Treatment of primary CTLA-4 checkpoint related immunodeficiencies with 1H-indole-3-carboxaldehyde or 1-methylindole-3-carboxylic acid

Abstract

The present disclosure is directed to a method of treating a CTLA-4 checkpoint associated immunodeficiency by administering a therapeutically effective amount of a pharmaceutical composition comprising 1H-indole-3-carboxaldehyde (3-IAld) or 1-methylindole-3-carboxylic acid.

Description

Cross-references to related applications

[0001] This international application claims priority to U.S. Provisional Application No. 63 / 509,223, filed June 20, 2023, which is incorporated herein by reference in its entirety. Technical Field

[0002] This disclosure relates to a method for treating CTLA-4 checkpoint-related immunodeficiency in patients. Background Technology

[0003] Primary CTLA-4 checkpoint-associated immunodeficiency is characterized by a variable combination of enteropathy, hypogammaglobulinemia, recurrent respiratory infections, granulomatous lymphocytic interstitial lung disease, lymphocytic infiltration of non-lymphoid organs (intestine, lung, brain, bone marrow, and kidney), autoimmune thrombocytopenic purpura or neutropenia, autoimmune hemolytic anemia, and lymphadenopathy. Heterozygous CTLA4 mutations in humans are associated with a severe immunomodulatory disorder known as CTLA-4 haploinemia with autoimmune infiltration (CHAI). Another type of primary CTLA-4 checkpoint-associated immunodeficiency has been reported in the literature in patients carrying a biallelic mutation in the lipopolysaccharide-responsive beige-like anchoring protein (LRBA) gene. Because LRBA deficiency also leads to secondary loss of CTLA-4, this recessive disorder is termed LRBA deficiency with autoantibodies, T-reg cell deficiency, autoimmune infiltration, and enteropathy (LATAIE).

[0004] Currently, hematopoietic stem cell transplantation (HSCT) is the only cure for patients with primary CTLA-4 checkpoint-related immunodeficiency. Apart from immunoglobulin replacement therapy, which corrects hypogammaglobulinemia and prevents infection, most other proposed therapies remain largely unsatisfactory.

[0005] Systemic corticosteroids can alleviate gastrointestinal symptoms in some patients, but only temporarily.

[0006] For all patients with CTLA-4-related CNS syndromes, abatacept or sirolimus is recommended to control lymphocyte activation. High-dose steroids remain recommended as first-line treatment, as advised for other demyelinating CNS conditions. Adding high-dose IVIG appears valuable if a rapid clinical response is not observed. As a second-line strategy, rituximab or cyclophosphamide should be considered while continuing first-line therapy, as recommended for autoimmune encephalitis.

[0007] In summary, systemic immunosuppressants and abatacept can provide partial control, but require continuous administration. Despite the significant risk of treatment-related death, allogeneic hematopoietic stem cell transplantation offers a potential cure for patients with CTLA-4 deficiency who are suitable for this approach. Another important issue is the need for multiple changes of immunosuppressants due to adverse reactions or steroid dependence. However, the development of further CTLA-4-related symptoms in untreated or undertreated patients indicates a progressive natural course of the disease, thus necessitating treatment.

[0008] Furthermore, despite the great success of cancer immunotherapy using immune checkpoint inhibitors, its beneficial therapeutic effects are hampered by various resistance mechanisms. Schoenfeld et al. , 2020 Limitations on the effects of gastrointestinal, endocrine, and skin toxicities, as well as fatal neurotoxicity and cardiotoxicity. Choi et al. , 2020 Therefore, novel treatment strategies that provide manageable side effects for existing immunotherapies will enhance and expand their therapeutic efficacy and application. Summary of the Invention

[0009] This disclosure provides, at least in part, a method for treating a patient with CTLA-4 checkpoint-related immunodeficiency, the method comprising administering to a patient in need a therapeutically effective amount of a pharmaceutical composition comprising 1H-indole-3-carboxaldehyde (3-IAld). In some aspects, CTLA-4 checkpoint-related immunodeficiency is selected from the group consisting of: CTLA-4 haploinemia with autoimmune infiltration (CHAI), lipopolysaccharide-responsive beige-like anchoring protein (LRBA) deficiency (LATAIE), regulatory T (Treg) cell deficiency, autoimmune infiltration, enteropathy, intestinal inflammation, immune-mediated colitis, gastrointestinal disorders, and gastric atrophy. In another aspect, CTLA-4 checkpoint-related immunodeficiency is immune-mediated colitis.

[0010] In one aspect, the pharmaceutical composition comprises a pharmaceutically acceptable carrier. In another aspect, the pharmaceutically acceptable carrier is at least one polymer. In another aspect, the pharmaceutically acceptable carrier is a group of polymers. In another aspect, this group of polymers is Eudragit® polymers. In another aspect, the pharmaceutical composition is formulated for enteral delivery. In another aspect, the pharmaceutical composition is administered orally. In another aspect, the pharmaceutical composition is in the form of capsules, tablets, gel tablets, gel capsules, gels, liquids, or gummies. In another aspect, the pharmaceutical composition is in the form of tablets or capsules.

[0011] In one aspect, the pharmaceutical composition is administered at every other day (qod). In another aspect, the pharmaceutical composition is administered at a 3-IAld dose of at least about 3 mg / kg, at least about 4 mg / kg, at least about 5 mg / kg, at least about 6 mg / kg, at least about 7 mg / kg, at least about 8 mg / kg, at least about 9 mg / kg, at least about 10 mg / kg, at least about 11 mg / kg, at least about 12 mg / kg, at least about 13 mg / kg, at least about 14 mg / kg, at least about 15 mg / kg, at least about 16 mg / kg, at least about 17 mg / kg, or at least about 18 mg / kg. In yet another aspect, the 3-IAld dose is about 18 mg / kg.

[0012] This disclosure further provides, at least in part, a method for treating a patient with CTLA-4 checkpoint-related immunodeficiency, the method comprising administering to a patient in need a therapeutically effective amount of a pharmaceutical composition comprising 1-methylindole-3-carboxylic acid. In some aspects, CTLA-4 checkpoint-related immunodeficiency is selected from the group consisting of: CTLA-4 haploid deficiency with autoimmune infiltration (CHAI), lipopolysaccharide-responsive beige-like anchoring protein (LRBA) deficiency (LATAIE), regulatory T (Treg) cell deficiency, autoimmune infiltration, enteropathy, intestinal inflammation, immune-mediated colitis, gastrointestinal disorders, and gastric atrophy. In another aspect, CTLA-4 checkpoint-related immunodeficiency is immune-mediated colitis.

[0013] In one aspect, the pharmaceutical composition comprises a pharmaceutically acceptable carrier. In another aspect, the pharmaceutically acceptable carrier is at least one polymer. In another aspect, the pharmaceutically acceptable carrier is a group of polymers. In another aspect, this group of polymers is Eudragit® polymers. In another aspect, the pharmaceutical composition is formulated for enteral delivery. In another aspect, the pharmaceutical composition is administered orally. In another aspect, the pharmaceutical composition is in the form of capsules, tablets, gel tablets, gel capsules, gels, liquids, or gummies. In another aspect, the pharmaceutical composition is in the form of tablets or capsules. Attached Figure Description

[0014] Figure 1 It is a representation of the chemical structure of 1H-indole-3-carboxaldehyde (3-IAld, molecular formula: C9H7NO, IUPAC name: 1H-indole-3-carboxaldehyde), also known as indole-3-aldehyde or 3-formylindole.

[0015] Figure 2 A to Figure 2H is a graphical representation and photographic image showing the protection provided by 3-IAld against inflammatory pathology in DSS+ anti-CTLA-4 induced colitis. (See experimental schedule diagram.) Figure 2 As described in A), C57BL / 6 mice were treated with DSS in drinking water for one week, followed by a one-week recovery period and administration of 100 µg of anti-CTLA-4 mAb and 3-IAld. The mice were then assessed for (…). Figure 2 B) % Survival, ( Figure 2 C) % weight change, ( Figure 2 D) Disease activity index, ( Figure 2 E) Rectal bleeding, ( Figure 2 F) Colon histology (by PAS staining), ( Figure 2 G) Histological score and ( Figure 2 H) Epithelial barrier function (ZO-1 and Ki-67 staining). Images were taken using a high-resolution microscope at 20x magnification (scale bar, 200 µm). White arrows indicate rectal bleeding. Yellow arrows indicate inflammatory cell recruitment. Each in vivo experiment included four to six mice per group (16–24 mice per experiment). Data are presented as mean ± SEM. H2O, untreated mice. ***P<0.001, ****P<0.0001.

[0016] Figure 3 A to Figure 3 H is a graphical representation showing that 3-IAld provides epithelial barrier integrity and promotes an anti-inflammatory state in rat colitis. Assessment as follows: Figure 2 C57BL / 6 mice that underwent DSS plus anti-CTLA-4 induced colitis and were administered 3-IAld as described Figure 3 A) Epithelial function markers; serum ( Figure 3 B) Glucan-FITC levels and ( Figure 3 C) sCD14 levels; in colon homogenate ( Figure 3 D) Cytokine levels and ( Figure 3 E) Calprotectin levels; and ( Figure 3 F, Figure 3 G, Figure 3 H)AhR-dependent gene expression. Data are expressed as mean ± SEM. H2O, untreated mice. *P<0.05, **P<0.01, ***P<0.001. ns, not significant.

[0017] Figure 4 A to Figure 4F is a graphical representation and photographic image showing the protection of mice from immune-induced colitis by 3-IAld. NSG mice were infused with hPBMCs and treated with anti-CTLA-4 mAb and 3-IAld. Mice were sacrificed at day 21, and the effects were evaluated. Figure 4 A) % Survival, ( Figure 4 B) % weight change, ( Figure 4 C) Colon histology (periodic acid-Schiff staining) and ( Figure 4 D) Histological score, ( Figure 4 E) ZO-1 protein expression and ( Figure 4 F) Expression of inflammatory cytokines. Images were taken using a high-resolution microscope (Olympus BX51) at 20x magnification (scale bar, 200 µm). Each in vivo experiment included four to six mice per group (16–24 mice per experiment). Data are presented as mean ± SEM. *P<0.05, **P<0.01, ****P<0.0001. Two-way ANOVA, Bonferroni post-hoc test. H2O, untreated mice.

[0018] Figure 5 A to Figure 5 D is a graphical representation and photographic image of how 3-IAld restricts the progression of immune-mediated colitis in RAG1-deficient mice. CD4 infusions were treated with αCTLA-4 mAb and 3-IAld. + T cells Rag1 – / – Mice. Mice were sacrificed at 21 days and their condition was assessed. Figure 5 A) % weight change, ( Figure 5 B) Gross histology, ( Figure 5 C) Histological scoring and ( Figure 5 D) Colonic histology (periodic acid-Schiff staining). Images were taken using a high-resolution microscope at 20x magnification (scale bar, 200 μm). For histology, data represent two independent experiments. Each in vivo experiment consisted of 4 mice per group (16 mice per experiment). Data are presented as mean ± SEM. **p < 0.01, ****p < 0.0001. Two-way ANOVA, Bonferroni post-hoc test. H2O, untreated mice.

[0019] Figure 6 A to Figure 6 C is a graphical representation and photographic image showing that 3-IAld provides protection against DSS+ anti-CTLA-4 induced colitis in IL-10- / - mice. (See image.) Figure 2As described in section A, mice were treated with DSS in drinking water for one week, followed by a one-week recovery period and administration of 100 µg of anti-CTLA-4 mAb and 3-IAld. The mice were then assessed for (…). Figure 6 A) % weight change, ( Figure 6 B) Colon histology (PAS staining) and ( Figure 6 C) Histological scoring. Images were taken using a high-resolution microscope at 20x magnification (scale bar, 200 µm). Data are expressed as mean ± SEM. H2O, untreated mice. *P<0.05, **p < 0.01.

[0020] Figure 7 A to Figure 7 B is a graphical representation and photographic image showing how 3-IAld provides protection against pathology in IL-10- / - mice after acute treatment with anti-CTLA-4 monoclonal antibody. Mice were administered 100 µg of anti-CTLA-4 mAb and 3-IAld as described above, and the efficacy was evaluated. Figure 7 A) % weight change and ( Figure 7 B) Colonic histology (PAS staining). Photographs were taken using a high-resolution microscope at 20x magnification (scale bar, 200 µm).

[0021] Figure 8 A to Figure 8 H is a graphical representation and photographic image showing how 3-IAld protects IL-10-deficient mice from pathological effects after chronic treatment with anti-CTLA-4 monoclonal antibody. (See experimental schedule diagram.) Figure 8 As described in A), 100 µg of anti-CTLA-4 mAb and 3-IAld were administered to control C56BL / 6 and IL-10 deficient mice. The mice were assessed for (…). Figure 8 B) Weight change, ( Figure 8 C) Disease activity index, ( Figure 8 D) Clinical laboratory testing, ( Figure 8 E) colon and ( Figure 8 F) Histology (PAS staining) (ZO-1 and BrdU staining in the illustration), ( Figure 8 G) Calprotectin levels in intestinal homogenate and ( Figure 8 H) Cytokine and defense gene expression in the gut. Images were taken using a high-resolution microscope at 20x magnification (scale bar, 200 µm). Data are presented as mean ± SEM (pooled data from both experiments). H2O, untreated mice. *P<0.05, **p<0.01.

[0022] Figure 9 A to Figure 9B is a graphical representation and photographic image showing how 3-IAld reduces lymphocyte infiltration in IL-10-deficient mice after chronic treatment with anti-CTLA-4 monoclonal antibody. Figure 8 The illustration in Figure A shows mouse treatments, and histological changes and lymphocyte infiltration in different organs were assessed by anti-CD3 staining (quantified using ImageJ software). DAPI (4′,6-diamidinyl-2-phenylindole) was used for DNA staining.

[0023] Figure 10 A to Figure 10 D is a graphical representation and photographic image showing how 3-IAld slows disease progression in 16-week-old IL-10-deficient mice. (e.g.) Figure 10 Mice were treated with 3-IAld and 1% DSS as described in (A) and their ( ) were evaluated Figure 10 B) % body weight, ( Figure 10 C) Histological scoring and ( Figure 10 D) Histopathology of inflammatory colon and epithelial renewal (by PAS and Ki-67 staining, respectively).

[0024] Figure 11 A to Figure 11 F is a graphical representation and photographic image showing how 3-IAld-modified microbiota provides protection against DSS+-induced colitis. C57BL / 6 mice were subjected to DSS colitis and subjected to ( Figure 11 E) or not to perform ( Figure 11 A to Figure 11 D) Anti-CTLA-4 treatment, and transplantation of fresh fecal particles (FMT) from control or 3-IAld-treated mice 1 day before and 2 days after colitis induction. 7 days after colitis induction ( Figure 11 A to Figure 11 D) or 14 days ( Figure 11 E) The mice were euthanized and evaluated. Figure 11 A, Figure 11 E) % weight change, ( Figure 11 B) Gross pathology, ( Figure 11 C) Histological score, ( Figure 11 D) Colonic histopathology (periodic acid-Schiff staining), ( Figure 11F) Methylation / demethylation status of the Foxp3 promoter in mesenteric lymph nodes. Images were taken using a high-resolution microscope at magnifications of 10x and 20x (scale bars, 500 and 200 µm). Data represent three independent experiments. Each in vivo experiment consisted of 3 mice per group (6–12 mice per experiment). Data are expressed as mean ± SD. H2O, untreated mice. *P<0.05, **P<0.01, ***P<0.001.

[0025] Figure 12 A to Figure 12 J is a graphical representation and photographic image showing how 3-IAld does not interfere with the development of antitumor immunity. For Figure 12 A to Figure 12 E. C57BL / 6 mice were subcutaneously injected with B16 tumor cells and administered 100 µg of anti-CTLA-4 mAb or an allotype control intraperitoneally four times at 3-day intervals, up to a maximum of 16 days. 3-IAld was administered intragastrically every other day. The mice were assessed for (…). Figure 12 A) Tumor growth, ( Figure 12 B) Histology (periodic acid-Schiff staining), ( Figure 12 C) Immunohistochemistry of CD8+ and CD4+ tumor-infiltrating lymphocytes (TILs), Figure 12 D) The number of positive TILs per high-power field (HPF) and ( Figure 12 E) Cxcl9 and perforin expression. Figure 12 F to Figure 12 J) C57BL / 6 mice were orally injected with LLC cells and administered 200 µg of anti-PD-1 mAb or an allotype control intraperitoneally five times at 3-day intervals, up to a maximum of 18 days. 3-IAld was administered intragastrically every other day. The mice were assessed for ( Figure 12 F) % Survival, ( Figure 12 G) Lung weight, ( Figure 12 H) Gross pathology of the lungs, ( Figure 12 I, 12J) CD4+CD25+Foxp3+ cells. Images were taken using a high-resolution microscope at 40x magnification (scale bar, 100 µm). Each in vivo experiment included three mice per group (nine mice per experiment). Data are presented as mean ± SEM. One-way ANOVA, Bonferroni post-hoc test. *P<0.05, **P<0.01, ***P<0.001.

[0026] Figure 13 A to Figure 13 E is presented graphically as a pharmacodynamic study, including analysis of unlabeled 3-IAld, which detected minute changes in baseline endogenous levels of 3-IAld at different time points. Figure 13 A), Analysis of labeled 3-IAld ( Figure 13 B), Analysis of potential metabolites of unlabeled 3-IAld ( Figure 13 C), the level of methylated forms of indole-3-carboxylic acid (i.e., 1-methylindole-2-carboxylic acid and methyl indole-3-carboxylic acid ester) ( Figure 13 D), and the presence of indole-3-carboxylic acid was also confirmed by the labeled 3-IAld. Figure 13 E).

[0027] Figure 14 The fold increase in Cyp1a1 mRNA levels with the addition of 3-IAld or Ox-3-IAld is graphically represented. Human cell line HepG2 (hepatocellular carcinoma cell line) was exposed to different concentrations of the ligand indole-3-carboxylic acid or Ox-3-IAld, and the results showed that the AhR activation marker Cyp1A1 was induced in a dose-dependent manner between 1 and 100 µM. Figure 14 ).

[0028] Figure 15 The histopathology of different organs after blind examination following hematoxylin and eosin (H&E) staining is illustrated in the figure.

[0029] Figure 16 It is a representation of the chemical structure of 1-methylindole-3-carboxylic acid (molecular formula: C10H9NO2, IUPAC name: 1-methyl-1H-indole-3-carboxylic acid), also known as 1-methyl-3-indole-carboxylic acid, 1-methyl-1H-indole-3-carboxylic acid, or 1ME3CA.

[0030] Figure 17 A to Figure 17 F is a graphical representation and photographic image showing the protective effect of 1-methylindole-3-carboxylic acid against inflammatory pathology in DSS+ anti-CTLA-4 induced colitis. As depicted in the experimental schedule diagram, C57BL / 6 mice were treated with DSS in drinking water for one week, followed by a one-week recovery period and administration of 100 µg of anti-CTLA-4 mAb and 1-methylindole-3-carboxylic acid, wherein 1-methylindole-3-carboxylic acid was administered every other day at doses of 0.09, 0.18, or 0.36 mg / mouse. Figure 17 A). Assess the mice's ( Figure 17 B) Weight change, ( Figure 17 C) Disease activity index, ( Figure 17 D) Clinical onset and rectal bleeding, ( Figure 17 E to Figure 17F) Histology of the colon and ileum (by PAS staining), epithelial barrier function (ZO-1 staining), and lymphocyte infiltration in the colon and ileum (CD3 staining). Images were taken using a high-resolution microscope at 20x magnification (scale bar, 200 µm). Each in vivo experiment included four to six mice per group (16–24 mice per experiment). Data are presented as mean ± SEM. H2O, untreated mice. One-way ANOVA, Bonferroni post-hoc test. ***P<0.001, ****P<0.0001.

[0031] Figure 18 Photographs showing how 1-methylindole-3-carboxylic acid reduces lymphocyte infiltration in mice treated with anti-CTLA-4. (See attached image.) Figure 17 As described in the experimental schedule diagram for A, C57BL / 6 mice were treated with DSS in drinking water for one week, followed by a one-week recovery period and administration of 100 µg of anti-CTLA-4 mAb and 1-methylindole-3-carboxylic acid. Histological changes and lymphocyte infiltration in the mouse lungs were assessed by anti-CD3 staining. DAPI (4′,6-diamidinyl-2-phenylindole) was used for DNA staining.

[0032] Figure 19 Represented graphically Il1b , Il10 , Cyp1a1 , Reg3g and Il22 The fold increase in mRNA levels indicates that the addition of 1-methylindole-3-carboxylic acid promotes an anti-inflammatory state and AhR activation in mice with colitis. Assessments such as... Figure 17 Cytokine and AhR-dependent gene expression in C57BL / 6 mice subjected to DSS+ anti-CTLA-4 induced colitis and administered 1-methylindole-3-carboxylic acid, as described in the experimental schedule diagram in A. Data are presented as mean ± SEM. H2O, untreated mice. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. One-way ANOVA, Bonferroni post-hoc test.

[0033] Figure 20A The pharmacokinetics of 3-IAld in the intestine, serum, lung, liver, brain, and kidney over a 24-hour period are shown. Figure 20B The pharmacokinetics of 1-methylindole-3-carboxylic acid in the intestine, serum, lung, liver, brain, and kidney over a 24-hour period are shown.

[0034] Figure 21 The AhR agonist activity of 3-IAld and 1ME3CA in vitro was demonstrated in the H1L6.1c3 cell line.

[0035] Figure 22 This shows the effects of 3-IAld and 1ME3CA in vitro on A549 alveolar epithelial cells ( Figure 22 A) Calu-3 bronchial epithelial cells ( Figure 22 B), Caco-2 colon cancer cells ( Figure 22 C) and HepG2 liver cancer cells ( Figure 22 AhR activity in D). ITE and FICZ are reference AhR agonists. Assays were performed 4 hours after incubation.

[0036] Figure 23 The results showed that MECA provided protection against inflammatory pathology in a dose-dependent manner in DSSS+ anti-CTLA-4 induced colitis. (See experimental schedule diagram). Figure 23 As described in A), C57BL / 6 mice were treated with DSS in drinking water for one week, followed by a one-week recovery period and administration of 100 µg of anti-CTLA-4 mAb and MECA. The mice were then assessed for (…). Figure 23 B) Weight change, ( Figure 23 C) Disease activity index, ( Figure 23 D) Clinical onset and rectal bleeding, ( Figure 23 E) Colon and ileum histology (PAS staining), magnified 40x (scale bar, 100 μm). Each in vivo experiment included four to six mice per group (20 mice per experiment).

[0037] Figure 24 The study showed that MECA reduced lymphocyte infiltration in anti-CTLA-4 treated mice in a dose-dependent manner. Lung, spleen, liver, and kidney tissue samples demonstrated a dose-dependent ability to reduce lymphocyte infiltration in mice that had been treated with MECA for anti-CTLA-4.

[0038] Figure 25A and Figure 25B The toxicological effects of 3-IAld and 1ME3CA are shown on PAS-stained tissue sections from naïve C57BL / 6 mice. Images were taken using a high-resolution Olympus DP71 microscope with a 10x objective lens. Scale bar, 400 μm. Naïve, untreated mice. Detailed Implementation

[0039] Some aspects of this disclosure relate to a method for treating a patient with CTLA-4 checkpoint-related immunodeficiency, the method comprising administering to a patient in need a therapeutically effective amount of a pharmaceutical composition comprising 1H-indole-3-carboxaldehyde (3-IAld).

[0040] Another aspect of this disclosure relates to a method for treating a patient with CTLA-4 checkpoint-related immunodeficiency, the method comprising administering to a patient in need a therapeutically effective amount of a pharmaceutical composition comprising 1-methylindole-3-carboxylic acid.

[0041] Before describing this disclosure in more detail, it should be understood that this disclosure is not limited to the specific compositions or method steps described, and therefore such compositions or method steps are of course subject to variation. It will be apparent to those skilled in the art upon reading this disclosure that the various individual aspects described and illustrated herein have discrete components and features that can be readily separated from or combined with features of any other aspects without departing from the scope or spirit of this disclosure. Any of the listed methods can be implemented in the order of the enumerated events or in any other logically possible order.

[0042] The headings provided herein are not intended to limit any aspect of this disclosure, which can be defined by referring to the entire specification. It should also be understood that the terminology used herein is for the purpose of illustrating particular aspects only and is not intended to be limiting.

[0043] I. Terminology To make this disclosure more readily understandable, certain terms are first defined. As used in this application, each of the following terms shall have the meaning described below unless expressly provided otherwise herein. Further definitions are set forth throughout the application.

[0044] As stated herein, unless otherwise indicated, any concentration range, percentage range, ratio range, or integer range shall be understood to include any integer within the listed range and (where appropriate) its fractional value (such as one-tenth and one-hundredth of an integer).

[0045] Throughout this disclosure, the term "an" or "a" entity refers to one or more of the aforementioned entities; for example, "chimeric polypeptide" should be understood to represent one or more chimeric polypeptides. Therefore, the terms "an(a)" or ("an(an)"), "one or more," and "at least one" are used interchangeably herein.

[0046] Furthermore, the term “and / or” as used herein should be considered as a specific disclosure of each of the two specified features or components, with or without the other. Thus, the term “and / or” as used in phrases such as “A and / or B” herein is intended to include “A and B”, “A or B”, “A” (alone), and “B” (alone). Similarly, the term “and / or” as used in phrases such as “A, B, and / or C” is intended to cover each of the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone). Additionally, “or” is used to indicate an open list of components. For example, “where X contains A or B” means that X contains A, X contains B, X contains A and B, or X contains A or B and any other component.

[0047] The terms “about” or “substantially comprise” mean that a value or composition is within an acceptable margin of error for a particular value or composition as determined by a person skilled in the art, which will depend in part on the method of measuring or determining the value or composition, i.e., the limitations of the measurement system. For example, “about” or “substantially comprise” may mean within or greater than one standard deviation according to practice in the art. Alternatively, “about” or “substantially comprise” may mean a range up to 10%. Furthermore, particularly for biological systems or processes, the term may mean up to an order of magnitude or up to five times the value. When a particular value or composition is provided in the application and claims, unless otherwise stated, the meaning of “about” or “substantially comprise” should be assumed to be within an acceptable margin of error for the particular value or composition.

[0048] As used herein, when applied to one or more target values, the term "approximately" refers to a value similar to the stated reference value. In some respects, the term "approximately" refers to a range of values ​​falling within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction of the stated reference value, unless otherwise stated or otherwise apparent from the context (except where such a number would exceed 100% of the possible value).

[0049] As used in this article, patients with a specific gene mutation who are "homozygous" have the same mutation on each allele.

[0050] As used in this article, a patient with a specific gene mutation who is "heterozygous" has the mutation in one allele and a different mutation in another allele.

[0051] The term "antibody" can include, for example, monoclonal antibodies (mAbs), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), recombinant antibodies, human antibodies, chimeric antibodies, and humanized antibodies. Furthermore, the term "antibody" can also encompass recombinantly expressed antigen-binding proteins and antigen-binding synthetic peptides.

[0052] An "antigen" is any molecule, such as a peptide, that elicits an immune response or can be bound by a TCR. An immune response may involve antibody production, activation of specific immune cells, or a combination thereof. Those skilled in the art will readily understand that any macromolecule (including almost all proteins or peptides) can be used as an antigen. Antigens can be expressed endogenously, i.e., through genomic DNA, or can be recombinantly expressed. Antigens and / or epitopes may be specific to a particular tissue, such as cancer cells, or they may be widely expressed. Furthermore, fragments of larger molecules can act as antigens. In one respect, an antigen is a tumor antigen.

[0053] As used in this article, "antitumor effect" refers to biological effects that can manifest as a reduction in tumor volume, a decrease in the number of tumor cells, a reduction in tumor cell proliferation, a decrease in the number of metastases, an increase in overall survival or progression-free survival, an increase in life expectancy, or an improvement in various physiological symptoms associated with cancer. Antitumor effect can also refer to the prevention of cancer development, such as through vaccines.

[0054] “Indole-3-aldehyde” or “3-IAld” is a metabolite derived from the microbial degradation of the amino acid tryptophan. The identification of 3-IAld is further described in U.S. Patent Application Publication No. 2016 / 0206595, which is incorporated herein by reference. 3-IAld is an agonist of the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor involved in a wide range of physiological activities, including maintaining mucosal homeostasis at barrier organs (Stockinger B et al., Ann. Rev. Immunol. 2014;32:403-32). By activating AhR in type 3 innate lymphocytes, 3-IAld induces the production of IL-22, a key cytokine in host defense and tissue repair at mucosal surfaces and in the regulation of microbial composition (Zelante T et al., Immunity. 2013;39(2):372-85; Borghi M et al., Immunol. 2019;10:2364). Therefore, the 3-IAld-AhR-IL-22 axis represents a functional signaling unit for enhancing barrier function, exhibiting good activity in pathological conditions characterized by epithelial damage, mucosal alterations, and excessive inflammatory responses. The formula of indole-3-aldehyde (or indole-3-carboxaldehyde, 3-IAld, molecular formula: C9H7NO, IUPAC name: IH-indole-3-carboxaldehyde) is given below and Figure 1As shown in the figure. 3-IAld is expected to be well tolerated in humans as it has no potential side effects.

[0055] Endogenous tryptophan (Trp) metabolites play a crucial role in mammalian gut immune homeostasis. In the gastrointestinal tract, dietary AhR ligands promote the production of localized IL-22 by innate lymphocytes (ILCs) (Qiu et al., 2012) (now referred to as group 3 ILCs (ILC3) (Spits et al., 2013)) (Lee et al., 2011). Metabolomics analyses have revealed that gut bacteria influence host metabolism and immunity through a variety of chemically distinct metabolites, including amino acid metabolites (Wikoff et al., 2009). Specifically, dietary Trp deficiency impairs gut immunity and alters the gut microbiota in mice (Hashimoto et al., 2012), suggesting that mucosal homeostasis is a multifactorial phenomenon in which Trp metabolism is a key regulatory component. However, the source and nature of any such AhR ligands, any effects of microbial dysbiosis on AhR and IL-22-driven mucosal reactivity, and whether microbiota-derived metabolites also activate AhR remain unclear. Although the enzyme Trp 2,3-dioxygenase, primarily expressed in the liver (Opitz et al., 2011), regulates Trp concentrations under normal conditions following nutrient Trp uptake, the high expression levels of IDO1 at mucosal sites during immune activation (Dai et al., 2010) indicate that IDO1 is a key enzyme regulating local amino acid nutrient levels, the size and metabolic activity of the gut microbiota, and mucosal immunoreactivity resulting from immunomodulatory activity mediated by the host via L-kynurenine production. Therefore, these data suggest that IDO1 is a crucial molecule determining host-microbiota symbiotic relationships and their integration within the adaptive immunity of vertebrate hosts.

[0056] The microbiota-AhR axis may represent an important strategy for co-evolutionary symbiosis to fine-tune host mucosal reactivity dependent on Trp catabolism. To this end, the drug 3-IAld can offer a more significant advantage than any other existing treatment by improving host immune responsiveness, epithelial barrier function, and pathogen colonization. Indole plays multiple roles, including bidirectional communication with the microbiome to fine-tune host immunity, tolerance, and metabolism by acting as a ligand for AhR (a transcription factor that controls the biodegradation of endogenous and exogenous toxins), preventing inflammatory damage and providing barrier integrity (Hubbard et al., 2015 b). Studies have shown that administration of indole to germ-free mice increases the expression of epithelial tight junction proteins and attenuates indicators of inflammatory colitis (Shimada et al., 2013). Similarly, the therapeutic efficacy of indole-3-aldehyde (3-IAld) in a mouse model of dextran-induced colitis has been reported (Zelante et al., 2013).

[0057] As used herein, the terms “active pharmaceutical ingredient” or “therapeutic agent” (“API”) refer to bioactive compounds.

[0058] The terms “patient” and “subject” are used interchangeably and refer to animals, including humans.

[0059] Those skilled in the art will recognize that when the amount of "a compound or a pharmaceutically acceptable salt thereof" is disclosed, the amount of the pharmaceutically acceptable salt form of the compound is equal to the concentration of the free base of the compound. It should be noted that the amounts of the compound or its pharmaceutically acceptable salt disclosed herein are based on its free base form.

[0060] As used herein, the term "in combination with" when referring to two or more compounds, agents or other active pharmaceutical ingredients means that the two or more compounds, agents or active pharmaceutical ingredients are administered to the patient before, simultaneously or after each other.

[0061] "CTLA-4 checkpoint-associated immunodeficiency" or "primary CTLA-4 checkpoint-associated immunodeficiency" or immunodeficiency is a condition or disease characterized in patients by a variable combination of enteropathy, hypogammaglobulinemia, recurrent respiratory infections, granulomatous lymphocytic interstitial lung disease, lymphocytic infiltration of non-lymphatic organs (intestine, lung, brain, bone marrow, and kidney), autoimmune thrombocytopenic purpura or neutropenia, autoimmune hemolytic anemia, and lymphadenopathy. CTLA-4 checkpoint-associated immunodeficiency is diagnosed based on clinical symptoms, laboratory findings, and genetic testing. Patients with only one functional copy of the gene encoding CTLA-4 suffer from severe autoimmunity. These heterozygous mutations result in a novel phenotype with overactive T and B cell infiltration of non-lymphatic organs (such as the intestine, lung, and brain) and more typical autoimmune signs. Some clinical manifestations of CTLA-4 haploinemia are similar to those observed in biopsies of inflamed organs. Patients with primary CTLA-4 checkpoint-related immunodeficiency may have recurrent respiratory infections, hypogammaglobulinemia, autoimmune cytopenia, autoimmune enteropathy, and granulomatous infiltrative lung disease.

[0062] Primary CTLA-4 checkpoint-associated immunodeficiency is diagnosed based on clinical symptoms, laboratory findings, and genetic testing. Patients with only one functional copy of the gene encoding CTLA-4 suffer from severe autoimmunity. These heterozygous mutations lead to a new phenotype with overactive T and B cell infiltration of non-lymphoid organs, such as the intestine, lung, and brain, and more typical autoimmune signs. Some clinical manifestations of CTLA-4 haploinemia are similar to those observed in biopsies of inflamed organs from patients receiving anti-CTLA-4 therapy. Patients with primary CTLA-4 checkpoint-associated immunodeficiency may present with recurrent respiratory infections, hypogammaglobulinemia, autoimmune cytopenia, autoimmune enteropathy, and granulomatous infiltrative lung disease.

[0063] CTLA-4 is a key negative regulator of T-cell-mediated immune responses. It contributes to the rapid development of lethal, destructive, multi-organ lymphocyte infiltration. ctla4Knockout mice clearly demonstrate the crucial role of CTLA-4 in lymphocyte homeostasis and tolerance. Autoimmunity is caused by self-reactive T and / or B lymphocytes and is the root cause of a wide range of conditions, from endocrine disorders to hematologic cytopenia. Genetic epidemiological studies have long demonstrated that many autoimmune conditions have a genetic component. Haploid deficiency is defined as the presence of a phenotype in heterozygotes despite the lack of negative dominance of the mutant allele relative to its wild-type counterpart. Heterozygous CTLA-4 germline mutations impair the suppressive function of T cells and lead to an immune dysregulation syndrome characterized by an activated immune system with autoimmune features and organ pathological manifestations caused by infiltrating effector T cells. Therefore, having a single working copy of CTLA-4 is insufficient to produce enough CTLA-4 protein for a normal immune system. These heterozygous mutations result in novel phenotypes with overactive T and B cell infiltration of the intestine, lungs, and brain, as well as more typical autoimmune symptoms.

[0064] Located sequentially on the long arm of chromosome 2 are paralogs encoding the immunoglobulin family coactivator receptors CD28 and CTLA-4. CD28 and CTLA-4 share the same B7 ligand but each delivers opposing proliferative signals to T cells. CD28 delivers a positive signal to T cells, leading to T cell activation and effector cell differentiation, while CTLA-4, linked to CD80 and CD86, delivers a negative signal to T cells, thereby limiting IL-2 production, T cell proliferation, and survival. CTLA-4 is inducibly expressed on activated CD4+ Foxp3 conventional T cells (Tconv) and constitutively expressed on CD4+ Foxp3+ regulatory T cells (Treg). The critical inhibitory function of CTLA-4 has been demonstrated by spontaneous massive T cell expansion accompanied by multi-organ lymphocyte infiltration and tissue destruction. Ctla4 - / - The rapid-onset, fatal inflammatory phenotype in mice was revealed (Tivol et al., 1995). The similarity of this phenotype to systemic autoimmune diseases prompted research into the role of CTLA-4 in T cell tolerance and autoimmunity. CTLA-4 acts on both Tconv and Treg cells and is a mediator of Treg cell inhibitory function.

[0065] Human CTLA4 haploid deficiency leads to dysregulation of FoxP3+ regulatory T (Treg) cells, overactivation of effector T cells, and lymphocytic infiltration of target organs (such as the intestine, lung, and brain), as well as more typical autoimmune symptoms. The adaptive immune response must strike a balance between the response to foreign antigens and the need to avoid damage to self-antigens and host tissues. On the one hand, insufficient activation of the immune response leads to pathology resulting from infection, while overactivation can drive autoimmune responses. It is expected that different gene mutations are the underlying causes of these seemingly contradictory outcomes; paradoxically, however, it is generally believed that autoimmunity and immunodeficiency can coexist in the same individual.

[0066] Since CTLA-4 inhibits the CD28 pathway (which plays a role in T cell helper responses to B cells), CTLA-4 deficiency is expected to enhance CD28 function and promote humoral immunity. One possible explanation is that excessive T cell activation may lead to infiltration and disruption of the bone marrow niche, thereby impairing B cell development. This is consistent with the disruption of B cell lymphocyte production in Treg-deficient mice. Alternatively, increased CD28-dependent follicular helper T (TFH) cell differentiation may lead to chronic stimulation of B cells, resulting in exhaustion.

[0067] CTLA-4 deficiency is characterized by the infiltration of immune cells into the intestines, lungs, bone marrow, central nervous system, kidneys, and possibly other organs. Most people with CTLA-4 deficiency experience diarrhea or intestinal problems. Enlarged lymph nodes, liver, and spleen are also common, as are respiratory infections. People with CTLA-4 deficiency often experience autoimmune problems that can affect various organs and tissues, including the blood, thyroid, skin, and joints. The condition may also slightly increase the risk of lymphoma (a type of immune cell cancer).

[0068] The increased cancer risk, combined with observed viral associations, leads us to hypothesize that deficiencies in the immune surveillance of chronically viral-infected cells and reduced elimination of oncogenic viruses result in dysregulation of cell growth. Decreased CTLA-4 expression leads to uncontrolled T cell proliferation and may result in the overgrowth of self-reactive clones relative to, for example, EBV-specific T cell clones, as is known in individuals with HIV infection.

[0069] It is noteworthy that autosomal dominant CTLA-4 deficiency is characterized by incomplete penetrance, as some heterozygous individuals are asymptomatic. A delicate balance is known to exist between self-tolerance and autoimmunity, a balance at least in part governed by quantitative changes in CTLA-4 expression. As predicted by preclinical models, complex interactions between genetics and environment may determine the evolution of different phenotypes associated with CTLA-4 deficiency, or conversely, the maintenance of an asymptomatic state. Kuehn et al. , 2014 ).

[0070] CTLA-4 checkpoint-associated immunodeficiency leads to immune dysregulation syndromes with a wide range of clinical manifestations. In many cases, the onset of clinical manifestations is not observed until adulthood. Some patients suffer from severe hematologic cytopenia. Patients with immune thrombocytopenic purpura are at risk of bleeding complications, and patients with immune hemolytic anemia may be at risk of complications secondary to hypoxia, including acute renal failure and acute vascular events. In patients with pulmonary lymphocytic infiltration, recurrent pulmonary infections may occur, and the presence of concomitant immunosuppressive therapy established to reduce lymphocytic infiltration and autoinflammation sometimes exacerbates this condition. Some patients also suffer from gastrointestinal disorders. Those with gastric atrophy are at risk of ulcers, also because the tendency to develop recurrent Helicobacter pylori infections can progress to lymphoproliferative disorders (such as MALT lymphoma). Serious complications that can alter the quality and quantity of life include colitis, pancreatitis, and central nervous system infiltration. Schubert et al. , 2014; Kuehn et al. , 2014 ).

[0071] The report by Kuehn et al., describing the consequences of decreased CTLA-4 expression, shares striking similarities with, but also differs from, reports of inflammatory conditions associated with anti-CTLA-4 cancer therapy. In 540 melanoma patients receiving intermittent CTLA-4 blockade with ipilimumab, approximately 60% experienced immune-related adverse events, and 11% had severe symptoms, the most common being skin (rash and vitiligo), gastrointestinal (enteritis), and endocrine (hypothyroidism and hypopituitarism) symptoms. (Hodi et al. , 2010 Less common inflammatory events include hepatitis, uveitis, neurological disorders, and pneumonia. (Attia et al. , 2005While most immune-related toxicities can be easily managed with immunosuppressive drugs, some are fatal. Biopsies of inflamed organs show mixed CD4+ and CD8+ T cell infiltrations. Increased serum titers of autoantibodies observed in some patients are against thyroid tissue, acetylcholine receptors, the pituitary gland, and other targets.

[0072] A significant correlation has been described between severe immune-related toxicity and major tumor regression. Attia et al. , 2005 This indicates a shared biological mechanism and highlights the unstable balance between self-tolerance and autoimmunity in both malignant and normal tissues. Cancer susceptibility in CTLA-4 deficiency appears to be due to immune activation accompanied by chronic inflammation, the inability to control oncogenic viruses or neoplastic cells through immunological means, coupled with intrinsic T-cell damage due to underlying genetic defects. Salavoura et al. , 2008 For these reasons, patients with CTLA-4 haploid deficiency are more susceptible to malignant tumors, particularly gastric cancer and lymphoma. Dhalla et al. , 2011 - Cunningham-Rundles et al. , 1999 Lymphoma is known to occur due to lymphopenia in most patients. Gastric cancer is usually caused by chronic inflammatory tissue, which is a well-known risk factor. Compare et al. , 2010 ) 。 Recent research by Egg et al. indicates that carriers of affected CTLA4 mutations have a 12.9% increased risk of developing malignant tumors. Furthermore, comparing cancer risk in the general population with that of carriers of affected CTLA4 mutations, carriers of affected CTLA4 mutations have a higher annual cancer rate. Egg et al. , 2018 ).

[0073] Therefore, patients with CTLA-4 checkpoint-related immunodeficiency or primary CTLA-4 checkpoint-related immunodeficiency may present with a number of different gastrointestinal diagnoses, syndromes or symptoms, including but not limited to CHAI, LRBA, LATAIE, regulatory T cell deficiency, enteropathy, intestinal inflammation, immune-mediated colitis (IMC), gastrointestinal disorders, gastric atrophy, colitis, and other diseases that may not yet be fully recognized.

[0074] As used in this article, “CTLA-4 haploinemia with autoimmune infiltration (CHAI)” is a CTLA-4 checkpoint-related immunodeficiency disorder characterized by heterozygous CTLA4 mutations in humans associated with severe immunomodulatory disorders. CHAI is characterized primarily by lymphoproliferation, autoimmune cytopenia, enteropathy, interstitial lung disease, and recurrent infections.

[0075] As used in this article, "lipopolysaccharide-responsive beige-like anchoring protein (LRBA)" is a CTLA-4 checkpoint-related immunodeficiency disorder characterized by a genetic biallelic mutation in the patient's gene. Because LRBA deficiency also leads to secondary loss of CTLA-4, this recessive disorder is known as LRBA deficiency with autoantibodies, T-reg cell deficiency, autoimmune infiltration, and enteropathy (LATAIE). LATAIE is also commonly characterized by enteropathy, autoimmune disorders, lymphoproliferation, and respiratory infections. In fact, while granulomas, malignancies, atopic diseases, skin conditions, and neurological disorders are more common in patients with CHAI, LATAIE patients are more frequently complicated by life-threatening infections, pneumonia, ear, nose, and throat conditions, organ enlargement, autoimmune enteropathy, and growth retardation. Although CHAI and LATAIE share similar characteristics, a significant difference is that LATAIE typically has a younger age of onset, with disease onset often evident in preschool children, while CHAI occurs in older children or young adults.

[0076] As used in this article, “regulatory T (Treg) cell deficiency” is a CTLA-4 checkpoint-related immunodeficiency disorder, which refers to any defective immune surveillance response that leads to reduced clearance of cells infected with chronic viruses and oncoviruses, resulting in dysregulation of cell growth. One example is decreased CTLA-4 expression, which leads to uncontrolled T cell proliferation and may result in an overgrowth of autoreactive clones relative to, for example, EBV-specific T cell clones.

[0077] As used in this article, "enteropathy" is a CTLA-4 checkpoint-associated immunodeficiency disorder, which refers to persistent damage or irritation and swelling of the small intestine.

[0078] As used in this article, "intestinal inflammation" is a symptom of CTLA-4 checkpoint-related immunodeficiency, referring to inflammation of the gastrointestinal (GI) tract. Chronic inflammation leads to gastrointestinal damage.

[0079] As used in this article, “immune-mediated colitis (IMC)” is a common immune-related adverse event associated with immune checkpoint inhibitors, including abdominal pain, mucus or blood in the stool, and fever. Immune checkpoint inhibitors are immunotherapeutic drugs that work by blocking the binding of checkpoint proteins to their chaperone proteins. This prevents the “switching off” signal from being emitted, thus allowing T cells to kill cancer cells. Dysregulation of T cells can also contribute to IMC, in which T cells attack gastrointestinal cells and tissues.

[0080] As used herein, “autoimmune infiltration” or “infiltration” is the diffusion or accumulation of foreign substances (in tissues or cells) in amounts exceeding the normal range. Infiltration can include lymphocyte infiltration in non-lymphoid organs such as the intestine (as in CHA). Thinning of the intestinal lining can also lead to the infiltration of other infiltrates into the intestine.

[0081] As used in this article, "gastric atrophy" is a condition characterized by thinning of the inner lining of the stomach and / or intestine, and loss of glandular cells (which release substances that aid digestion) within the lining. This can be caused by Helicobacter pylori infection or certain autoimmune conditions.

[0082] "Cancer" refers to a broad group of diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division and growth lead to the formation of malignant tumors that invade adjacent tissues and can also metastasize to distant parts of the body via the lymphatic system or bloodstream. "Cancer" or "cancer tissue" can include tumors. Examples of cancers that can be treated by the methods of the present invention include, but are not limited to, cancers of the immune system, including lymphoma, leukemia, and other white blood cell malignancies. In some embodiments, the methods of the present invention can be used to reduce the size of tumors derived from cancers such as bone cancer, pancreatic cancer, skin cancer, head and neck cancer, malignant melanoma of the skin or eye, uterine cancer, ovarian cancer, rectal cancer, anal cancer, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, breast cancer, prostate cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC)), Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small bowel cancer, endocrine system cancers, thyroid cancer, and thyroid cancer. Parathyroid carcinoma, adrenal carcinoma, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, childhood solid tumors, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal pelvis cancer, central nervous system (CNS) tumors, primary CNS lymphoma, tumor angiogenesis, spinal cord axis tumors, brainstem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, including those induced by asbestos, or any combination thereof. A particular cancer may respond to chemotherapy or radiation therapy, or the cancer may be refractory. Refractory cancer is defined as cancer for which surgical intervention is not applicable, and said cancer initially did not respond to chemotherapy or radiation therapy, or said cancer has become unresponsive over time.

[0083] It should be understood that whenever the term “contains” is used in this document to describe an aspect, other similar aspects described by “consisting of” and / or “substantially consisting of” are also provided.

[0084] As used herein, “cytokine” refers to a non-antibody protein released by a cell in response to contact with a specific antigen, wherein the cytokine interacts with a second cell to mediate the response in the second cell. Cytokines can be expressed endogenously by cells, added to cultured cells, or administered to a subject, or any combination thereof. Immune cells (including macrophages, B cells, T cells, and mast cells) can release cytokines to propagate an immune response. Cytokines can induce a variety of responses in recipient cells. Cytokines can include homeostatic cytokines, chemokines, pro-inflammatory cytokines, effectors, and acute-phase proteins. For example, homeostatic cytokines, including interleukin (IL) 7 and IL-15, promote immune cell survival and proliferation, and pro-inflammatory cytokines can promote inflammatory responses. Examples of homeostatic cytokines include, but are not limited to, IL-2, IL-4, IL-5, IL-7, IL-10, IL-12p40, IL-12p70, IL-15, IL-21, and interferon (IFN)γ. Examples of pro-inflammatory cytokines include, but are not limited to, IL-1a, IL-1b, IL-6, IL-13, IL-17a, tumor necrosis factor (TNF)-α, TNF-β, fibroblast growth factor (FGF) 2, granulocyte-macrophage colony-stimulating factor (GM-CSF), soluble intercellular adhesion molecule-1 (sICAM-1), soluble vascular adhesion molecule-1 (sVCAM-1), vascular endothelial growth factor (VEGF), VEGF-C, VEGF-D, and placental growth factor (PLGF). Examples of effectors include, but are not limited to, granzyme A, granzyme B, soluble Fas ligand (sFasL), and perforin. Examples of acute-phase proteins include, but are not limited to, C-reactive protein (CRP) and serum amyloid A (SAA).

[0085] Chemokines are a type of cytokine that mediates cellular chemotaxis or directed movement. Examples of chemokines include, but are not limited to, IL-8, IL-16, eosinophil chemokine, eosinophil chemokine-3, macrophage-derived chemokines (MDC or CCL22), monocyte chemoattractant protein 1 (MCP-1 or CCL2), MCP-4, macrophage inflammatory protein 1a (MIP-1a, MIP-1a), MIP-Ib (MIP-1b), γ-inducible protein 10 (IP-10), and thymus activation-regulated chemokines (TARC or CCL17).

[0086] Other examples of cytokines include, but are not limited to, chemokine (CC motif) ligands (CCL1), CCL5, monocyte-specific chemokine 3 (MCP3 or CCL7), monocyte chemoattractant protein 2 (MCP-2 or CCL8), CCL13, IL-1, IL-3, IL-9, IL-11, IL-12, IL-14, IL-17, IL-20, IL-21, granulocyte colony-stimulating factor (G-CSF), leukemia inhibitory factor (LIF), oncokinase M (OSM), CD154, lymphotoxin (LT) β, 4-IBB ligand (4-1BBL), proliferation-inducing ligand (APRIL), CD70, CD153, CD178, glucocorticoid-induced TNFR-associated ligand (GITRL), tumor necrosis factor superfamily member 14 (TNFSF14), OX40L, and TNF and ApoL-associated leukocyte expression ligand 1. (TALL-1) or TNF-associated apoptosis-inducing ligand (TRAIL).

[0087] "Immune response," as understood in the art, and generally refers to a biological response within a vertebrate to foreign agents or abnormalities such as cancerous cells, which protects the organism against these agents and the diseases they cause. An immune response is mediated by the action of one or more cells of the immune system (e.g., T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells, or neutrophils) and soluble macromolecules (including antibodies, cytokines, and complement) produced by any of these cells or the liver, resulting in the selective targeting, binding, damage, destruction, or elimination of invading pathogens, pathogen-infected cells or tissues, cancerous or other abnormal cells, or (in cases of autoimmune or pathological inflammation) normal human cells or tissues and / or their elimination from the vertebrate body. Immune responses include, for example, T cells (e.g., effector T cells, Th cells, CD4+). + Cells or CD8 + The activation or suppression of T cells (or Treg cells), or any other cell of the immune system (e.g., NK cells). In some respects, an immune response refers to NK cell-mediated killing of foreign cells, such as allogeneic T cell therapy.

[0088] "Immunotherapy" refers to the treatment of a subject who has a disease or is at risk of infection or disease recurrence by means of methods including inducing, enhancing, suppressing or otherwise altering the immune system or immune response.

[0089] As used herein, the terms “inactivating” or “inactivation,” for example, when referring to a gene or protein, refer to measures that induce a reduction in protein expression. In some aspects, inactivation can be achieved by deleting or mutating all or part of the coding region of a gene or all or part of the non-coding region of a gene, thereby resulting in a reduction in the expression of the gene or the protein encoded by the gene. In some aspects, inactivation is achieved by the deletion of the entire coding region of a gene. In some aspects, inactivation is achieved by the partial deletion of the coding region of a gene. In some aspects, inactivation is achieved by the deletion of one or more regulatory elements that promote gene expression. In some aspects, inactivation is achieved by mutations in one or more regulatory elements that result in a reduction or loss of gene expression. In some aspects, inactivation is achieved by one or more nucleic acid mutations that result in the expression of a nonfunctional protein. In some aspects, inactivation is achieved by missense mutations that result in the expression of a nonfunctional protein. In some aspects, inactivation is achieved by gene transcriptional or translational interference that results in a reduction in protein expression. In some aspects, the reduction in expression is relative to the expression of the target gene in the cell prior to modification (e.g., deletion or mutation). In some cases, gene expression is measured before modification, then the cells are modified, and then gene expression is measured after modification.

[0090] As used herein, the term "lymphocyte" includes natural killer (NK) cells, T cells, or B cells. NK cells are a type of cytotoxic lymphocyte that represents a major component of the innate immune system. NK cells eliminate tumor cells and virus-infected cells by inducing apoptosis or programmed cell death in target cells. They are called "natural killers" because NK cells do not require activation to kill target cells. T cells play a major role in cell-mediated immunity. The T cell receptor (TCR) expressed on the surface of T cells distinguishes them from other lymphocyte types. The thymus is a specialized organ of the immune system, primarily responsible for the maturation of T cells. There are six types of T cells: helper T cells (e.g., CD4+ cells); cytotoxic T cells (also known as TC, cytotoxic T lymphocytes, CTL, T killer cells, cytolytic T cells, CD8+ T cells, or killer T cells); memory T cells (i) stem memory TCM cells, which, like immature cells, are CD45RO-, CCR7+, CD45RA+, CD62L+ (i) L-selectin, CD27+, CD28+ and IL-7Ra+, but they also express large amounts of CD95, IL-2R.p, CXCR3 and LFA-1 and exhibit many functional properties unique to memory cells); (ii) central memory TCM cells express L-selectin and CCR7, they secrete IL-2 but do not secrete IFNy or IL-4; and (iii) effector memory TEM cells, however, do not express L-selectin or CCR7 but produce effector cytokines such as IFNy and IL-4; regulatory T cells (Treg, suppressor T cells or CD4+CD25+ regulatory T cells); natural killer T cells (NKT); and γδ T cells.

[0091] B cells play a major role in humoral immunity (with the participation of antibodies). B cells produce antibodies and antigens, and act as antigen-presenting cells (APCs), transforming into memory B cells upon activation by antigen-antigen interactions. In mammals, immature B cells form in the bone marrow, from which they derive their name.

[0092] Regulatory T cells (Tregs) are specialized subsets of T cells that suppress immune responses, thereby maintaining homeostasis and self-tolerance. Studies have shown that Tregs can suppress T cell proliferation and cytokine production and play a crucial role in preventing autoimmunity. Different Treg cell subsets exist with various functions. Tregs are typically identified by flow cytometry. The most specific marker for these cells is FoxP3, which is located intracellularly. Selected surface markers, such as CD25high (high molecular weight) and CD127low (low molecular weight), can be used as surrogate markers for detecting Tregs in routine clinical practice. Dysregulation of Treg cell frequency or function can contribute to the development of autoimmune diseases.

[0093] As used herein, "pharmaceutically acceptable carriers" include any and all aqueous solvents (e.g., water, alcohol / aqueous solutions, saline solutions, parenteral media such as sodium chloride, Ringer's glucose, etc.), non-aqueous solvents (e.g., propylene glycol, polyethylene glycol, vegetable oils, and injectable organic esters such as ethyl oleate), dispersion media, polymers, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial or antifungal agents, antioxidants, chelating agents, and inert gases), isotonic agents, absorption delay agents, salts, pharmaceuticals, pharmaceutical stabilizers, gels, binders, excipients, disintegrants, lubricants, sweeteners, flavorings, dyes, liquids, and nutritional supplements, as well as similar materials and combinations thereof, as known to those skilled in the art. The pH and precise concentration of the various components in a pharmaceutical composition are adjusted according to well-known parameters.

[0094] Pharmaceutically acceptable carriers can comprise one polymer, multiple polymers, or mixtures of polymers. An exemplary polymer mixture is Eudrgit® (Evonik Industries AG, Essen, Germany). EUDRAGIT® polymers are easily handled and processed at any scale and are available in a variety of forms, such as aqueous dispersions, granules, organic solutions, powders, or ready-to-use powders. EUDRAGIT® polymers are compatible with all relevant process technologies, including film coating, melt, wet or dry granulation, hot melt extrusion, microencapsulation, and spray drying. Furthermore, all our polymers are manufactured to consistently high quality in well-established, audited facilities, helping to ensure global supply security. EUDRAGIT® polymers can be used alone or in combination to match virtually any target release profile, including immediate release, delayed release, sustained release, pulsatile release, accelerated release, and zero-order release. Standard options for each coating layer include using a single EUDRAGIT® polymer, or multiple EUDRAGIT® polymers or combinations with other polymers, as well as certain other oral excipients and pharmaceutical substances. Evonik's proprietary AEMP® (Advanced Excipient Manufacturing Process) technology can also be used to combine the functional benefits of different EUDRAGIT® polymers to create new combinatorial polymers that can further improve functionality and create new opportunities in formulation development and drug product design. Pharmaceutically acceptable carriers can be designed for immediate, delayed, or sustained release.

[0095] As used herein, the term “pharmaceutical acceptable” specifically indicates that a “pharmaceutical acceptable” compound or “pharmaceutical acceptable” composition is suitable for administration to a subject to achieve treatment and / or prevention of a disease, condition or symptom, particularly at least one CTLA-4 checkpoint-related immunodeficiency.

[0096] The pharmaceutical compositions of the present invention may be in solid or liquid form and may be, in particular, in the form of one or more powders, one or more tablets, one or more liquids (especially one or more solutions) or one or more aerosols. The pharmaceutical compositions of the present invention may also contain one or more additional bioactive agents, such as active agents, such as 3-IAld, for the treatment and / or prevention of at least one CTLA-4 checkpoint-related immunodeficiency. Administration of the pharmaceutical compositions of the present invention may be, for example, administration selected from the group consisting of: intraperitoneal, intravenous, parenteral, intrarenal, subcutaneous, topical, intrabronchial, intrapulmonary, and intranasal administration, as well as intralesional administration (if local treatment is desired). Enteric administration may be, for example, oral administration or other means of delivering 3-IAld to the intestines and gastrointestinal tract. The compositions of the present invention may also be applied directly to target sites, for example, by biological ballistic delivery to target sites, such as specific organs affected by disease, symptom, or condition.

[0097] Specifically, the administration can be carried out by injection and / or infusion and / or delivery, such as intravenous or intraperitoneal injection or infusion. The pharmaceutical composition may be in the form of an injectable dosage form or a dosage form for administration by infusion, particularly in the form of an injectable dosage form for intravenous or intraperitoneal injection or an infusion dosage form for intravenous or intraperitoneal administration, or in the form of a solid dosage form (such as enteral delivery by oral administration, such as by tablets, capsules or other solid or semi-solid dosage forms, such as dry chewable or aqueous gel or soft gum or gummies).

[0098] The pharmaceutical composition according to the invention can be administered to a subject in a suitable dose. The dosage regimen can be determined, for example, by the attending physician. As is well known in the art, the dosage for a patient can depend on a variety of factors, such as the patient's body size, body surface area, age, weight, whether the administration is for prophylactic or therapeutic purposes, the target indication, the specific compound to be administered, general health condition, and other concurrently administered medications. According to one embodiment, at least one antibody of the invention...

[0099] According to one aspect, the pharmaceutical composition of the present invention may be a pharmaceutical composition comprising 3-IAld and optionally at least one pharmaceutically acceptable carrier.

[0100] Furthermore, doses of 3-IAld and optionally at least one pharmaceutically acceptable carrier of the present invention may be administered at levels below or above the exemplary range indicated above, for example, to treat and / or prevent at least one CTLA-4 checkpoint-related immune deficiency. The pharmaceutical compositions of the present invention can be formulated as short-acting, rapid-release, long-acting, or sustained-release.

[0101] Furthermore, depending on the intended use of the pharmaceutical composition, the pharmaceutical compositions of the present invention may contain additional bioactive agents.

[0102] As used herein, the terms “decreased expression” and “increased expression” refer to the expression of a specific gene or protein in a cell relative to a control, for example, the expression of a specific gene in modified cells compared to the expression of that gene in wild-type (unmodified) cells. Relative expression can be based on mRNA and / or protein levels. Any means of measuring mRNA and / or protein levels can be used to determine whether gene or protein expression is decreased or increased, including but not limited to immunohistochemistry and PCR-based techniques.

[0103] As used herein, the terms “subject” and “patient” are used interchangeably and refer to humans or non-humans, such as primates, mammals, and vertebrates. In a particular context, a subject is a human.

[0104] As used herein, the terms "beneficial therapeutic effect" or "effective therapeutic effect" refer to anything that promotes or enhances the health of the subject in relation to medical treatment for the condition. This includes, but is not limited to, a reduction in the frequency or severity of the signs or symptoms of the disease.

[0105] The terms "effective amount," "therapeutic dose," or "therapeutic effective dose" refer to a drug dose or concentration that produces a positive biological response. For the purposes of this disclosure, one aspect of an effective amount can produce reduced symptoms of inflammation, infiltration, weight loss, irritation, diarrhea, hematochezia, mucus in stool, and tissue and cell damage.

[0106] As used herein, the term “treating” or “treatment” for a disease or condition refers to the implementation of a program that may include administering one or more therapies to a patient to alleviate the signs or symptoms of the disease. In some respects, treatment slows the rate of disease progression, improves or alleviates the disease state, and / or promotes remission or improved prognosis. Alleviation can occur before or after the signs or symptoms of the disease or condition appear. Therefore, in some respects, “treating” or “treatment” includes “preventing” or “prevention” of the disease or undesirable condition. However, “treating” or “treatment” does not necessarily require complete relief of all signs and / or symptoms, does not require a cure, and particularly includes programs that have only a minor effect on the patient.

[0107] In various aspects, it can be used to treat subjects in need for the treatment of diseases associated with CTLA-4 checkpoint-associated immunodeficiency or to alleviate their symptoms.

[0108] As used in this article, the terms “ug” and “uM” are used interchangeably with “μg” and “μM”, respectively.

[0109] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd edition, 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd edition, 1999, Academic Press; and the Oxford Dictionary of Biochemistry and Molecular Biology, Revised, 2000, Oxford University Press provide a general dictionary of many terms used in this disclosure for those skilled in the art.

[0110] Units, prefixes, and symbols are represented in their form accepted by the International System of Units (SI). Numerical ranges include the numbers that define the range. As stated herein, unless otherwise indicated, any concentration range, percentage range, ratio range, or integer range shall be understood to include any integer within the listed range and (where appropriate) its fractions (such as tenths and hundredths of an integer).

[0111] The abbreviations used herein are defined throughout this disclosure. Various aspects of this disclosure are further elaborated in the following sections.

[0112] The various aspects described in this article are described in more detail in the following sections.

[0113] II. The compositions disclosed herein Some aspects of this disclosure relate to 3-IAld. Other aspects of this disclosure relate to a pharmaceutical composition comprising 3-IAld and a pharmaceutically acceptable carrier.

[0114] 1H-Indole-3-carboxaldehyde, also known as indole-3-aldehyde or 3-formylindole (3-IAld, molecular formula: C9H7NO, IUPAC name: 1H-Indole-3-carboxaldehyde), belongs to the class of organic compounds known as indole. Indole is a compound containing an indole moiety, consisting of a pyrrole ring formed by fusion with benzene to form 2,3-benzopyrrole. Figure 1 1H-Indole-3-carboxaldehyde exists as a solid, slightly soluble in water, and very weakly acidic compound (based on its pKa). 3-IAld is a metabolite of dietary L-tryptophan, synthesized by human gastrointestinal bacteria, particularly species of the genus *Lactobacillus*. (Zelante et al. , 2013 - Lamas et al. , 2016 ).

[0115] 3-IAld is an agonist of the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor involved in a wide range of physiological activities, including maintaining mucosal homeostasis at barrier organs (Z). elante et al. , 2013 - Zhang et al. , 2016 - Stockinger et al. , 2014 ).

[0116] AhR is a ligand-dependent basic helical-loop-helical transcription factor that is highly conserved evolutionarily and expressed in most immune cell types and human tissues (Stockinger et al., 2014). Traditionally, AhR has been considered to metabolize harmful toxins by activating cytochrome P450 drug-metabolizing enzymes, but its functional diversity, including developmental biology and interactions with the microbiome, is increasingly recognized for its role in modulating host immunity, tolerance, and metabolism. Specifically, AhR's ability to bind to Th17 cells to exert antimicrobial activity, induce IL-22 production to repair and protect epithelial cells, and activate regulatory T cells to control inflammation makes the gut and respiratory barriers highly sensitive to AhR activity and activation. Esser et al. , 2015 Therefore, AhR ligands (such as 3-IAlD) are promising compounds for drug development to treat inflammatory pathologies at the mucosal surface. Several studies have highlighted AhR's ability to respond to indole and indole metabolites, thus positioning AhR as a candidate indole receptor. Hubbard et al. , 2015a Indole represents a large group of compounds derived from gut bacteria and produced from tryptophan, which exert significant biological effects and may contribute to the pathogenesis of cardiovascular, metabolic, and mental illnesses. Konopelski et al. , 2018 However, most research in this field has been limited to experimental studies, likely due to AhR's environment- and ligand-dependent activity. (Safe et al. , 2020Therefore, activating AhR via 3-IAld holds immense therapeutic potential, provided that ligands with optimal efficacy / safety profiles (such as 3-IAld) are precisely delivered to the target organ through appropriate biopharmaceutical formulations. This appears to be the case with 3-IAld, which has been formulated to improve inflammatory histopathology and barrier function through release in the gut via intestinal microparticles, such as through IL-22 production, increased expression of tight junction atresia small band protein (ZO) 1, and proliferation of intestinal Lrg5+ cells (containing leucine-rich G protein-coupled receptor 5, a stem cell marker in intestinal crypts). Kumar et al. , 2014 ), reduction of intestinal leakage, and Nfil3 (a transcription factor that guides the development of intrinsic lymphocytes 3, known to maintain epithelial barrier integrity). Puccetti et al. , 2021 The expression of )) Yu et al. , 2014 What is revealed by )

[0117] Some aspects of this disclosure relate to 1-methylindole-3-carboxylic acid. Other aspects of this disclosure relate to a pharmaceutical composition comprising 1-methylindole-3-carboxylic acid and a pharmaceutically acceptable carrier.

[0118] 1-Methylindole-3-carboxylic acid, also known as 1-methyl-3-indolecarboxylic acid or 1-methyl-1H-indole-3-carboxylic acid (MECA) (1ME3CA) (molecular formula: C10H9NO2, IUPAC name: 1-methyl-1H-indole-3-carboxylic acid), belongs to the class of organic compounds called methylindole. 1-Methylindole-3-carboxylic acid contains a methylindole moiety, which consists of a 1-methylpyrrole ring that fused with benzene to form 1-methylindole. Figure 16 ).

[0119] III. Treatment Methods Some aspects of this disclosure relate to methods for treating a disease or symptom in a subject in need, the methods comprising administering to the subject the compositions disclosed herein. In some aspects, the methods comprise administering 3-IAld alone or in combination with a pharmaceutically acceptable carrier. In one aspect, the invention relates to methods for using indole-3-aldehyde (3-IAld) to prevent and / or treat inflammation and other symptoms associated with patients suffering from CTLA-4 checkpoint-associated immunodeficiency. In another aspect, the invention relates to methods for using AhR ligands with biosimilar activity to prevent and / or treat inflammation associated with patients suffering from CTLA-4 checkpoint-associated immunodeficiency, increase the intestinal health and gastrointestinal lining health of said patients, reduce cellular infiltration of their intestinal lining, and other gastrointestinal, stomach, and intestinal symptoms. In some aspects, the AhR ligand is indole-3-acetaldehyde (IAAld), indole-3-acetic acid (IAA, indoleacetic acid), or indole-3-lactic acid (ILA, indolelactic acid).

[0120] In some aspects, the method includes administering 1-methylindole-3-carboxylic acid alone or in combination with a pharmaceutically acceptable carrier. In one aspect, the invention relates to a method for using 1-methylindole-3-carboxylic acid to prevent and / or treat inflammation and other symptoms associated with patients suffering from CTLA-4 checkpoint-associated immunodeficiency. In another aspect, the invention relates to a method for using an AhR ligand with biosimilar activity to prevent and / or treat inflammation associated with patients suffering from CTLA-4 checkpoint-associated immunodeficiency, increase the intestinal health and gastrointestinal lining health of said patients, reduce cellular infiltration of their intestinal lining, and other gastrointestinal, stomach, and intestinal symptoms. In some aspects, the AhR ligand is 1-methylindole-3-carboxylic acid.

[0121] In some aspects, the disease or symptom includes CTLA-4 checkpoint-associated immunodeficiency, characterized by a variable combination of enteropathy, hypogammaglobulinemia, recurrent respiratory infections, granulomatous lymphocytic interstitial lung disease, lymphocytic infiltration of non-lymphatic organs (intestine, lung, brain, bone marrow, kidney), autoimmune thrombocytopenic purpura or neutropenia, autoimmune hemolytic anemia, and lymphadenopathy. CTLA-4 checkpoint-associated immunodeficiency is diagnosed based on clinical symptoms, laboratory findings, and genetic testing. Patients with only one functional copy of the gene encoding CTLA-4 suffer from severe autoimmunity. These heterozygous mutations result in a novel phenotype with overactive T and B cell infiltration of non-lymphatic organs (such as the intestine, lung, and brain) and more typical autoimmune signs. Some clinical manifestations of CTLA-4 haploinemia are similar to those observed in biopsies of inflamed organs. Patients with primary CTLA-4 checkpoint-related immunodeficiency may have recurrent respiratory infections, hypogammaglobulinemia, autoimmune cytopenia, autoimmune enteropathy, and granulomatous infiltrative lung disease.

[0122] One example of CTLA-4 checkpoint-related immunodeficiency is CTLA-4 haploinemia with autoimmune infiltration (CHAI), a CTLA-4 checkpoint-related immunodeficiency disorder characterized by heterozygous CTLA4 mutations in humans associated with severe immunomodulatory disorders. Another example of CTLA-4 checkpoint-related immunodeficiency is lipopolysaccharide-responsive beige-like anchoring protein (LRBA), a disorder characterized by a genetically inherited biallelic mutation carried by the patient. Because LRBA deficiency also leads to secondary loss of CTLA-4, this recessive disorder is known as LRBA deficiency with autoantibodies, T-reg cell deficiency, autoimmune infiltration, and enteropathy (LATAIE).

[0123] Description of asymptomatic adults with heterozygous CTLA4 deficiency (Kuehn et al. , 2014 - Schubert et al. , 2014 The wide variation in onset age among symptomatic individuals suggests that additional interacting factors are required to overcome the autoimmune threshold. These factors may include other genetic or epigenetic events and environmental influences (microbes or others). Exposure to the microbiome in some frequently affected organs (skin and gut) suggests that such environmental factors may promote the development of autoimmunity in patients receiving the CTLA4 blocker ipilimumab. Interestingly, even considering… CTLA4 - / - Mice reintroduced into a sterile environment also died within 2 weeks of age, suggesting the importance of self-antigens in driving inflammatory phenotypes. Tivol et al. , 1995 ).

[0124] Microbial indoles are very attractive metabiotics because they have been shown to extend the healthy lifespan of a wide variety of evolutionarily diverse species from different phyla. (Descamps et al. , 2019 The gastrointestinal tract carries many species capable of synthesizing indole and indole-containing compounds, which may be associated with dysbiosis by affecting host immune reactivity, epithelial barrier function, and pathogen colonization. Roager et al. , 2018 By acting as a ligand for AhR (a transcription factor that controls the biodegradation of endogenous and exogenous toxins), it prevents inflammatory damage and provides barrier integrity. Hubbard et al. , 2015 b Indole has multiple functions, including bidirectional communication with the microbiome to fine-tune host immunity, tolerance, and metabolism. Stockinger, 2014 Previous studies have shown that administration of indole to germ-free mice increases the expression of epithelial tight junction proteins and attenuates markers of inflammatory colitis. (Shimada et al. , 2013) Similarly, the therapeutic efficacy of 3-IAld in a mouse model of dextran-induced colitis has been reported. Zelante et al. , 2013 ).

[0125] By activating type 3 innate lymphocytes to produce IL-22, 3-IAld enhances barrier integrity and antimicrobial peptide production in mouse models of colitis, gastrointestinal and vaginal candidiasis. Zelante et al. , 2013; Borghi et al. , 2019; Puccetti et al. , 2021 It has also been shown that treatment with 3-IAld limits intestinal epithelial damage, reduces transepithelial bacterial translocation, and decreases the production of inflammatory cytokines in a mouse model of graft-versus-host disease (GvHD), a systemic inflammatory state induced by donor T cells leading to colitis. 3-IAld treatment also induced recipient strain-specific tolerance to transplanted T cells. Transcriptional profiling and gene ontology analysis revealed that 3-IAld administration upregulated genes associated with type I interferon responses known to provide protection against radiation-induced intestinal damage. (Swimm et al. , 2018 Therefore, by acting via different downstream effector pathways, 3-IAld can limit intestinal inflammation and damage, and can provide a treatment option for patients at risk of gastrointestinal inflammatory damage. This opens the possibility of developing microbiome-based therapeutic agents of indole or its derivatives to promote human epithelial barrier function.

[0126] Given the similarity in mechanism of action between CTLA-4 checkpoint-associated immunodeficiency and anti-CTLA-4 therapy ( Bakacs et al. , 2015 This suggests that 3-IAld has therapeutic potential not only in CTLA-4 checkpoint-related immunodeficiency, but also in the prevention of adverse immune events (including colitis and enteritis) associated with cancer treatment using immune checkpoint inhibitors. Karamchandani et al. , 2018; Marin-Acevedo et al. , 2018 Despite the tremendous success of cancer immunotherapy using immune checkpoint inhibitors, its beneficial therapeutic effects are hampered by various resistance mechanisms. Schoenfeld et al. , 2020 Limitations on the effects of gastrointestinal, endocrine, and skin toxicities, as well as fatal neurotoxicity and cardiotoxicity. Choi et al. , 2020 Therefore, novel treatment strategies that provide manageable side effects for existing immunotherapies will enhance and expand their efficacy and application. Studies have shown that the effectiveness of immunotherapy against various tumors requires the presence of symbiotic bacteria. Iida et al. , 2013 Clinical studies have corroborated these findings with compelling evidence that the richness and diversity of the gut microbiota are associated with durable responses to immunotherapy, and that gut microbiota characteristics predict toxicity associated with combined CTLA-4 and PD-1 blockade. Andrews et al. , 2021 ).

[0127] Another example of CTLA-4 checkpoint-related immunodeficiency is regulatory T (Treg) cell deficiency, which refers to any defective immune surveillance response that leads to reduced elimination of cells infected with chronic viruses and oncoviruses, resulting in dysregulation of cell growth. One example is decreased CTLA-4 expression, which leads to uncontrolled T cell proliferation and may result in an overgrowth of self-reactive clones relative to, for example, EBV-specific T cell clones.

[0128] Another example of CTLA-4 checkpoint-related immunodeficiency is enteropathy, which refers to persistent damage, irritation, and swelling of the small intestine. Another example of CTLA-4 checkpoint-related immunodeficiency is intestinal inflammation, which refers to inflammation of the gastrointestinal (GI) tract. Chronic inflammation leads to gastrointestinal damage.

[0129] Another example of CTLA-4 checkpoint-related immunodeficiency is immune-mediated colitis (IMC), a common immune-related adverse event associated with immune checkpoint inhibitors, including abdominal pain, mucus or blood in the stool, and fever. Immune checkpoint inhibitors are immunotherapeutic drugs that work by blocking the binding of checkpoint proteins to their chaperone proteins. This prevents the "shutdown" signal from being emitted, thus allowing T cells to kill cancer cells. Dysregulation of T cells can also contribute to IMC, in which T cells attack gastrointestinal cells and tissues.

[0130] Therapeutic compositions (e.g., 3-IAld) can be administered based on the weight or mass of the individual to whom the therapeutic composition is administered. Weight-adjusted doses of the therapeutic composition can be administered from about 1 mg / kg to about 25 mg / kg. Weight-adjusted doses of the therapeutic composition can be administered from about 3 mg / kg to about 25 mg / kg, from about 10 mg / kg to about 25 mg / kg, from about 15 mg / kg to about 20 mg / kg, or from about 15 mg / kg to about 18 mg / kg. The dosage of 3-IAld and / or the 3-IAld pharmaceutical composition can be delivered in the form of an immediate-release, delayed-release, or sustained-release formulation.

[0131] Therapeutic compositions (e.g., 1-methylindole-3-carboxylic acid) can be administered based on the weight or mass of the individual to whom the therapeutic composition is administered. Weight-adjusted doses of the therapeutic composition can be administered from about 1 mg / kg to about 25 mg / kg. Weight-adjusted doses of the therapeutic composition can be administered from about 3 mg / kg to about 25 mg / kg, from about 10 mg / kg to about 25 mg / kg, from about 15 mg / kg to about 20 mg / kg, or from about 15 mg / kg to about 18 mg / kg. The dosage of 1-methylindole-3-carboxylic acid and / or the 1-methylindole-3-carboxylic acid pharmaceutical composition can be delivered in the form of an immediately released, delayed-release, or sustained-release formulation.

[0132] IV. Pharmaceutical Compositions The compounds of the present invention can be incorporated into pharmaceutical compositions suitable for administration to a subject. Such compositions typically contain an active compound and a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, isotonic agents, and absorption delay agents that are compatible with drug administration. The use of such media and agents for pharmaceutically active substances is well known in the art. Unless any conventional media or agent is incompatible with the active compound, its use in the composition is contemplated. Complementary active compounds (such as those described above) may also be incorporated into the composition.

[0133] The pharmaceutical compositions of the present invention are formulated to be compatible with their intended route of administration. Examples of routes of administration include parenteral (e.g., intravenous), intradermal, subcutaneous, oral (e.g., swallowing), transdermal (topical), mucosal, and rectal administration. In one aspect, the active compound is prepared together with a carrier that will prevent rapid elimination of the compound from the body, such as a controlled-release formulation comprising implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Methods for preparing such formulations will be apparent to those skilled in the art.

[0134] The pharmaceutical composition may be included in a container, package, or dispenser along with the instructions for use.

[0135] Exemplary aspects provided in this article In one aspect (Aspect 1; A1), this article provides a method for treating a patient with CTLA-4 checkpoint-related immunodeficiency, the method comprising administering to the patient in need a therapeutically effective amount of a pharmaceutical composition comprising 1H-indole-3-carboxaldehyde (3-IAld).

[0136] In one aspect of A1, namely A2, the CTLA-4 checkpoint-related immunodeficiency is selected from the group consisting of: CTLA-4 haploid deficiency with autoimmune infiltration (CHAI), lipopolysaccharide-responsive beige-like anchoring protein (LRBA) deficiency (LATAIE), regulatory T (Treg) cell deficiency, autoimmune infiltration, enteropathy, intestinal inflammation, immune-mediated colitis, gastrointestinal disorders, and gastric atrophy.

[0137] In one aspect of A1 or A2, namely A3, the CTLA-4 checkpoint-associated immunodeficiency is immune-mediated colitis.

[0138] In any one of A1 to A3, namely A4, the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

[0139] In one aspect of A4, namely A5, the pharmaceutically acceptable carrier is at least one polymer.

[0140] In one aspect of A4, namely A6, the pharmaceutically acceptable carrier is a group of polymers.

[0141] In one aspect of A6, namely A7, the polymer of the group is Eudragit® polymer.

[0142] In any one of A1 to A7, namely A8, the pharmaceutical composition is formulated for enteral delivery.

[0143] In any one of A1 to A8, namely A9, the pharmaceutical composition is administered orally.

[0144] In any one aspect of A1 to A9, namely A10, the pharmaceutical composition is in the form selected from capsules, tablets, gel sheets, gel capsules, gels, liquids and gummies.

[0145] In any aspect of A10, namely A11, the pharmaceutical composition is in the form of tablets or capsules.

[0146] In any one of A1 to A11, namely A12, the pharmaceutical composition is administered at an interval of every other day (qod).

[0147] In any one aspect of A1 to A12, namely A13, the pharmaceutical composition is administered at a dose of at least about 3 mg / kg, at least about 4 mg / kg, at least about 5 mg / kg, at least about 6 mg / kg, at least about 7 mg / kg, at least about 8 mg / kg, at least about 9 mg / kg, at least about 10 mg / kg, at least about 11 mg / kg, at least about 12 mg / kg, at least about 13 mg / kg, at least about 14 mg / kg, at least about 15 mg / kg, at least about 16 mg / kg, at least about 17 mg / kg, or at least about 18 mg / kg of 3-IAld.

[0148] In one aspect of A13, namely A14, the 3-IAld dose is approximately 18 mg / kg.

[0149] In one aspect, namely A15, this article provides a method for treating a patient with CTLA-4 checkpoint-related immunodeficiency, the method comprising administering to the patient in need a therapeutically effective amount of a pharmaceutical composition comprising 1-methylindole-3-carboxylic acid.

[0150] In one aspect of A15, namely A16, the CTLA-4 checkpoint-related immunodeficiency is selected from the group consisting of: CTLA-4 haploid deficiency with autoimmune infiltration (CHAI), lipopolysaccharide-responsive beige-like anchoring protein (LRBA) deficiency (LATAIE), regulatory T (Treg) cell deficiency, autoimmune infiltration, enteropathy, intestinal inflammation, immune-mediated colitis, gastrointestinal disorders, and gastric atrophy.

[0151] In one aspect of A15 or A16, namely A17, the CTLA-4 checkpoint-associated immunodeficiency is immune-mediated colitis.

[0152] In any one aspect of A15 to A17, namely A18, the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

[0153] In one aspect of A18, namely A19, the pharmaceutically acceptable carrier is at least one polymer.

[0154] In one aspect of A18, namely A20, the pharmaceutically acceptable carrier is a group of polymers.

[0155] In one aspect of A20, namely A21, the polymer of the group is Eudragit® polymer.

[0156] In any one aspect of A15 to A21, namely A22, the pharmaceutical composition is formulated for enteral delivery.

[0157] In any one aspect of A15 to A22, namely A23, the pharmaceutical composition is administered orally.

[0158] In any one aspect of A15 to A23, namely A24, the pharmaceutical composition is in the form selected from capsules, tablets, gel sheets, gel capsules, gels, liquids and gummies.

[0159] In one aspect of A24, namely A25, the pharmaceutical composition is in the form of tablets or capsules.

[0160] In any one of A15 to A25, namely A26, the pharmaceutical composition is administered at an interval of every other day (qod).

[0161] In any one aspect of A15 to A23, namely A27, the pharmaceutical composition is administered at a dose of 1-methylindole-3-carboxylic acid of at least about 2 mg / kg, at least 3 mg / kg, at least about 4 mg / kg, at least about 5 mg / kg, at least about 6 mg / kg, at least about 7 mg / kg, at least about 8 mg / kg, at least about 9 mg / kg, at least about 10 mg / kg, at least about 11 mg / kg, at least about 12 mg / kg, at least about 13 mg / kg, at least about 14 mg / kg, at least about 15 mg / kg, at least about 16 mg / kg, at least about 17 mg / kg, or at least about 18 mg / kg.

[0162] In any one aspect of A15 to A23, namely A28, the pharmaceutical composition is administered at a dose of about 2.25 mg / kg of 1-methylindole-3-carboxylic acid.

[0163] The invention is further illustrated by the following embodiments, which should not be construed as further limitations. All references cited throughout this application are expressly incorporated herein by reference.

[0164] Unless otherwise indicated, the practice of this disclosure will employ conventional techniques of cell biology, cell culture, molecular biology, transgenic biology, microbiology, recombinant DNA, and immunology that are within the skill of those skilled in the art. Such techniques are described in detail in the literature. See, for example, Sambrook et al., eds. (1989) Molecular Cloning: A Laboratory Manual (2nd ed.; Cold Spring Harbor Laboratory Press); Sambrook et al., eds. (1992) Molecular Cloning: A Laboratory Manual, (Cold Springs Harbor Laboratory, NY); DN Glover, ed. (1985) DNA Cloning, Volumes I and II; Gait, ed. (1984) Oligonucleotide Synthesis; Mullis et al., U.S. Patent No. 4,683,195; Hames and Higgins, eds. (1984) Nucleic Acid Hybridization; Hames and Higgins, eds. (1984) Transcription and Translation; Freshney (1987) Culture of Animal Cells (Alan R. Liss, Inc.); Immobilized Cells and Enzymes (IRL Press) (1986); Perbal (1984) A Practical Guide to Molecular Cloning; the treatise, Methods In Enzymology (AcademicPress, Inc., NY); Miller and Calos, eds. (1987) Gene Transfer Vectors For Mammalian Cells, (Cold Spring Harbor Laboratory); Wu et al., eds., Methods In Enzymology, Vol. 154 and 155; Mayer and Walker, eds. (1987) Immunochemical Methods In Cell And Molecular Biology (Academic Press, London); Weir and Blackwell, eds. (1986) Handbook Of Experimental Immunology, Vol. I-IV; Manipulating the Mouse Embryo, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, (1986); Crooks, Antisensedrug Technology: Principles, strategies and applications, 2nd ed., CRC Press (2007); and Ausubel et al. (1989) Current Protocols in Molecular Biology (John Wiley and Sons, Baltimore, Md.).

[0165] All references cited above, as well as all references cited in this paper, are incorporated into this paper in their entirety through citation.

[0166] The following examples are provided by way of illustration rather than limitation.

[0167] Example Example 1 3-IAld prevents intestinal inflammatory pathology in a mouse model of dextran sulfate sodium (DSS) + anti-CTLA-4 induced colitis. Given the similarity in mechanism of action between CTLA-4 checkpoint-associated immunodeficiency and anti-CTLA-4 therapy ( Bakacs et al. , 2015 It is hypothesized that 3-IAld may have therapeutic potential not only in CTLA-4 checkpoint-related immunodeficiency, but also in the prevention of adverse immune events associated with cancer treatment using immune checkpoint inhibitors, including colitis and enteritis such as enteropathy or gastroenteritis. Karamchandani et al. , 2018; Marin-Acevedoet al. , 2018 ). Pathology caused by CTLA4 blockade is known in mouse models of colitis (e.g., C57BL / 6 mice). Scott et al. , 2020 ) The pathogenic role of hereditary human CTLA-4 haploid deficiency in the gut ( Bakacs et al. , 2015 ).

[0168] In the experiment, C57BL / 6 mice received sodium dextran sulfate (DSS) (3%) in drinking water and were intraperitoneally administered 100 µg of anti-CTLA-4 monoclonal antibody (mAb) or isotype control antibody on days 0, 4, and 8 after DSS administration. Figure 2 As indicated in A. DSS is a water-soluble, negatively charged sulfated polysaccharide with a highly variable molecular weight ranging from 5 to 1400 kDa. Sulfated polysaccharides do not directly induce intestinal inflammation, but rather act as direct chemical toxins on the colonic epithelium, leading to epithelial cell damage. (For example...) Figure 2 As shown in A, Eudragit-prepared 3-IAld was administered intragastric at a dose of 18 mg / kg every other day. Puccetti et al. , 2018 Monitor animals daily for the occurrence of diarrhea, bloody stools, weight loss, and survival. One week after DSS treatment (14 days after treatment initiation), the time it takes for the model to reproduce human disease is recorded. Manicassamy et al. , 2014 Surviving mice were sacrificed, their colons were removed, and macroscopic damage and local immune parameters were assessed. Treatment with 3-IAld improved survival rate. Figure 2 B) and weight ( Figure 2 C) and reduce the disease activity index ( Figure 1 D). Surviving mice showed improved gross pathology ( Figure 2 E) and the restoration of the following: i) the normal structure of the colon ( Figure 2 F and Figure 2 G) ii) Epithelial barrier function, as revealed by ZO-1 expression ( Figure 2 H), and iii) epithelial cell proliferation and renewal, as revealed by Ki-67 staining ( Figure 2 The results showed that 3-IAld had the ability to prevent intestinal inflammatory pathology caused by CTLA4 blockade in a mouse colitis model.

[0169] 3-IAld also promotes Lgr5 , Nfil3 and Muc1 Expression of (a cell surface mucin that serves as an infection barrier and inflammation regulator).

[0170] After confirmation Figure 2 After functional recovery in C57BL / 6 mice, 3-IAld was found to also promote Lgr5 , Nfil3 and Muc1 Expression of (a cell surface mucin used as an infection barrier and inflammatory regulator) Dhar et al. , 2019 ()( Figure 3 A). Therefore, the amount of dextran-FITC crossing the intestinal barrier is reduced ( Figure 3 B), and the level of soluble CD14 (a marker of intestinal permeability) was also reduced. Figure 3 C). These changes are accompanied by a shift towards an anti-inflammatory profile with decreased levels of TNF-α, IL-1β, and IL-17A and increased levels of IL-10. Figure 3 D). Consistent with this anti-inflammatory spectrum, the level of calprotectin was also reduced ( Figure 3 E). Considering the defect of 3-IAld in mice with colitis ( Alexeev et al. , 2018 These results indicate that 3-IAld supplementation provides protection against DSS+ anti-CTLA-4 induced colitis by maintaining epithelial barrier integrity and suppressing the inflammatory response. This activity is consistent with its AhR agonist activity. Zelante et al. , 2013 - Puccetti et al. 2021 ), such as through AhR-dependent genes Cyp1a1 and Ahrr Induced expression ( Figure 3 F), induction of antimicrobial peptide Reg3γ expression ( Figure 3 F) and, importantly, the induction of IL-22 expression ( Figure 3 G, gene and protein expression (a key mediator of mucosal functional activity in response to 3-IAld) (Renga et al. 2022) .

[0171] Example 2 3-IAld in an immune-mediated colitis model The activity of 3-IAld was evaluated in an immune-mediated colitis model, which, unlike chemically induced colitis models, was considered the best way to reproduce immune-dependent colitis (such as in patients with CTLA-4 haploid deficiency). Constant et al. , 2022 ) or patients treated with checkpoint inhibitors ( Westdorp et al. , 2021The pathological mechanism of colitis in mice. Due to the absence of CTLA-4 in mice, massive lymphocyte proliferation and fatal multi-organ tissue destruction occur, leading to early death. Tivol et al. , 1995 Therefore, an alternative reference model for immune-mediated colitis is used. Kiesler et al. , 2015 ).

[0172] Humanized immunodeficient NSG mice were injected with human peripheral blood mononuclear cells from healthy donors and treated with anti-CTLA-4 monoclonal antibody. 8-10 week old NOD.Cg-Prkdcscid Il2rgtm1Wjl / SzJ (NSG) mice were intraperitoneally injected with freshly isolated human peripheral blood mononuclear cells from healthy donors. 7 ), and intraperitoneal treatment with 100 µg anti-CTLA-4 mAb or human IgG as an antibody control was performed on days 0, 4, 8, 12 and 16.

[0173] Starting from the day of cell infusion, 3-IAld (18 mg / kg) was administered orally every other day. Overall mortality and weight loss in mice were monitored daily, and they were sacrificed on day 21. Results showed that treatment with 3-IAld improved survival rate. Figure 4 A), reducing weight loss ( Figure 4 B), improves inflammatory tissue pathology ( Figure 4 C) and histological score ( Figure 4 D), restoring barrier function (as revealed by ZO-1 expression), Figure 4 E), and reduce the expression of inflammatory cytokine genes ( Figure 4 F).

[0174] 3-IAld restricts the progression of immune-mediated colitis in RAG1-deficient mice Using a childish CD4 + T-cell regeneration of immunodeficiency Rag1 – / – Mice were treated with anti-CTLA-4 monoclonal antibody. Rag1– / – Mice, specifically those homozygous for the Rag1tm1Mom mutation, do not produce mature T cells or B cells. Their phenotype can be described as "non-leaky" immunodeficiency. Rag1– / – 8–10 week old mice were injected intraperitoneally with 4 × 10 5 Naïve CD4 purified from the spleen of C57BL / 6 mice + T cells were treated with anti-CTLA-4 mAb or control IgG on the second day after T cell regeneration and subsequently on days 0, 4, 8, 12, and 16.

[0175] Starting from the day of cell infusion, 3-IAld (18 mg / kg) was administered orally every other day. Overall mortality and weight loss were monitored daily in mice, and they were sacrificed on day 21. Similar to what was observed in NSG mice, treatment with 3-IAld reduced weight loss ( Figure 5 A) and improve gross pathology ( Figure 5 B) and colon histopathology ( Figure 5 C and Figure 5 D).

[0176] IL-10 defects ( Il10 - / - Mice were treated acutely or chronically with anti-CTLA-4 monoclonal antibody (with and without DSS).

[0177] Spontaneous onset of post-weaning intestinal inflammation (the earliest pathological changes observed in 3-week-old mice) leads to IL-10 deficiency in mice. Il10 - / - This serves to simulate the progression of inflammatory lesions through discontinuous and transmural processes, including cellular infiltrations (primarily lymphocytes) into the lamina propria and submucosa, epithelial hyperplasia, mucin depletion, ulceration, and intestinal wall thickening. Berg et al. , 1996 ; Gomes-Santos et al. , 2012 An excellent model of the pathogenesis of immune-mediated colitis (characterized by) Keubler et al. , 2015 By 10 weeks of age, Il10 - / - Mice developed mild colitis, the severity of which stabilized at 16 weeks of age. Gomes-Santos et al. , 2012 We evaluated the effects of 3-IAld on patients of different ages after acute or chronic treatment with anti-CTLA-4 mAb (with and without 1% DSS). Il10 - / - Activity in mice (i.e., 8-10 weeks or 16 weeks old). The model and results are described below.

[0178] 8-10 weeks old Il10 - / - DSS+ anti-CTLA-4 induced colitis in mice.

[0179] Il10 - / -Mice were given 1% DSS in drinking water and intraperitoneally administered 100 µg of anti-CTLA-4 monoclonal antibody on days 0, 4, and 8 after DSS administration. Throughout the experiment, 3-IAld-Eudragit (18 mg / kg 3-IAld) was administered intragastric every other day, starting 2 days prior to this treatment. Animal body weight and inflammatory pathology in the colon were monitored daily until day 14 after DSS administration. Results showed weight loss in 8–10 week old IL-10 deficient mice. Figure 6 A) and increased epithelial thickening ( Figure 6 B and Figure 6 C); Both of these characteristics were exacerbated after DSS+ anti-CTLA-4 treatment, with further weight loss and worsening colonic pathology, showing signs of epithelial destruction and transmural infiltration. 3-IAld treatment prevented weight loss ( Figure 6 A) and reduce inflammatory pathology and disease activity indices ( Figure 6 B and Figure 6 C).

[0180] 8-10 weeks old Il10 - / - Acute anti-CTLA-4 treatment in mice As mentioned above ( Figure 6 Treatment with anti-CTLA-4 monoclonal antibody alone and 3-IAld (DSS-free) Il10 - / - Mice. Animal body weight and inflammatory pathology in the colon were monitored daily, and mice were sacrificed 14 days after the first administration of anti-CTLA-4 monoclonal antibody. Results showed that treatment with anti-CTLA-4 alone reduced body weight in 8-10 week old IL-10 deficient mice compared to wild-type mice. Figure 7 A) and promotes epithelial thickening associated with extensive infiltration in the submucosa. Figure 7 B). 3-IAld treatment prevents weight loss ( Figure 7 A) and improve inflammatory pathology, particularly by reducing large amounts of cellular infiltrates ( Figure 7 B).

[0181] 16 weeks old Il10 - / - Chronic anti-CTLA-4 treatment in mice like Figure 8 As indicated in section A, treatment with 100 µg of anti-CTLA-4 monoclonal antibody alone and 3-IAld (DSS-free) Il10 - / -Mice. Animal body weight, disease activity index, clinical examination, inflammatory pathology in the ileum and colon, epithelial barrier function, and immunological parameters of inflammation were monitored daily. Results showed that long-term treatment with anti-CTLA-4 alone reduced body weight ( Figure 8 B) Disease Activity Index ( Figure 8 C), increasing clinical activity in the ileum and colon ( Figure 8 D) and histopathology ( Figure 8 E and Figure 8 F) promotes epithelial damage (as revealed by decreased ZO-1 expression) and dysfunction (as revealed by staining with bromodeoxyuridine (BrdU) (a thymidine analog that is incorporated into the DNA of dividing cells during the S phase of the cell cycle), and increases inflammatory calprotectin ( Figure 8 G) and IL-6 Figure 8 The level of H) was reduced, while the expression of the antimicrobial peptide Reg3γ was decreased. Figure 8 H). Treatment with 3-IAld effectively antagonized CTLA-4 blockade-induced cytotoxicity. This is consistent with the widespread T cell infiltration observed in various organs in CTLA-4 haploid deficiency. Kuehn et al. , 2014; Schubert et al. , 2014; Schwab et al. , 2018 We found that, during anti-CTLA-4 treatment, in the ileum and colon of mice ( Figure 9 A) Lungs and liver ( Figure 9 B) contains a wide range of CD3 + T cell infiltration, which was significantly reduced by simultaneous administration of 3-IAld, such as Figure 9 The data was quantified in the literature. No lymphocyte infiltration or immunopathology was detected in organs such as the spleen and kidneys (data not shown).

[0182] 3-IAld treatment slowed disease progression in 16-week-old IL-10-deficient mice. like Figure 10 As shown in A, 16-week-old IL-10-deficient mice were treated with 3-IAld every other day. Il10 - / - Mice were treated with 3-IAld for 4 weeks. As a mimicry of disease exacerbation, mice were given 1% DSS after 3-IAld treatment, and their weight changes, intestinal pathology, and epithelial integrity were subsequently assessed. The results showed that treatment with 3-IAld significantly slowed weight loss. Figure 10 B) and histopathology of inflammatory colon ( Figure 10 D) and maintain epithelial integrity and renewal ( Figure 10 D).

[0183] Example 3 3-IAld promotes beneficial microbiota Given the role of the AhR / IL-22 axis in maintaining microbiome homeostasis, 3-IAld was evaluated for its impact on fecal microbiome composition. Fecal microbiota transplantation (FMT) was used to assess the activity of 3-IAld-modified microbiota in colitis. Feces were collected from untreated or 3-IAld-treated mice for 6 consecutive days and transplanted into recipient mice at the onset of colitis. Unlike transfers of feces from untreated mice, FMT from 3-IAld-treated mice prevented weight loss in DSS-treated mice. Figure 11 A), and improves gross and histopathological findings of the colon ( Figure 11 B to Figure 11 D). Similar results were obtained in DSS+ anti-CTLA-4 treated mice, where FMT from 3-IAld treated mice also prevented weight loss ( Figure 11 E) and induce the production of IL-10 in regulatory T cells (Tregs), as revealed by the reversal of DNA hypermethylation at the Foxp3 promoter. Figure 11 F). In summary, these results suggest that the beneficial activities of 3-IAld can occur through various pathways, including enhancing the intestinal barrier via the AhR / IL-22 axis, altering the composition and function of the microbiome, and controlling inflammation via Treg cells.

[0184] Example 4 3-IAld does not interfere with the development of anti-tumor immunity. The potential applications of 3-IAld require that its immunomodulatory activity not affect tumor immune surveillance. For this purpose, the role of 3-IAld was evaluated in an anti-CTLA-4 reactive B16 melanoma model. Renga et al. , 2022 3-IAld neither alters tumor growth nor interferes with the therapeutic efficacy of anti-CTLA-4 antibodies. Figure 12 A to Figure 12 B), and does not affect CD4. + and CD8 + Recruitment of tumor-infiltrating lymphocytes ( Figure 12 C to Figure 12 D), which is consistent with the increased expression of leukocyte recruitment chemokine Cxcl9 and effector perforin. Figure 12 E). Similarly, in a Lewis lung carcinoma (LLC) model, 3-IAld did not interfere with the therapeutic efficacy of anti-PD-1 antibodies. In fact, 3-IAld did not prevent anti-PD-1 antibodies from improving survival rates. Figure 12 F) Reduce tumor growth ( Figure 12 G), Improve gross pathology ( Figure 12 H) and reducing lung Foxp3 + CD25 + Treg cell recruitment ( Figure 12 I arrive Figure 12 Therefore, the beneficial activity of 3-IAld does not interfere with tumor immune surveillance.

[0185] This study is a proof-of-concept demonstration of the therapeutic potential of bacterial metabolites (such as 3-IAld) as biologics capable of mitigating anti-CTLA-4-induced intestinal toxicity without interfering with tumor surveillance.

[0186] This indicates that 3-IAld is a unique molecule capable of disrupting the dynamic feedback loop of intestinal inflammation (where the tissue-destructive effects of infiltrative lymphocytes in CTLA-4 deficiency symptoms lead to microbial dysbiosis, thereby further promoting inflammation and pathology). By acting simultaneously on both the host and the microbe, 3-IAld is particularly well-suited to breaking this vicious cycle of pathogenesis and inhibiting inflammation.

[0187] Example 5 Given the similarity in the mechanisms of action between inherited human CTLA-4 haploinsufficiency and anti-CTLA-4 therapy (Bakacs et al., 2015), representative models used to demonstrate the efficacy of 3-IAld are mouse models of colitis induced by immune checkpoint inhibitors when treated with both anti-CTLA-4 and sodium dextran sulfate (DSS) (Wang et al., 2018–Wang et al., 2019; Perez Riuz et al., 2019). To further support the medical plausibility of our findings, the activity of 3-IAld was evaluated in immune-mediated colitis models, which, unlike chemically induced colitis models, are considered to best reproduce the pathogenesis of immune-dependent colitis (such as colitis occurring in patients with CTLA-4 haploinsufficiency (Constant et al., 2022) or in patients treated with checkpoint inhibitors (Westdorp et al., 2021).

[0188] Pharmacology Preliminary pharmacokinetic studies were conducted. The mode of action of 3-IAld occurs upon binding to AhR, both in vitro and in vivo. In vitro, 3-IAld induced luciferase activity in the H1L1.1c2 cell line containing stably transfected AhR-responsive firefly luciferase at doses ranging from 0.1 to 100 mM. Zelante et al. , 2013 In vivo, intragastric administration of 3-IAld to a mouse model of mucosal candidiasis and DSS-induced colitis induced IL-22 production in an AhR-dependent manner. (Zelante et al. , 2013 Preliminary pharmacokinetic data were obtained by administering 3-IAld formulated with Eudragit in a composition of 90% 3-IAld / 10% 13C8-labeled 3-IAld, and performing targeted / untargeted analysis of serum at different time points (30 min, 1 h, 2 h, 4.5 h, 6 h, 24 h) using mass spectrometry. Analysis of unlabeled 3-IAld at different time points showed minor changes compared to baseline endogenous levels. Figure 13 As shown in A. However, analysis of the labeled 3-IAld revealed that it peaked between 30 minutes and 1 hour after application, declined sharply at 2 hours, and was detected in trace amounts at subsequent time points. Figure 13 B). These data demonstrate the rapid metabolic transformation of 3-IAld. Further analysis of potential metabolites of unlabeled 3-IAld revealed that the oxidized form of 3-IAld (indole-3-carboxylic acid or Ox-3-IAld) peaked at 30 min with an average of 80 µM, rapidly decreased to 20 µM after 1 hour, and was detectable in trace amounts at subsequent time points. Figure 13 C). Lower levels of the methylated forms of indole-3-carboxylic acid, namely 1-methylindole-2-carboxylic acid and methyl indole-3-carboxylic acid, were also detected, reaching peaks of approximately 150 and 25 nM, respectively, at 30 minutes, and decreasing at 1 hour, thus following the same kinetics as indole-3-carboxylic acid. Figure 13 D). The presence of indole-3-carboxylic acid was also confirmed by labeled 3-IAld. Figure 13 E), showing similar kinetics, reached a peak level of about 25 µM at 30 min.

[0189] Further in vitro experiments showed that indole-3-carboxylic acid possesses pharmacological activity and may therefore contribute to the pharmacokinetics of 3-IAld. To demonstrate this, we exposed the human cell line HepG2 (a hepatocellular carcinoma cell line) to different concentrations of the ligands indole-3-carboxylic acid or Ox-3-IAld. Figure 14 As shown, indole-3-carboxylic acid and / or Ox-3-IAld can induce the AhR activation marker Cyp1A1 in a dose-dependent manner between 1 and 100 µM.

[0190] toxicology Continue administering the specified dose to naïve C57BL / 6 and naïve C57BL / 6 every other day for 3 weeks. AhR - / - Mice were administered 3-IAld formulated with Eudragit intragastric administration. Puccetti et al. , 2018At the end of the treatment, the mice were euthanized, and their histopathology was examined blinded in different organs after staining with hematoxylin and eosin (H&E). Figure 15 The results showed that no visible histopathology was found in any of the examined organs at both different concentrations.

[0191] It should be understood that the claims are intended to be interpreted using the detailed description section rather than the summary and abstract section. The summary and abstract section may set forth one or more, but not all, exemplary embodiments of this disclosure as contemplated by the inventors, and is therefore not intended to limit this disclosure and the appended claims in any way.

[0192] The foregoing description of specific embodiments so fully reveals the general nature of this disclosure that, without departing from the general conception of this disclosure, others can readily modify and / or adapt various applications of such specific embodiments by applying knowledge well known in the art, without much experimentation. Therefore, based on the teachings and guidance presented herein, such modifications and adaptations are intended within the meaning and scope of equivalents of the disclosed embodiments. It should be understood that the phrases or terms used herein are for descriptive rather than limiting purposes, and that the terminology or phrases in this specification should be interpreted by one of skill in the art based on the teachings and guidance.

[0193] The breadth and scope of this disclosure should not be limited by any of the above exemplary embodiments, but should be defined only by the appended claims and their equivalents.

[0194] All references cited throughout this application (including bibliographic references, U.S. or foreign patents or patent applications, and websites) are expressly incorporated herein by reference as if their entirety were included for any purpose. In the event of any inconsistency, the material disclosed herein shall prevail.

[0195] While various specific aspects have been shown and described, the foregoing description is not restrictive. It should be understood that various changes can be made without departing from the spirit and scope of the invention. Many changes will become apparent to those skilled in the art upon reading this description.

[0196] Example 6 1-Methylindole-3-carboxylic acid is used to prevent intestinal pathologies in cases of CTLA-4 blockade or deficiency. As presented in Example 5, a rapid metabolic transformation of 3-IAld occurs after administration. Specifically, after administration of the labeled 3-IAld, both the oxidized form (indole-3-carboxylic acid) and the methylated form of indole-3-carboxylic acid were detected. For example, 1-methylindole-3-carboxylic acid was detected with the same kinetics as indole-3-carboxylic acid, reaching peak levels at 30 min. Figure 13 C and Figure 13 D). Few studies have evaluated the bioactivity of methylindole and methoxyindole as aryl hydrocarbon receptor (AhR) activators. Preliminary in vitro data obtained in cell lines suggest that 1-methylindole-3-carboxylic acid activates AhR-dependent genes in a dose-dependent manner (data not shown). Given that 3-IAld can induce AhR-dependent genes and protect the mucosal barrier from damage, it is hypothesized that 1-methylindole-3-carboxylic acid may have similar therapeutic potential in preventing intestinal inflammatory pathology caused by CTLA-4 blockade in a mouse model of colitis, a model known to mimic the pathogenicity of hereditary human CTLA-4 haploinsufficiency in the intestine. Bakacs et al. , 2015 ).

[0197] In the experiment, C57BL / 6 mice received sodium dextran sulfate (DSS) (3%) in drinking water and were intraperitoneally administered 100 µg of anti-CTLA-4 monoclonal antibody (mAb) or isotype control antibody on days 0, 4, and 8 after DSS administration. Figure 17 As indicated in A. Eudragit-formulated 1-methylindole-3-carboxylic acid was administered orally every other day at a scaled dose of 0.09, 0.18, or 0.36 mg / mouse. Figure 17 A). Animals were monitored daily for the occurrence of diarrhea, bloody stools, and weight loss. One week after DSS treatment (14 days after treatment initiation), the rate of reproducing human disease in the mouse model was monitored. Manicassamy et al. , 2014 Surviving mice were sacrificed, their colons were removed, and macroscopic damage and local immune parameters were assessed. Mice treated with 1-methylindole-3-carboxylic acid showed a significant reduction in weight loss. Figure 17 B) and a reduced disease activity index ( Figure 17 C), and is protected from clinical onset and rectal bleeding ( Figure 17 D). Positive effects were observed at every administered dose of 1-methylindole-3-carboxylic acid (including a dose of 0.09 mg / mouse). Mice treated with 1-methylindole-3-carboxylic acid showed restoration of the following: i) normal structure of the colon and ileum ( Figure 17 E to Figure 17 F); ii) Maintenance of epithelial barrier function, as revealed by ZO-1 expression ( Figure 17 E to Figure 17 F). CTLA-4 haploid deficiency is typically characterized by widespread infiltration of CD3+ T cells in various organs (F). Kuehn et al. , 2014 - Schubert et al. , 2014 - Schwab et al. , 2018 Extensive CD3+ T cell infiltration in the ileum and colon of mice treated with anti-CTLA-4 was significantly reduced by co-administration of 1-methylindole-3-carboxylic acid. Figure 17 E to Figure 17 F). Administration of 1-methylindole-3-carboxylic acid also effectively reduced CD3+ T cell infiltration in the lungs (F). Figure 18 These results indicate that the 3-IAld metabolite 1-methylindole-3-carboxylic acid has the ability to prevent intestinal inflammatory pathology induced by CTLA4 blockade in a mouse colitis model, similar to... Figure 2 The processing was performed using 3-IAld.

[0198] 1-Methylindole-3-carboxylic acid is a potent activator of AhR-dependent genes. After confirmation Figure 17 After functional recovery in C57BL / 6 mice, 1-methylindole-3-carboxylic acid was found to promote the reduction of [a certain condition] in the colon. Il1b level of expression and increase Il10 Anti-inflammatory spectrum of expression level ( Figure 19 These results indicate that 1-methylindole-3-carboxylic acid supplementation provides protection against DSS+ anti-CTLA-4 induced colitis by maintaining epithelial barrier integrity and suppressing the inflammatory response. Consistent with its in vitro AhR agonist activity (data not shown), 1-methylindole-3-carboxylic acid induces AhR-dependent gene... Cyp1a1 Antimicrobial peptides Reg3g and Il22 Expression of (a key mediator of AhR-dependent mucosal protective activity) Figure 19 ) ( Stockinger et al. , 2021 These results indicate that 1-methylindole-3-carboxylic acid has effective activity against intestinal pathogenesis associated with CTLA-4 deficiency during CTLA-4 blockade, at least in part by activating the protective AhR / IL-22-dependent pathway.

[0199] Example 7 Pharmacology of 3-IAld / 1ME3CA Pharmacokinetics A single-dose pharmacokinetic (PK) study was performed on healthy C57BL / 6 mice to compare 1ME3CA and 3-IAld at a dose of 0.36 mg / topo. Untreated juvenile mice served as controls. Four animals were sacrificed at 0.5, 2, 6, and 24 hours. Blood was obtained by cardiac puncture, collected in EDTA-containing tubes, and the organs (brain, lung, intestine, liver, and kidney) were evaluated. All samples were stored at -80°C until use. To assess the functional activity of 1ME3CA and 3-IAld, the expression of AhR downstream genes in these mice was also evaluated.

[0200] Table 1: Pharmacokinetics

[0201] In C57BL / 6 mice, after a single oral administration of 18 mg / kg of 1ME3CA or 3-IAld (both formulated as intestinal microparticles), pharmacokinetic data were obtained by targeting the intestine, serum, lung, liver, brain, and kidney at different time points (30°C, 2 hours, 6 hours, and 24 hours) using mass spectrometry. Figure 20A As shown, 3-IAld disappears rapidly in most tissues (after 2 hours), and the terminal half-life depends on the tissue and ranges from 3 to 5 hours, as reported ( Puccetti et al. , Int J Pharm. 2021, 602:120610 Analysis of potential metabolites of 3-IAld revealed that indole-3-carboxylic acid was present as the major metabolite in serum and other tissues, followed by 1ME3CA, with a tmax of 30 minutes observed in every tissue tested, including the brain, and rapid clearance was observed from all organs studied. Compared to 3-IAld, 1ME3CA concentrations were at basal levels in the intestine, serum, and kidney, but significantly higher in the lung, liver, and brain, ranging from 103 to 105 nmol / kg. These observations suggest that 1ME3CA is formed as a secondary metabolite of 3-IAld and / or indole-3-carboxylic acid. Due to the rapid disappearance of 3-IAld in most tissues (after 2 hours) and the identical terminal half-life of 3 minutes for all compounds, it can be inferred that the biotransformation of 3-IAld and indole-3-carboxylic acid is extremely rapid.

[0202] Analysis of 1ME3CA PK after transenteral administration ( Figure 20BThis confirmed the above considerations. Indeed, 3-IAld and indole-3-carboxylic acid concentrations were at basal levels in all organs, confirming the role of 1ME3CA as a metabolite of 3-IAld and / or indole-3-carboxylic acid. The tmax values ​​(30 min) were identical in all organs and serum, and the Cmax values ​​were very similar. Even though 1ME3CA was undetectable in serum, lung, and kidney at 6 hours, its characteristic profile was similar. Furthermore, 1ME3CA levels remained higher than controls at 24 hours post-administration (at least in some organs, intestine, liver, and brain), by which time 1ME3CA had disappeared in mice administered 3-IAld. Systemic levels of 1ME3CA appeared to be more persistent after oral administration and, as expected, significantly higher than those observed after oral administration of 3-IAld.

[0203] Pharmacokinetics—In vitro activity 3-IAld has been shown to induce luciferase activity in H1L1.1c2 cell lines containing stably transfected AhR-responsive firefly luciferase at doses ranging from 0.1 to 100 µM. Zelante et al. Immunity. 2013;39:372- 85 The ability of 1ME3CA to induce luciferase activity was comparatively evaluated in mouse hepatocellular carcinoma cells (H1L6.1c3), courtesy of Allison K. Ehrlich (Meyer Hall, University of California, Davis, United States), containing a stably integrated AhR xenobiotic responsive element driven by the firefly luciferase reporter plasmid pGudLuc6.167. Cells were seeded in 24-well plates in MEMA (Gibco) supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin solution and stimulated with different concentrations of 3-IAld or 1ME3CA. Luciferase activity, calculated as relative light units (RLU) per microgram of protein and expressed as fold induction, was determined at 2 or 24 hours of exposure. The results clearly demonstrate that 1ME3CA is a more potent inducer of luciferase activity than 3-IAld, as observed by the higher RLU observed at 100 mM concentration for 2 h and continuing up to 24 h. Figure 21 ).

[0204] To further characterize the AhR activating potential of 1ME3CA, the activation of downstream AhR target genes by 3-IAld and 1ME3CA in the A549 cell line derived from adenocarcinoma human alveolar basal epithelial cells, the Calu-3 cell line derived from adenocarcinoma human bronchial epithelial cells, the CaCo-2 cell line derived from human colon cancer, and the HepG32 human hepatocellular carcinoma cell line was comparatively evaluated. (Cyp1a1, Cyp2a1) and AhRR) The ability of cells to express AhR-dependent genes was assessed by exposing them to 1, 10, 100, and 1000 μM of 1ME3CA or DMSO at 37°C for 4 hours or overnight (continued). The results showed that i) 1ME3CA promoted AhR-dependent expression in the tested cell lines. Cyp1a1, Cyp2a1 and AhRR Gene expression at levels comparable to, or even better than, reference AhR ligands ITE or FICZ and similar concentrations (i.e., 100 μM) of 3-IAld; ii) both molecules at 10-100 It functions within the optimal concentration range of M; results obtained at higher concentrations (1000 μM) are inconsistent, with AhR activity increasing in CaCo-2 and HepG32 cell lines but not in Calu-3 cells; iii) this activity first appears at 4 hours of exposure and appears to remain thereafter. Figure 22 A to Figure 22 D). Its ability to activate AhR was not evaluated. IDO1 and IDO2 Activity was observed in gene expression. Taken together, these data suggest that the AhR agonist activity of 1ME3CA is comparable to, if not superior to, that of 3-IAld.

[0205] Pharmacodynamics—Dose-dependent in vivo activity In DSS+ anti-CTLA-4 induced colitis, mice were administered intragastric treatment (Eudragit formulation) every other day, starting 4 days before DSS treatment and continuing until sacrifice. Disease activity in mice was assessed in terms of histopathology, barrier permeability parameters, and intestinal inflammation.

[0206] In a mouse model of immune-mediated colitis (DSS+ anti-CTLA-4), scaled doses of 0.09–0.045–0.022 mg / mouse (corresponding to 4.5, 2.25, and 1.12 mg / kg) were evaluated, as shown in Table 2 below.

[0207] Table 2: Colitis / Intestinal Inflammation

[0208] like Figure 23As depicted in the experimental schedule diagram in A, C57BL / 6 mice were treated with DSS in drinking water for one week, followed by a one-week recovery period and administration of 100 µg of anti-CTLA-4 mAb and 1ME3CA. Weight changes in the mice were assessed. Figure 23 B) Disease Activity Index ( Figure 23 C) Clinical onset and rectal bleeding ( Figure 23 D) Histology of the colon and ileum ( Figure 23 E)(PAS staining), magnification 40x (scale bar, 100 μm). Each in vivo experiment included four to six mice per group (20 mice per experiment).

[0209] To confirm the effect of 1ME3CA on lymphocyte infiltration, mice were treated as before and their histological changes (PAS staining) were assessed. High-resolution microscopy was used to photograph the lungs and spleen at 10x magnification (scale bar, 400 µm) and the liver and kidneys at 20x magnification (scale bar, 200 µm). Figure 24 As shown, 1ME3CA reduced lymphocyte infiltration in anti-CTLA-4 treated mice in a dose-dependent manner, with an effective dose of 3-IAld of 18 mg / kg and an effective dose of 1ME3CA of 2.25 mg / kg.

[0210] toxicology To assess toxicology, C57BL / 6 mice were administered 0.36–0.18–0.09 mg / mouse in escalating intragastric doses of 3-IAld or 1ME3CA (prepared as intestinal microparticles) for up to 3 weeks, as shown in Table 3 below. At the end of treatment, mice were sacrificed, and histopathology was performed blinded on different sections from each organ after PAS staining. Sections from organs from 4 mice in each group were evaluated individually. Results ( Figure 25A and Figure 25B The results showed that no visible histopathology was found in any of the examined organs at the three different concentrations.

[0211] Table 3: Toxicology

[0212] References

Claims

1. A method for treating a patient with CTLA-4 checkpoint-related immunodeficiency, the method comprising administering to the patient in need a therapeutically effective amount of a pharmaceutical composition comprising 1H-indole-3-carboxaldehyde (3-IAld).

2. The method of claim 1, wherein the CTLA-4 checkpoint-related immunodeficiency is selected from the group consisting of: CTLA-4 haploid deficiency with autoimmune infiltration (CHAI), lipopolysaccharide-responsive beige-like anchoring protein (LRBA) deficiency (LATAIE), regulatory T (Treg) cell deficiency, autoimmune infiltration, enteropathy, intestinal inflammation, immune-mediated colitis, gastrointestinal disorders, and gastric atrophy.

3. The method of any one of claims 1 or 2, wherein the CTLA-4 checkpoint-related immunodeficiency is immune-mediated colitis.

4. The method of any one of claims 1-3, wherein the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

5. The method of claim 4, wherein the pharmaceutically acceptable carrier is at least one polymer.

6. The method of claim 4, wherein the pharmaceutically acceptable carrier is a group of polymers.

7. The method of claim 6, wherein the polymer of the group is Eudragit® polymer.

8. The method of any one of claims 1-7, wherein the pharmaceutical composition is formulated for enteral delivery.

9. The method of any one of claims 1-8, wherein the pharmaceutical composition is administered orally.

10. The method of any one of claims 1-9, wherein the pharmaceutical composition is in the form of capsules, tablets, gel sheets, gel capsules, gels, liquids, and gummies.

11. The method of claim 10, wherein the pharmaceutical composition is in tablet or capsule form.

12. The method of any one of claims 1-11, wherein the pharmaceutical composition is administered at an interval of every other day (qod).

13. The method of any one of claims 1-12, wherein the pharmaceutical composition is administered at a dose of at least about 3 mg / kg, at least about 4 mg / kg, at least about 5 mg / kg, at least about 6 mg / kg, at least about 7 mg / kg, at least about 8 mg / kg, at least about 9 mg / kg, at least about 10 mg / kg, at least about 11 mg / kg, at least about 12 mg / kg, at least about 13 mg / kg, at least about 14 mg / kg, at least about 15 mg / kg, at least about 16 mg / kg, at least about 17 mg / kg, or at least about 18 mg / kg.

14. The method of claim 13, wherein the 3-IAld dose is about 18 mg / kg.

15. A method for treating a patient with CTLA-4 checkpoint-related immunodeficiency, the method comprising administering to the patient in need a therapeutically effective amount of a pharmaceutical composition comprising 1-methylindole-3-carboxylic acid.

16. The method of claim 15, wherein the CTLA-4 checkpoint-related immunodeficiency is selected from the group consisting of: CTLA-4 haploid deficiency with autoimmune infiltration (CHAI), lipopolysaccharide-responsive beige-like anchor protein (LRBA) deficiency (LATAIE), regulatory T (Treg) cell deficiency, autoimmune infiltration, enteropathy, intestinal inflammation, immune-mediated colitis, gastrointestinal disorders, and gastric atrophy.

17. The method of any one of claims 15 or 16, wherein the CTLA-4 checkpoint-associated immunodeficiency is immune-mediated colitis.

18. The method of any one of claims 15-17, wherein the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

19. The method of claim 18, wherein the pharmaceutically acceptable carrier is at least one polymer.

20. The method of claim 18, wherein the pharmaceutically acceptable carrier is a group of polymers.

21. The method of claim 20, wherein the polymer of the group is an Eudragit® polymer.

22. The method of any one of claims 15-21, wherein the pharmaceutical composition is formulated for enteral delivery.

23. The method of any one of claims 15-22, wherein the pharmaceutical composition is administered orally.

24. The method of any one of claims 15-23, wherein the pharmaceutical composition is in the form selected from capsules, tablets, gel sheets, gel capsules, gels, liquids and gummies.

25. The method of claim 24, wherein the pharmaceutical composition is in tablet or capsule form.

26. The method of any one of claims 15-25, wherein the pharmaceutical composition is administered at every other day (qod).

27. The method of any one of claims 15-23, wherein the pharmaceutical composition is administered at a dose of 1-methylindole-3-carboxylic acid of at least about 2 mg / kg, at least 3 mg / kg, at least about 4 mg / kg, at least about 5 mg / kg, at least about 6 mg / kg, at least about 7 mg / kg, at least about 8 mg / kg, at least about 9 mg / kg, at least about 10 mg / kg, at least about 11 mg / kg, at least about 12 mg / kg, at least about 13 mg / kg, at least about 14 mg / kg, at least about 15 mg / kg, at least about 16 mg / kg, at least about 17 mg / kg, or at least about 18 mg / kg.

28. The method of any one of claims 15-23, wherein the pharmaceutical composition is administered at a dose of about 2.25 mg / kg of 1-methylindole-3-carboxylic acid.

Images