixamod is administered once every two weeks.

In another general aspect, the present invention relates to a method for inducing an immune response against HIV in a human individual who has received antiretroviral therapy (ART) infected with human immunodeficiency virus (HIV), the method comprising: ( i) Intramuscularly administer the Ad26 vaccine to a human individual at a total dose of about 5×10 ⁹ to about 1×10 ¹¹ viral particles (vp), preferably about 5×10 ^{10 vp of Ad26 vector per time.} The Ad26 vaccine includes one or more adenovirus 26 (Ad26) vectors that together encode four HIV antigens with the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4 And a pharmaceutically acceptable carrier; (ii) a total dose of Ad26 vector of ^{about 5×10 9} to about 1×10 ¹¹ viral particles (vp), preferably about 5×10 ^{10 vp per administration} The Ad26 vaccine is administered to human individuals intramuscularly, wherein the Ad26 vaccine is administered 10 to 14 weeks after the Ad26 vaccine is administered in step (i), preferably 12 weeks; (iii) about 1×10 each administration ⁷ to about 5×10 ⁸ infection units (IU), preferably about 2×10 ⁸ IU. The total dose of one or more MVA vectors is administered intramuscularly to a human individual with an MVA vaccine. The MVA vaccine contains four codes. HIV antigen one or more MVA carriers (preferably one or more MVA-BN carriers) and a pharmaceutically acceptable carrier; optionally combined with the MVA vaccine to give about 125 μg to 350 μg each time, more The total dose of about 250 μg of the glycoprotein of the two isolated HIV gp140 mantle polypeptides is further administered to include two isolated HIV gp140 mantles with amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 10 The polypeptide gp140 vaccine, aluminum adjuvant and pharmaceutically acceptable carrier, wherein 22 to 26 weeks after the Ad26 vaccine is administered in step (i), preferably 24 weeks, the MVA vaccine and optionally the gp140 vaccine are administered ; And (iv) a total dose of about 3 mg to about 15 mg per administration, such as about 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg The total dose of 12 mg, 13 mg, 14 mg or 15 mg of visamod is orally administered to a human individual with visamod or a pharmaceutically acceptable salt thereof, wherein Ad26 is administered in step (i) For 26 to 34 weeks after the vaccine, administer Vixamod or its pharmaceutically acceptable salt once every two weeks.

In certain embodiments, the method further comprises: administering about 1×10 ⁷ to about 5×10 ⁸ infectious units (IU), preferably about 2×10 ⁸ IU of one or more MVA at a time The total dose of the carrier is then administered to human subjects by intramuscular MVA vaccine; optionally combined with the MVA vaccine to administer about 125 μg to 350 μg, preferably about 250 μg, of two isolated HIV gp140 mantle polypeptides each time The total dose of glycoprotein is further administered to the human individual with gp140 vaccine, aluminum adjuvant and pharmaceutically acceptable carrier, wherein 34 to 38 weeks, preferably 36 weeks after the Ad26 vaccine is administered in step (i) Re-administer the MVA vaccine and optionally the gp140 vaccine.

In certain embodiments, the method further comprises: each administration in a total dose of about 3 mg to about 15 mg, such as about 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 The total dose of mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, or 15 mg of visamod is orally administered to a human individual with visamod or a pharmaceutically acceptable salt thereof, wherein in the step (i) 38 to 46 weeks after the Ad26 vaccine was administered in China, the administration of Vixamod or its pharmaceutically acceptable salt once every two weeks.

According to an embodiment of the present invention, it is preferable to administer a total dose of about 3 mg to about 15 mg, such as about 3 mg, 4 mg, 26 to 34 weeks and 38 to 46 weeks after the initial administration of Ad26 vaccine. The total dose of, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, or 15 mg visamod is administered orally to humans once every two weeks With Vixamod or its pharmaceutically acceptable salt.

In certain embodiments, the method comprises administering a total dose of 4 mg visamod at each time 26 and 28 weeks after the initial administration of the Ad26 vaccine to the human individual and 30, 32, For 34, 38, 40, 42, 44, and 46 weeks, a total dose of 6 mg of visamod per administration was administered to a human individual orally with visamod or a pharmaceutically acceptable salt thereof.

In certain embodiments, the method comprises administering a total dose of 4 mg of visamod at 26, 28, and 30 weeks after the initial administration of the Ad26 vaccine to the human individual and 32, after the initial administration of the Ad26 vaccine. For 34, 38, 40, 42, 44, and 46 weeks, a total dose of 6 mg of visamod per administration was administered to a human individual orally with visamod or a pharmaceutically acceptable salt thereof.

In certain embodiments, the method comprises administering a total dose of 4 mg visamod at 26, 28, 30, and 32 weeks after the initial administration of the Ad26 vaccine to the human individual and after the initial administration of the Ad26 vaccine For 34, 38, 40, 42, 44, and 46 weeks, a total dose of 6 mg of visamod per administration was administered to a human individual orally with visamod or a pharmaceutically acceptable salt thereof.

In certain embodiments, 26, 28, 30, 32, 34, 38, 40, 42, 44, and 46 weeks after the initial administration of Ad26 vaccine is administered in a total dose of about 6 mg visamod per administration With Vixamod or its pharmaceutically acceptable salt.

In certain embodiments, at 26 and 28 weeks after the initial administration of the Ad26 vaccine, a total dose of about 6 mg visamod is administered per administration and 34, 38, 40, 42, 44 after the initial administration of the Ad26 vaccine. And for 46 weeks, the total dose of 8 mg of visamod per administration was administered to visamod or its pharmaceutically acceptable salt.

In certain embodiments, 26, 28, and 30 weeks after the initial administration of the Ad26 vaccine, a total dose of about 10 mg visamod is administered each time and 32, 34, 38, 40 after the initial administration of the Ad26 vaccine. Administer Vesalimod or its pharmaceutically acceptable salt at a total dose of 12 mg of visamod per administration at 42, 44, and 46 weeks.

In some embodiments, the individual is an individual chronically infected with HIV. In some embodiments, the individual initiates ART outside the acute phase of HIV infection. In a preferred embodiment, the individual initiates ART during the acute phase of HIV infection.

The present invention also relates to a vaccine combination, which is used to induce an immune response against human immunodeficiency virus (HIV) in HIV-infected individuals receiving antiretroviral therapy (ART); and relates to the vaccine combination used in manufacturing Use in an agent for inducing an immune response against human immunodeficiency virus (HIV) in HIV-infected individuals receiving antiretroviral therapy (HIV).

In one embodiment, the vaccine combination includes: (i) Ad26 vaccine, which includes the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 One of the four HIV antigens or multiple adenovirus 26 (Ad26) vectors and pharmaceutically acceptable carriers; (ii) MVA vaccine, which includes one or more modified Ankara acne vectors encoding the four HIV antigens together (Preferably, Bavarian Nordic modified Ankara acne (MVA-BN) carrier) and a pharmaceutically acceptable carrier; and (iii) VES or a pharmaceutically acceptable salt thereof.

In another embodiment, the vaccine combination further comprises: a gp140 vaccine comprising at least one of the isolated HIV branch C having the amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 10 and the mosaic gp140 mantle polypeptide One; aluminum adjuvant; and pharmaceutically acceptable carrier. Preferably, the gp140 vaccine contains both HIV branch C and mosaic gp140 mantle polypeptides.

In certain embodiments, the present invention relates to a method for inducing an immune response against HIV-1 in human individuals infected with HIV-1 treated with antiretroviral therapy (ART), wherein ART has successfully inhibited HIV in the individual. The replication in the bloodstream allows the individual to discontinue ART and maintain control of virus replication in the bloodstream for at least 24 weeks after discontinuation of ART. In view of the present invention, methods known in the art can be used, such as measuring the control of virus replication by measuring HIV viral load (eg, HIV RNA content).

In certain embodiments, the present invention relates to a method for providing HIV relief in HIV-infected human subjects receiving antiretroviral therapy (ART), which comprises inducing an anti-HIV immune response in the subject to thereby improve The HIV infection is controlled after the ART is stopped, wherein the method according to the embodiment of the present invention is used to induce an immune response against HIV. Preferably, after the method of the present invention is used to treat and terminate ART, in at least 20%, preferably at least 30%, 40%, 50% of human individuals, for example, control the viral load of HIV patients (and possibly reduce the HIV viral load) The ART-free viremia control in the reservoir) lasts for at least 24 weeks, preferably longer.

Another general aspect of the present invention relates to a method of treating human immunodeficiency virus (HIV) infection in a human individual in need, the method comprising: (i) Treating human individuals with antiretroviral therapy (ART); and (ii) Using the method of the present invention to induce an immune response against HIV in human individuals.

In certain embodiments, the treatment method further comprises discontinuing the ART treatment of step (i), preferably after the method of the present invention induces an anti-HIV immune response in a human individual. Preferably, the human individual maintains viral suppression for at least 24 weeks after discontinuing ART.

Cross-reference of sequence table submitted electronically

This application claims the benefits of U.S. Provisional Application No. 62/851,258 filed on April 22, 2019, and the entire content of the application is hereby incorporated by reference in its entirety.

Various publications, articles, and patents are cited or described throughout the prior art and this specification; each of these references is incorporated herein by reference in its entirety. The discussion of documents, operations, materials, devices, articles or the like included in this specification is for the purpose of providing the content of the present invention. Such discussion is not an admission that any or all of these content forms part of the prior art regarding any disclosed or claimed invention.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention belongs. In addition, certain terms used herein have the meanings as described in this specification. All patents, published patent applications and publications cited in this article are incorporated by reference as if they were fully described in this article.

It must be noted that unless the context clearly dictates otherwise, as used in the scope of this document and the accompanying application, the singular forms "一(a)", "一(an)" and "the (the)" include plural references .

Unless otherwise indicated, the term "at least" preceding a series of elements should be understood to refer to each element in the series. Those skilled in the art will recognize or be able to ascertain many equivalents of the invention described herein with regard to specific embodiments using only routine experimentation. The present invention is intended to cover such equivalents.

Throughout this specification and the scope of subsequent patent applications, unless the context requires otherwise, the words "comprise" and variations such as "comprises" and "comprising" should be understood as implying the inclusion of a The integer or step or a group of integers or steps, but does not exclude any other integers or steps or groups of integers or steps. When used herein, the term "comprising" can be replaced with the term "containing" or "including", or sometimes when used herein, the term "having" is substituted.

When used in this article, "consisting of" excludes any element, step or ingredient not specified in the claimed element. When used in this article, "essentially composed of" does not exclude materials or steps that do not materially affect the basic and novel features of the technical solution. Whenever used herein in the context of an aspect or embodiment of the present invention, any of the aforementioned terms "comprising", "containing", "including" and "having" may use the term "consisting of" Or "substantially consist of" to change the scope of the present invention.

As used herein, the conjunctive term "and/or" between a plurality of said elements is understood to cover individual and combined options. For example, when two elements are linked by "and/or", the first option refers to the applicability of the first element, excluding the second element. The second option refers to the applicability of the second element, excluding the first element. The third option refers to the applicability of the first element and the second element together. Any one of these options should be understood to be within the scope of this meaning, and therefore meet the requirements of the term "and/or" as used herein. The simultaneous applicability of more than one of these options should also be understood as being within the scope of the meaning, and therefore satisfying the requirements of the term "and/or".

As used herein, "individual" means a human being who will be or has been treated by a method according to an embodiment of the present invention.

The terms "adjuvant" and "immunostimulant" are used interchangeably herein and are defined as one or more substances that stimulate the immune system. In some embodiments, adjuvants are used to enhance the immune response to the HIV antigens and antigenic HIV polypeptides of the present invention.

As used herein, in the context of administering two or more therapies or components to an individual, the terms and phrases "combination", "combination with", "co-delivery" and "administration with" refer to Refers to the simultaneous administration of two or more therapies or components, such as viral expression vectors and isolated antigenic polypeptides. "Simultaneous administration" can mean that at least two components are administered on the same day. When two components are "administered together" or "administered in combination with", they can be administered within a short period of time such as 24, 20, 16, 12, 8 or 4 hours, or within 1 hour, or 30 Within minutes, or within 10 minutes, or within 5 minutes, or within 2 minutes, they can be administered as separate compositions sequentially, or they can be administered as a single composition at the same time. In a typical embodiment, the two components or therapies are administered as separate compositions. The use of the term "in combination with" does not limit the order in which the therapy or components are administered to the individual. For example, the first therapy or component (e.g., viral expression vector) can be administered before (e.g. 5 minutes to one hour before), simultaneously or simultaneously or after the second therapy (e.g., isolated HIV antigenic polypeptide). For example, after 5 minutes to an hour) administration.

The present invention relates to a method for inducing an immune response against human immunodeficiency virus (HIV) in HIV-infected human individuals receiving antiretroviral therapy (ART). According to an embodiment of the present invention, the Ad26 vaccine comprises one or more adenovirus 26 (Ad26) vectors, which encode the amine groups of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4 Acid sequence of four HIV antigens. In some embodiments of the present invention, the immunogenically effective amount of Ad26 vaccine is administered to the individual more than once. According to the present invention, the MVA vaccine comprises one or more modified Ankara acne carriers, preferably one or more Bavarian Nordic modified Ankara acne (MVA-BN) vectors, which encode four HIV antigens, that is, have SEQ ID NO: 1. The HIV antigen of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 amino acid sequence. In some embodiments of the invention, the immunogenically effective amount of MVA vaccine is administered to the individual more than once. In certain embodiments of the present invention, another vaccine is administered with the Ad26 vaccine, or preferably with the MVA vaccine. Preferably, the other vaccine is a gp140 vaccine containing one or more isolated HIV gp140 mantle polypeptides. Preferably, the gp140 vaccine comprises one or more of isolated HIV branch C and mosaic gp140 protein having the amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 10, respectively. More preferably, the gp140 vaccine contains both HIV branch C and mosaic gp140 protein. In all the embodiments of the present invention, the human subject is further administered with Vixamod or its pharmaceutically acceptable salt. "HIV vaccine" as used herein includes the use and administration of Ad26 vaccine and MVA vaccine as described herein, and optionally further includes the use and administration of gp140 vaccine as described herein.

Human immunodeficiency virus (HIV)

Human immunodeficiency virus (HIV) is a member of the Lentivirinae family, which is part of the Retroviridae family. Two types of HIV infect humans: HIV-1 and HIV-2. HIV-1 is the most common strain of HIV virus and is known to be more pathogenic than HIV-2. As used herein, the terms "human immunodeficiency virus" and "HIV" refer to but are not limited to HIV-1 and HIV-2. In a preferred embodiment, the mantle protein described herein refers to the mantle protein present on HIV-1.

HIV is classified into multiple branches with a high degree of genetic diversity. As used herein, the term "HIV branch" or "HIV subtype" refers to related human immunodeficiency viruses classified according to their degree of genetic similarity. There are currently three groups of HIV-1 isolates: M, N and O. Group M (main strain) consists of at least ten branches (A to J). Group O (external strains) can consist of a similar number of branches. Group N is a new HIV-1 isolate that has not yet been classified in Group M or Group O.

The terms "chronic set point", "chronic HIV infection set point", "viral load set point" and "chronic HIV infection viral set point" refer to the steady-state HIV viral load established in the blood of HIV-infected humans. The chronic set point may refer to the infection after the introduction of antiretroviral therapy or treatment (including administration of ART, TLR7 agonist and/or HIV vaccine as described herein) or after the interruption of antiretroviral therapy or treatment The value of steady-state HIV viral load. The chronic set point can be determined in a single HIV-infected human or it can be determined as the 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, the first chronic set point of 100 copies of HIV-1 RNA/mL is 10% of the second chronic set point of 1000 copies of HIV-1 RNA/mL, and may alternatively be described as being more chronic than the first The second chronic set point that is 10 times higher than the set point.

"Viral rebound" refers to the observation that the undetectable HIV viral load in HIV-infected humans virologically suppressed after ART treatment often returns to detectable pre-treatment HIV viral load after ART is interrupted. Viral rebound can occur within days or weeks (eg, 4 weeks) after the interruption of ART. "Delay of viral rebound" refers to the expected observed result of viral rebound after interruption of ART (for example, the 4th week) and interruption of another therapy (for example, administration of ART, TLR7 agonist and HIV vaccine according to the method described herein) Later (for example, the 12th week) the time period between actual observed virus rebounds. In the above hypothetical example, the delay of viral rebound is 8 weeks after treatment of ART, TLR7 agonist and HIV vaccine. The delay of viral rebound can be measured in a single HIV-infected human or as the median delay of viral rebound in a group of HIV-infected humans.

According to embodiments of the present invention, the method described herein can be used to induce an immune response against one or more branches of HIV.

HIV antigen

As used herein, the terms "HIV antigen", "HIV antigenic polypeptide", "HIV antigenic polypeptide", "HIV antigen protein", "HIV immunogenic polypeptide" and "HIV immunogen" all refer to the A polypeptide that induces an immune response against HBV, such as a humoral and/or cell-mediated response. The HIV antigen can be an HIV protein, fragments or epitopes thereof, or a combination of multiple HIV proteins or parts thereof, which can induce an immune response against HIV in an individual. The HIV antigen can produce a protective immune response in the host, such as inducing an immune response to a viral disease or infection, and/or making an individual immune to a viral disease or infection (ie, vaccination), thereby protecting the individual from Affected by a viral disease or infection. For example, HIV antigens may include proteins or fragments from HIV, such as HIV gag , pol, and env gene products.

According to an embodiment of the present invention, the HIV antigen may be HIV-1 or HIV-2 antigen or fragments thereof. Examples of HIV antigens include, but are not limited to, gag , pol and env gene products encoding structural proteins and essential enzymes. Gag , pol, and env gene products are synthesized into polymerized proteins, which are further processed into a variety of other protein products. The primary protein product of the gag gene is the viral structural protein gag polymerized protein, which is further processed into MA, CA, SP1, NC, SP2 and P6 protein products. The pol gene encodes a viral enzyme (Pol, polymerase), and the primary protein product is further processed into RT, RNase H, IN and PR protein products. The env gene encodes a structural protein, specifically the glycoprotein of the viral particle mantle. The primary protein product of the env gene is gp160, which is further processed into gp120 and gp41. The heterologous nucleic acid sequence according to the present invention preferably encodes gag , env and/or pol gene products or parts thereof. According to a preferred embodiment, the HIV antigen comprises HIV Gag, Env or Pol antigen, or any part or combination thereof, more preferably HIV-1 Gag, Env or Pol antigen, or any part or combination thereof.

According to a preferred embodiment of the present invention, the HIV antigen is a mosaic HIV antigen. As used herein, "mosaic antigen" refers to a recombinant protein assembled from fragments of a natural sequence. "Mosaic antigen" can be generated by computer and optimized using genetic algorithm. Mosaic antigens are similar to natural antigens, but are optimized to maximize the coverage of potential T cell epitopes found in the natural sequence, thereby increasing the breadth and coverage of the immune response.

Examples of embedded HIV Gag-Pol-Env antigens include antigens described in, for example, US20120076812, Barouch et al., Nat Med 2010, 16:319-323, Barouch et al., Cell 155:1-9, 2013 and WO 2017/102929 All of these references are incorporated herein by reference in their entirety.

Preferably, the mosaic HIV antigen encoded by the vector according to the present invention comprises one or more amino acid sequences selected from the group consisting of SEQ ID NO: 1-4 and SEQ ID NO: 10. Alternative and/or additional HIV antigens can be encoded by the primary vaccine and/or booster vaccine of the present invention in certain embodiments, for example to further amplify the immune response.

In view of the present invention, methods known in the art can be used to generate mosaic HIV antigens. See, for example, US20120076812, Fischer et al., Nat Med , 2007. 13(1): p. 100-6, Barouch et al., Nat Med 2010, 16:319-323, all of which are incorporated herein by reference in their entirety in.

Mantle polypeptide

As used herein, the terms "mantle polypeptide", "mantle glycoprotein", "env polypeptide", "env glycoprotein" and "Env" each refer to, but are not limited to, a set of HIV virus particles. The glycoproteins on the surface of the membrane and the surface of the plasma membrane of HIV-infected cells and fragments that can induce anti-HIV immune responses or produce anti-HIV immunity in individuals in need.

The env gene encodes gp160, which is cleaved into gp120 and gp41 in a proteolytic manner. More specifically, gp160 tripolymerizes to (gp160) ₃ and then undergoes cleavage into two non-covalently associated fragments, gp120 and gp41. Subsequently, the virus entry is mediated by the trimer of the gp120/gp41 heterodimer. gp120 is a receptor-binding fragment and binds to the CD4 receptor on target cells that have such receptors, such as T helper cells. The gp41 non-covalently bound to gp120 is a fusion fragment and provides a second step for HIV to enter the cell. gp41 was originally embedded in the viral mantle, but when gp120 binds to the CD4 receptor, gp120 changes its configuration so that gp41 becomes exposed, and gp41 can help to fuse with host cells. gp140 is the uncleaved extracellular domain of the trimer gp160 (ie (gp160) ₃ ), which is used as a substitute for the natural state of the split virus spike.

According to certain embodiments of the present invention, isolated HIV mantle polypeptides (for example, gp160, gp140, gp120 or gp41) can be administered, preferably gp140 protein and more preferably stabilized trimeric gp140 protein to enhance A single expression vector (such as adenovirus 26 and/or MVA vector) induces immunity.

As used herein, each of the terms "stabilized trimer gp140 protein" and "stabilized trimer of gp140" refers to a trimer of gp140 polypeptides, which include increased stability of the trimer structure Sexual peptide sequence. The gp140 polypeptide may have or may be modified to include a trimerizing domain that stabilizes the trimer of gp140. Examples of trimerizing domains include, but are not limited to, the "foldable" trimerizing domain of T4-fibrin; the coiled-coil trimerizing domain derived from GCN4; and the E. coli aspartate transaminase as a trimer marker The catalytic subunit of enzymes.

An example of an isolated antigenic polypeptide is the stabilized trimer gp140, such as Nkolola et al., 2010, J. Virology 84(7): 3270-3279, Kovacs et al., PNAS 2012, 109(30): 12111- 6. The trimer gp140 described in WO 2010/042942 and WO 2014/107744, these documents are all incorporated by reference in their entirety.

In some embodiments of the present invention, "mantle polypeptide" or "mantle glycoprotein" is a mosaic of multiple epitopes containing one or more of the Env polymer protein sequences derived from one or more branches of HIV Mantle protein. For example, as used herein, "gp140 protein" may be a "mosaic gp140 protein" containing multiple epitopes derived from one or more gp140 protein sequences of one or more HIV branches. Preferably, the mosaic gp140 protein is a stabilized trimeric gp140 protein.

In a preferred embodiment, the mosaic gp140 protein is a stabilized trimer of the mosaic gp140 protein comprising the amino acid sequence of SEQ ID NO: 10.

In some embodiments of the present invention, "mantle polypeptide" or "mantle glycoprotein" is a mantle protein derived from a specific HIV branch (such as HIV branch A, B, or C). For example, as used herein, "gp140 protein" may be a "branched C gp140 protein" containing a mantle protein sequence derived from branch C of HIV. Preferably, the branched C gp140 protein is a stabilized trimeric branched C gp140 protein.

In a preferred embodiment, the branched C gp140 protein is a stabilized trimer of the branched C gp140 protein comprising the amino acid sequence of SEQ ID NO: 9.

According to certain embodiments of the present invention, gp140 polypeptides (such as stabilized trimeric gp140 protein) can be administered with viral expression vectors (such as adenovirus 26 and/or MVA vectors).

In certain embodiments of the present invention, two gp140 proteins are administered to the same individual, preferably branched C gp140 having the amino acid sequence of SEQ ID NO: 9 and the amino acid sequence of SEQ ID NO: 10 The inlay gp140. The two gp140 proteins can be administered together in one pharmaceutical composition, preferably together with an adjuvant (such as an aluminum phosphate adjuvant). The preferred dose for administering the total amount of gp140 to humans is between about 125 and 350 µg, such as 125, 150, 175, 200, 225, 250, 275, 300, 325, 350 µg or any amount in between , Preferably about 250 µg. If both branched C gp140 and mosaic gp140 are administered, a suitable dose will be, for example, about 125 µg of each protein to provide a total dose of 250 µg of gp140 protein for administration to a human individual. As used herein, unless otherwise specified, the amount of gp140 polypeptide refers to the amount of gp140 polypeptide measured as a glycoprotein.

The isolated gp140 protein can be co-delivered with an adenovirus (for example, Ad26) expression vector or MVA expression vector or administered in combination with an adenovirus (for example, Ad26) expression vector or MVA expression vector. According to a preferred embodiment, gp140 protein and Ad26 or MVA vector are administered separately in the form of two different formulations. Alternatively, the gp140 protein can be administered in a single formulation together with Ad26 or MVA. Simultaneous administration or co-delivery can take place at the same time, within an hour, or on the same day. In addition, gp140 protein can be administered as an adjuvanted formulation. Suitable adjuvants can be, for example, aluminum phosphate or saponin-based adjuvants, preferably aluminum phosphate adjuvants.

In view of the present invention, any method known in the art can be used to produce and isolate antigenic polypeptides, such as gp140. For example, the antigenic polypeptide can be expressed from a host cell, preferably a recombinant host cell optimized to produce the antigenic polypeptide. According to an embodiment of the present invention, the recombinant gene is used to express the mutant gp140 protein to eliminate the cleavage and fusion activity. Preferably, the optimized gp140 protein is immunized after the individual (e.g., human) (e.g., when When incorporated into the composition of the present invention (e.g., the vaccine of the present invention), the antiviral immune response (e.g., cellular or humoral immune response) produced by the composition is increased in breadth, intensity, depth, or lifespan. The optimized gp140 protein may also include cleavage site mutations, factor Xa sites and/or foldable trimerization domains. The leader/signal sequence is operably linked to the N-terminus of the optimized gp140 protein for maximum protein performance. The leader/signal sequence is usually cleaved from the nascent polypeptide during delivery into the endoplasmic reticulum lumen. Any leader/signal sequence suitable for the relevant host cell can be used. An exemplary leader/signal sequence includes the amino acid sequence of SEQ ID NO: 11.

Adenovirus vector

The primary vaccine and booster vaccine used in the method of the present invention include one or more adenovirus vectors, in particular, human adenovirus 26 vectors (Ad26) encoding one or more mosaic HIV antigens. The adenovirus according to the present invention belongs to the family of Adenoviridae and is preferably a virus belonging to the genus Mastadenovirus . As used herein, the symbol "rAd" means a recombinant adenovirus, for example "rAd26" means a recombinant human adenovirus 26.

According to an embodiment of the present invention, the adenovirus is human adenovirus serotype 26 (Ad26). So far, the advantage of human adenovirus serotype 26 is that significant experience has been gained in clinical trials with such vectors, and this does not reveal that the pre-existing neutralizing antibody response to such vectors will respond to the desired vaccine-induced response ( For example, it causes substantial interference with the encoded antigen in such vectors. Preferably, the adenovirus vector is a replication-defective recombinant virus vector, such as a replication-defective recombinant adenovirus 26 vector.

In some embodiments, the recombinant adenovirus vector suitable for the present invention is mainly or completely derived from Ad26 (that is, the vector is rAd26). In some embodiments, the adenovirus line is replication-defective, for example because it contains a deletion in the E1 region of the genome. For the adenovirus derived from the Ad26 used in the present invention, the E4-orf6 coding sequence of the adenovirus is typically exchanged with the E4-orf6 of the human subgroup C adenovirus (such as Ad5). This allows such adenoviruses to multiply in well-known complementary cell lines (such as 293 cells, PER.C6 cells and the like) that express the E1 gene of Ad5 (see, for example, Havenga et al., 2006, J Gen Virol 87: 2135-43 ; WO 03/104467). However, such adenoviruses will not be able to replicate in non-supplementary cells that do not express the El gene of Ad5. Therefore, in certain embodiments, the adenovirus is a human adenovirus of serotype 26, which has a deletion in the E1 region that has been colonized with nucleic acid encoding one or more mosaic HIV antigens, and has the E4 or f6 region of Ad5.

The preparation of recombinant adenovirus vectors is well known in the art. The preparation of the rAd26 vector is described in, for example, WO 2007/104792 and Abbink et al. (2007) Virol 81(9):4654-63, both of which are incorporated herein by reference in their entirety. The exemplary genome sequence of Ad26 can be found in GenBank deposit EF 153474 and SEQ ID NO: 1 of WO 2007/104792, which are incorporated herein by reference in their entirety. Generally, the adenoviral vector system that can be used in the present invention is prepared using nucleic acid (such as plastid, muxid or baculovirus vector) that constitutes the complete recombinant adenoviral genome.

The adenovirus vectors that can be used in the present invention are usually replication-defective. In these embodiments, the virus exhibits replication defects by deletion or inactivation of regions (such as the E1 region) that are essential for virus replication. These regions can be substantially deleted or inactivated by, for example, inserting related genes in the region, such as genes encoding HIV antigens (usually linked to a promoter). In some embodiments, the vector of the present invention may contain deletions in other regions (such as the E3 region), or insert heterologous genes linked to the promoter in such regions. This cell line does not need to supplement the mutations in the E3 region of adenovirus, because E3 is not required for replication.

Packaging cell strains are usually used to produce a sufficient amount of adenovirus vector for use in the present invention. The packaging cell line contains cells that have deleted or inactivated genes in the replication-defective vector, thereby allowing the virus to replicate in the cell. Suitable packaging cell lines include, for example, PER.C6, 911 and HEK293.

According to an embodiment of the present invention, the HIV antigen can be expressed in the adenovirus 26 vector described herein. Optionally, the heterologous gene encoding the mosaic HBV antigen can be codon-optimized to ensure proper performance in the treated host (e.g., human). Codon optimization is a widely used technique in this technology. Generally, heterologous genes encoding mosaic HIV antigens are cloned into the E1 and/or E3 regions of the adenovirus genome. The non-limiting examples of the codon-optimized nucleotide sequence encoding the HIV antigen with SEQ ID NO: 1-4 are respectively improved herein as SEQ ID NO: 5-8.

According to an embodiment of the present invention, one or more adenovirus 26 (Ad26) vectors comprise nucleic acid encoding one or more HIV antigens. In particular, one or more Ad26 vectors together encode SEQ ID NO: 1 and SEQ ID NO: 2. , SEQ ID NO: 3 and SEQ ID NO: 4 amino acid sequences of four mosaic HIV antigens. In certain embodiments, the Ad26 vaccine used in the present invention includes: a first Ad26 vector encoding the HIV antigen of SEQ ID NO: 1, a second Ad26 vector encoding the HIV antigen of SEQ ID NO: 2, and a second Ad26 vector encoding the HIV antigen of SEQ ID NO: : The third Ad26 vector of the HIV antigen of 3 and the fourth Ad26 vector of the HIV antigen of SEQ ID NO: 4. In certain embodiments, these vectors are present in a single composition in a 1:1:1:1 ratio (based on viral particles).

The heterologous gene encoding the mosaic HIV antigen can be under the control of an adenovirus-derived promoter (such as a major late promoter) (that is, it can be operably linked to an adenovirus-derived promoter), or it can be under the control of a heterologous promoter Under control. Examples of suitable heterologous promoters include the cell megavirus immediate early (CMV) promoter and the Rous sarcoma virus (RSV) promoter. Preferably, the promoter is located upstream of the heterologous gene encoding the mosaic HIV antigen in the expression cassette. In a preferred embodiment, the heterologous promoter is a CMV promoter.

MVA Carrier

In some embodiments, the MVA vaccine used in the method of the present invention comprises one or more modified Ankara acne (MVA) vectors, which together encode four mosaic HIV antigens, in particular, SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 HIV antigens. The MVA vector suitable for use in the present invention utilizes an attenuated virus derived from the MVA virus, which is characterized by the loss of its ability to multiply and replicate in human cell lines.

MVA has been produced by the dermal pox virus strain Ankara (Chorioallantoic Ankara pox virus, CVA; see Mayr et al. (1975) Infection 3, 6-14) on chicken embryonic fibroblasts for more than 570 consecutive passages The dermal pox virus strain Ankara has been maintained for many years in the Ankara Vaccination Institute in Turkey and is used as the basis for human vaccination. In 1976, MVA derived from MVA-571 seed stock (corresponding to the 571th generation) was registered in Germany as the primary vaccine in the two-stage parenteral smallpox vaccination program. As a result of the subcultures used to attenuate MVA, there are many different virus strains or isolates, depending on the number of subcultures performed in the CEF cells. For example, during the smallpox eradication program, MVA-572 was used as a pro-vaccine in Germany in small doses, and MVA-575 was widely used as a veterinary vaccine. Compared with the previous generation CVA virus, MVA and MVA-BN lack about 15% (31 kb from six regions) genome. The deletion affects many toxic and host-range genes, as well as genes for type A inclusion bodies. The MVA-575 series was deposited in the European Animal Cell Culture Collection (ECACC) on December 7, 2000, with the deposit number V00120707.

MVA virus strains with an enhanced safety profile for the development of safer negative reviews (such as vaccines or drugs) have been developed, for example, by Bavarian Nordic. MVA was further succeeded by Bavarian Nordic and named MVA-BN. A representative sample of MVA-BN was deposited at the European Animal Cell Culture Collection (ECACC) on August 30, 2000, with the deposit number V00083008. MVA-BN is further described in WO 02/42480 (US 2003/0206926) and WO 03/048184 (US 2006/0159699), both of which are incorporated herein by reference in their entirety.

A "derivative" or "variant" of MVA refers to a virus that exhibits substantially the same replication characteristics as MVA as described herein, but exhibits differences in one or more parts of its genome. For example, MVA-BN and derivatives or variants of MVA-BN failed to replicate in humans and mice, even in severely immunosuppressed mice. More specifically, the derivatives or variants of MVA-BN or MVA-BN preferably also have the ability to reproduce and replicate in chicken embryonic fibroblasts (CEF), but do not have the ability to reproduce and replicate in the following: human keratinocytes Strain HaCat (Boukamp et al. (1988), J. Cell Biol. 106: 761-771), human osteosarcoma cell line 143B (ECACC deposit number 91112502), human embryonic kidney cell line 293 (ECACC deposit number 85120602) and human progeny Cervical adenocarcinoma cell line HeLa (ATCC deposit number CCL-2). In addition, the virus amplification rate of derivatives or variants of MVA-BN is at least two times lower than that of MVA-575 in Hela cells and HaCaT cell lines, and more preferably three times lower. The testing and analysis of these characteristics of MVA variants are described in WO 02/42480 (US 2003/0206926) and WO 03/048184 (US 2006/0159699).

The term "inability to reproduce" or "not capable of reproduction" is described in WO 02/42480, for example, which also teaches how to obtain MVA with the required characteristics as mentioned above. This term applies to viruses that have a virus amplification ratio of less than 1 4 days after infection using the analysis method described in WO 02/42480 or U.S. Patent No. 6,761,893, both of which are incorporated herein by reference in their entirety. in.

The term "failed to replicate" refers to a virus that has a virus amplification ratio of less than 1 4 days after infection. The analytical method described in WO 02/42480 or US Patent No. 6,761,893 is suitable for determining the virus amplification ratio.

The advantages of MVA-based vaccines include its safety profile and availability in large-scale vaccine manufacturing. In addition, in addition to its efficacy, the feasibility of industrial-scale manufacturing can also be beneficial. In addition, MVA-based vaccines can deliver multiple heterologous antigens and allow simultaneous induction of humoral and cellular immunity.

The MVA vector that can be used in the present invention can be prepared using methods known in the art, such as the methods described in WO/2002/042480, WO/2002/24224, US20110159036, US 8197825, etc., the relevant disclosures of which are incorporated by reference Incorporated into this article.

In another aspect, replication-deficient MVA virus strains are also applicable to the present invention, such as virus strains MVA-572 and MVA-575, or any other similarly attenuated MVA virus strains. Mutant MVA may also be suitable, such as deletion of chorioallantoic Ankara pox virus (dCVA). dCVA contains the deletion sites of del I, del II, del III, del IV, del V, and del VI of the MVA genome. These sites are particularly suitable for inserting multiple heterologous sequences. dCVA can multiply and replicate in human cell lines (such as human 293, 143B and MRC-5 cell lines) (amplification ratio greater than 10), which then enables optimization by further mutations suitable for virus-based vaccination strategies (See, for example, WO 2011/092029).

In a preferred embodiment of the present invention, the MVA carrier system MVA-BN vector, such as the MVA-BN vector described in WO 2018/229711, is incorporated herein by reference.

According to an embodiment of the present invention, the MVA vector comprises a nucleic acid encoding one or more HIV antigens, the one or more HIV antigens having an amino acid sequence selected from the group consisting of SEQ ID NO: 1-4. Preferably, one or more MVA vectors together encode four mosaic HIV antigens with amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4. A particularly suitable but non-limiting example of a MVA vaccine that can be used in the present invention is MVA-mBN414, as described in Example 7 of WO 2018/229711.

The nucleic acid sequence encoding the mosaic HIV antigen can be inserted into one or more intergenic regions (IGR) of MVA. In certain embodiments, the IGR is selected from IGR07/08, IGR 44/45, IGR 64/65, IGR 88/89, IGR 136/137, and IGR 148/149. In certain embodiments, fewer than 5, 4, 3, or 2 IGRs of the recombinant MVA comprise heterologous nucleotide sequences encoding HIV antigens (such as mosaic HIV antigens). The heterologous nucleotide sequence may additionally or alternatively be inserted into one or more naturally-occurring deletion sites in the MVA genome, in particular the major deletion sites I, II, III, IV, or VI. In certain embodiments, fewer than 5, 4, 3, or 2 naturally occurring deletion sites of the recombinant MVA comprise heterologous nucleotide sequences encoding mosaic HIV antigens.

The number of insertion sites for MVA containing heterologous nucleotide sequences encoding HIV antigens can be 1, 2, 3, 4, 5 or more. In certain embodiments, the heterologous nucleotide sequence is inserted into 4, 3, 2 or fewer insertion sites. Preferably, two insertion sites are used. In certain embodiments, three insertion sites are used. Preferably, the recombinant MVA contains at least 2, 3, 4, 5, 6 or 7 genes inserted into 2 or 3 insertion sites.

The recombinant MVA virus provided herein can be produced by conventional methods known in the art. The method of obtaining recombinant poxvirus or inserting foreign coding sequences into the poxvirus genome is well known to those familiar with the art. For example, standard molecular biology techniques such as DNA selection, DNA and RNA isolation, Western blot analysis, RT-PCR and PCR amplification techniques are described in Molecular Cloning, A laboratory Manual (2nd Edition) (J Sambrook et al., Cold Spring Harbor Laboratory Press (1989)), and techniques for processing and manipulating viruses are described in the Virology Methods Manual (BWJ Mahy et al. (eds), Academic Press (1996)). Similarly, the techniques and know-how for the processing, manipulation and genetic engineering of MVA are described in Molecular Virology: A Practical Approach (AJ Davison & RM Elliott (eds), The Practical Approach Series, Oxford University Press’ IRL Press, Oxford , UK (1993) (see e.g. Chapter 9: Expression of genes by Vaccinia virus vectors) and Current Protocols in Molecular Biology (John Wiley & Son, Inc. (1998) (see e.g. Chapter 16, Section IV: Expression of proteins in mammalian cells using vaccinia viral vector)).

Different methods can be applied to produce the various recombinant MVAs disclosed herein. The DNA sequence to be inserted into the virus can be placed in the E. coli plastid construct that has inserted DNA homologous to a piece of DNA of MVA. The DNA sequence to be inserted can be separately linked to the promoter. The promoter-gene linkage can be positioned in the plastid construct so that the promoter-gene linkage is flanked on both ends by DNA that is homologous to the DNA sequence of the MVA DNA region containing the non-essential locus. The resulting plastid construct can be amplified and isolated by propagation in Escherichia coli bacteria. The isolated plastids containing the DNA gene sequence to be inserted can be transfected into a cell culture such as chicken embryonic fibroblasts (CEF), and the culture is infected with MVA at the same time. The recombination between the respective homologous MVA DNA in the plastid and the viral genome can produce MVA modified by the presence of foreign DNA sequences.

According to a preferred embodiment, cells suitable for cell culture, such as CEF cells, can be infected with poxvirus. The infected cell can then be transfected with a first plastid vector containing one or more exogenous or heterologous genes, which are preferably under the transcriptional control of the poxvirus expression control element. As explained above, the plastid vector also contains sequences capable of directing the insertion of foreign sequences into selected parts of the poxvirus genome. Optionally, the plastid vector also contains a cassette containing a marker and/or selection gene operably linked to a poxvirus promoter.

Suitable markers or selection genes are, for example, genes encoding green fluorescent protein, β-galactosidase, neomycin-phosphoribosyltransferase or other markers. The use of selection or marker cassettes makes it easy to identify and isolate the recombinant poxvirus produced. However, PCR technology can also be used to identify recombinant poxviruses. Subsequently, another cell can be infected with the recombinant poxvirus obtained as described above and transfected with a second vector containing one or more second foreign or heterologous genes. In this case, the gene should be introduced into a different insertion site of the poxvirus genome, and the second vector is also guiding the integration of the one or more second foreign genes or genes into the poxvirus homologous sequence in the poxvirus genome different. After homologous recombination has occurred, a recombinant virus containing two or more foreign or heterologous genes can be isolated. For the introduction of additional foreign genes into the recombinant virus, the recombinant virus isolated in the previous step can be used for infection and another vector containing another or more foreign genes can be used for transfection to reinfect and Transfection step.

Alternatively, the infection and transfection steps as described above can be interchanged, that is, suitable cells can be first transfected with a plastid vector containing the foreign gene and then infected with a poxvirus. As another alternative, it is also possible to introduce each foreign gene into different viruses, co-infect cells with all the obtained recombinant viruses, and screen for recombinants that include all the foreign genes. The third alternative is to use a helper virus to connect the DNA genome and foreign sequences in vitro and reconstruct the recombinant pox virus DNA genome. The fourth alternative is the homologous recombination between the pox virus genome selected as a bacterial artificial chromosome (BAC) in Escherichia coli or another bacterial species and a linear exogenous sequence flanked by DNA sequence homologous to the sequence of the desired integration site in the pox virus genome.

The heterologous nucleic acid encoding one or more mosaic HIV antigens can be under the control of one or more poxvirus promoters (ie, operably linked to one or more poxvirus promoters). In certain embodiments, the poxvirus promoter is Pr7.5 promoter, mixed early/late promoter, or PrS promoter, PrS5E promoter, synthetic or natural early or late promoter, or pox virus ATI promoter.

In certain embodiments of the present invention, the MVA vector suitable for the present invention exhibits an HIV antigen having the amino acid sequence of SEQ ID NO: 1 to SEQ ID NO: 4.

Immunogenic composition

The immunogenic composition is a composition containing an immunogenically effective amount of the purified or partially purified adenovirus 26 or MVA vector used in the present invention. According to the present invention, the adenovirus 26 and MVA vector can encode any mosaic HIV antigen, and preferably encode one or more HIV antigens selected from the group consisting of SEQ ID NO: 1 to 4. The one or more mosaic HIV antigens encoded by the adenovirus 26 vector may be different from the one or more mosaic HIV antigens encoded by the MVA vector, but are preferably the same as the one or more mosaic HIV antigens. The immunogenic composition can be formulated as a vaccine according to methods well known in the art. Such compositions may include adjuvants to enhance the immune response. According to the present invention, the optimal ratio of each component in the formulation can be determined by techniques well known to those skilled in the art.

As used herein, "immunogenically effective amount" or "immunely effective amount" means an amount of a composition or carrier sufficient to induce a desired immune effect or immune response in an individual in need. In one embodiment, the immunogenically effective amount means an amount sufficient to induce an immune response in an individual in need, preferably a safe and effective immune response in a human individual in need. In another embodiment, the immunogenically effective amount means an amount sufficient to generate immunity in an individual in need, for example, to provide a therapeutic effect against diseases such as HIV infection. The immunogenic effective amount can vary depending on various factors, such as the individual's physical condition, age, weight, and health status. Those skilled in the art can easily determine the immunogenicity effective amount according to the present invention.

The immunogenically effective amount can be administered in a single step (such as a single injection) or multiple steps (such as multiple injections) or in the form of a single composition or multiple compositions. It is also possible to administer an immunogenically effective amount to an individual according to the so-called first-plus-beat regimen, and then administer another immunogenically effective amount to the same individual. The general concept of this initial shot-plus shot program is well-known to those skilled in the field of vaccines. If necessary, other boosters can be added to the scheme as appropriate.

As a general guide, when used with reference to recombinant viral vectors, an immunogenically effective amount may range from about ¹⁰⁶ viral particles (VPS), plaque forming units (PFUS) or infectious units (IU) to about ¹⁰¹² viral particles Or within the range of infection units (for example, 10 ⁶ , 10 ⁷ , 10 ⁸ , 10 ⁹ , 10 ¹⁰ , 10 ¹¹ or 10 ¹² virus particles or infection units).

In one embodiment, the immunogenic composition is an Ad26 vaccine for initial administration to induce an immune response. According to an embodiment of the present invention, the Ad26 vaccine comprises an immunogenically effective amount of one or more adenovirus 26 (Ad26) vectors, which together encode SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: Four HIV antigens with 4 amino acid sequences, and a pharmaceutically acceptable carrier. The HIV antigen can be encoded by the same Ad26 vector or different Ad26 vectors (such as one, two, three, four or more Ad26 vectors).

The immunogenic effective amount of one or more Ad26 vectors can be about 10 ⁸ , 10 ⁹ , 10 ¹⁰ , 10 ¹¹ or 10 ¹² virus particles (vp), preferably about 10 ⁹ to 10 ¹¹ virus particles, and more preferably About 10 ¹⁰ virus particles, such as about 0.5×10 ¹⁰ , 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 10×10 ¹⁰ virus particles. In certain embodiments, the immunogenically effective amount is about 5×10 ⁹ to about 1×10 ¹¹ virus particles, preferably about 5×10 ¹⁰ virus particles, so that one or more Ad26 carrier systems are immunized with each Step administer a total dose of about 5×10 ⁹ to about 1×10 ^{11 virus particles.}

The immunogenically effective amount can be derived from one Ad26 vector or multiple Ad26 vectors. For example, about 5×10 ⁹ to about 1×10 ¹¹ virions in the Ad26 vaccine, such as about 5×10 ¹⁰ virions, the total dose can come from four Ad26 vectors each encoding different mosaic HIV antigens. Such as the vectors shown in SEQ ID NO: 1, 2, 3, and 4.

In a specific embodiment, the immunogenically effective amount of Ad26 vector encoding SEQ ID NO: 1, 2, 3, and 4 together consists of four adenovirus vectors, that is, the first of the HIV antigen encoding SEQ ID NO: 1. Ad26 vector, the second Ad26 vector encoding the HIV antigen of SEQ ID NO: 2, the third Ad26 vector encoding the HIV antigen of SEQ ID NO: 3, and the fourth Ad26 vector encoding the HIV antigen of SEQ ID NO: 4.

In such embodiments where the Ad26 vaccine contains more than one Ad26 vector, the Ad26 vector may be included in the composition in any ratio to achieve the desired immunogenicity effective amount. Preferably, when the immunogenically effective amount of Ad26 vector is composed of four Ad26 vectors, the first, second, third and fourth Ad26 vector systems have viral particles (vp) at a ratio of 1:1:1:1 Contribute.

The MVA vaccine suitable for the present invention comprises: an immunogenically effective amount of one or more MVA vectors, which together encode an amine having SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4 Four HIV antigens based on the base acid sequence; and a pharmaceutically acceptable carrier. The HIV antigen expressed by the MVA vector can be encoded by a single MVA vector or multiple MVA vectors (such as one, two or more MVA vectors). In certain advantageous embodiments, the HIV antigen line is expressed by a single MVA vector.

One or more MVA vaccine immunogenically effective amount of the MVA vector may be about ^{106, 107, 108, 109} or ¹⁰¹⁰ infectious units (IU), preferably from about ¹⁰⁷ to ¹⁰⁹ IU, And more preferably about 2×10 ⁸ IU, such as about 0.5×10 ⁸ , 1×10 ⁸ , 2×10 ⁸ , 3×10 ⁸ , 4×10 ⁸ or 5×10 ⁸ IU. In certain embodiments, the immunogenically effective amount is about 1×10 ⁷ to about 5×10 ⁸ IU, preferably about 2×10 ⁸ IU, so that one or more MVA carrier systems are A total dose of about 1×10 ⁷ to about 5×10 ⁸ IU, preferably about 2×10 ^{8 IU is administered.}

The immunogenically effective amount can be derived from one MVA vector or multiple MVA vectors. For example, in some embodiments, about 1×10 ⁷ to about 5×10 ⁸ IU in the MVA vaccine, such as about 1×10 ⁷ , 5×10 ⁷ , 1×10 ⁸ , 2×10 ⁸ , 5 ×10 ⁸ IU or any dose in between can be from two MVA vectors encoding four HIV antigens together, the four HIV antigens have SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO : 3 and SEQ ID NO: 4 amino acid sequences, such as the first MVA vector encoding the HIV antigen of SEQ ID NO: 1 and 3 and the second encoding the HIV antigen of SEQ ID NO: 2 and SEQ ID NO: 4 MVA carrier, wherein preferably, the first and second MVA carrier systems are administered at a ratio of 1:1 IU. In a more preferred embodiment, about 1×10 ⁷ to about 5×10 ⁸ IU in the MVA vaccine, such as about 1×10 ⁷ , 5×10 ⁷ , 1×10 ⁸ , 2×10 ⁸ , 5×10 ⁸ The total administered dose of one IU or any dose in between can be derived from a single MVA vector encoding four HIV antigens having the amino acid sequences of SEQ ID NOs: 1, 2, 3, and 4.

In some embodiments of the present invention, the Ad26 vaccine or MVA vaccine administered after the initial administration of the Ad26 vaccine is administered in combination with one or more isolated HIV gp140 mantle polypeptides. According to an embodiment of the present invention, when referring to the gp140 vaccine administered as part of the immunization (such as the isolated HIV gp140 mantle polypeptide having the amino acid sequence of SEQ ID NO: 9 (branched C gp140 polypeptide) and having SEQ ID NO When the total amount of isolated HIV gp140 mantle polypeptides (at least one of the mosaic gp140 polypeptides) of 10 amino acid sequence) is used, the immunogenicity effective amount can be, for example, about 125 μg to 350 μg, such as about 125 , 150, 200, 250, 300, or 350 μg of one or more isolated HIV mantle polypeptides. In certain embodiments, a vaccine composition comprising one or more Ad26 vectors or one or more MVA vectors is administered in combination with a gp140 vaccine composition, the gp140 vaccine composition comprising two isolated HIV mantle gp140 polypeptides, one A branched C gp140 polypeptide having the amino acid sequence of SEQ ID NO: 9 and a mosaic gp140 polypeptide having the amino acid sequence of SEQ ID NO: 10, each of which is present at about 125 µg per administration, for a total of about 250 µg , All are measured in the form of glycoprotein.

The preparation and use of immunogenic compositions are well known to those skilled in the art. Liquid pharmaceutical compositions generally include a liquid carrier, such as water, petroleum, animal or vegetable oil, mineral oil, or synthetic oil. It may also include physiological saline solution, dextrose or other sugar solutions or glycols (such as ethylene glycol, propylene glycol or polyethylene glycol). In view of the present invention, the immunogenic composition used in the present invention can be formulated for administration according to any method known in the art, and is preferably formulated for intramuscular administration.

The vaccine composition of the present invention may contain other antigens. Other antigens used in combination with adenovirus 26 and/or MVA vector and/or gp140 polypeptide can be, for example, other HIV antigens and nucleic acids expressing these HIV antigens.

The immunogenic composition suitable for use in the present invention may further contain an adjuvant as appropriate. The adjuvant suitable for co-administration according to the present invention should be a potentially safe, well-tolerated and effective adjuvant for humans. Non-limiting examples include QS-21, Detox-PC, MPL-SE, MoGM-CSF, TiterMax-G, CRL-1005, GERBU, TERamide, PSC97B, Adjumer, PG-026, GSK-I, GcMAF, B-A Lixin (B-alethine), MPC-026, Adjuvax, CpG ODN, Betafectin, aluminum salts (such as aluminum phosphate (eg AdjuPhos) or aluminum hydroxide), and MF59.

For example, the preferred adjuvant for administration with isolated HIV mantle polypeptides is aluminum phosphate. According to an embodiment of the present invention, when used with reference to the total amount of aluminum phosphate in the gp140 vaccine composition containing one or more HIV mantle polypeptides, the total amount of aluminum phosphate administered can be, for example, about 10 µg to about 1000 µg , For example, about 200 µg to 650 µg, such as about 200, 250, 300, 350, 400, 425, 450, 475, 500, 550 or 600 µg, preferably about 425 µg of aluminum.

The immunogenic composition for generating an immune response according to an embodiment of the present invention comprises a pharmaceutically acceptable carrier, such as a pharmaceutically acceptable excipient, carrier, buffer, stabilizer or familiar item Other materials familiar to the skilled person. These materials should be non-toxic and should not interfere with the effectiveness of the active ingredients. The exact nature of the carrier or other material may depend on the route of administration, such as intramuscular, subcutaneous, oral, intradermal, skin, transmucosal (e.g., intestinal), intranasal, or intraperitoneal routes. Preferably, the pharmaceutically acceptable carrier included in the composition of the present invention is suitable for intramuscular administration.

TLR 7 Vixamod

Toll-like receptor 7 (TLR 7) is one of the protein members of the receptor (toll-like receptor) family, which recognizes pathogen-associated molecular patterns (PAMP) on infectious agents and is expressed by the human gene HGNC:15631 .

In a specific embodiment, the TLR agonist suitable for use in the present invention is Vixamod (VES), also known as GS-9620, which has the IUPAC name 4-amino-2-butoxy-8- [[3-(Pyrrolidin-1-ylmethyl)phenyl]methyl]-5,7-dihydropteridine-6-one is a small molecule or a pharmaceutically acceptable salt thereof. Vixamod is an agonist of TLR 7 receptors, which induces IFN responses, interleukins and chemokines (National Center for Biotechnology Information. PubChem Database. Vesatolimod, CID=46241268, https://pubchem.ncbi. nlm.nih.gov/compound/46241268 (accessed April 23, 2019)).

。Visamod has the following structure:

.

Vixamod can be formulated with conventional carriers and excipients, and these carriers and excipients will be selected according to common practice. The lozenge will contain excipients, gliding agents, fillers, binding agents and the like. Aqueous formulations are prepared in a sterile form, and are generally isotonic when they are intended to be delivered by means other than oral administration. All formulations will optionally contain excipients, such as those described in Handbook of Pharmaceutical Excipients (1986), which are incorporated herein by reference in their entirety. Excipients include ascorbic acid and other antioxidants, chelating agents (such as EDTA), carbohydrates (such as dextrin), hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearic acid and the like. The pH of the formulation is in the range of about 3 to about 11, but usually about 7 to 10.

Formulations include formulations suitable for the aforementioned administration routes. The formulation may conveniently be presented in unit dosage form and may be prepared by any method well known in the pharmaceutical technology. The techniques and formulations are generally found in Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, Pa.), which is incorporated herein by reference in its entirety. These methods include the step of bringing the active ingredient into association with a carrier that constitutes one or more accessory ingredients. Generally speaking, formulations are prepared by uniformly and intimately combining the active ingredient with a liquid carrier or a finely powdered solid carrier or both, and then, if necessary, shaping the product.

The formulations suitable for oral administration with Vixamod or its pharmaceutically acceptable salt can be presented as discrete units, such as capsules, cachets or lozenges each containing a predetermined amount of active ingredient; presented as a powder Or particles; in the form of a solution or suspension in an aqueous or non-aqueous liquid; or in the form of an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient can also be administered in the form of a bolus, lick or paste.

According to an embodiment of the present invention, suitable routes for administering Vixamod or its pharmaceutically acceptable salt include oral, rectal, nasal, topical (including buccal and sublingual), and vaginal And parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural) and similar routes. It should be understood that the preferred route may vary with, for example, the condition of the recipient. The advantage of the Vixamod compound used in the present invention is that it is orally bioavailable and can be administered orally.

The effective dose of Vixamod depends at least on the nature and toxicity of the condition to be treated, whether the compound is used prophylactically (lower dose) or used for active diseases or conditions, delivery methods and pharmaceutical formulations, and will be determined by clinicians Physicians use conventional dose escalation studies to determine. The effective dose can be expected to be about 0.0001 to about 10 mg per kilogram of body weight per day, usually about 0.001 to about 1 mg per kilogram of body weight per day, more usually about 0.01 to about 1 mg per kilogram of body weight per day, and even more usually about 0.05 per kilogram of body weight per day To about 0.5 mg. For example, a daily candidate dose for an adult weighing about 70 kg will be in the range of about 0.05 mg to about 100 mg, or between about 0.1 mg and about 25 mg, or between about 0.4 mg and about 15 mg, And can be in the form of single or multiple doses.

According to an embodiment of the present invention, Vesalimod or its pharmaceutically acceptable salt is administered at a dose of about 3 mg to about 15 mg, such as about 3 mg, 4 mg, 5 mg, 6 mg, 7 mg each time The total dose of, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, or 15 mg visamod is administered orally to a human individual once every two weeks.

An effective amount of Vixamod or its pharmaceutically acceptable salt can be orally administered to a human subject using any suitable dosage form. For example, 4 mg of visamod or its equivalent pharmaceutically acceptable salt can be administered orally together with a single 4 mg lozenge or two 2 mg lozenges; 6 mg of visamod or its equivalent The equivalent of a pharmaceutically acceptable salt can be administered orally together with two 3 mg lozenges or three 2 mg lozenges; etc.

In some embodiments, VES or its pharmaceutically acceptable salt can be administered in 10 doses, preferably every 14 days using one of the following dosage forms and regimens: (a) The first to second doses are 1×4 mg tablets and the 3rd to 10th doses are 3×2 mg tablets; (b) The first to third doses are 1×4 mg tablets and the 4th to 10th doses It is 3×2 mg tablets; (c) The first to fourth doses are 1×4 mg tablets and the 5th to 10th doses are 3×2 mg tablets; (d) The first to 10th doses are 3×2 mg Tablets; (e) The first and second doses are 3×2 mg tablets and the third to tenth doses are 2×4 mg tablets; (f) The first to third doses are 5×2 mg tablets and The 4th to 10th doses are 6×2 mg tablets; (g) The 1st to 3rd doses are 4×2 mg tablets and the 4th to 10th doses are 5×2 mg tablets; or (h) other suitable dosage forms And the plan.

Induced resistance HIV Methods of immune response to infection

The aforementioned Ad26 and MVA vaccine compositions can be used in the methods of the invention described herein. The method of the present invention relates to inducing an immune response against human immunodeficiency virus (HIV) in HIV-infected individuals receiving antiretroviral therapy. The method of administering Ad and MVA vaccines according to an embodiment of the present invention effectively induces an immune response against one or more branches of HIV.

The present invention relates to a method for inducing an immune response against human immunodeficiency virus (HIV) in HIV-infected human subjects receiving antiretroviral therapy (ART), the method comprising: (i) administer an immunogenically effective amount of Ad26 vaccine to the human individual, and the Ad26 vaccine includes an amine that encodes SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4. One or more adenovirus 26 (Ad26) vectors and pharmaceutically acceptable carriers of four HIV antigens based on the base acid sequence; (ii) Depending on the situation, re-administer an immunogenically effective amount of Ad26 vaccine to the human individual; (iii) administer an immunogenically effective amount of MVA vaccine to the human individual, the MVA vaccine comprising one or more of four HIV antigens (ie, the HIV antigens of SEQ ID NO: 1, 2, 3, and 4) that are encoded together A modified Ankara acne (MVA) carrier and a pharmaceutically acceptable carrier; (iv) administering an effective amount of Vexamod (VES) or its pharmaceutically acceptable salt to the human individual; and (v) Re-administer the MVA vaccine to the human individual as appropriate.

In some embodiments of this aspect of the present invention, the method further comprises a gp140 vaccine comprising one or more isolated HIV gp140 envelope polypeptides in steps (ii) and/or (iii) and/or (v) Combination with Ad26 vaccine or MVA vaccine is administered to human individuals. In such embodiments, the gp140 vaccine is preferably administered in step (iii) and step (v). In such an embodiment, the method preferably further comprises: in steps (ii) and/or (iii) and/or (v), preferably in steps (iii) and (v), selecting from At least one isolated HIV gp140 mantle polypeptide of the group consisting of two trimers of the amino acid sequence of SEQ ID NO: 9 and SEQ ID NO: 10 is administered to humans in combination with the Ad26 vaccine or MVA vaccine individual. In such embodiments, the gp140 vaccine preferably includes two HIV gp140 polypeptides having the amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 10, respectively.

In some embodiments, the method of inducing an immune response against human immunodeficiency virus (HIV) in HIV-infected human subjects receiving antiretroviral therapy (ART) comprises: (i) administering about 5 ×10 ⁹ to about 1×10 ¹¹ viral particles (vp), preferably about 5×10 ¹⁰ vp. The total dose of Ad26 vector is preferably administered intramuscularly to the human individual with the Ad26 vaccine, and the Ad26 vaccine contains a code One or more adenovirus 26 (Ad26) vectors of four HIV antigens with the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 and pharmaceutically acceptable Accepted carrier; (ii) A total dose of Ad26 vector of ^{about 5×10 9} to about 1×10 ¹¹ viral particles (vp) per administration, preferably about 5×10 ^{10 vp, preferably transmuscular} Inwardly administer the Ad26 vaccine to the human individual, wherein the Ad26 vaccine is administered 10 to 14 weeks, preferably 12 weeks after the administration of the Ad26 vaccine in step (i); (iii) about 1 dose per dose ×10 ⁷ to about 5×10 ⁸ infection units (IU), preferably about 2×10 ⁸ IU of one or more of the total dose of MVA-BN vector, preferably intramuscularly administer the MVA vaccine to the human individual The MVA vaccine contains one or more MVA vectors encoding four HIV antigens (preferably one or more MVA-BN vectors) and a pharmaceutically acceptable carrier; it can be combined with the MVA vaccine as appropriate for each administration The total dose of the glycoprotein of two isolated HIV gp140 mantle polypeptides of about 125 μg to 350 μg, preferably about 250 μg, is further administered to include the amino acid sequence having SEQ ID NO: 9 and SEQ ID NO: 10 The two isolated HIV gp140 mantle polypeptide gp140 vaccines, aluminum adjuvants and pharmaceutically acceptable carriers, wherein the Ad26 vaccine is administered 22 to 26 weeks, preferably 24 weeks after the administration of the Ad26 vaccine in step (i) MVA vaccine and optional gp140 vaccine; (iv) A total dose of about 3 mg to about 15 mg per administration, such as about 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 The total dose of mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg or 15 mg of vesamod is preferably orally administered to a human individual with vesamod or a pharmaceutically acceptable salt thereof, Wherein 26 to 34 weeks after the Ad26 vaccine is administered in step (i), vexamod or its pharmaceutically acceptable salt is administered once every two weeks; (v) about 1×10 ^{7 per administration} To about 5×10 ⁸ infection units (IU), preferably about 2×10 ⁸ IU, the total dose of one or more MVA carriers, preferably intramuscularly The human individual is then administered the MVA vaccine; as appropriate, combined with the MVA vaccine to give a total dose of about 125 μg to 350 μg, preferably about 250 μg, of the glycoprotein of two isolated HIV gp140 mantle polypeptides each time, The gp140 vaccine, aluminum adjuvant and pharmaceutically acceptable carrier are further administered to human individuals, wherein the MVA vaccine and video are administered 34 to 38 weeks after the Ad26 vaccine is administered in step (i), preferably 36 weeks. The gp140 vaccine selected under circumstances; and (vi) a total dose of about 3 mg to about 15 mg per administration, such as about 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 The total dose of mg, 11 mg, 12 mg, 13 mg, 14 mg or 15 mg of visamod is preferably orally administered to a human individual again with visamod or a pharmaceutically acceptable salt thereof, wherein In step (i), from 38 to 46 weeks after the Ad26 vaccine is administered, vexamod or its pharmaceutically acceptable salt is administered once every two weeks.

In a preferred embodiment, step (iii) comprises administering the gp140 vaccine, an aluminum adjuvant, and a pharmaceutically acceptable carrier in combination with the MVA vaccine.

In another preferred embodiment, step (v) comprises combining gp140 vaccine, aluminum adjuvant and pharmaceutically acceptable carrier with MVA vaccine before administering.

In certain embodiments, the method comprises administering a total dose of 4 mg visamod at each time 26 and 28 weeks after the initial administration of the Ad26 vaccine to the human individual and 30, 32, For 34, 38, 40, 42, 44, and 46 weeks, a total dose of 6 mg of visamod per administration was administered to a human individual orally with visamod or a pharmaceutically acceptable salt thereof.

In certain embodiments, the method comprises administering a total dose of 4 mg of visamod at 26, 28, and 30 weeks after the initial administration of the Ad26 vaccine to the human individual and 32, after the initial administration of the Ad26 vaccine. For 34, 38, 40, 42, 44, and 46 weeks, a total dose of 6 mg of visamod per administration was administered to a human individual orally with visamod or a pharmaceutically acceptable salt thereof.

In certain embodiments, the method comprises administering a total dose of 4 mg visamod at 26, 28, 30, and 32 weeks after the initial administration of the Ad26 vaccine to the human individual and after the initial administration of the Ad26 vaccine For 34, 38, 40, 42, 44, and 46 weeks, a total dose of 6 mg of visamod per administration was administered to a human individual orally with visamod or a pharmaceutically acceptable salt thereof.

In certain embodiments, 26, 28, 30, 32, 34, 38, 40, 42, 44, and 46 weeks after the initial administration of Ad26 vaccine is administered in a total dose of about 6 mg visamod per administration With Vixamod or its pharmaceutically acceptable salt.

In certain embodiments, at 26 and 28 weeks after the initial administration of the Ad26 vaccine, a total dose of about 6 mg visamod is administered per administration and 34, 38, 40, 42, 44 after the initial administration of the Ad26 vaccine. And for 46 weeks, the total dose of 8 mg of visamod per administration was administered to visamod or its pharmaceutically acceptable salt.

In certain embodiments, 26, 28, and 30 weeks after the initial administration of the Ad26 vaccine, a total dose of about 10 mg visamod is administered each time and 32, 34, 38, 40 after the initial administration of the Ad26 vaccine. Administer Vesalimod or its pharmaceutically acceptable salt at a total dose of 12 mg of visamod per administration at 42, 44, and 46 weeks.

In certain embodiments, 26, 28, and 30 weeks after the initial administration of the Ad26 vaccine at a total dose of about 8 mg visamod per administration and 32, 34, 38, 40 after the initial administration of the Ad26 vaccine Administer Vesalimod or a pharmaceutically acceptable salt thereof at a total dose of 10 mg visamod per administration at 42, 44, and 46 weeks.

Any of the vaccine compositions described herein can be used in the methods according to the invention. Examples of Ad26 vaccines, MVA vaccines, Ad26 vectors, MVA vectors, HIV antigens encoded by Ad26 and MVA vectors, isolated gp140 polypeptides, etc., which can be used in the method of the present invention are discussed in detail in the above and below illustrative examples in.

According to an embodiment of the present invention, "inducing an immune response" when used with reference to the methods described herein encompasses inducing a desired immune response or effect against HIV infection (preferably for therapeutic purposes) in an individual in need. "Inducing an immune response" also covers the provision of therapeutic immunity against the pathogen, that is, HIV for treatment. As used herein, the term "therapeutic immunity" or "therapeutic immune response" means that a vaccinated HIV-infected individual is able to control the infection of the pathogen (ie, HIV) against which the vaccination is directed. In one embodiment, "inducing an immune response" means generating immunity in an individual in need, for example, to provide a therapeutic effect against a disease, such as HIV infection. In certain embodiments, "inducing an immune response" refers to inducing or improving cellular immunity against HIV, such as a T cell response. In certain embodiments, "inducing an immune response" refers to inducing or improving a humoral immune response against HIV. In certain embodiments, "inducing an immune response" refers to inducing or improving cellular and humoral immune responses against HIV. Generally, administration of the Ad26 and MVA vaccine compositions according to the embodiments of the present invention will have a therapeutic purpose to generate an immune response against HIV after HIV infection or the development of symptoms specific to HIV infection. In certain embodiments, ART has successfully inhibited the immune response induced in individuals with HIV replication in the bloodstream so that the individual can discontinue ART and maintain control of virus replication in the bloodstream for at least 24 weeks after discontinuation of ART.

The patient population to be treated according to the method of the present invention described herein is HIV-infected human individuals, especially HIV-infected human individuals receiving retroviral therapy (ART). As used herein, the terms "HIV infection" and "HIV infection" refer to the invasion of the human host by HIV. As used herein, a "human subject infected with HIV" refers to a human subject who has invaded and subsequently replicated and reproduced in a human host, thereby causing the human host to be infected with HIV or have HIV infection or its symptoms. A "human individual infected with HIV" has been diagnosed with HIV infection, that is, for example, any analysis approved by the US FDA will test positive in the screening for HIV infection.

As used herein, "receiving antiretroviral therapy" refers to human individuals who are being administered or have started treatment with antiretroviral drugs, especially human individuals who are infected with HIV. According to an embodiment of the present invention, antiretroviral therapy (ART) is started before the first administration of Ad26 vaccine, for example about 2 to 6 weeks before, such as about 2, 3, 4, 5 or 6 weeks before , Or 2 to 48 months before, such as about 2, 3, 5, 6, 8, 12, 16, 20, 24, 30, 36, 42, or 48 months or more before. In certain embodiments, ART starts about 44 to 52 weeks before the first administration of the Ad26 vaccine, preferably earlier than about 48 weeks. In individuals receiving antiretroviral therapy, antiretroviral therapy continues during the administration of the regimen of the invention. If an individual has a plasma HIV RNA content of less than 50 copies/ml for a certain period of time (including possible spikes), then ART is regarded as "inhibitory" as used herein. As used herein, the term "stable suppression" ART means that the inhibitory ART regimen is not modified for a certain period of time.

In certain embodiments, a human subject receiving antiretroviral therapy has undergone a current stable inhibitory ART within at least twenty-four weeks, meaning that when receiving the same ART regimen, the subject is at least 24 hours before starting the regimen according to the present invention. Weeks have a plasma HIV ribonucleic acid (RNA) content of less than 50 copies/ml. However, during this period of time, such as a period of 24 weeks before starting the regimen, a human individual may have one or more deviations (blip) of plasma HIV RNA from more than 50 copies/ml to less than 200 copies/ml. (That is, a case), the restriction is that the screening immediately before the start of the program is less than 50 copies/ml.

Individuals infected with HIV can initiate ART during the acute phase of HIV infection or outside the acute phase of HIV infection. In a preferred embodiment, the individual initiates ART during the acute phase of HIV infection. The term "acute HIV infection" refers to the initial HIV infection period. Generally speaking, there are three HIV infection periods: (1) acute HIV infection, (2) clinical incubation period, and (3) acquired immune deficiency syndrome (AIDS). During acute HIV infection, the host usually produces symptoms such as fever, gland swelling, sore throat, skin rash, muscle and joint pain, headache, etc. based on the body's natural response to HIV infection. During the acute infection period, a large amount of HIV virus is produced in the host, and the CD4 content can be rapidly reduced because HIV uses CD4 to replicate and subsequently destroy CD4. Once the host's natural immune response increases the host's HIV content to a stable level (also known as the viral set point), the CD4 count begins to increase, but it may not reach the pre-infection level. Acute HIV infection is also characterized by Fiebig I, II, III and IV stages, such as Fiebig et al. "Dynamics of HIV viremia and antibody seroconversion in plasma donors: implications for diagnosis and staging of primary HIV infection", AIDS (London, England) (2003) 17(13) 1871-1879, which is incorporated herein by reference in its entirety. In certain embodiments, the HIV-infected human subject receiving the protocol of the present invention is characterized as Fiebig stage I, Fiebig stage II, Fiebig stage III, or Fiebig stage IV.

Acute HIV infection usually occurs within two to four weeks after the host is exposed to and infected with HIV, and lasts for another two to four weeks. The acute HIV infection period lasts until the host produces its own antibodies against HIV, at which time the clinical incubation period begins. During the clinical incubation period, HIV survives or develops in the host without causing any symptoms or only mild symptoms. HIV reproduces at very low levels during the clinical incubation period, although HIV is still active. The clinical incubation period is sometimes referred to as "chronic HIV infection" or "asymptomatic HIV infection". Chronic HIV infection is characterized by Fiebig VI. In certain embodiments, the HIV-infected human subject receiving the protocol of the present invention is characterized as Fiebig V stage or Fiebig VI.

Chronic HIV infection (i.e., Fiebig VI) usually begins about 100 days (i.e., about 14 weeks) after the host is exposed to HIV and infected with HIV. Individuals infected with HIV who have progressed to stage VI of Fiebig can be called or described as "individuals chronically infected", "individuals chronically infected with HIV" or "individuals with chronic HIV infection". Individuals who initiated the ART system outside of the acute or early stages of HIV infection and did not start ART before entering Fiebig VI. Whether an individual has initiated ART before entering Fiebig VI of HIV infection can be determined by the clinician based on the individual's available medical history and laboratory data at the time of HIV diagnosis.

As used herein, individuals who initiate ART outside the acute phase of HIV infection (that is, during chronic HIV infection) are exposed to HIV at the earliest about 12 to 16 weeks after being infected with HIV (such as about 12 to about 12 weeks after exposure and infection with HIV). , 13, 14, 15, or 16 weeks or after 16 weeks) start treatment with antiretroviral drugs. In contrast, individuals who initiate ART during acute HIV infection are usually about 2 weeks to about 8 weeks after exposure to HIV and HIV infection (such as about 1, 2, 3, 4, 5, 6, 6 after exposure and infection). 7 or 8 weeks) or before the start of treatment with antiretroviral drugs. Therefore, it is believed that chronic HIV infection is more difficult to treat than acute HIV infection, at least because chronically infected individuals usually have a larger HIV virus reservoir than acutely infected individuals due to a longer period of infection before any treatment is initiated. Individuals who started ART during acute HIV infection and whose plasma HIV RNA content is less than 50 copies/ml for at least 24 weeks, preferably at least 48 weeks, have a lower HIV virus reservoir and/or its viral reservoir compared with HIV Low participation, and therefore a higher probability of maintaining viral suppression in the absence of ART, that is, HIV remission. However, due to the difference in the progression of acute and chronic HIV infection, it is uncertain whether therapies that are effective in treating acute HIV infection will be equally effective in treating chronic HIV infection.

In some embodiments of the present invention, the individual infected with HIV is an individual chronically infected with HIV. Individuals chronically infected with HIV can initiate ART during any period of infection, such as during the acute phase of HIV infection or outside the acute phase of HIV infection. Preferably, individuals who are chronically infected with HIV initiate ART during the acute phase of HIV infection. In a preferred embodiment of the present invention, human individuals infected with HIV are in the acute phase of HIV infection.

Individuals receiving ART can be administered or treated with any antiretroviral drugs known in the art according to the present invention. ART is a drug therapy for the treatment of HIV, but the drug does not kill the virus or remove the virus from the body. However, when used in combination, it can prevent virus growth. When the virus slows down, the HIV disease also slows down. Antiretroviral drugs are called ARVs. Combination ARV therapy (cART) is called highly active ART (HAART). Generally, the ART regimen includes at least three antiviral compounds, such as two different reverse transcriptase inhibitors plus non-nucleoside reverse transcriptase inhibitors or protease inhibitors or integrase inhibitors.

Those who are generally familiar with the technology will be able to determine the appropriate retroviral therapy, the frequency of administration, the dosage of ART, etc., so as to be compatible with the simultaneous administration of the present invention. Examples of antiretroviral drugs used in ART include (but are not limited to) nucleoside reverse transcriptase inhibitors (NRTIs, non-limiting examples thereof include zidovudine, didanosine, and Tavudine (stavudine), lamivudine (lamivudine), abacavir (abacavir), tenofovir (tenofovir), combivir (combivir) (combination of zidovudine and lamivudine), Trizine Trizivir (combination of zidovudine, lamivudine and abacavir], emtricitabine, truvada (combination of andrositabine and tenofovir) And epzicom (a combination of abacavir and lamivudine"), non-nucleoside reverse transcriptase inhibitors (NNRTI, non-limiting examples of which include nevirapine, delavirdine) , Efavirenz, etravirine and rilpivirine), protease inhibitors (PI, non-limiting examples thereof include saquinavir, indinavir ( indinavir, ritonavir, nelfinavir, amprenavir, lopinavir/ritonavir, atazanavir , Fosamprenavir (fosamprenavir), tipranavir (tipranavir), darunavir (darunavir), integrase inhibitors (INSTI, non-limiting examples include raltegravir (raltegravir), eltegravir) (elvitegravir) and dolutegravir, and fusion inhibitors, entry inhibitors and/or chemokine receptor antagonists (FI, CCR5 antagonists; non-limiting examples include enfuvirtide, Aplaviroc, Maraviroc, Vicriviroc and Cenicriviroc).

According to an embodiment of the present invention, Ad26 vaccine or MVA vaccine is administered after the initial administration of Ad26 vaccine. In certain embodiments of the present invention, about 12 to 52 weeks after the initial administration of Ad26 vaccine, such as about 8 to 15 weeks, about 21 to 27 weeks, and/or about 33 to 39 weeks, and then administration or administration Ad26 vaccine or MVA vaccine. Generally, those who are familiar with this technology will be able to change the exact timing of the vaccine, its frequency of administration, and its dosage based on the teachings and clinical experience in this article.

According to an embodiment of the present invention, the Ad26 vaccine is administered at least once and the MVA vaccine is administered at least once. According to an embodiment of the present invention, the Ad26 vaccine is administered about 10 to 14 weeks after the initial administration of the Ad26 vaccine, and 22 to 26 weeks after the initial administration of the Ad26 vaccine, such as 22, 23, 24, 25, or 26 weeks. MVA vaccine was administered once. In certain embodiments, the Ad26 vaccine is administered approximately 12 weeks after the initial administration of the Ad26 vaccine, and the MVA vaccine is administered for the first time approximately 24 weeks after the initial administration of the Ad26 vaccine. In certain embodiments, the MVA vaccine is administered about 32 to 40 weeks after the initial administration of the Ad26 vaccine, such as about 36 weeks. In certain embodiments, the first administration of Vixamod or its pharmaceutically acceptable salt is after the first administration of the MVA vaccine. In other embodiments, after the second administration of the MVA vaccine, additional visamod or a pharmaceutically acceptable salt thereof is administered. In certain embodiments, Vixamod or a pharmaceutically acceptable salt thereof is administered multiple times in a series of administrations at approximately bi-weekly intervals. In certain embodiments, the first administration of such a series is about two weeks after the first administration of the MVA vaccine and/or the second administration of the MVA vaccine.

In some preferred embodiments, the Ad26 vaccine is administered after the initial administration of the Ad26 vaccine, and in such embodiments, it is preferably administered before the first administration of the MVA vaccine. For example, about 10 to 14 weeks after the initial administration of the Ad26 vaccine, such as about 10, 11, 12, 13 or 14 weeks after the initial administration of the Ad26 vaccine, preferably about 12 weeks after the initial administration of the Ad26 vaccine Vaccine with Ad26.

In other preferred embodiments, the MVA vaccine is administered after the Ad26 vaccine is re-administered. In certain embodiments, about 22 to 26 weeks after the initial administration of the Ad26 vaccine, such as 22, 23, 24, 25, or 26 weeks, preferably about 24 weeks after the initial administration of the Ad26 vaccine, the first administration of MVA vaccine. In certain embodiments, the MVA vaccine can be administered, for example, about 32 to 40 weeks after the initial administration of the Ad26 vaccine, such as 32, 33, 34, 35, 36, 37, 38, 39, or 40 weeks. In certain preferred embodiments, the MVA vaccine is administered approximately 36 weeks after the initial administration of the Ad26 vaccine.

Further administration is possible, and the embodiments of the disclosed method also encompass administration of such additional immunizations using immunogenic compositions containing Ad26 vectors, MVA vectors, and/or HIV gp140 polypeptides. Any of the Ad26 vector, MVA vector, and HIV gp140 polypeptide described herein can be used for additional immunization.

The vaccine composition can be administered according to the present invention by any method known in the art, and the administration is usually via intramuscular, intradermal or subcutaneous administration, preferably intramuscular administration. Intramuscular administration can be achieved by using a needle to inject a suspension or solution of adenovirus and/or MVA vector and/or gp140 polypeptide. An alternative is to use a needle-free injection device to administer the composition (using, for example, Biojector ^™ ) or a lyophilized powder containing the vaccine.

Other modes of administration are also envisioned, such as intravenous, skin, intradermal, oral, intratracheal, or nasal. For intravenous, dermal or subcutaneous injection, the carrier will be in the form of a parenterally acceptable aqueous solution, which does not contain pyrogens and has suitable pH, isotonicity and stability. Those who are generally familiar with this technique are better able to use, for example, isotonic vehicles (such as sodium chloride injection, Ringer's injection and lactated Ringer's injection (to prepare suitable solutions. If necessary, preservatives and stabilizers can be included) , Buffers, antioxidants and/or other additives. Sustained release formulations can also be used.

In some embodiments, the method for inducing an immune response according to the present invention further comprises administering a latent virus reservoir scavenger. Cells latently infected with HIV carry an integrating virus, which is transcriptionally silent, making it difficult to effectively eradicate HIV infection in the individual being treated. As used herein, "reservoir scavenger" and "latent virus reservoir scavenger" refer to substances that reduce the latent area of HIV by reactivating the HIV reservoir (such as by inducing the performance of resting HIV) . Examples of latent virus reservoir scavengers suitable for use in the present invention include, but are not limited to, histone deacetylase (HDAC) inhibitors and modulators of toll-like receptors (for example, TLR7), such as WO2016/007765 And those described in WO2016/177833, which are incorporated herein by reference in their entirety. The latent virus reservoir scavenger can be administered before, after, or co-administered (ie, administered in combination) with one or more of the vaccines described herein. The combination of adenovirus 26 vector encoding Gag, Pol and Env antigens with TLR7 stimulation combination vaccine was used as the first shot, followed by vaccination with the MVA vector encoding such antigens as an extra shot, which has been shown in the rhesus monkey model study to stop the anti- The retroviral therapy caused improved virological control and delayed viral rebound, indicating the potential of the combination of therapeutic vaccination and innate immune stimulation to achieve the functional cure of HIV infection (Borducchi EN, et al., 2016, Nature 540: 284 -287 (doi: 10/1038/nature20583)), the content of which is incorporated herein by reference in its entirety.

Therefore, another general aspect of the present invention relates to a method of treating human immunodeficiency virus (HIV) infection in a human individual in need, the method comprising: (i) Treating human individuals with antiretroviral therapy (ART); and (ii) Use the method according to an embodiment of the present invention to induce an anti-HIV immune response in the human individual.

In some embodiments, the treatment method further comprises suspending the ART treatment of step (i), preferably after the immune response is induced by the method of the present invention. Preferably, the human individual maintains viral suppression for at least 24 weeks after discontinuing ART.

In some embodiments, the individual discontinues (also used interchangeably herein) ART after completing the therapy according to an embodiment of the present invention. In some embodiments, the individual may undergo anti-retroviral assay treatment interruption (ARV ATI) after completing the therapy according to an embodiment of the present invention. As used in the present invention, "interruption of antiretroviral analysis therapy" and "ARV ATI" refer to the discontinuation of antiretroviral drug treatment in the absence of sustained ART to evaluate viral suppression and viremia control. Generally, for example, when an individual has a plasma HIV RNA content of less than 50 copies/ml for at least about 52 weeks, the individual can receive ARV ATI, that is, ART can be discontinued, but even if the individual has plasma HIV RNA greater than 50 copies/ml during this period When one or more deviations (ie, conditions) are less than 200 copies/ml, the individual can still accept ARV ATI, and the restriction is that the screening immediately before ARV ATI is less than 50 copies/ml plasma HIV RNA copies. The known method in view of the present invention can be used to measure the HIV viral load, such as the plasma HIV RNA content, using Abbott RealTime HIV-1 viral load analysis or Roche Cobas Taqman HIV-1 viral load analysis.

According to an embodiment of the present invention, ART can be stopped about 10 to 14 weeks after the last MVA vaccine is administered, such as 10, 11, 12, 13, or 14 weeks. In certain embodiments, the final MVA vaccine is administered about 34 to 38 weeks after the initial administration of the Ad26 vaccine. In these embodiments, ART can be stopped about 46 to 50 weeks after the initial administration of the Ad26 vaccine, such as 46, 47, 48, 59, or 60 weeks, and preferably about 60 weeks after the initial administration of the Ad26 vaccine. In other embodiments, for individuals undergoing non-nucleoside reverse transcriptase inhibitor (NNRTI)-based ART, a boosted protease inhibitor can be administered about 1 to 2 weeks before stopping NNRTI to reduce the development of NNRTI resistance. risk. It is also possible to administer activators (e.g., histone deacetylase inhibitors or TLR7 modulators) during the ATI phase to activate any (e.g., latent) HIV reservoirs and thereby increase the immune response.

Individuals receiving ARV ATI can be monitored, for example, by measuring the plasma HIV RNA content. As a non-limiting example, monitoring after initiation of ARV ATI can be performed at most twice a week for the first six weeks when rebound viremia is most likely to occur. "Rebound viremia" is defined as, for example, the plasma HIV RNA content after ARV ATI is greater than 1,000 copies/ml. ART can be restarted in individuals with rebound viremia. Preferably, individuals treated in accordance with the methods of the present invention will maintain viremia control after interruption of ART. As used herein, "maintain viremia control" is defined in the exemplary embodiment as plasma HIV RNA after ARV ATI is less than 50 copies/ml for at least 24 weeks. If there are one or more cases of plasma HIV RNA greater than 50 copies/ml to less than 1000 copies/ml, as long as the individual’s plasma HIV RNA content exceeds 1000 copies/ml in two consecutive measurements separated by at least one week, then Certain exemplary embodiments are still considered to meet the "maintenance of viremia control" criteria.

Normally (without using the method of the present invention), human HIV-infected individuals recover from viremia 2 to 3 weeks after the interruption of ART. Without wishing to be bound by any theory, it is believed that the use of the vaccine composition according to the embodiments of the present invention and the therapy of vexamod or its pharmaceutically acceptable salt in an individual who completely inhibits HIV will cause measurable immunity Respond and maintain viremia control after ARV ATI. In some embodiments, the individual can discontinue ART after treatment according to the methods of the invention. ART suspension can last for a long period of time (e.g. at least 24 weeks, preferably longer, e.g. at least about 28, 32, 36, 40, 44, 48, 52 weeks, 16 months, 18, 20, 22, 24 months Or even longer). Such time periods during which ART is stopped or suspended are called "vacations" or "ART holidays" or "treatment holidays". In other embodiments, vaccines and TLR7 therapies according to the methods of the present invention can provide HIV relief, meaning that viral suppression is maintained in the absence of ART. In certain embodiments of the invention, a human individual receiving a vaccine according to the invention and Vixamod or a pharmaceutically acceptable salt thereof discontinues ART and maintains viral suppression for at least 24 weeks after discontinuation of ART.

In an exemplary embodiment of the present invention, the Ad26 vaccine containing one or more adenovirus 26 vectors contains about 10 ⁸ to 10 ¹² virus particles/ml at a concentration of about 100 µl to about 2 ml, preferably about 0.5 ml. The amount of the solution is administered (for example, intramuscularly). Ad26 initial vaccination optionally repeated, and after about 100 μl to about 2 ml, preferably about 0.5 ml of an amount of about 10 ⁶ to 10 ⁹ pfu solution concentration / ml comprising the administration (e.g., intramuscular administration And) MVA vaccines containing one or more MVA vectors. After the administration of the MVA vaccine, administer visamod or its pharmaceutically acceptable salt, preferably, then further administer the MVA vaccine, and further administer visamod or its pharmaceutically acceptable salt salt.

In another exemplary embodiment of the present invention, the Ad26 vaccine containing one or more adenovirus 26 vectors contains about 10 ⁸ to 10 ¹² virus particles/ml in a volume of about 100 µl to about 2 ml, preferably about 0.5 ml. The amount of concentrated solution is administered (for example, intramuscularly). Ad26 vaccine after initial inoculation and then administered as a vaccine comprising Ad26 26 or more adenoviral vectors of the one or more adenoviral vector system 26 about 100 μl to about 2 ml, preferably containing of from about 0.5 ml to about ¹⁰⁸ 10 ¹² viral particles/ml concentration of the solution is administered (for example, intramuscular administration). After that, about 100 μl to about 2 ml, preferably of about 0.5 ml containing about ¹⁰⁶ to ¹⁰⁹ concentration of the solution of pfu / ml of MVA with an amount of vaccine administered (e.g., intramuscularly administered), in In some embodiments, a solution of about 100 µl to about 2 ml, preferably about 0.5 ml, is administered in an amount of about 250 mg polypeptide and aluminum phosphate adjuvant (425 micrograms (µg) aluminum per dose) with one or Multiple isolated HIV gp140 polypeptides are administered in combination. After the MVA vaccine is administered, the administration of Vixamod or its pharmaceutically acceptable salt is repeated. In a preferred embodiment, thereafter is another administration of MVA vaccine, and thereafter further administration of Vixamod or its pharmaceutically acceptable salt is repeated.

Those skilled in the art (for example, physicians) will understand that certain factors may affect the dosage required to effectively treat an individual, including (but not limited to) the severity of the disease or condition, previous treatments, the individual’s general health and/or Age and other diseases.

The present invention also relates to a combination of vaccine and visamod or its pharmaceutically acceptable salt, which is used to induce resistance in human subjects infected with human immunodeficiency virus (HIV) receiving antiretroviral therapy (ART) The immune response of HIV, wherein the combination of the vaccine and vexamod or its pharmaceutically acceptable salt comprises the Ad26 vaccine and MVA vaccine according to the embodiments of the present invention and vexamod or its pharmaceutically acceptable salt . The present invention further relates to the combination of a vaccine and Vixamod or its pharmaceutically acceptable salt in the manufacture for inducing human immunodeficiency virus (HIV) infected human subjects receiving antiretroviral therapy (ART) Use of an anti-HIV immune response medicament, wherein the combination of a vaccine and vexamod or its pharmaceutically acceptable salt comprises the Ad26 vaccine and MVA vaccine according to the embodiments and vexamod or its pharmaceutically acceptable salt salt. As described herein, all aspects and embodiments of the present invention can be applied to vaccines as described herein in relation to the method for inducing an immune response against HIV in human individuals infected with human immunodeficiency virus (HIV) receiving antiretroviral therapy (ART) And vexamod or its pharmaceutically acceptable salt combination for use and/or a vaccine and vexamod or its pharmaceutically acceptable salt combination are manufactured for use in HIV-infected human subjects receiving ART The use of agents for inducing immune response against HIV.

The clinical improvement of the treated humans infected with HIV is expected to be better than that of the comparators (humans infected with HIV under standard care treatment). Clinical improvement may include one or more of lower peak viral load, lower chronic set point, or increased viral rebound delay.

In some embodiments, the methods as described herein are effective for treating HIV infection, for example, as determined by a lower peak viral load compared to standard therapies (for example, ART only). As generally understood in the art, the comparison of the first peak viral load in the first HIV-infected human and the second peak viral load in the second HIV-infected human is measured in the same time period. In some embodiments, the measurement is performed after all antiviral therapies are discontinued. In some embodiments, after treatment with ART, TLR7 agonist, and HIV vaccine, the viral load in the first human infected with HIV is maintained at an undetectable level.

In some embodiments, the first peak viral load of the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine is lower than the second peak viral load of the second HIV-infected human after treatment with ART alone load. In some embodiments, the second peak viral load of the second HIV-infected human after treatment with only ART is greater than the first peak viral load of the first HIV-infected human after treatment with ART, TLR7 agonist, and HIV vaccine High, for example, about 1.2 times to about 10000 times, about 2 times to about 10000 times, about 5 times to about 10000 times, about 10 times to about 10000 times higher. In some embodiments, the second peak viral load of the second HIV-infected human after treatment with only ART is greater than the first peak viral load of the first HIV-infected human after treatment with ART, TLR7 agonist, and HIV vaccine 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, or about 10000 times. In some embodiments, the second peak viral load of the second HIV-infected human after treatment with only ART is greater than the first peak viral load of the first HIV-infected human after treatment with ART, TLR7 agonist, and HIV vaccine About 1000 times higher.

In some embodiments, the first peak viral load of the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine is lower than that of the second HIV-infected human after treatment with ART and TLR7 agonist The second peak viral load. In some embodiments, the second peak viral load of the second HIV-infected human after treatment with ART and TLR7 agonist is higher than the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine. A peak viral load is high, 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. In some embodiments, the second peak viral load of the second HIV-infected human after treatment with ART and TLR7 agonist is higher than the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine. A peak viral load 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, or about 10000 times. In some embodiments, the second peak viral load of the second HIV-infected human after treatment with ART and TLR7 agonist is higher than the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine. A peak viral load is about 20 times higher.

In some embodiments, the first peak viral load of the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine is lower than that of the second HIV-infected human after treatment with ART and HIV vaccine. Peak viral load. In some embodiments, the second peak viral load of the second HIV-infected human after treatment with ART and HIV vaccine is higher than the first peak after the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine The viral load is high, 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. In some embodiments, the second peak viral load of the second HIV-infected human after treatment with ART and HIV vaccine is higher than the first peak after the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine Viral load 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, or about 10000 times. In some embodiments, the second peak viral load of the second HIV-infected human after treatment with ART and HIV vaccine is higher than the first peak after the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine The viral load is about 100 times higher.

In some embodiments, the methods as described herein are effective for treating HIV infection, for example, as determined by a lower chronic set point compared to standard therapies (e.g., ART). As generally understood in the art, the comparison of the first chronic set point in the first HIV-infected human and the second chronic set point in the second HIV-infected human is measured at the same time point. In some embodiments, the measurement is performed after all antiviral therapies are discontinued.

In some embodiments, the first chronic setting point of the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine is lower than the second chronic setting of the second HIV-infected human after treatment with ART alone point. In some embodiments, the second chronic set point of a second HIV-infected human after treatment with only ART is greater than the first chronic set point of a first HIV-infected human after treatment with ART, TLR7 agonist, and HIV vaccine High, for example, about 1.2 times to about 10000 times, about 2 times to about 10000 times, about 5 times to about 10000 times, about 10 times to about 10000 times higher. In some embodiments, the second chronic set point of a second HIV-infected human after treatment with only ART is greater than the first chronic set point of a first HIV-infected human after treatment with ART, TLR7 agonist, and HIV vaccine 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, or about 10000 times. In some embodiments, the second chronic set point of a second HIV-infected human after treatment with only ART is greater than the first chronic set point of a first HIV-infected human after treatment with ART, TLR7 agonist, and HIV vaccine About 10 times higher.

In some embodiments, the first chronic set point of the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine is lower than that of the second HIV-infected human after treatment with ART and TLR7 agonist Second chronic set point. In some embodiments, the second chronic set point of the second HIV-infected human after treatment with ART and TLR7 agonist is lower than the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine. A chronic set point is high, such as about 1.2 times to about 10000 times, about 2 times to about 10000 times, about 5 times to about 10000 times, about 10 times to about 10000 times higher. In some embodiments, the second chronic set point of the second HIV-infected human after treatment with ART and TLR7 agonist is lower than the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine. A chronic set point 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, or about 10000 times. In some embodiments, the second chronic set point of the second HIV-infected human after treatment with ART and TLR7 agonist is lower than the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine. A chronic set point is about 2 times higher.

In some embodiments, the first chronic set point of the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine is lower than the second HIV-infected human after treatment with ART and HIV vaccine. Chronic set point. In some embodiments, the second chronicity set point of the second HIV-infected human after treatment with ART and HIV vaccine is higher than the first chronicity of the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine The set point is high, for example, about 1.2 times to about 10000 times, about 2 times to about 10000 times, about 5 times to about 10000 times, about 10 times to about 10000 times higher. In some embodiments, the second chronicity set point of the second HIV-infected human after treatment with ART and HIV vaccine is higher than the first chronicity of the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine Set point is about 1.2 times higher, 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, or about 10000 times. In some embodiments, the second chronicity set point of the second HIV-infected human after treatment with ART and HIV vaccine is higher than the first chronicity of the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine The set point is about 10 times higher.

After all antiviral therapies are discontinued, the method of the present invention can increase the delay of viral rebound compared with standard therapies. In some embodiments, the viral load in HIV-infected humans does not rebound after treatment with ART, TLR7 agonist, and HIV vaccine. In the absence of a rebound, humans previously infected with HIV must be at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months after the discontinuation of antiretroviral therapy , 9 months, 10 months, 11 months, 1 year, 1.5 years, 2 years, 3 years, 5 years or at least 10 years or longer to maintain the undetectable viral load after the antiviral therapy is stopped.

In some embodiments, the first delay in viral rebound after treatment with ART, TLR7 agonist, and HIV vaccine in the first HIV-infected human is than the second in HIV-infected human after treatment with ART alone. Second, the delay is long. In some embodiments, the first delay in viral rebound after treatment with ART, TLR7 agonist, and HIV vaccine in the first HIV-infected human is than the second in HIV-infected human after treatment with ART alone. 2. The delay is about 1 day to about 10 years, such as 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, compared to the second delay of viral rebound in a second HIV-infected human after treatment with only ART, the virus in a first HIV-infected human after treatment with ART, TLR7 agonist, and HIV vaccine The first delay of the rebound exceeds 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 longer. In some embodiments, compared to the second delay of viral rebound in a second HIV-infected human after treatment with only ART, the virus in a first HIV-infected human after treatment with ART, TLR7 agonist, and HIV vaccine The first delay of the rebound 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. In some embodiments, the first delay in viral rebound after treatment with ART, TLR7 agonist, and HIV vaccine in the first HIV-infected human is than the second in HIV-infected human after treatment with ART alone. Second, the delay is about 3 months.

In some embodiments, the first delay of viral rebound after treatment with ART, TLR7 agonist, and HIV vaccine in a first HIV-infected human is greater than that of a second HIV-infected human after treatment with ART and TLR7 agonist The second delay of the virus rebound is long. In some embodiments, the first delay of viral rebound after treatment with ART, TLR7 agonist, and HIV vaccine in a first HIV-infected human is greater than that of a second HIV-infected human after treatment with ART and TLR7 agonist The second delay of viral rebound is about 1 day to about 10 years, such as 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 first HIV-infected humans are using ART, TLR7 agonists and HIV vaccines compared to the second delay of viral rebound after treatment with ART and TLR7 agonists in second HIV-infected humans The first delay of viral rebound after treatment exceeds 1, 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. In some embodiments, the first HIV-infected humans are using ART, TLR7 agonists and HIV vaccines compared to the second delay of viral rebound after treatment with ART and TLR7 agonists in second HIV-infected humans The first delay of viral rebound after treatment 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, and about 6 Month, 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. In some embodiments, the first delay of viral rebound after treatment with ART, TLR7 agonist, and HIV vaccine in a first HIV-infected human is greater than that of a second HIV-infected human after treatment with ART and TLR7 agonist The second delay of the virus rebound is about 3 months.

In some embodiments, the first delay of viral rebound after treatment with ART, TLR7 agonist, and HIV vaccine in a first HIV-infected human is greater than the viral rebound in a second HIV-infected human after treatment with ART and HIV vaccine The second delay is long. In some embodiments, the first delay of viral rebound after treatment with ART, TLR7 agonist, and HIV vaccine in a first HIV-infected human is greater than the viral rebound in a second HIV-infected human after treatment with ART and HIV vaccine The second delay is about 1 day to about 10 years, such as 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 first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine is compared to the second delay of viral rebound after treatment with ART and HIV vaccine in a second HIV-infected human The first delay of the viral rebound of the virus exceeds 1, 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 more. In some embodiments, the first HIV-infected human after treatment with ART, TLR7 agonist and HIV vaccine is compared to the second delay of viral rebound after treatment with ART and HIV vaccine in a second HIV-infected human The first delay of the virus rebound 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. In some embodiments, the first delay of viral rebound after treatment with ART, TLR7 agonist, and HIV vaccine in a first HIV-infected human is greater than the viral rebound in a second HIV-infected human after treatment with ART and HIV vaccine The second delay is about 1 month long.

The following examples of the present invention further illustrate the nature of the present invention. It should be understood that the following examples do not limit the present invention and the scope of the present invention is determined by the scope of the attached patent application. Example

The present invention also provides the following non-limiting examples.

Example 1 is a method for inducing an immune response against HIV in human subjects infected with human immunodeficiency virus (HIV) receiving antiretroviral therapy (ART), the method comprising: (i) administer an immunogenically effective amount of Ad26 vaccine to the human individual, and the Ad26 vaccine includes an amine that encodes SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4. One or more adenovirus 26 (Ad26) vectors and pharmaceutically acceptable carriers of four HIV antigens based on the base acid sequence; (ii) administer an immunogenically effective amount of MVA vaccine to the human individual, the MVA vaccine comprising a modified Ankara acne (MVA) vector encoding one or more of the four HIV antigens and a pharmaceutically acceptable carrier Agent; and (iii) Administer an effective amount of Vixamod or its pharmaceutically acceptable salt (VES) to the human individual.

Embodiment 1a is the method of embodiment 1, wherein step (i) is performed before step (ii).

Embodiment 1b is the method of embodiment 1 or 1a, wherein step (ii) is performed before step (iii).

Embodiment 1c is the method of any one of Embodiments 1 to 1b, wherein an immunogenically effective amount of MVA vaccine is administered about 22 to 26 weeks, preferably 24 weeks after the initial administration of Ad26 vaccine.

Embodiment 1d is the method of any one of Embodiments 1 to 1c, wherein the effective amount of Vixamod or its pharmaceutically acceptable salt is administered about 26 to 34 weeks after the initial administration of the Ad26 vaccine.

Example 1e is the method of Example 1d, wherein 26, 28, 30, 32, and 34 weeks after the initial administration of the Ad26 vaccine, an effective amount of Vixamod or its pharmaceutically acceptable dose is administered once every two weeks Of salt.

Embodiment 2 is the method of any one of Embodiments 1 to 1e, which further comprises re-administering to the human individual an immunogenically effective amount of Ad26 vaccine.

Example 2a is the method of Example 2, wherein about 10 to 14 weeks, preferably 12 weeks after the initial administration of the Ad26 vaccine, an immunogenically effective amount of the Ad26 vaccine is administered again.

Embodiment 3 is the method of any one of Embodiments 1 to 2a, which further comprises re-administering an immunogenically effective amount of MVA vaccine to the human individual.

Example 3a is the method of Example 3, wherein the MVA vaccine is administered about 34 to 38 weeks after the initial administration of the Ad26 vaccine, preferably 36 weeks.

Embodiment 4 is the method of any one of embodiments 1 to 3a, wherein the Ad26 vaccine comprises a first Ad26 vector encoding the HIV antigen of SEQ ID NO: 1 and a second Ad26 vector encoding the HIV antigen of SEQ ID NO: 2 Vector, the third Ad26 vector encoding the HIV antigen of SEQ ID NO: 3 and the fourth Ad26 vector encoding the HIV antigen of SEQ ID NO: 4.

Embodiment 5 is the method of any one of embodiments 1 to 4, wherein the MVA vaccine is composed of a single MVA vector encoding four HIV antigens.

Embodiment 5a is the method as in embodiment 5, wherein the single MVA vector is an MVA-BN vector.

Embodiment 6 is the method of any one of Embodiments 1 to 5a, in which one or more Ad26 carrier systems are administered together at one time of about 5×10 ⁹ to about 1×10 ¹¹ viral particles (vp) or The total dose of multiple Ad26 vectors is administered.

Embodiment 6a is the method of embodiment 6, wherein the one or more Ad26 carrier systems are administered together in a ^{total dose of about 5×10 10} vp of one or more Ad26 carriers per administration.

Embodiment 7 is the method of any one of embodiments 1 to 6a, wherein the one or more MVA carrier systems are administered together at a rate of about 1×10 ⁷ to about 5×10 ⁸ infectious units (IU). The total dose of one or more MVA vectors is administered.

Embodiment 7a is the method of embodiment 7, wherein the one or more MVA carrier systems are administered together in ^{a total dose of about 2×10 8} IU of the one or more MVA carriers per administration.

Embodiment 8 is the method of any one of embodiments 1 to 7a, wherein visamod or a pharmaceutically acceptable salt thereof is administered at about 3 to about 15 mg visamod per administration, such as about The total dose of 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, or 15 mg visamod is administered.

Example 8a is the method of Example 8, wherein the vexamod or a pharmaceutically acceptable salt thereof is administered in a total dose of about 4 mg to about 6 mg of visamod per administration.

Example 8b is the method of Example 8, wherein the vexamod or a pharmaceutically acceptable salt thereof is administered in a total dose of about 6 mg to about 8 mg of visamod per administration.

Embodiment 8c is the method of embodiment 8, wherein the vexamod or a pharmaceutically acceptable salt thereof is administered in a total dose of about 8 mg to about 10 mg of visamod per administration.

Example 8d is the method of Example 8, wherein the vexamod or a pharmaceutically acceptable salt thereof is administered in a total dose of about 10 mg to about 12 mg of visamod per administration.

Example 8e is the method of Example 8, wherein the vexamod or a pharmaceutically acceptable salt thereof is administered in a total dose of about 12 mg to about 14 mg of visamod per administration.

Embodiment 9 is the method of any one of embodiments 1 to 8e, wherein about 26 to 46 weeks after the initial administration of the Ad26 vaccine, preferably about 26, 28, 30, 32, 34, 38, 40, 42 , 44 and 46 weeks multiple times to administer Vixamod or its pharmaceutically acceptable salt.

Example 9a is the method of Example 9, wherein a total dose of 4 mg visamod is administered at a total dose of 4 mg per administration at 26 and 28 weeks after the initial administration of the Ad26 vaccine to the human individual and 30, after the initial administration of the Ad26 vaccine For 32, 34, 38, 40, 42, 44, and 46 weeks, the total dose of 6 mg of visamod per administration was administered orally to visamod or its pharmaceutically acceptable salt.

Example 9b is the method of Example 9, wherein a total dose of 4 mg of visamod is administered at a total dose of 4 mg visamod at 26, 28 and 30 weeks after the initial administration of the Ad26 vaccine to the human individual and the Ad26 vaccine is initially administered After 32, 34, 38, 40, 42, 44, and 46 weeks, the total dose of 6 mg of vesamod per administration was administered orally to vesamod or its pharmaceutically acceptable salt.

Example 9c is the method of Example 9, wherein a total dose of 4 mg visamod is administered at 26, 28, 30, and 32 weeks after the initial administration of the Ad26 vaccine to the human individual and the Ad26 is initially administered. Vesalimod or its pharmaceutically acceptable salt was administered orally at 34, 38, 40, 42, 44, and 46 weeks after the vaccine, in a total dose of 6 mg of vexamod per administration.

Example 9d is the method of Example 9, wherein about 6 mg of vesamox is administered at 26, 28, 30, 32, 34, 38, 40, 42, 44, and 46 weeks after the initial administration of the Ad26 vaccine The total dose of Dezhi is administered orally to Vixamod or its pharmaceutically acceptable salt.

Example 9e is the method of Example 9, wherein a total dose of about 10 mg of visamod is administered at 26, 28 and 30 weeks after the initial administration of the Ad26 vaccine and after the initial administration of the Ad26 vaccine 32, For 34, 38, 40, 42, 44, and 46 weeks, the total dose of 12 mg of visamod per administration was administered orally to visamod or its pharmaceutically acceptable salt.

Example 9f is the method of Example 9, wherein a total dose of about 8 mg of visamod is administered at 26, 28, and 30 weeks after the initial administration of the Ad26 vaccine and after the initial administration of the Ad26 vaccine 32, For 34, 38, 40, 42, 44, and 46 weeks, the total dose of 10 mg of Vixamod per administration was orally administered with Vixamod or its pharmaceutically acceptable salt.

Embodiment 10 is the method of any one of embodiments 1 to 9, wherein about 26, 28, 30, 32, 34, 38, 40, 42, 44, and 46 weeks after the initial administration of the Ad26 vaccine in each administration Vissamod or its pharmaceutically acceptable salt was administered orally with a total dose of 6 mg.

Embodiment 11 is the method of any one of embodiments 1 to 9, wherein a total dose of 6 mg per administration is administered at about 26 and 28 weeks after the initial administration of the Ad26 vaccine and 30 after the initial administration of the Ad26 vaccine. , 32, 34, 38, 40, 42, 44, and 46 weeks with a total dose of 8 mg per administration orally administered Vixamod or its pharmaceutically acceptable salt.

Embodiment 12 is the method of any one of Embodiments 1 to 11, which further comprises administering to the human individual an immunogenically effective amount of a gp140 vaccine comprising one or more isolated HIV gp140 mantle polypeptides.

Example 12a is the method of Example 12, wherein the gp140 vaccine is administered in combination with the Ad26 vaccine, and the Ad26 vaccine is preferably administered again.

Example 12b is the method of Example 12, wherein the gp140 vaccine is administered in combination with the MVA vaccine.

Example 12c is the method of Example 12b, wherein the administration of the gp140 vaccine is combined with the initial administration of the MVA vaccine.

Example 12d is the method of Example 12b, wherein the administration of the gp140 vaccine is combined with the re-administration of the MVA vaccine.

Example 12e is the method of Example 12b, wherein the administration of the gp140 vaccine is combined with the initial administration of the MVA vaccine and the re-administration of the MVA vaccine.

Embodiment 13 is the method of any one of embodiments 12 to 12e, wherein the gp140 vaccine comprises two isolated HIV gp140 mantle polypeptides having the amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 10, respectively.

Embodiment 14 is the method of any one of embodiments 12 to 13, wherein the one or more isolated HIV gp140 mantle polypeptides are administered with about 125 to 350 μg of the total glycoprotein of HIV gp140 mantle polypeptides each time The total dose is administered.

Embodiment 14a is the method of any one of embodiments 12 to 13, wherein the one or more isolated HIV gp140 mantle polypeptides are administered at a time of about 250 μg of HIV gp140 mantle polypeptide total glycoprotein Dosage administration.

Example 15 is a method for inducing an immune response against HIV in a human subject infected with human immunodeficiency virus (HIV) receiving antiretroviral therapy (ART), the method comprising: (i) about each administration 5×10 ⁹ to about 1×10 ¹¹ viral particles (vp), preferably about 5×10 ¹⁰ vp of the total dose of Ad26 vector to the human individual to administer the Ad26 vaccine, the Ad26 vaccine comprising a code with SEQ ID NO : 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 amino acid sequence of four HIV antigens one or more adenovirus 26 (Ad26) carrier and pharmaceutically acceptable carrier (Ii) The total dose of Ad26 vector of about 5×10 ⁹ to about 1×10 ¹¹ viral particles (vp), preferably about 5×10 ¹⁰ vp, is administered to the human individual per administration. Ad26 vaccine, wherein the Ad26 vaccine is administered 10 to 14 weeks after the Ad26 vaccine is administered in step (i), preferably 12 weeks; (iii) about 1×10 ⁷ to about 5×10 are administered each time ^{A total dose of 8} infectious units (IU), preferably about 2×10 ⁸ IU or more of the MVA vector is administered to the human individual with an MVA vaccine, the MVA vaccine comprising one or more of the four HIV antigens One MVA carrier (preferably one or more MVA-BN carriers) and a pharmaceutically acceptable carrier; optionally combined with the MVA vaccine to give about 125 μg to 350 μg each time, preferably about 250 μg The total dose of the glycoprotein of the two isolated HIV gp140 mantle polypeptides is further administered to the human individual comprising two isolated HIV gp140 sets of amino acid sequences having SEQ ID NO: 9 and SEQ ID NO: 10, respectively Membrane polypeptide gp140 vaccine, aluminum adjuvant and pharmaceutically acceptable carrier, wherein 22 to 26 weeks after the Ad26 vaccine is administered in step (i), preferably 24 weeks, the MVA vaccine is administered and optionally selected Gp140 vaccine; and (iv) administering visamod or a pharmaceutically acceptable salt thereof to a human individual in a total dose of about 3 mg to about 15 mg of visamod per administration, wherein in step ( i) 26 to 34 weeks after the Ad26 vaccine is administered, preferably 26, 28, 30, 32 and 34 weeks after the Ad26 vaccine is administered in step (i), and Vixamod is administered once every two weeks Or its pharmaceutically acceptable salt.

Example 15a is the method of Example 15, wherein the Ad26 vaccine comprises the first Ad26 vector encoding the HIV antigen of SEQ ID NO: 1, the second Ad26 vector encoding the HIV antigen of SEQ ID NO: 2, and the second Ad26 vector encoding the HIV antigen of SEQ ID NO: The third Ad26 vector of the HIV antigen of 3 and the fourth Ad26 vector of the HIV antigen of SEQ ID NO: 4 are encoded.

Example 15b is the method of Example 15 or 15a, wherein the MVA vaccine is composed of a single MVA vector encoding four HIV antigens.

Embodiment 15c is the method of embodiment 15b, wherein the single MVA vector is an MVA-BN vector.

Embodiment 15d is the method of any one of Embodiments 15 to 15c, wherein: (i) the Ad26 vaccine is initially administered at a total dose of ^{about 5×10 10 vp of Ad26 vector per administration; (ii)} ) The Ad26 vaccine was re-administered 12 weeks after the initial administration of the Ad26 vaccine at ^{a total dose of approximately 5×10 10} vp of Ad26 vector per administration; (iii) The MVA vaccine was initially administered after the initial administration of the Ad26 vaccine The total dose of one or more MVA carriers of about 2×10 ⁸ IU is administered for 24 weeks; and (iv) Vesalimod or its pharmaceutically acceptable salt is in step (i) The Ad26 vaccine was administered 26, 28, 30, 32, and 34 weeks after the administration.

Embodiment 15e is the method of any one of Embodiments 15 to 15d, wherein each of the Ad26 vaccine and the MVA vaccine is administered intramuscularly, and visamod or its pharmaceutically acceptable salt is Orally administered.

Embodiment 15f is the method of any one of embodiments 15 to 15e, wherein the MVA vaccine is administered in combination with a gp140 vaccine, an aluminum adjuvant, and a pharmaceutically acceptable carrier.

Example 15g is the method of Example 15f, wherein the gp140 vaccine is administered intramuscularly.

Example 15h is the method of Example 15f or 15g, wherein the gp140 vaccine is administered in a total dose of about 250 μg of two glycoproteins of isolated HIV gp140 mantle polypeptides per administration.

Embodiment 16 is the method of any one of embodiments 15 to 15h, which further comprises: administering about 1×10 ⁷ to about 5×10 ⁸ infectious units (IU), preferably about 2×10 ^{A total dose of 8} IU of the one or more MVA vectors is then administered to the human individual with an MVA vaccine; optionally combined with the MVA vaccine, each administration is about 125 μg to 350 μg, preferably about 250 μg. The total dose of the glycoprotein of the isolated HIV gp140 mantle polypeptide is further administered to the human individual with gp140 vaccine, aluminum adjuvant and pharmaceutically acceptable carrier, wherein after the Ad26 vaccine is administered in step (i) 34 to 38 weeks, preferably 36 weeks before administration of MVA vaccine and optionally gp140 vaccine.

Example 16a is the method of Example 16, wherein 36 weeks after the initial administration of the Ad26 vaccine, the MVA vaccine is administered at a total dose of ^{about 2×10 8 IU of one or more MVA vectors per administration.}

Embodiment 16b is the method of embodiment 16 or 16a, wherein the MVA vaccine is combined with gp140 vaccine, aluminum adjuvant and pharmaceutically acceptable carrier before administration.

Example 16c is the method of Example 16b, wherein the gp140 vaccine is administered intramuscularly.

Example 16d is the method of Example 16b or 16c, wherein the gp140 vaccine is re-administered at a total dose of about 250 μg two glycoproteins of isolated HIV gp140 mantle polypeptides per administration.

Example 17 is the method of any one of Examples 15 to 16d, which further comprises: 38 to 46 weeks after the initial administration of the Ad26 vaccine at a total dose of about 3 mg to about 15 mg per administration to the human individual Re-administer Vixamod or its pharmaceutically acceptable salt.

Embodiment 17a is the method of embodiment 17, wherein visamod or its pharmaceutically acceptable salt is administered orally.

Embodiment 17b is the method of embodiment 17a, wherein Vixamod or its pharmaceutically acceptable salt is about 26 to 46 weeks after the initial administration of Ad26 vaccine, preferably about 26, 28, 30, 32, 34, 38, 40, 42, 44 and 46 weeks to cast multiple times.

Example 17c is the method of Example 17b, wherein a total dose of 4 mg visamod per administration is 26 and 28 weeks after the initial administration of the Ad26 vaccine to the human individual and 30, after the initial administration of the Ad26 vaccine For 32, 34, 38, 40, 42, 44, and 46 weeks, the total dose of 6 mg of visamod per administration was administered orally to visamod or its pharmaceutically acceptable salt.

Example 17d is the method of Example 17b, wherein a total dose of 4 mg visamod per administration is 26, 28 and 30 weeks after the initial administration of the Ad26 vaccine to the human individual and after the initial administration of the Ad26 vaccine For 32, 34, 38, 40, 42, 44, and 46 weeks, the total dose of 6 mg of visamod per administration was administered orally to visamod or its pharmaceutically acceptable salt.

Example 17e is the method of Example 17b, wherein a total dose of 4 mg visamod is administered at 26, 28, 30, and 32 weeks after the initial administration of the Ad26 vaccine to the human individual and the Ad26 is initially administered. Vesalimod or its pharmaceutically acceptable salt was administered orally at 34, 38, 40, 42, 44, and 46 weeks after the vaccine, in a total dose of 6 mg of vexamod per administration.

Example 17f is the method of Example 17b, wherein about 6 mg of vesamox is administered at 26, 28, 30, 32, 34, 38, 40, 42, 44, and 46 weeks after the initial administration of Ad26 vaccine The total dose of Dezhi is administered orally to Vixamod or its pharmaceutically acceptable salt.

Example 17g is the method of Example 17b, wherein a total dose of about 10 mg visamod is administered at 26, 28 and 30 weeks after the initial administration of the Ad26 vaccine and after the initial administration of the Ad26 vaccine 32, For 34, 38, 40, 42, 44, and 46 weeks, the total dose of 12 mg of visamod per administration was administered orally to visamod or its pharmaceutically acceptable salt.

Example 17h is the method of Example 17b, wherein about 26, 28, 30, 32, 34, 38, 40, 42, 44, and 46 weeks after the initial administration of the Ad26 vaccine, a total dose of 6 mg is administered each time Vixamod or its pharmaceutically acceptable salt is administered orally.

Example 17i is the method of Example 17b, wherein a total dose of 6 mg is administered at a total dose of 6 mg each time about 26 and 28 weeks after the initial administration of the Ad26 vaccine and 30, 32, 34, 38, and 38 after the initial administration of the Ad26 vaccine. Vesalimod or its pharmaceutically acceptable salt was administered orally in a total dose of 8 mg per administration for 40, 42, 44, and 46 weeks.

Example 17j is the method of Example 17b, wherein a total dose of about 8 mg visamod is administered at 26, 28, and 30 weeks after the initial administration of the Ad26 vaccine and after the initial administration of the Ad26 vaccine 32, For 34, 38, 40, 42, 44, and 46 weeks, the total dose of 10 mg of Vixamod per administration was orally administered with Vixamod or its pharmaceutically acceptable salt.

Embodiment 18 is the method of any one of embodiments 1 to 17i, wherein the human individual initiates ART during the acute phase of HIV infection.

Embodiment 18a is the method of any one of embodiments 1 to 17i, wherein the human individual initiates ART outside the acute phase of HIV infection.

Embodiment 18b is the method of any one of embodiments 1 to 18a, wherein the human individual has chronic HIV infection.

Embodiment 19 is the method of any one of embodiments 1 to 18b, wherein the human subject is undergoing inhibitory ART during treatment.

Example 20 is the method of Example 19, wherein the inhibitory ART is discontinued after the treatment.

Embodiment 21 is a method for treating human immunodeficiency virus (HIV) infection in a human individual in need, which comprises: (i) Treating human individuals with antiretroviral therapy (ART); and (ii) Use the method as in any one of Examples 1 to 20 to induce an anti-HIV immune response in the human individual.

Embodiment 22 is the method of embodiment 21, which further comprises suspending the ART treatment of step (i) after step (ii).

Embodiment 23 is the method of embodiment 22, wherein the human individual maintains viral suppression for at least 24 weeks after discontinuing ART.

Example 24 is a combination for inducing an immune response against HIV in human individuals infected with human immunodeficiency virus (HIV) receiving antiretroviral therapy (ART), the combination comprising: (i) Ad26 vaccine, which includes one or more adenoviruses that together encode four HIV antigens with the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4 26 (Ad26) carrier and pharmaceutically acceptable carrier; (ii) MVA vaccine, which includes one or more modified Ankara acne (MVA) vectors encoding the four HIV antigens, preferably one or more MVA-BN vectors, and a pharmaceutically acceptable carrier; and (iii) A composition comprising Vixamod or its pharmaceutically acceptable salt (VES).

Embodiment 25 is the combination of embodiment 24, which further comprises a gp140 vaccine, which comprises two isolated HIV gp140 mantle polypeptides having the amino acid sequence of SEQ ID NO: 9 and SEQ ID NO: 10, respectively At least one of and a pharmaceutically acceptable carrier.

Example 25a is the combination as in Example 24, wherein the MVA vaccine consists of a single MVA vector encoding four HIV antigens, preferably a single MVA-BN vector.

Example 25b is the combination as in Example 25, wherein the gp140 vaccine contains two isolated HIV gp140 mantle polypeptides.

Embodiment 26 is a combination as any one of Embodiments 25 to 25b, which further comprises an adjuvant, preferably an aluminum adjuvant.

Embodiment 27 is a combination as in embodiment 26, wherein the adjuvant is formulated with the gp140 vaccine or co-administered with the gp140 vaccine.

Example 28 is a kit comprising a combination as in any of Examples 24 to 27 and using the combination to induce resistance in human subjects infected with human immunodeficiency virus (HIV) receiving retroviral therapy (ART) Instructions for the immune response to HIV. Instance

Instance 1 : Research is receiving antiretroviral therapy (ART) Of HIV Infect humans HIV Vaccine program

A clinical study was conducted in humans to study the effects of two different vaccine regimens and TLR7 agonists in adults infected with HIV 1 who started ART during acute HIV infection. In addition to ART, research vaccination (for example, vaccination with Ad26 vector, MVA vector, and adjuvanted isolated HIV gp140 polypeptide for vaccination) and TLR7 agonist (for example, Vixamod) will be administered.

the goal

This study will evaluate whether the combination of 2 different vaccine regimens and TLR7 agonist will be safe and immunogenic in HIV-1 infected adults who start ART during acute HIV infection, and will be the initial virus after ART is interrupted The rebound led to continued virological control.

Research group

The screening of eligible participants will be conducted within 70 days prior to the administration of the research intervention. The inclusion criteria for recruiting participants in this study include, for example 1. Must have confirmed HIV-1 infection (including the route and estimated duration of HIV infection, pre-ART virus set point and ARV history). 2. ART must be initiated within 14 days of a documented acute HIV-1 infection as defined below: -Confirm that HIV can be detected by PCR and negative or uncertain EIA; or -Positive 4th generation HIV analysis and negative/uncertain HIV conventional Ab test or negative/uncertain Western blot method; or -Confirm that HIV can be detected by PCR within 21 days of negative HIV NAAT; or -Recording standards for Fiebig I-IV period. 3. Inhibitory ART must be performed for at least 48 weeks before screening. ART is defined as a combination therapy regimen that includes more than 2 compounds, such as 2x nucleoside reverse transcriptase inhibitors plus non-nucleoside reverse transcriptase inhibitors or integrase inhibitors. Permitted ARTs that are part of the current ARV protocol include nucleoside reverse transcriptase inhibitors (NRTI), letgevir, dululamir, ripivirine, bictegravir, and maravirin. 4. Must have plasma HIV RNA <50 cps/mL at the time of screening, and at least one recorded plasma HIV RNA <50 cps/mL evidence after the last ART change. ● Within 12 weeks before screening, a deviation of HIV RNA ≥50 and <200 cps/mL is acceptable, and the restriction is that the latest HIV RNA <50 cps/mL. In the case of changes in ART during screening, one deviation of HIV RNA ≥50 and <200 cps/mL is allowed 6 weeks after the new ART protocol, and the restriction condition is that for repeated tests, HIV RNA <50 cps/mL.

Vaccination and experimental design

This is a multi-center, randomized, parallel group, placebo-controlled, double-blind, phase 1/2a clinical study to study two different vaccine regimens and TLR7 promotion in HIV 1-infected adults who started ART during acute HIV infection. The safety, tolerability, efficacy, immunogenicity and effect of the effective agent ARV ATI on the control of HIV viremia afterwards. In addition to ART, research vaccination and TLR7 administration will be carried out.

The study will enroll 90 participants, who are randomly divided into 2 vaccine groups and 1 matched placebo group at a ratio of 1:1:1. The study population will include suppressive ART for at least 48 weeks prior to screening and maintenance of undetectable viremia (HIV RNA <50 cps/mL) ≥ 12 weeks and maintenance of CD4 counts before the initial vaccine/placebo administration> Adults infected with HIV with 450 cells/mm ^3.

The study included a 10-week screening period, a 48-week treatment period (Phase 1), and a 48-week ATI period (Phase 2 and Phase 3). Based on the 72-week efficacy results, a long-term extension period (Phase 4, 104 weeks [approximately 2 years], after the 96th week) was added to the participants in the active treatment group. The details of the evaluation during this optional LTE phase will be defined at that time. As used herein, "week X" refers to weeks X after the initial administration of the Ad26 vaccine at week 0.

Dosage and administration

Individuals received four doses of the study vaccine: the adenovirus 26 vector (Ad26.Mos4.HIV) encoding the embedded HIV antigen or placebo was administered at weeks 0 and 12; and it was administered at weeks 24 and 36 (i) MVA vector encoding embedded HIV antigen (MVA-BN-HIV) (or placebo) or (ii) a combination of a mixture of MVA-BN-HIV and adjuvanted HIV gp140 polypeptide (or placebo). At 26, 28, 30, 32, 34, 38, 40, 42, 44 and 46 weeks at a dose of 6 mg (3×2 mg tablets) per administration or at 26 and 28 weeks A dose of 6 mg (3×2 mg tablets) and a dose of 8 mg (2×4 mg tablets) per administration at 30, 32, 34, 38, 40, 42, 44 and 46 weeks, after Oral vote with Visamod. Alternatively, Vixamod is administered orally in the following doses: (a) One dose of 4 mg (1×4 mg lozenge) is administered at the 26th and 28th weeks and the 30th, 32nd, 34th, and 38th , 40, 42, 44 and 46 weeks, one dose of 6 mg (3×2 mg tablets); (b) One dose of 4 mg (1×4 mg) in weeks 26, 28 and 30 Lozenges) and one dose of 6 mg (3×2 mg lozenges) at the 32, 34, 38, 40, 42, 44 and 46 weeks; (c) at the 26, 28, 30, and 32 weeks One dose of 4 mg (1×4 mg lozenge) each time and one dose of 6 mg (3 x 2 mg lozenges) at the 34th, 38th, 40th, 42, 44 and 46 weeks; (d) One dose of 10 mg (5×2 mg lozenge) was administered at weeks 26, 28, and 30, and a dose of 12 mg was administered at weeks 32, 34, 38, 40, 42, 44, and 46 ( 6×2 mg tablets); or (e) One dose of 8 mg (4×2 mg tablets) was administered on the 26th, 28th and 30th weeks and the 32nd, 34th, 38th, 40th, 42nd and 44th And one dose of 10 mg (5×2 mg lozenge) was administered every 46 weeks.

The research vaccines (Ad26 _mos and MVA-BN-HIV and adjuvanted HIV gp140 polypeptide), VES and placebo with the administered doses are as follows: (i) Ad26.Mos4.HIV is pre-mixed in the same vial as follows: The four vaccine products supplied are composed of virus particles (vp) at a ratio of 1:1:1:1: HIV mosaic Env1 (SEQ ID NO: 1), mosaic Env2S (SEQ ID NO: 2), Ad26.Mos1Env, Ad26.Mos2SEnv, Ad26.Mos1Gag-Pol and Ad26.Mos2Gag-Pol with mosaic GagPol1 (SEQ ID: NO 3) and mosaic GagPol2 (SEQ ID NO: 4) genes; ×10 ¹⁰ viral particles (vp) total dose administration; (ii) MVA-BN-HIV is expressed as Mos1.Gag-Pol (SEQ ID NO: 3), Mos2.Gag-Pol (SEQ ID NO: 4) , Mos1.Env (SEQ ID NO: 1) and Mos2S.Env (SEQ ID NO: 2) MVA-BN; in 0.5 mL injection with ^{a total dose of about 2×10 8} infection units (IU) ; MVA-BN-HIV has been described in more detail as MVA-mBN414 in Example 7 of WO 2018/229711; (iii) gp140 HIV bivalent vaccine, which is adjuvanted and contains 0.16 mg/mL branch C gp140 ( SEQ ID NO: 9), 0.15 mg/mL mosaic gp140 (SEQ ID NO: 10) and 0.85 mg/mL aluminum (Al); 125 micrograms (mcg) branch C gp140 glycoprotein, 125 mcg mosaic gp140 per 0.5 mL dose Glycoprotein (corresponding to 80 mcg and 75 mcg protein respectively) and aluminum adjuvant (425 mcg Al) were administered; (iv) The vaccine placebo was 0.9% sodium chloride (0.5 mL injection); (v) Participants will Receive 10 doses of VES orally, once every 14 days as follows: a. At 26, 28, 30, 32, 34, 38, 40, 42, 44, and 46 weeks, each dose of 6 mg (3×2 mg tablets) Dose) the total dose of VES; b. In the 26th and 28th weeks, each administration of 6 mg (3×2 mg tablets) the total dose of VES and on the 30th, 32nd, 34th, 38th, 40th, 42nd, 44th And 46 weeks, each administration of the total dose of 8 mg (2×4 mg tablets) VES; c. in the 26th and 28th 4 mg (1×4 mg tablet) per week for the total dose of VES and 6 mg (3×2 mg tablet) at 30, 32, 34, 38, 40, 42, 44 and 46 weeks. Dose) the total dose of VES; d. 4 mg (1×4 mg lozenge) per administration at the 26th, 28th, and 30th weeks of the total dose of VES and the total dose at the 32nd, 34th, 38th, 40th, 42nd, 44th and The total dose of 6 mg (3×2 mg tablets) of VES per administration at 46 weeks; e. The total dose of 4 mg (1×4 mg tablets) VES per administration at weeks 26, 28, 30 and 32 Dose and the total dose of 6 mg (3×2 mg lozenge) VES per administration on the 34th, 38th, 40th, 42, 44 and 46 weeks; f. 10 per administration on the 26th, 28th and 30th weeks mg (5×2 mg tablets) total dose of VES and 12 mg (6×2 mg tablets) total dose of VES at 32, 34, 38, 40, 42, 44 and 46 weeks; or g. The total dose of 8 mg (4×2 mg lozenge) VES per administration at 26, 28 and 30 weeks and 10 per administration at 32, 34, 38, 40, 42, 44 and 46 weeks mg (5×2 mg tablets) total dose of VES (ii) VES placebo is lactose, which is administered in a total dose corresponding to the same number of tablets used for the treatment of VES.

Individuals received the study vaccine by intramuscular administration and oral administration of VES or placebo according to the schedule in Table 1 below: Table 1 : Schedule of administration of the study vaccine group N Week 0 Week 12 Week 24 Week 26, 28, 30, 32, 34 Week 36 Weeks 38, 40, 42, 44, 46 Test group 1 30 Ad26.Mos4.HIV Ad26.Mos4.HIV MVA-BN-HIV + placebo VES MVA-BN-HIV + placebo VES Test group 2 30 Ad26.Mos4.HIV Ad26.Mos4.HIV MVA-BN-HIV + branch C gp140, mosaic gp140, adjuvanted VES MVA-BN-HIV + branch C gp140, mosaic gp140, adjuvanted VES Control group 30 Vaccine placebo Vaccine placebo Vaccine placebo VES placebo Vaccine placebo VES placebo

Individuals in the test group and the control group continue to receive standard ART during the study period (e.g. at least three antiviral compounds, such as two nucleoside reverse transcriptase inhibitors plus nucleoside reverse transcriptase inhibitors or protease inhibitors or integrase inhibitors ) For HIV treatment. Collect blood and optionally genital secretions during specific clinical visits to evaluate immune response (cellular and humoral immune response) and viremia control throughout the study.

SEQ ID NO: 2 (Mos2SEnv) 711 aa

SEQ ID NO:3 (Mos1.Gag-Pol) 1350 aa:

SEQ ID NO:4 (Mos2.Gag-Pol) 1341 aa:

SEQ ID NO: 5 (Mos1.Env DNA)

SEQ ID NO: 6 (Mos2SEnv DNA)

SEQ ID NO: 7 (Mos1.Gag-Pol DNA)

SEQ ID NO: 8 (Mos2.Gag-Pol DNA)

SEQ ID NO: 9 ( 分支 C gp140 蛋白 )- 679 個胺基酸

SEQ ID NO: 10 ( 鑲嵌 gp140 蛋白 )- 695 個胺基酸

SEQ ID NO: 11 ( 例示性前導序列 ) - 29 個胺基酸 MRVRGIQRNC QHLWRWGTLI LGMLMICSA 參考文獻 1.  US20120076812 2.  Barouch等人，Nat Med 2010, 16:319-323 3.  Barouch等人，Cell 155:1-9, 2013 4.  Fischer等人，Nat Med, 2007. 13(1): p. 100-6 5.  Havenga,等人，2006, J Gen Virol 87: 2135-43 6.  WO 03/104467 7.  WO 2007/104792 8.  Abbink等人，(2007) Virol 81(9): 4654-63 9.  Mayr等人(1975) Infection 3, 6-14 10.      Mayr, A. & Danner, K. (1978), Dev. Biol. Stand. 41: 225-234 11.      Stickl (1974), Prev. Med. 3: 97-101 12.      Stickl及Hochstein-Mintzel (1971), Munch. Med. Wochenschr. 113: 1149-1153 13.      Mayr等人(1978), Zentralbl. Bacteriol. (B) 167:375-390 14.      WO 02/42480 (US 2003/0206926) 15.      WO 03/048184 (US 2006/0159699) 16.      US20110159036 17.      US 8197825 18.      WO 2011/092029 19.      Nkolola等人2010,J. Virology 84(7): 3270-3279 20.      Kovacs等人，PNAS 2012, 109(30):12111-6 21.      WO 2010/042942 22.      WO 2014/107744 23.      National Center for Biotechnology Information. PubChem Database. Vesatolimod, CID=46241268, https:// pubchem.ncbi.nlm.nih.gov /compound/46241268 (2019年4月23日存取)It should be understood that the examples and embodiments described herein are for illustrative purposes only, and changes can be made to the above-described embodiments without departing from the broad concept of the invention. Therefore, it should be understood that the present invention is not limited to the specific embodiments disclosed, but is intended to cover modifications within the spirit and scope of the present invention as defined by the scope of the appended patent application. Sequence Listing SEQ ID NO:1 (Mos1.Env) 685 aa:

SEQ ID NO: 2 (Mos2SEnv) 711 aa

SEQ ID NO: 3 (Mos1.Gag-Pol) 1350 aa:

SEQ ID NO: 4 (Mos2.Gag-Pol) 1341 aa:

SEQ ID NO: 5 (Mos1.Env DNA)

SEQ ID NO: 6 (Mos2SEnv DNA)

SEQ ID NO: 7 (Mos1.Gag-Pol DNA)

SEQ ID NO: 8 (Mos2.Gag-Pol DNA)

SEQ ID NO: 9 ( branched C gp140 protein )-679 amino acids

SEQ ID NO: 10 ( inlaid gp140 protein )-695 amino acids

SEQ ID NO: 11 ( exemplary leader sequence )-29 amino acids MRVRGIQRNC QHLWRWGTLI LGMLMICSA Reference 1. US20120076812 2. Barouch et al., Nat Med 2010, 16:319-323 3. Barouch et al., Cell 155:1 -9, 2013 4. Fischer et al., Nat Med, 2007. 13(1): p. 100-6 5. Havenga, et al., 2006, J Gen Virol 87: 2135-43 6. WO 03/104467 7. WO 2007/104792 8. Abbink et al. (2007) Virol 81(9): 4654-63 9. Mayr et al. (1975) Infection 3, 6-14 10. Mayr, A. & Danner, K. (1978) , Dev. Biol. Stand. 41: 225-234 11. Stickl (1974), Prev. Med. 3: 97-101 12. Stickl and Hochstein-Mintzel (1971), Munch. Med. Wochenschr. 113: 1149-1153 13. Mayr et al. (1978), Zentralbl. Bacteriol. (B) 167:375-390 14. WO 02/42480 (US 2003/0206926) 15. WO 03/048184 (US 2006/0159699) 16. US20110159036 17. US 8197825 18. WO 2011/092029 19. Nkolola et al. 2010, J. Virology 84(7): 3270-3279 20. Kovacs et al., PNAS 2012, 109(30): 12111-6 21. WO 2010/042942 22 . WO 2014/107744 23. National Center for Biotechnology Information. PubChem Database. Vesatolimod, CID=46241268, http s:// pubchem.ncbi.nlm.nih.gov /compound/46241268 (accessed April 23, 2019)

Claims (23)

A method for inducing an immune response against HIV in human individuals infected with human immunodeficiency virus (HIV) receiving antiretroviral therapy (ART), the method comprising: (i) administer an immunogenically effective amount of Ad26 vaccine to the human individual, and the Ad26 vaccine includes an amine encoding SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4. One or more adenovirus 26 (Ad26) vectors and pharmaceutically acceptable carriers of four HIV antigens based on the base acid sequence; (ii) Administer an immunogenically effective amount of MVA vaccine to the human individual, the MVA vaccine comprising a modified Ankara acne (MVA) vector encoding one or more of the four HIV antigens and a pharmaceutically acceptable carrier Agent; and (iii) Administer an effective amount of vesatolimod or its pharmaceutically acceptable salt (VES) to the human individual. The method of claim 1, further comprising administering to the human individual an immunogenically effective amount of the Ad26 vaccine, preferably about 10 to 14 weeks, more preferably 12 weeks after the initial administration of the Ad26 vaccine. The method of claim 1 or 2, further comprising re-administering to the human individual an immunogenically effective amount of the MVA vaccine, preferably, wherein the MVA vaccine is initially administered about 22 to about 22 to after the Ad26 vaccine is initially administered 26 weeks, preferably 24 weeks, and preferably, the MVA vaccine is administered approximately 34 to 38 weeks, preferably 36 weeks after the initial administration of the Ad26 vaccine. The method according to any one of claims 1 to 3, wherein the Ad26 vaccine comprises a first Ad26 vector encoding the HIV antigen of SEQ ID NO: 1, a second Ad26 vector encoding the HIV antigen of SEQ ID NO: 2, The third Ad26 vector encoding the HIV antigen of SEQ ID NO: 3 and the fourth Ad26 vector encoding the HIV antigen of SEQ ID NO: 4. The method according to any one of claims 1 to 4, wherein the MVA vaccine is composed of a single MVA vector encoding the four HIV antigens. Such as the method of any one of claims 1 to 5, wherein the one or more Ad26 vectors are administered together at about 5×10 ⁹ to about 1×10 ¹¹ viral particles (vp), preferably about 5× The total dose of the one or more Ad26 vectors of 10 ^{10 vp is administered.} Such as the method of any one of claims 1 to 6, wherein the one or more MVA vectors are administered together at about 1×10 ⁷ to about 5×10 ⁸ infectious units (IU), preferably about 2× 10 ⁸ IU of the total dose of the one or more MVA vectors is administered. The method according to any one of claims 1 to 7, wherein the vexamod or a pharmaceutically acceptable salt thereof is administered at about 3 to 15 mg of vexamod per time, preferably about 4 to 12 mg of visamod, for example, a total dose of about 6 to 8 mg of visamod is administered. The method according to any one of claims 1 to 8, wherein Vixamod or its pharmaceutically acceptable salt is about 26 to 46 weeks after the initial administration of the Ad26 vaccine, preferably about 26, 28, 30, 32, 34, 38, 40, 42, 44 and 46 weeks to cast multiple times. The method according to any one of claims 1 to 9, wherein the total dose of Vixamod or its pharmaceutically acceptable salt is about 26, 28 after the initial administration of the Ad26 vaccine in a total dose of 6 mg per administration , 30, 32, 34, 38, 40, 42, 44 and 46 weeks. The method according to any one of claims 1 to 9, wherein a total dose of 6 mg is administered at a time about 26 and 28 weeks after the initial administration of the Ad26 vaccine, and 30, 32, 30, 32, 30, 32, and 30 after the initial administration of the Ad26 vaccine. Vesalimod or its pharmaceutically acceptable salt was administered at a total dose of 8 mg per administration for 34, 38, 40, 42, 44 and 46 weeks. The method according to any one of claims 1 to 11, which further comprises administering to the human subject an immunogenically effective amount of a gp140 vaccine comprising one or more isolated HIV gp140 envelope polypeptides, which is preferably the same as the Ad26 vaccine Or the MVA vaccine combination, more preferably combined with the MVA vaccine. The method of claim 12, wherein the gp140 vaccine comprises two isolated HIV gp140 mantle polypeptides having the amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 10, and the gp140 vaccine system and the MVA vaccine Portfolio investment. The method of claim 12 or 13, wherein the one or more isolated HIV gp140 mantle polypeptides are administered at a time of about 125 to 350 μg, preferably about 250 μg, of the total glycoprotein of the HIV gp140 mantle polypeptides The total dose is administered. A method for inducing an immune response against HIV in human individuals infected with human immunodeficiency virus (HIV) receiving antiretroviral therapy (ART), the method comprising: (i) about 5×10 ^{9 doses per time} The total dose of Ad26 vector to about 1×10 ¹¹ viral particles (vp), preferably about 5×10 ¹⁰ vp, is administered intramuscularly to the human individual with the Ad26 vaccine, and the Ad26 vaccine contains a code with SEQ ID NO: 1 , SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 amino acid sequences of four HIV antigens of one or more adenovirus 26 (Ad26) vectors and pharmaceutically acceptable carriers; ( ii) The total dose of the Ad26 vectors of about 5×10 ⁹ to about 1×10 ¹¹ viral particles (vp), preferably about 5×10 ¹⁰ vp, is administered to the human subject intramuscularly each time The Ad26 vaccine, wherein the Ad26 vaccine is administered 10 to 14 weeks, preferably 12 weeks after the Ad26 vaccine is administered in step (i); (iii) about 1×10 ⁷ to about 5× is administered each time 10 ⁸ infectious units (IU), preferably one of about 2 × 10 ⁸ IU or more of th total dose of MVA vector intramuscularly administered to a human subject with MVA vaccine, the vaccine comprising MVA encoding one of four kinds of HIV antigens Or more MVA carriers (preferably one or more MVA-BN carriers) and a pharmaceutically acceptable carrier; as appropriate, combine with the MVA vaccine to give about 125 μg to 350 μg each time, preferably about The total dose of 250 μg of the glycoproteins of the two isolated HIV gp140 mantle polypeptides is further administered to the human individual comprising two isolated HIV comprising the amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 10, respectively gp140 mantle polypeptide gp140 vaccine, aluminum adjuvant and pharmaceutically acceptable carrier, wherein 22 to 26 weeks after the Ad26 vaccine is administered in step (i), preferably 24 weeks, the MVA vaccine and the video are administered The gp140 vaccine selected under the circumstances; and (iv) administering the human individual orally to the human individual at a total dose of about 3 mg to about 15 mg of visamod or its pharmaceutically acceptable Salt, wherein 26 to 34 weeks after the Ad26 vaccine is administered in step (i), vexamod or a pharmaceutically acceptable salt thereof is administered once every two weeks; preferably, in step (i) 26, 28, 30, 32, and 34 weeks after administration of the Ad26 vaccine, the total dose is 6 mg of Vixamod per administration, or 26 and 28 weeks after administration of the Ad26 vaccine in step (i), the total dose The dose is 6 mg visamod per administration, and 30, 32, and 34 weeks after the Ad26 vaccine is administered in step (i), the total dose is 8 mg visamod per administration. The method of claim 15, which further comprises: administering about 1×10 ⁷ to about 5×10 ⁸ infectious units (IU), preferably about 2×10 ⁸ IU of the one or more MVAs each time The total dose of the vector is intramuscularly administered to the human individual with the MVA vaccine; optionally combined with the MVA vaccine, each administration of about 125 μg to 350 μg, preferably about 250 μg, two isolated HIV gp140 mantles The total dose of the glycoprotein of the polypeptide is further administered to the human individual with the gp140 vaccine, aluminum adjuvant and pharmaceutically acceptable carrier, wherein 34 to 38 weeks after the Ad26 vaccine is administered in step (i), It is preferable to administer the MVA vaccine and optionally the gp140 vaccine in 36 weeks. Such as the method of claim 15 or 16, which further includes: In a total dose of about 3 mg to about 15 mg per administration, the human individual is orally re-administered to the human individual with Vixamod or a pharmaceutically acceptable salt thereof, wherein the Ad26 vaccine is administered in step (i) After 38 to 46 weeks, administer Vixamod or its pharmaceutically acceptable salt once every two weeks; preferably, 38, 40, 42, 44 after administering the Ad26 vaccine in step (i) And for 46 weeks, the total dose is 6 or 8 mg of Vixamod per administration. The method of any one of claims 1 to 17, wherein the human individual initiates the ART during the acute phase of HIV infection. The method of any one of claims 1 to 18, wherein the human individual undergoes inhibitory ART during the treatment. The method of claim 19, wherein the inhibitory ART is stopped after the treatment. A method for treating human immunodeficiency virus (HIV) infection in human individuals in need, which comprises: (i) Treat the human individual with antiretroviral therapy (ART); and (ii) Use the method as in any one of Claims 1 to 20 to induce an immune response against the HIV in the human individual. The method of claim 21, further comprising: suspending the ART treatment of step (i) after step (ii). The method of claim 22, wherein the human individual maintains viral suppression for at least 24 weeks after discontinuing the ART.