Methods and compositions for clinical evaluation of therapeutic agents

Abstract

To provide new technologies and approaches to enable multidimensional analysis of immune responses that will strengthen precision medicine approaches in clinical studies.SOLUTION: A method of identifying an antigen that elicits an immune perturbation in a healthy subject reflecting an immune response signature of a disease comprises: (1) administering to the healthy subject an effective amount of a composition comprising an antigen; (2) collecting a biological sample from the healthy subject before the administering step (1) and at multiple time points thereafter; (3) measuring longitudinal changes from the baseline (i.e., before the administering of step (1)) in immune cell populations and / or inflammatory mediators in the biological sample; (4) comparing the measured longitudinal changes with the immune response signature of the disease to thereby detect an effect of an immune perturbation induced by the antigen in the healthy subject; and (5) identifying the antigen that elicits the immune perturbation in the healthy subject reflecting the immune response signature of the disease.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to the field of systems approaches to immune responses and therapeutic efficacy. Specifically, the present invention provides a method for the treatment of a disease that is characterized by the immune response signature of the disease. This paper concerns methods for the clinical evaluation of therapeutic drugs in elephants. [Background technology]

[0002] Currently, most first-in-human trials are conducted to evaluate the safety and pharmacokinetics of new drugs. The aim is to evaluate the efficacy and safety of drugs, with less emphasis on pharmacodynamic drug effects. In healthy human volunteers, the lack of acute or chronic (e.g., autoimmune) immune stimulation As a result, many disease-specific molecules are not expressed, and therefore targeting these molecules is The effects of drugs like these cannot be easily measured.

[0003] Vaccines have been included in early clinical studies of therapeutic agents. For example, vedolizumab (α4β In one clinical trial of a 7-integrin monoclonal antibody, healthy participants He received intravenous rituximab, followed by intramuscular HBV vaccine and cholera vaccine. The subjects were given oral administration. (Wyant et al.,Gut.2015 Jan;64(1 ):77-83). Vedolizumab alters the response to intramuscularly administered HBV antigen. Although it did not have any clinical benefit, it was shown to reduce the response to orally administered cholera antigens. A gut-selective mechanism of action was demonstrated. However, this study was not performed in patients with HBV vaccine or cholesterol. The vaccine is being developed to treat patients with moderate to severe active ulcerative colitis, the disease targeted by vedolizumab. It does not indicate whether the results induced immune perturbation, including immune response signatures reflecting inflammation. Although antigenic challenges exist that can induce activation of immunological targets, These biological responses in the context of floor testing are poorly understood.

[0004] Current methods for evaluating disease pathology and drug mechanisms at the cellular level have limitations. For example, existing methods for assessing immune status measure immunoglobulin levels, regardless of functional status or specificity. IgG, IgM, IgA) and general immune cell populations (e.g., CD4 Previous studies have relied on qualitative or quantitative measurements of IgG4, CD8, neutrophils, and monocytes. Biomarkers measuring immunological changes in the setting of inflammatory responses include soluble inflammatory factors and upregulation of gene expression. The phenotypic and functional properties of immune cell populations have been limited. A comprehensive analysis incorporating the characterization of functional immune response parameters has been performed to characterize immune-mediated pathogenic processes. This could be part of an essential strategy towards understanding the mechanisms of immune responses and measuring the efficacy of different immune-modulating therapeutic modalities. However, measurements of immune cell profiles are currently being standardized for use in clinical studies. It hasn't been done yet. Summary of the Invention [Problem to be solved by the invention]

[0005] Enabling multidimensional analysis of immune responses to enhance precision medicine approaches in clinical research New technologies and approaches are needed to [Means for solving the problem]

[0006] According to an embodiment of the present application, immune perturbation, including immune response signatures of disease, is administered to healthy subjects. The therapeutic agents of interest are then clinically evaluated in healthy subjects with immune perturbations.

[0007] In one general aspect, the present application provides a method for treating immune disorders that are associated with immune perturbations that reflect immune response signatures of disease. The present invention relates to a method for identifying an antigen that induces a cellular response in a subject, the method comprising: (1) administering to a healthy subject an effective amount of a composition comprising an antigen; (2) administering to a healthy subject a biological sample prior to and at multiple time points thereafter; Collecting fees and (3) baseline (i.e., quantification of immune cell populations and / or inflammatory mediators in biological samples) That is, measuring long-term changes from before administering step (1); (4) By comparing the measured changes over time with immune response signatures of disease, Detecting the effects of immune perturbation induced in healthy subjects by (5) Identify antigens that induce immune perturbations in healthy subjects that reflect the immune response signature of disease. This includes determining the

[0008] In another general aspect, the present application relates to a method for predicting an effect of a therapeutic agent, comprising: The method is: (1) They are known to induce immune perturbations in healthy subjects that mirror the immune response signatures of disease. administering to a healthy subject an effective amount of a composition comprising the antigen of interest; (2) administering a pharmaceutical composition comprising a Therapeutic Agent to a healthy subject having immune perturbation; and (3) measuring the effect of therapeutic agents on immune perturbation in healthy subjects; and (4) Treatment of a disease in a subject in need of treatment based on the impact measured in (3). and predicting the efficacy of the drug.

[0009] In one embodiment of the present application, the method comprises: (5) administering a pharmaceutical composition comprising the therapeutic agent to a subject in need of treatment for a disease; (6) evaluating the efficacy of a therapeutic agent in a subject in need of treatment for a disease. nothing.

[0010] In a general aspect, the present application relates to another method for predicting efficacy of a therapeutic agent. The method is as follows: (1) They are known to induce immune perturbations in healthy subjects that mirror the immune response signatures of disease. administering to a subject in need of treatment for a disease an effective amount of a composition comprising the antigen having been identified; , (2) administering to a subject in need of treatment before and at multiple time points thereafter in step (1); collecting a biological specimen from the subject; (3) baseline (i.e., quantification of immune cell populations and / or inflammatory mediators in biological samples) That is, measuring long-term changes from before administering step (1); (4) detecting the effects of antigen-induced immune disruption in a subject in need of treatment; and comparing the measured changes over time with immune response signatures of disease to determine the pathogenesis of the disease. (5) A method for treating a subject in need of treatment based on antigen-induced immune perturbation. and predicting the efficacy of a therapeutic agent.

[0011] In one embodiment of the present application, the method comprises: (6) administering a pharmaceutical composition comprising the therapeutic agent to a subject in need of treatment for a disease; (7) evaluating the efficacy of a therapeutic agent in a subject in need of treatment for a disease. nothing.

[0012] In another embodiment of the present application, the method comprises: (8) A therapeutic agent for antigen-induced immune disruption in a subject in need of such treatment. The method further includes assessing the efficacy of the therapeutic agent in the subject by measuring the effect of the therapeutic agent on the subject.

[0013] In one embodiment, the method of the present application provides a first infusion of a therapeutic agent for the treatment of a disease. Used in clinical evaluations.

[0014] In one embodiment, the present invention relates to a method for the treatment of a disease comprising administering to a subject, preferably a human subject, in need of treatment for a disease. The effect can include, but is not limited to, the biochemical, physiological, and / or therapeutic effect of a therapeutic agent on a subject. The effect includes a pharmacodynamic effect, including a molecular effect. Preferably, the effect is a therapeutic effect in a subject. Clinical pharmacodynamic effects selected from at least one of the clinical efficacy and safety of the therapeutic agent The effect can also include the pharmacodynamics of the therapeutic agent in a subject.

[0015] Any suitable antigen can be used in the present invention to induce the desired immune perturbation. Preferably, the antigen is a vaccine already approved for use in humans. In some embodiments, the antigen may be a smallpox vaccine (e.g., Imvamunn® / Immun). vanex®), shingles vaccine (e.g., Shingrix® ), Zostar vaccine (e.g., Zostavax®), tetanus vaccine, L PS endotoxin, and antigen skin challenge (CANDIN®, Candida albicans albicans, etc.

[0016] Further aspects, features, and advantages of the present invention are set forth in the detailed description of the invention, as well as in the preferred embodiments thereof. These and other aspects of the present invention will become apparent from the following disclosure, including the embodiments and appended claims.

[0017] The above Summary of the Invention and the following Detailed Description of the Invention should be read in conjunction with the accompanying drawings. The invention will be better understood by reading the embodiments shown in the drawings. It should be understood that the present invention is not limited to the above.

[0018] The drawings are as follows: [Brief description of the drawings]

[0019] [Figure 1] We show that vaccines can highlight immune targets / pathways of disease development targeted for drug modulation. [Diagram 2] 1 shows a study design for a Phase 0 clinical trial to provide a multilevel analysis of immune responses in healthy human subjects administered a known vaccine. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] Various publications, articles and patents are cited or referenced in the Background and throughout this specification. Each of these references is incorporated herein by reference in its entirety. The discussion of documents, operations, materials, devices, articles and the like which has been included in the specification is provided for the purpose of providing a context for the invention. Such discussion is provided without prejudice to the inventive concept or practice of the present invention, even if any or all of these matters are disclosed or claimed. No admission is made that any of the above references constitutes prior art against any invention contained herein.

[0021] Unless otherwise defined, all technical and scientific terms used herein refer to the It has the same meaning as commonly understood by one of ordinary skill in the art to which it pertains. Certain terms used herein have the meanings set forth herein. All patents, published patent applications and publications cited herein are hereby incorporated by reference. The entire contents of which are incorporated herein by reference as if set forth herein. As used in the claims, the singular forms "a," "an," and "the" are used interchangeably with the Please note that plural referents are included unless otherwise clear from the context.

[0022] Unless otherwise indicated, all numerical values, such as concentrations or concentration ranges, described herein are total values. In all cases, the above statements should be understood as being modified by the word "about." Thus, numerical values ​​typically include ±10% of the stated value. For example, 100 A dose of μg includes 90 μg to 110 μg. The use includes integers and fractions of values ​​within the range, unless the context clearly indicates otherwise. Including all the possible subranges, and explicitly including all individual numerical values ​​within that range.

[0023] Throughout this specification and the claims that follow, unless the context otherwise requires, the term "Comprise", "comprises", "comprising", etc. Variations include the specified integer or step or group of integers or steps, but may include any and that it does not exclude any other integer or step or group of integers or steps. As used herein, the term "comprises" is understood to mean the term "containing "including" or "including" may be substituted or are sometimes used herein. When used in this way, it can be replaced with the term "having."

[0024] As used herein, "consisting of" refers to any element of a claim that is specified in the claim. As used herein, "excludes any element, step, or ingredient not included in the present invention." "Qualitative" means material that does not materially affect the basic and novel characteristics of the claim. As used herein in the context of an aspect or embodiment of the invention, Whenever "comprising," "containing," or "including" is used to vary the scope of the disclosure, Any of the above terms, "including," "having," and "including" may be used interchangeably with the term "or "comprising" or "consisting essentially of."

[0025] As used herein, the conjunctive term "and / or" between multiple listed elements means Both individual and combined options are understood to be encompassing. For example, When elements are connected by "and / or," the first option may be used without the second option. The first element is applicable. The second option is that the second element can be applied without the first element. The third option refers to the first and second elements being applicable together. Any one of these options is within the meaning and therefore the As used in the specification, it is understood that the requirement of the term "and / or" is met. The simultaneous applicability of two or more of the following is also included within the meaning, and thus the term "and / or" It is understood that the above requirements are met.

[0026] As used herein, a "subject" refers to an immunogenic component according to an embodiment of the present invention. Any animal, preferably a mammal, to which the composition and / or the composition may be administered or to which the composition may be administered. As used herein, the term "mammal" refers to a mammal, most preferably a human. Any mammal may be included. Examples of mammals include, but are not limited to, However, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, and non-human primates (NHPs), such as monkeys or apes, and humans, more preferably humans. can be.

[0027] As used herein, an "effective amount" refers to an amount sufficient to induce a desired immune effect or immune response. An effective amount refers to an amount of antigen sufficient to induce a desired effect in a subject. An effective amount will depend on a variety of factors. For example, immune perturbations that reflect the immune response signature of disease in healthy subjects. With respect to an "effective amount" of a composition containing an antigen used to induce The particular composition containing the antigen used, the immunological characteristics of the subject, such as the physical condition, age, weight, and health of the subject, Factors such as specific diseases whose response signatures are reflected by immune perturbations in healthy subjects The effective amount can be determined by one of skill in the art in light of the present disclosure.

[0028] In healthy subjects, the body maintains the stability and stability necessary to maintain a normal immune balance. Vaccination maintains the homeostasis of the immune system, allowing the maintenance and regulation of sexual function. Induces immune disruption, such as a slower and less robust primary immune response. A subset of B lymphocytes detects the antigen in the vaccine and secretes specific antibodies against that antigen. Thus, the vaccine is administered to the infected cells, which then multiply to produce more clones that can be used for vaccination. Species increase the levels of circulating antibodies against the antigen. This primary response is amplified to maximum strength. Over time, the primary vaccine-induced response gradually fades away. During infection with a pathogen, the secondary response is lost, but memory cells remain. A strong and more rapid response is induced, which is stronger and more rapid than the primary response. The response dissipates over time. do.

[0029] As shown in Figure 1, vaccines target disease-initiating immune targets / targets for drug modulation. For example, systemic lupus erythematosus (SLE) is a disease that affects the CD4 + T cells, B Cells, antibody (Ab) response, plasmacytoid dendritic cells (pDC), NK cells, CD8 + T cells, Dysregulation of various immune targets and / or pathways, including type I interferon (T1-IFN) and / or known to be associated with abnormal activity or function. These are known to induce immune perturbation of various relevant immune targets and / or pathways, Thus, immune perturbation that reflects one or more immune response signatures of SLE can be induced in healthy subjects. For example, the use of varicella zoster vaccine can be used to inhibit the expression of T1-IFN. Induce immune perturbations in healthy subjects that reflect immune response signatures in SLE, including levels or activity. However, the use of a complete vaccinia vaccine can induce the induction of pDCs, NK cells, and CD8 + Immune perturbation reflects the immune response signature of SLE, including one or more characteristics of T cells In addition, the use of tetanus vaccines can induce CD4 + The immune response signature of SLE, which includes one or more characteristics of T cell, B cell, and Ab responses, Immune perturbation reflecting the vasculitis can be induced in healthy subjects. Clinical evaluation of a drug may involve testing it against one or more of the immune perturbations induced by the vaccine in healthy subjects. The effects of therapeutic agents on one or more of the vaccine-induced immune perturbations may be investigated. The results of clinical studies of therapeutic agents in healthy subjects may be used to support the treatment of SLE in subjects in need of treatment. This will enable them to lead clinical research into potential therapeutic drugs.

[0030] As used herein, "immune perturbation" refers to an immune response induced by administration of a composition containing an antigen. The response of the immune system of a subject to a disease or the immune system of a subject induced by or associated with a disease. For example, immune perturbation includes the primary response induced by the administration of an antigen. Immune perturbation can also be used to treat inflammation-induced immune system disorders, such as chronic inflammation associated with disease. Immune perturbation can also include alterations in one or more immune responses that are dysregulated in a subject with a disease. It may further include alterations in the pathway.

[0031] According to an embodiment of the present application, immune stimulation in healthy subjects induced by administration of an antigen is The perturbations may reflect the immune response signature of the disease.

[0032] As used herein, an "immune response signature" refers to an immune response signature obtained after administration of an antigen to a subject or after administration of an antigen to a patient. Innate immunity and during the effector and / or memory phases of disease-associated immune responses in and / or the cellular and / or molecular signatures of adaptive immunity. may be induced by a single antigen or associated with a single disease.

[0033] As used herein, "immune response signature of a disease" refers to a signature of an immune response of a patient or The present invention provides a method for identifying disease-associated immune response signatures observed in subjects in need of treatment for a disease. For example, the immune response signature of a disease can be determined by (1) determining the acute immune response associated with the disease in a patient; (2) immune responses that arise during infection or chronic inflammatory conditions, or The disease is defined by one or more of the following: local and / or systemic soluble and cellular immune responses This can be done.

[0034] As used herein, "immune response signatures in healthy subjects that reflect disease" "Immune perturbation" refers to a healthy immune response signature that corresponds to a disease immune response signature. Immune perturbation in a healthy subject refers to immune disruption in a subject by the use of known vaccines or other immune-modulating agents. Immune perturbation that reflects the immune response signature of a disease can be induced by administration of a specific antigen. The total duration of immune perturbation ( For example, such an induced immune response may be near or substantially below baseline levels. According to an embodiment of the present application, the antigen may Immune perturbation induced in healthy subjects may reflect immune response signatures of one or more diseases In other embodiments of the present application, a disease may have one or more immune response signatures. Using the above antigens or vaccines, one or more immune response signatures of one or more diseases are identified. In one embodiment, immune perturbation reflecting the effects of IL-19 can be induced in healthy subjects.

[0035] Antigens that induce immune perturbations in healthy subjects that reflect the immune response signature of a disease are disclosed herein. The method can be used to identify the (1) administering to a healthy subject an effective amount of a composition comprising an antigen; (2) administering to a healthy subject a biological sample prior to and at multiple time points thereafter; Collecting fees and (3) baseline (i.e., quantification of immune cell populations and / or inflammatory mediators in biological samples) That is, measuring long-term changes from before administering step (1); (4) By comparing the measured changes over time with immune response signatures of disease, Detecting the effects of immune perturbation induced in healthy subjects by (5) Identify antigens that induce immune perturbations in healthy subjects that reflect the immune response signature of disease. This includes determining the

[0036] Upon administration, the therapeutic agent may result in alteration of immune perturbation induced by an antigen or disease. For example, a disease-causing immune response induced in a healthy subject by an antigen, such as a known vaccine. The immune perturbation that reflects the signature is altered upon administration of a therapeutic agent to treat the disease. The efficacy of a therapeutic agent may be based on its effect on immune perturbation induced in healthy subjects by an antigen. For example, a therapeutic agent may be used to predict or evaluate the duration of an immune-perturbing response. a reduction in the amplitude of the immune-perturbed response, and / or resolution of the immune-perturbed response or otherwise enhancing the immune-perturbed response. If the alteration results in a decrease in the expression of the agonist, the alteration may be useful in treating the disease in a patient in need of such treatment. Alternatively, the therapeutic agent may be administered to healthy subjects. have no effect or adversely affect the resolution or response of induced immune perturbation in In such cases, it may be suggested that the therapeutic agent is ineffective against the patient's disease.

[0037] Thus, in another general aspect, the present application provides therapeutic approaches based on studies in healthy subjects. The present invention relates to a method for predicting the efficacy of a therapeutic drug, the method comprising: (1) They are known to induce immune perturbations in healthy subjects that mirror the immune response signatures of disease. administering to a healthy subject an effective amount of a composition comprising the antigen of interest; (2) administering a pharmaceutical composition comprising a Therapeutic Agent to a healthy subject having immune perturbation; and (3) measuring the effect of therapeutic agents on immune perturbation in healthy subjects; and (4) Treatment of a disease in a subject in need of treatment based on the impact measured in (3). and predicting the efficacy of the drug.

[0038] According to one embodiment of the present application, the method comprises administering different amounts of a pharmaceutical composition comprising a therapeutic agent to an immunosuppressant. administering the therapeutic agent to a healthy subject having the disorder, thereby determining the dose-response relationship of the therapeutic agent. This includes obtaining.

[0039] A therapeutic drug that is predicted to be effective against a disease based on tests in healthy subjects is considered to be effective against the disease. The present invention can be further evaluated in subjects in need of treatment for the disease. In an embodiment, the method comprises: (1) administering a pharmaceutical composition comprising a Therapeutic Agent to a subject in need of treatment for a disease; (2) evaluating the effect of the therapeutic agent in the subject.

[0040] According to an embodiment of the present application, the method comprises administering different amounts of a pharmaceutical composition comprising a therapeutic agent to a patient. and administering the therapeutic agent to a subject in need of the therapeutic agent to obtain a dose-response relationship in the subject. Preferably, the different amounts of the pharmaceutical composition are administered using a method according to an embodiment of the present application. The therapeutic agent is selected based on the dose-response relationship of the therapeutic agent in healthy subjects obtained using a 10-mg / kg / day, 10 ...50-mg / kg / day dose-response relationship of the therapeutic agent in healthy subjects obtained using a 10-mg / kg / day dose-response relationship of the therapeutic agent in healthy subjects.

[0041] Therapeutic agents may be used to treat one or more immune-perturbing disorders induced or associated with a disease. The effect of a therapeutic agent can be determined by targeting or modulating an immune pathway involved in the For example, immune pathways may be constitutively downregulated in a patient. When this occurs, the patient may fail to respond to treatment with therapeutic agents that target the upregulated immune pathway. It may be possible to induce immune perturbations in healthy subjects that reflect immune response signatures of disease. The responsiveness of subjects to antigens known to be involved in immune perturbation is used to target immune pathways involved in immune perturbation. The responsiveness of a subject to treatment with a targeted therapeutic agent can be predicted. For example, However, they suffer from immune perturbation that matches the immune response signature observed in healthy subjects upon antigen challenge. When induced in a subject, or when the antigen enhances immune perturbation, including the immune response signature of the disease. When immune perturbation is induced in a patient, the induced immune perturbation may be sufficient to prevent or reduce the effectiveness of the therapeutic agent in treating the patient's disease. Alternatively, the antigen may suggest or predict that the disease is in need of treatment. or fail to affect desired immune perturbation in patients with If it does not, it suggests that the therapeutic agent is not effective against the patient's disease. It is possible.

[0042] Accordingly, another aspect of the present application is to provide a method for the treatment of a disease comprising administering to a subject in need thereof a therapeutic agent. A method for predicting (1) They are known to induce immune perturbations in healthy subjects that mirror the immune response signatures of disease. administering to a subject in need of treatment an effective amount of a composition comprising the antigen to be treated; (2) administering to a subject in need of treatment before and at multiple time points thereafter in step (1); collecting a biological specimen from the subject; (3) baseline (i.e., quantification of immune cell populations and / or inflammatory mediators in biological samples) That is, measuring long-term changes from before administering step (1); (4) detecting the effects of antigen-induced immune disruption in a subject in need of treatment; and comparing the measured changes over time with immune response signatures of disease to determine the pathogenesis of the disease. (5) A method for treating a subject in need of treatment based on antigen-induced immune perturbation. and predicting the efficacy of a therapeutic agent.

[0043] The predicted effect of a therapeutic agent may be tested by administering the therapeutic agent to a subject. Thus, in one embodiment of the present application, the method comprises: (1) administering a pharmaceutical composition comprising a Therapeutic Agent to a subject in need of treatment for a disease; (2) evaluating the efficacy of a therapeutic agent in a subject in need of treatment for a disease. nothing.

[0044] According to one embodiment of the present application, the method comprises administering to a subject in need of treatment an antigen. The effect of the therapeutic agent on induced immune perturbation is measured to determine whether the therapeutic agent is effective in treating a subject. and evaluating the effect of said method on the efficacy of said method.

[0045] Using techniques or methods known in the art, the immune response after vaccination can be evaluated. To identify key molecular mechanisms of innate and adaptive immunity during effector and memory stages. and thereby, the immune response phenotype of a disease in a healthy subject administered an antigen or vaccine. The signal can be obtained by using the munol. 2017; 8: 1563. published online 2 For example, dual-color reverse transcription multiplex ligation-dependent probe amplification ( Long-term multiplex transcriptome profiling assays such as dcRT-MLPA have been Transcriptomic signatures of innate, adaptive, regulatory, inflammatory, and memory responses Biologically probing human immune responses following vaccination against human infectious diseases, with a special emphasis on Optimal response and high density dynamic transcriptomes can be used to file Determination of immune response measures will aid in the optimization of novel immunization strategies. Global transcriptome gene expression profiling (mRNA and We will analyze novel mechanisms using transcriptome assays such as genomics (e.g., genomics ... and microRNAs) to Correlate immunity with safety, with special emphasis on age and time since vaccination. The basis for this is hydrogen-deuterium exchange (HDX) in combination with mass spectrometry (MS). Using quantitative proteomic profiling techniques, It is possible to define functionally active epitopes and antibodies in the polyclonal sera of Antigen-specific memory T and B cells and their ability to migrate to the appropriate tissue site In light of the present disclosure, methods known in the art may be used for analysis. Multidisciplinary approaches (antigen stimulation, TCR deep sequencing, and Th1, Th2, and and cloning of Th17 memory subsets) to determine the T cell subset responses. The assay can be used to analyze T cell proliferation in healthy and diseased mucosal sites. Raphishing, chemokines (CCL20, CXCL10, and CXCL12, etc.) We aim to clarify the molecular mechanisms regulating T cell migration in response to IL-1, and their role in T cell migration, particularly in individuals with chronic immune activation. We also evaluate the ability of Th cells to reach the mucosal niche and support cell maturation and functional activity in It is possible.

[0046] In one embodiment of the present application, induced immune perturbation in a healthy subject is These include alterations in one or more immune pathways known to be dysregulated in the immune system.

[0047] In another embodiment of the present application, induced immune perturbation in a healthy subject is Alterations in one or more immune cell populations known to be associated with disease in Immune cell profiles can be determined by any method known in the art in light of the present disclosure. can be used to test and monitor.

[0048] In yet another embodiment of the present application, the induced immune perturbation in a healthy subject is one or more inflammatory mediators known to be associated with disease in a subject. The levels and / or activity of inflammatory mediators may be determined in light of the present disclosure. This can be studied and monitored using methods known in the art.

[0049] In a further embodiment of the present application, induced immune perturbation in healthy subjects is Expression levels of one or more biomarkers known to be associated with disease in elephants Methods known in the art can be used to detect changes in the markers of interest and / or their activity. It can be used to test and monitor biomarkers.

[0050] Any suitable antigen can be used to generate immune responses in healthy subjects that reflect the immune response signature of the disease. Preferably, the vaccine or antigen challenge is capable of inducing perturbation in a mammalian cell. 1) approved uses and a safety record; 2) the ability to activate multiple immune pathways at once; the ability to stimulate specific immune cells and targeted pathways; and / or ) their ability to activate responses at peripheral or mucosal sites.

[0051] In one embodiment, a method is provided for inducing immune perturbation in a healthy subject that reflects an immune response signature of a disease. To combat these diseases, vaccines against human infectious diseases are used. Examples of such vaccines include , vaccinia (smallpox) vaccines (e.g., Imvamunn® / Imvan ex ), shingles vaccines (e.g., Shing rix®), shingles vaccine (e.g., Zostavax®), and tetanus vaccines, but are not limited to.

[0052] In another embodiment, immune perturbation is induced in healthy subjects that reflects an immune response signature of a disease. To prevent this, endotoxin chaperones such as lipopolysaccharide (LPS) endotoxin are used. Endotoxin challenge can be used during acute infection or in chronic inflammatory states. It has been safely used for decades to elicit immune responses similar to those that occur during (Dillingh et al, J of Inflammation 201 4, Kiers et al, Sci Reports 2017).

[0053] In another embodiment, immune perturbation is induced in healthy subjects that reflects an immune response signature of a disease. To achieve this, we use skin antigens such as CANDIN® (Candida albicans) A range of antigenic skin challenges are used to stimulate local and systemic soluble and cellular immunity. These types of immune system challenges have been used to assess tissue-initiated antigenic stimulation of responses. These ranges may provide insight into the expected range of normal immune responses in healthy individuals. and therefore in autoimmune diseases or with drugs that target specific immune pathways. Thus, it can serve as a standard for comparing the functional antigen-specific immunity that can be altered (L ynn et al, J of Pharm and Exp Ther, 2004 ).

[0054] In one embodiment of the present application, the biological sample collected from a healthy subject is a peripheral blood sample. Preferably, the biological sample comprises peripheral blood mononuclear cells (PBMCs). Any known method may be used to collect a biological sample from a healthy subject. It can be done.

[0055] Biological samples were collected from healthy subjects prior to and at various time points after administration of the antigen or vaccine. Preferably, the biological sample is collected from a healthy subject prior to administration of an antigen or vaccine. Collected on day 0, and on days 1, 3, 7, and 14 after administration of antigen or vaccine. , 21st, 28th, and up to 90 days later.

[0056] Long-term perturbations in cellular immune populations may result in changes in activation state and other functional attributes. These include phenotypic changes measured by cell surface antigens associated with sex. Immune cells include, but are not limited to, cells of the innate immune system as well as cells of the adaptive immune system. Examples of immune cells include natural killer (NK) cells, eosinophils, and dendritic cells (D cells). C) include, but are not limited to, neutrophils, macrophages, B cells, and T cells. stomach.

[0057] Changes in inflammatory mediators include any biochemical mediators released during inflammation. These include, but are not limited to, changes in the amount and / or activity of inflammatory mediators. Examples include cytokines, chemokines, vasoactive amines (histamine and serotonin) etc.), kinins (bradykinin etc.), eicosanoids (thrombocytopenic acid, leukotrienes etc.) These include complement and complement-derived peptides, Not limited to these.

[0058] According to an embodiment of the present application, the vaccine or antigen induces an immune response that reflects the pathophysiology of the disease. The antibody is used to induce an immune response containing a target immune signature in a healthy subject, thereby preventing a disease of interest. The effect of a therapeutic agent, preferably an immunotherapy, for treating immune perturbations in healthy subjects The effect of a therapeutic agent on the response to a given drug is evaluated clinically by measuring the effect of the therapeutic agent on the response to a given drug in healthy subjects. Including a vaccine in early clinical trials to treat patients with the disease would be a step toward ensuring treatment in those in need. Provides information on the response to drugs (e.g., immunotherapy), provides information on the mechanism of action of therapeutic drugs, and During early development stages before assessing immune response thresholds or conducting larger clinical trials Define individual baseline and immune response signatures to support decision-making It helps.

[0059] Therefore, we focus on the safety and pharmacokinetics of new drugs without placing much emphasis on pharmacodynamic drug effects. Unlike most first-in-human trials, which are intended to evaluate The method according to the embodiment includes: determining the pharmacodynamics of a therapeutic agent by first-in-human testing of the therapeutic agent; For example, the therapeutic effect of the present invention can be evaluated in patients or humans in need of treatment for a disease. Before further testing in mice, we will first identify immune response signatures that reflect the pathophysiology of the disease. To study the dose-response relationship of therapeutic drugs in healthy human subjects with induced immune perturbations It can be tested in in-human trials.

[0060] Vaccination or antigen challenge is used to establish an individual's immune response threshold or immunity score. (Tsang JS Trends in Immunology 36: , 2015, Furman D and Davis MM, Vacci ne 33:5271, 2015, Boyd, SD and Jackson, KJL Cell Host and Microbe, 17:301, 2015, Kaczorowski, KJ et al PNAS, 2017). Considering this, vaccination or antigen challenge can be used to induce immune responses to disease in healthy subjects. This induced immune perturbation can be induced by immunomodulators. In addition, it has potential predictive value for assessing responsiveness to a vaccine or antigen challenge. Practical use of ranges to assess biomarker responses in clinical trials of novel immunomodulators The effect of immunomodulators on their targets can be evaluated and proof of mechanism (POM) studies can be performed. ) has potential utility in collecting evidence.

[0061] Immune perturbation can be measured and / or assessed using methods known in the art in light of the present disclosure. or may be monitored. The ability of the Available techniques for assessing the development and activation of immune responses include: For a general description of Current Protocols in Immunology; ed. J Wiley & Sons Inc, National Institute of Cellular immunity was measured using CD4+ and CD8+ T cell derived immunoglobulins. Measurement of cytokine profiles secreted by activated effector cells, including (e.g., quantification of IL-10 or interferon gamma producing cells by ELISPOT) By determining the activation state of immune effector cells (e.g., classical 3 H] Chimiji T cell proliferation assays using cytometry-based assays or cytometry-based assays Thus, assaying for antigen-specific T lymphocytes in sensitized subjects (e.g., cell This may be performed by peptide-specific lysis in a toxicity assay, etc.

[0062] The ability of an antigen or vaccine to stimulate a cellular and / or humoral response is determined by antibody binding and / or This can be determined by competition in binding (e.g., Harlow, 1989, Ant ibodies, Cold Spring Harbor Press For example, the titer of antibodies produced in response to administration of a composition providing an immunogen may be increased by enzyme binding. The immune response can also be measured by antibody assay. The neutralization of the virus can be measured by neutralizing the virus with specific antibodies. Immune response is defined as the loss of infectivity of a virus by reaction / inhibition / neutralization. It can further be measured by an assay for advanced cellular phagocytosis (ADCP).

[0063] Any computational method can be used to monitor changes in the measured immune response over time. For example, a measure of an individual's in vivo immune response threshold may be analyzed, thereby identifying a particular immune perturbation. To provide a qualitative metric by which the likelihood of response and recovery to Lo may be assessed. The calculation method called oper is used (Rath et al., Predicting position along a looping immune response e trajectory, PLoS One, October 2018 According to an embodiment of the present application, Lo Using oper to analyze immune perturbation induced in healthy subjects by vaccination .

[0064] Individual biomarkers, including static biomarkers (e.g., in situ PD1 expression, IFN signature) immune threshold responses to systemic immune challenges, as well as functional responses to systemic immune challenges (e.g., anti-influenza Several parameters are important in determining the outcome of influenza infection or influenza vaccine response. In the field of systems immunology, there are many methods to measure the range of responses. The use of specific perturbations can be used to compare static biomarker readouts to define a descriptive “immunoscore.” Thus, an individual is able to measure their baseline physiological state or responsiveness. Depending on the outcome, the patient may have a high or low score. This immune score is then used to determine the treatment stratification. and monitoring of treatment response, or later consideration of clinical trial enrollment for subjects requiring treatment of the disease. (Farsaci, B et al., Cancer Immunol Res, 4:755-765, 2016). According to an embodiment of the present application, By analyzing immune cell subsets within peripheral blood mononuclear cells in healthy subjects, It is possible to include a "peripheral immune score" similar to that of Saci, B et al. in the immune response signature of a disease. You can determine whether it is possible.

[0065] According to embodiments of the present application, the present invention provides a method for evaluating cell populations and functional antigen-specific immune responses. PBMCs are then isolated from the subject before and after administration of the antigen. For each group (e.g., healthy or diseased), the immune challenge (e.g., vaccination or non-vaccine status) by type of vaccine administered) and by therapeutic agent being evaluated (e.g., targeted immunomodulatory agent and checkpoint inhibitors). PBMC samples were then split into or other questions specific to the immune target of therapeutics being evaluated in clinical trials. Healthy subjects may be included in future studies to be interrogated. (smallpox vaccine), Shingrix (herpes zoster vaccine) cutin), endotoxin (Escherichia coli LPS), Candida albicans, tetanus toxoid and keyhole limpet hemocyanin (KLH), A panel of biomarkers is measured in PBMCs. Markers include, but are not limited to: For example, multiparameter assays are used in addition to other types of functional assays that evaluate antibody and cytokine production. Immune cell-specific expression can be assessed by parametric flow cytometry or CyTof A panel of phenotypic and functional markers was included. Single-cell RNAseq from isolated PBMCs Transcriptional data may also be obtained, including using one or more immune response signals for a disease of interest. Necha measures markers and compares them to markers known to be associated with disease. The efficacy of a therapeutic agent against a disease can be identified based on the immune response signature of the disease. The evaluation is based on measuring the effect of the drug on the patient's condition.

[0066] The methods of the present application include any method for treating a target disease in a subject in need of such treatment. The present invention may be used in the clinical evaluation of suitable therapeutic agents for the treatment of cancer. Preferably, the therapeutic agent is an immunomodulatory agent. In some embodiments of the application, the therapeutic agent is a medicament for treating rheumatoid arthritis, juvenile idiopathic arthritis, ankylosing arthritis, For the treatment of spondylitis, psoriatic arthritis, Crohn's disease, ulcerative colitis, or plaque psoriasis and targeted immunomodulators such as those described herein.

[0067] Also provided herein is a kit for the clinical evaluation of a therapeutic agent. Antigens for use in inducing immune perturbation in healthy subjects that includes an immune response signature of disease - Patents.com and a pharmaceutical composition comprising a therapeutic agent. The kit also includes one or more additional The kit may include an antigen or vaccine, an adjuvant, or an immunostimulant of the present application. The present invention relates to the use of a composition comprising an antigen and a pharmaceutical composition in a clinical trial by a method Preferably, the composition comprising the antigen is a vaccine against a human infectious disease, Preferably, non-replicating smallpox vaccine, herpes zoster vaccine, non-live varicella zoster vaccine or a tetanus vaccine, an antigen skin challenge, preferably a Candida albicans antigen, and endotoxin, preferably lipopolysaccharide (LPS). Preferably, the pharmaceutical composition comprises an immunomodulatory agent, preferably a targeted immunomodulator, as an active ingredient. Includes modulators or checkpoint inhibitors.

[0068] The following examples of the present application are intended to further illustrate the principles of the present application. The examples do not limit the present application, and the scope of the present application is defined by the appended claims. It should be understood that the above is determined. EXAMPLES

[0069] Multilevel analysis of vaccine-induced human immune responses Potential physiological perturbations that could be used in future Phase 1 or Phase 2 human volunteer trials To determine the usefulness of a vaccine or other antigen / metabolic challenge An evaluation is performed to determine the time course and extent of specific responses, which may The study will identify one or more immune response signatures against the disease. It allows for an assessment of the types of analytical tools that might actually be considered for use in a study.

[0070] Materials and Methods This includes, where applicable, two vaccines and a vaccine directed at the systemic immune response and local tissues. A Phase 0 study in healthy adult volunteers to evaluate the efficacy of 10-fold increased creatinine and two antigen challenge agents This is a preliminary single-center study. Each subject will have baseline cell populations and inflammatory mediators. The end of the study will be determined at a fixed time for each cohort. This occurs at the defined time of the last sample taken (maximum 90 days) (see Figure 3).

[0071] As shown in Table 1, the commercially available vaccine Imvamune® / Imvan ex® (Bavarian Nordic) and Shingrix® ) (GSK) was used. A commercially available allergen test solution was applied to the skin of Candida albicans antigen. Commercially available LPS endotoxin is used for intravenous endotoxin injection and skin reaction induction. Used for the challenge.

[0072] [Table 1]

[0073] The objectives and endpoints of this study are described in Table 2 below.

[0074] [Table 2]

[0075] result Different challenge agents, including changes in antigen-specific antibodies, cellular responses, and inflammatory mediators Blood samples will be taken from participants for analysis of their immune response to the agent. The analyses performed include immunological, hematological, and other organ system and / or general immune system function. The study is intended to provide insight into the effects of interindividual variability in efficacy and response. The challenge drug-specific antibodies, cellular responses, and soluble inflammatory mediators were examined. Detection and characterization of antigens are performed using validated, specific, and sensitive immunoassay methods. DNA extraction of genes and epigenetic markers related to challenge-specific immune pathways Pharmacogenomic blood samples will be collected to allow for the extraction and analysis of This may include complete genome-wide assays and / or targeted sequencing.

[0076] The examples and embodiments described herein are for illustrative purposes only and are not intended to be limiting of the scope of the invention. Changes may be made to the embodiments of the present invention without departing from the broad inventive concept thereof. It is understood, therefore, that the invention is not limited to the particular embodiments disclosed, but rather to the specific embodiments set forth in the appended claims. and the like, including modifications within the spirit and scope of the invention as defined by the appended claims. It is understood that this is intended.

Claims

1. Identifying antigens that induce immune perturbations in healthy subjects that reflect immune response signatures of disease 1. A method comprising: (1) administering to the healthy subject an effective amount of a composition comprising an antigen; and (2) from the healthy subject at multiple time points before and after the administering step (1). collecting a biological sample; (3) determining whether the immune cell population and / or inflammatory mediators in the biological sample are at baseline; To measure long-term changes in these (4) comparing the measured longitudinal changes to the immune response signature of the disease. and detecting the effect of immune perturbation induced by said antigen in said healthy subject. And, (5) administering to the healthy subject the immune perturbation that reflects the immune response signature of the disease. and identifying said inducing antigen.

2. The immune perturbation is a process for disrupting one or more immune pathways that are dysregulated in a subject with the disease. Alterations in one or more immune cell populations associated with the disease in a subject having the disease. a change in one or more inflammatory mediators associated with the disease in a subject having the disease; and a change in a modulator associated with the disease in a subject having the disease. The present invention relates to a method for treating a pulmonary artery disease, comprising administering to a patient a therapeutically effective amount of a pulmonary artery disease, comprising administering to the patient a therapeutically effective amount of a pulmonary artery disease, and / or administering to the patient a therapeutically effective amount of a pulmonary artery disease, the method comprising ... The method according to

3. A method for predicting an effect of a therapeutic agent, comprising: (1) It is known to induce immune perturbations in healthy subjects that reflect the immune response signature of disease. administering to said healthy subject an effective amount of a composition comprising the antigen identified in said study; (2) administering a pharmaceutical composition comprising the therapeutic agent to the healthy subject having the immune perturbation. And, (3) measuring the effect of the therapeutic agent on the immune perturbation in the healthy subject. and, (4) Based on the effect measured in (3), a subject in need of treatment for the disease is and predicting the effect of the therapeutic agent in said patient.

4. (1) administering the pharmaceutical composition containing the therapeutic agent to the subject in need of the treatment of the disease. and administering (2) evaluating the effect of the therapeutic agent in the subject in need of the treatment of the disease. The method of claim 3 , further comprising:

5. The therapeutic agent reduces the immune perturbation in the healthy subject and reduces the immune perturbation in the healthy subject.

4. The method of claim 3, which is predicted to be effective in treating said disease in said subject. Method of posting.

6. The immune perturbation is a process for disrupting one or more immune pathways that are dysregulated in a subject with the disease. Alterations in one or more immune cell populations associated with the disease in a subject having the disease. a change in one or more inflammatory mediators associated with the disease in a subject having the disease; and a change in a modulator associated with the disease in a subject having the disease. The above-mentioned changes in expression and / or activity of biomarkers are selected from the group consisting of: The method according to

7. A method for predicting an effect of a therapeutic agent, comprising: (1) Inducing immune perturbation in a healthy subject that reflects an immune response signature of the disease. and administering to a subject in need of treatment for a disease an effective amount of a composition comprising an antigen known to be effective against the disease. And, (2) administering to a patient in need of said treatment at multiple time points prior to and after said administering step (1). collecting a biological sample from said subject; (3) determining whether the immune cell population and / or inflammatory mediators in the biological sample are at baseline; To measure long-term changes in these (4) comparing the measured longitudinal changes to the immune response signature of the disease. and thereby inhibiting immune disruption induced by said antigen in said subject in need of said treatment. Detecting the effects; and (5) determining whether the treatment is necessary based on the immune perturbation induced by the antigen; and predicting the efficacy of the therapeutic agent in the subject.

8. (1) administering a pharmaceutical composition containing the therapeutic agent to the subject in need of the treatment of the disease; Giving and (2) evaluating the effect of the therapeutic agent in the subject in need of the treatment of the disease. The method of claim 7 , further comprising:

9. to the immune perturbation induced by the antigen in the subject in need of said treatment and determining the effect of the therapeutic agent in the subject by measuring the effect of the therapeutic agent on the The method of claim 7, comprising: evaluating.

10. The immune perturbation is a process for disrupting one or more immune pathways that are dysregulated in a subject with the disease. Alterations in one or more immune cell populations associated with the disease in a subject having the disease. a change in one or more inflammatory mediators associated with the disease in a subject having the disease; and a change in a modulator associated with the disease in a subject having the disease. The above-mentioned changes in expression and / or activity of biomarkers are selected from the group consisting of: The method according to

11. A first-in-human trial of said therapeutic agent, preferably The results include the pharmacodynamic effects of said therapeutic agent in human subjects, and more preferably, the pharmacodynamic effects The method of claim 3 , wherein the results include a dose-response relationship of the therapeutic agent.

12. The immune response signature of the disease may comprise: (1) an acute infection or (2) the immune response that occurs during a chronic inflammatory condition and (3) the local and / or (3) a systemic soluble and cellular immune response to dietary components associated with the disease and a metabolic response associated with said at least one of the following: Method of posting.

13. The immune response signature of the disease may comprise: (1) an acute infection or (2) the immune response that occurs during a chronic inflammatory condition and (3) the local and / or (3) a systemic soluble and cellular immune response to dietary components associated with the disease and a metabolic response associated with said at least one of the following: Method of posting.

14. The immune response signature of the disease may comprise: (1) an acute infection or (2) the immune response that occurs during a chronic inflammatory condition and (3) the local and / or (3) a systemic soluble and cellular immune response to dietary components associated with the disease and a metabolic response associated with said at least one of the following: Method of posting.

15. The antigen is a vaccine against a human infectious disease and preferably the composition is a non-replicating Smallpox vaccine, herpes zoster vaccine, non-live varicella zoster vaccine, and tetanus vaccine 2. The method of claim 1 , wherein the compound is selected from the group consisting of:

16. The composition comprising the antigen is a vaccine against a human infectious disease, preferably The compositions include a non-replicating smallpox vaccine, a herpes zoster vaccine, a non-live varicella zoster vaccine, and a tetanus vaccine.

17. The composition comprising the antigen is a vaccine against a human infectious disease, preferably The compositions include a non-replicating smallpox vaccine, a herpes zoster vaccine, a non-live varicella zoster vaccine, 8. The method of claim 7, wherein the vaccine is selected from the group consisting of: a tetanus vaccine.

18. The antigen is selected from the group consisting of an antigen skin challenge and an endotoxin. Item 1. The method according to item 1.

19. A kit for clinical evaluation of a therapeutic agent, comprising: a. Use in inducing immune perturbation, including immune response signatures of disease, in healthy subjects and an antigen for b. A pharmaceutical composition comprising a therapeutic agent.

20. The composition comprising the antigen is a vaccine against a human infectious disease, preferably a non-replicating smallpox vaccine. Vaccine, Herpes Zoster vaccine, non-live varicella zoster vaccine, or tetanus vaccine, A primary cutaneous challenge, preferably a Candida albicans antigen, and an endotoxin, preferably or lipopolysaccharide (LPS).

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