# US 2024/0201187 A1 · Method of Tracking Maintenance of Immunological Tolerance

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- Number: US 2024/0201187 A1
- Document ID: US20240201187A1
- Status: Published application
- Kind: A1
- Application number: 18/287,065
- Filed: 2022-04-15
- Published: 2024-06-20
- Earliest claimed: 2021-04-16
- Assignees: Cour Pharmaceuticals Development Company, Inc.
- Applicants: Cour Pharmaceuticals Development Company Inc.
- Inventors: John Puisis; James Herrmann; Michael Boyne; Adam Elhofy; Joseph Podojil; Greta Wodarcyk
- Web page: https://patentgrep.com/patent/US20240201187A1

## CPC

- G01N 33/53: Immunoassay; Biospecific binding assay; Materials therefor
- G01N 33/56972: {White blood cells}
- G01N 33/68: involving proteins, peptides or amino acids {(involving lipoproteins G01N33/92)}
- G01N 33/6863: {Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors}
- G01N 2333/52: Assays involving cytokines
- G01N 2800/24: Immunology or allergic disorders
- G01N 2800/52: Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

The present application relates, in general, to methods for tracking the induction of and long-term maintenance of antigen specific immune tolerance during administration of an antigen specific immune tolerizing therapy, such as a TIMP.

## Claims

### Claim 1

A method for monitoring immune tolerance status of a subject undergoing immune tolerizing treatment for a disease or condition, the method comprising the steps of
(a) obtaining one or more biological samples from the subject prior to administration of treatment and determining the immune tolerance status of the subject by assaying said biological sample(s),
(b) obtaining one or more biological samples from the subject after administration of treatment and determining the immune tolerance status of the subject by assaying the biological sample(s), and
(c) obtaining one or more biological samples from the subject at regular intervals after administration of treatment and determining the immune tolerance status of the subject by assaying the biological sample(s),
(d) comparing the results from the assay of one or more biological samples in step (c) with the results of steps (a) and/or (b) to generate an immune tolerance signature, and
(e) re-administering the tolerizing treatment if the immune tolerance signature indicates a weakening, and/or loss of immunological tolerance.

### Claim 2 (depends on 1)

The method of claim 1, wherein the disease or condition is selected from the group consisting of an inflammatory condition, an autoimmune condition, an allergy, an abnormal immune response, a lysosomal storage disease, an enzyme deficiency, a protein deficiency, a genetic disorder, and/or is a transplant recipient.

### Claim 3 (depends on 2)

The method of claim 2, wherein the autoimmune condition is selected from the group consisting of atopic atopic dermatitis, multiple sclerosis, autoimmune myelitis, myelitis, transverse myelitis, neuromyelitis optica (NMO), neuromyelitis optica spectrum disorder (NMSOD), type-1 diabetes (T1D), type-2 diabetes (T2D), Celiac Disease (CD), Grave's Disease, Myasthenia Gravis, acute disseminated encephalomyelitis, Addison's Disease, alopecia, rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, autoimmune myocarditis, autoimmune neutropenia, autoimmune skin disease, autoimmune uveitis, ballus pemphigoid, Behcet's Syndrome, cerebral degeneration, chronic neuropathy, cicatrical pemphigoid, pemphigus vulgaris, Crohn's Disease, Inflammatory Bowel Disease (IBD), Inflammatory Bowel Syndrome (IBS), cryopathy, dermatitis hyperformis, Eaton Lambert's Disease, encephalomyelitis, epidermolysis bullosa acquisita, erythema nodosa, glomerulonephritis, Goodpasture's Disease, granulomatosis, Guillain-Barre Syndrome, Hashimoto Disease, Kawasaki Disease, hemolytic anemia, hypersensitivity vasculitis, lupus erythematosus, mixed connective tissue disease, mixed essential cryoglobulinemia, multifocal motor neuropathy, opsonoclonus-myoclonus, paraneoplastic pemphigus, pemphigoig gestationis, pemphigus folaceus, pernicious anemia, peripheral biliary cirrhosis, polyangiitis overlap syndrome, polyarteritis nodosa, polyglandular failure, polyglandular syndrome, polymyositis/dermatomyositis, psoriasis, eczema, retinopathy, Reynaud's Syndrome, sarcoidosis, Scleroderma Type 1, sclerosis cholangitis, Sjogren's Syndrome, Stiffman's syndrome, Takayasu arteritis, termporal arteritis, thyroiditis, ulcerative colitis, immune thrombocytopenia purpura, thrombotic thrombocytopenia purpura, autoimmune hepatitis, primary biliary cholangitis (PBC), ANCA diseases, Granulamatosis with Polyangiitis, and Microscopic Polyangiitis.

### Claim 4 (depends on 1, 2)

The method of claim 1 or 2, wherein the immune tolerizing treatment comprises an antigen selected from the group consisting of one or more autoimmune antigens, allergens, enzyme replacement therapies, protein therapies, transplant antigens, and gene therapy vectors.

### Claim 5 (depends on 4)

The method of claim 4, wherein the one or more autoimmune antigens are selected from the group consisting of myelin basic protein, acetylcholine receptor, endogenous antigen, myelin oligodendrocyte glycoprotein (MOG), myelin basic protein (MBP), proteolipid protein (PLP), myelin associated glycoprotein (MAG), cyclic nucleotide phosphohydrolase, pancreatic beta-cell antigen, insulin, proinsulin, islet-specific glucose-6-phophatase catalytic subunit-related protein (IGRP), glutamic acid decarboxylase (GAD), collagen type 11, human cartilage gp39, fp130-RAPS, fibrillarin, small nucleolar protein, thyroid stimulating factor receptor, histones, glycoprotein gp70, pyruvate dehydrogenase dehydrolipoamide acetyltransferase (PCD-E2), hair follicle antigen, aqua porin 4, Desmoglein 1, Desmoglein 3, nicotinic acetylcholine receptor, gliadin, ADAMTS13, GPIlb/GPIlla, CYP2D6, BP180, NC16, BP230, Ro60, MPO, thyroid stimulating hormone receptor, and human tropomyosin isoform 5.

### Claim 6 (depends on 4)

The method of claim 4, wherein the one or more allergens is selected from the group consisting of Bahia grass pollen (BaGP), peach allergen, milk allergens, celery allergens, nut allergens, bovine serum albumin, Hazelnut allergens, ovalbumin, egg allergen, peanut allergens, fish allergens, shellfish allergens, and Japanese cedar pollen.

### Claim 7 (depends on 1, 2, 3, 4, 5, 6)

The method of any one of claims 1-6, wherein said immune tolerizing treatment is antigen specific.

### Claim 8 (depends on 1, 2, 3, 4, 5, 6, 7)

The method of any one of claims 1-7, wherein the immune tolerizing treatment is selected from the group consisting of oral immunotherapy (OIT), subcutaneous immunotherapy (SCIT), sublingual immunotherapy (SLIT), and immune tolerizing nanomedicine.

### Claim 9 (depends on 8)

The method of claim 8, wherein the immune tolerizing nanomedicine comprises tolerizing immune modifying particles (TIMPs).

### Claim 10 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9)

The method of any one of claims 1-9, wherein said biological sample(s) of any one of steps (a)-(c) are obtained 1-7 days, 1-4 weeks, and/or 1-12 months prior to or after administration of the treatment.

### Claim 13 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)

The method of any one of claims 1-12, wherein the one or more biological samples are selected from the group consisting of whole-blood, peripheral blood, peripheral blood mononuclear cells (PBMCs), serum, plasma, urine, cerebrospinal fluid (CSF), stool, a tissue biopsy, and a bone-marrow biopsy.

### Claim 14 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)

The method of any one of claims 1-13, wherein said assay of the one or more biological samples is selected from the group consisting of analyzing cells, cell-surface proteins, extracellular proteins, intracellular proteins, nucleic acids, metabolites, and combinations thereof.

### Claim 15 (depends on 14)

The method of claim 14, wherein the cells are immune cells and/or non-immune cells.

### Claim 16 (depends on 15)

The method of claim 15, wherein the immune cells are innate immune cells and/or adaptive immune cells.

### Claim 17 (depends on 15, 16)

The method of claim 15 or 16, wherein the immune cells are selected from the group consisting of monocytes, macrophages, neutrophils, granulocytes, dendritic cells, mast cells, eosinophils, basophils, T-cells, B-cells, NK cells, and NK-T cells.

### Claim 18 (depends on 17)

The method of claim 17, wherein the T cells are T1 cells, T2a cells, Treg cells, Tr1 cells and/or Teff cells.

### Claim 19 (depends on 15)

The method of claim 15, wherein the non-immune cells are selected from the group consisting of epithelial cells, stromal cells, endothelial cells, fibroblasts, pericytes, adipocytes, mesenchymal stem cells, hematopoietic stem cell, hematopoietic progenitor cells, liver sinusoidal endothelial cells (LSECs), and/or Kupffer cells.

### Claim 20 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)

The method of any one of claims 1-19, wherein the assay of one or more biological samples in Step (a) is used to determine the immune tolerance status of the subject at Baseline.

### Claim 22 (depends on 21)

The method of claim 21, wherein the immune tolerance status of the subject at Baseline is determined by comparing the results from the assay of one or more biological samples in Step (a) to the assay of samples obtained from a healthy subject.

### Claim 23 (depends on 20, 21)

The method of claim 20 or 21, wherein the immune tolerance status of the subject at Baseline is used to determine whether the subject is administered a tolerizing treatment.

### Claim 24 (depends on 20, 21, 22, 23)

The method of any one of claims 20-23, wherein the immune tolerance status of the subject at Baseline indicates weak immune tolerance or absence of immune tolerance.

### Claim 25 (depends on 20, 21, 22, 23, 24)

The method of any one of claims 20-24, wherein the subject is administered treatment if the immune tolerance status of the subject at Baseline indicates weak immune tolerance or absence of immune tolerance.

### Claim 26 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)

The method of any one of claims 1-25, wherein the subject is re-administered treatment in Step (e) if the results of an assay in Step (c) compared to the results from the assay in Steps (a) and/or (b) indicate weakening and/or loss of immune tolerance.

### Claim 27 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)

The method of any one of claims 1 to 26, wherein the immune tolerance signature of a subject is generated using one or more of the following parameters assayed from one or more biological samples obtained from the subject and stimulated in vivo and/or ex vivo:
a. proportion of effector T cells in the total T cell population,
b. proportion of Treg cells in the total T cell population,
c. proportion of effector B cells in the total B cell population,
d. levels and/or ratios of specific IgG, IgA, IgM, and/or IgE,
e. levels of inflammatory cytokines and chemokines,
f. levels of anti-inflammatory cytokines and chemokines,
g. levels of inflammatory metabolites, and/or
h. levels of anti-inflammatory metabolites.

### Claim 28 (depends on 27)

The method of claim 27, wherein the immune tolerance signature is indicative of maintenance of immune tolerance if 2, 3, 4, 5, 6, 7, or 8 parameters listed in (a)-(h) indicate maintenance of immune tolerance. [

### Claim 29 (depends on 28)

The method of claim 28, wherein the subject is determined to not require treatment with TIMPs if at least 2/8 parameters listed in (a)-(h) above indicate maintenance of immune tolerance.

### Claim 30 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29)

The method of any one of claims 1-29, wherein the subject is re-administered treatment if immune tolerance is weakened and/or lost by about 5%-100% in at least one parameter of immune tolerance.

### Claim 31 (depends on 30)

The method of claim 30 wherein the at least one parameter of immune tolerance is selected form the group consisting of:
a. proportion of effector T cells in the total T cell population,
b. proportion of Treg cells in the total T cell population,
c. proportion of effector B cells in the total B cell population,
d. levels and/or ratios of specific IgG, IgA, IgM, and/or IgE,
e. levels of inflammatory cytokines and chemokines,
f. levels of anti-inflammatory cytokines and chemokines,
g. levels of inflammatory metabolites, and/or
h. levels of anti-inflammatory metabolites,
assayed from one or more biological samples obtained from the subject and stimulated in vivo and/or ex vivo.

### Claim 32 (depends on 30)

The method of claim 30, wherein immune tolerance is weakened and/or lost by 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%.

### Claim 33 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29)

The method of any one of claims 1-29, wherein the subject is re-administered treatment in Step (e) if the results of Step (d) indicate weakening and/or loss of immune tolerance by about 2-100-fold in at least one parameter of immune tolerance.

### Claim 34 (depends on 33)

The method of claim 33 wherein the at least one parameter of immune tolerance is selected form the group consisting of:
a. proportion of effector T cells in the total T cell population,
b. proportion of Treg cells in the total T cell population,
c. proportion of effector B cells in the total B cell population,
d. levels and/or ratios of specific IgG, IgA, IgM, and/or IgE,
e. levels of inflammatory cytokines and chemokines,
f. levels of anti-inflammatory cytokines and chemokines,
g. levels of inflammatory metabolites, and/or
h. levels of anti-inflammatory metabolites,
assayed from one or more biological samples obtained from the subject and stimulated in vivo and/or ex vivo.

### Claim 35 (depends on 33, 34)

The method of claim 33 or 34, wherein immune tolerance is weakened and/or lost by about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100-fold.

### Claim 36 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35)

The method of any one of claims 1-35, wherein the treatment administered in Step (e) is TIMPs.

### Claim 37 (depends on 36)

The method of claim 36, wherein the TIMPs are administered at a dose level of between about 0.1 to 12 mg/kg.

### Claim 38 (depends on 37)

The method of claim 37, wherein the TIMPs are administered at a dose level of about 0.1 mg/kg, 0.25 mg/kg, 0.5 mg/kg, 0.75 mg/kg, 1.0 mg/kg, 1.25 mg/kg, 1.5 mg/kg, 2.0 mg/kg, 2.5 mg/kg, 3 mg/kg, 3.5 mg/kg, 4.0 mg/kg, 4.5 mg/kg, 5 mg/kg, 5.5 mg/kg, 6 mg/kg, 6.5 mg/kg, 7 mg/kg, 7.5 mg/kg, 8.0 mg/kg, 8.5 mg/kg, 9 mg/kg, 9.5 mg/kg, 10 mg/kg, 10.5 mg/kg, 11 mg/kg, 11.5 mg/kg, or 12 mg/kg.

### Claim 39 (depends on 36)

The method of claim 36, wherein the TIMPs are administered at a dose level of between about 50 mg to 800 mg.

### Claim 40 (depends on 39)

The method of claim 39, wherein the TIMPs are administered at a dose level of about 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 375 mg, 400 mg, 425 mg, 450 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, or 800 mg.

### Claim 41 (depends on 36, 37, 38, 39, 40)

The method of any one of claims 36-40, wherein the TIMPs are administered in a single dose or multiple doses.

### Claim 42 (depends on 36, 37, 38, 39, 40, 41)

The method of any one of claims 36-41, wherein the TIMPs are administered once weekly, once every two weeks, once every three weeks, once every 4 weeks, once every two months, once every three months, once every 6 months, or once per year.

### Claim 43 (depends on 36, 37, 38, 39, 40, 41, 42)

The method of any one of claims 36-42, wherein the TIMPs are administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, or orally.

### Claim 44 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43)

The method of any one of claims 1-43, wherein said tolerance status is determined by assay of one or more biological samples after one or more stimuli.

### Claim 45 (depends on 44)

The method of claim 44, wherein the stimuli are provided in vivo and/or ex vivo.

### Claim 46 (depends on 44, 45)

The method of claim 44 or 45, wherein the one or more stimuli are selected from the group consisting of one or more antigens, allergens, and activating agents.

### Claim 47 (depends on 46)

The method of claim 46, wherein the antigens and allergens are associated with the disease or condition being treated.

### Claim 48 (depends on 47)

The method of claim 47, wherein the antigens and allergens are not associated with the disease or condition being treated.

### Claim 49 (depends on 46)

The method of claim 46, wherein the one or more activating agents are selected from the group consisting of an antibody, a chemical agent, a viral component, and a bacterial component.