Methods and compositions for preventing type 1 diabetes

Abstract

Methods for preventing or delaying the onset of clinical type 1 diabetes (T1D) are provided. [Solution] The method includes the steps of preparing a non-diabetic subject at risk of T1D, determining that the non-diabetic subject (1) substantially does not have antibodies against zinc transporter 8 (ZnT8), (2) is HLA-DR4+, and / or (3) is not HLA-DR3+, and administering a prophylactically effective amount of an anti-CD3 antibody to the non-diabetic subject.

Description

Technical Field

[0001] [Cross - Reference to Related Applications] This application claims priority and benefit of U.S. Provisional Application No. 62 / 847,466, filed May 14, 2019, and U.S. Utility Patent Application No. 15 / 931,685, filed May 14, 2020, the entire disclosures of which are incorporated herein by reference.

[0002] [Sequence Listing] The ASCII text file entitled "010701seq.txt", created on May 14, 2020, and having a size of 6,083 bytes, filed via EFS - Web on May 14, 2020, is hereby incorporated by reference in its entirety. [Field of the Invention]

[0003] The present disclosure generally relates to compositions and methods for preventing or delaying the onset of clinical type 1 diabetes (T1D) in subjects at risk of developing clinical T1D, and more particularly to the use of anti - CD3 antibodies.

Background Art

[0004] Type 1 diabetes (T1D) is caused by the autoimmune destruction of insulin - producing β - cells in the islets of Langerhans, leading to dependence on exogenous insulin injections for survival. Approximately 1.6 million Americans have type 1 diabetes, and T1D remains one of the most common childhood diseases after asthma. Despite improved care, individuals most affected by T1D consistently cannot achieve desired blood glucose target levels. 1 . 2 ​​​​​​​​For individuals with type 1 diabetes, there is a persistent concern regarding the increased risk of both morbidity and mortality. Two recent studies have shown that the number of years of survival is reduced by 17.7 years for children diagnosed before the age of 10, and by 11 years and 13 years for Scottish men and women diagnosed as adults, respectively. 3,4

[0005] In genetically susceptible individuals, T1D progresses through an asymptomatic stage characterized first by the appearance of autoantibodies (stage 1) and then by glucose abnormalities (stage 2), before overt hyperglycemia. At stage 2, the metabolic response to a glucose load is impaired, but other metabolic markers, such as glycosylated hemoglobin, are normal and insulin treatment is not required. 5 These immunological and metabolic features identify individuals at high risk of developing a clinical disease with overt hyperglycemia and requiring insulin treatment (stage 3). When testing newly diagnosed clinical T1D, several immune interventions have been shown to delay the decline in β-cell function. 6 One promising treatment is the FcR-non-binding anti-CD3 monoclonal antibody teplizumab, which has been shown in several trials to durably reduce β-cell function loss with short-term treatment, with observable effects for up to 7 years after diagnosis and treatment. 7~11 The drug modifies the function of CD8+ T lymphocytes, which are considered to be important effector cells that cause β-cell death. 12,13

[0006] To date, there has been no intervention initiated prior to clinical diagnosis (i.e., stage 1 or 2) that changes the progression to clinical stage 3 T1D. Therefore, there is a need for a treatment that prevents or delays the onset of clinical T1D in individuals at high risk of developing clinical T1D. SUMMARY OF THE INVENTION

[0007] In one aspect, a method for preventing or delaying the onset of clinical type 1 diabetes (T1D) is provided, the method comprising: preparing a non-diabetic subject at risk of T1D; determining that the non-diabetic subject is (1) substantially free of antibodies to zinc transporter 8 (ZnT8), (2) HLA-DR4+, and / or (3) not HLA-DR3+; administering a prophylactically effective amount of an anti-CD3 antibody to the non-diabetic subject.

[0008] In some embodiments, the non-diabetic subject is a blood relative of a T1D patient. In certain embodiments, the non-diabetic subject has two or more diabetes-related autoantibodies selected from islet cell antibodies (ICA), insulin autoantibodies (IAA), and antibodies to glutamic acid decarboxylase (GAD), tyrosine phosphatase (IA-2 / ICA512), or ZnT8.

[0009] In some embodiments, detection of autoantibodies related to T1D is performed by a point-of-care (POC) screening method in the general population or in blood relatives of T1D patients. These POC methods can be qualitative rapid lateral flow assays.

[0010] ​​​​​​​​​​​​​In some embodiments, the non - diabetic subject has an infection by coxsackievirus B (CVB) and / or one or more other beta - cell tropic viruses. In some embodiments this subject infected with a beta - cell tropic virus has HLA - DR4 and is highly reactive to teplizumab.

[0011] In various embodiments, the non - diabetic subject has an abnormality in glucose tolerance in an oral glucose tolerance test (OGTT). The abnormality in glucose tolerance in the OGTT is defined as a fasting glucose level of 110 - 125 mg / dL, or a plasma 2 - hour value of 140 mg / dL or more and less than 200 mg / dL, or a mid - glucose value at 30, 60, or 90 minutes during the OGTT that is greater than 200 mg / dL.

[0012] In some embodiments, the non - diabetic subject does not have antibodies against ZnT8. In certain embodiments, the non - diabetic subject is HLA - DR4+ and not HLA - DR3+.

[0013] In one embodiment, the anti - CD3 antibody is teplizumab.

[0014] In various embodiments, the prophylactically effective amount is an anti - CD3 antibody, such as teplizumab, given as a total dose of anti - CD3 / teplizumab of 6 - 15 milligrams, at 10 - 1000 micrograms per square meter (μg / m ) by subcutaneous (SC) injection or intravenous (IV) infusion daily for 10 - 2 14 days, preferably, teplizumab is IV infused at 51 μg / m on days 0 - 3 at 51 μg / m m 2 , 103 μg / m 2 , 207 μg / m 2 , and 413 μg / m 2 respectively, and on day 4 ​​​​​​On each of the days from day 0 to day 13, a single dose of 826 μg / m 2 is included in a 14-day course of intravenous infusion. In certain embodiments, a prophylactically effective amount of an anti-CD3 antibody, such as teprotumumab, delays the median time to clinical diagnosis of T1D by at least 50%, at least 80%, or at least 90%, or by at least 12 months, at least 18 months, at least 24 months, at least 36 months, at least 48 months, or at least 60 months, or longer.

[0015] In some embodiments, an anti-CD3 antibody, such as teprotumumab, otelixizumab, or fulolumab, is administered by an SC pump, or encapsulated in a sustained-release biomaterial, or orally. In other embodiments, an anti-CD3 antibody, such as teprotumumab, otelixizumab, or fulolumab, is encapsulated in a biomaterial encapsulating β-cell precursors or β-cells, or provided parenterally in combination with β-cell precursors or β-cells.

[0016] In some embodiments, an anti-CD3 antibody, such as teprotumumab, otelixizumab, or fulolumab, is administered in combination with other pharmacological agents, such as metabolic agents, B-cell inhibitors, or other immunomodulatory agents.

[0017] In some embodiments, an anti-CD3 antibody, such as teprotumumab, otelixizumab, or fulolumab, is administered in combination with antigen-specific immunotherapy and / or a vaccine.

[0018] In some embodiments, the method further comprises determining the frequency of TIGIT+KLRG1+CD8+ T cells in peripheral blood mononuclear cells of non-diabetic subjects by flow cytometry. ​​​​​​​ comprising, wherein an increased frequency after administration of an anti-CD3 antibody, such as teplizumab, otelixizumab or foralumab, indicates responsiveness to the anti-CD3 antibody, such as teplizumab is shown thereby

[0019] Another aspect is a method of predicting responsiveness to an anti-CD3 antibody, such as teplizumab, otelixizumab or foralumab, in the prevention or delay of the onset of type 1 diabetes (T1D), comprising preparing a non-diabetic subject at risk of T1D, administering to the non-diabetic subject a prophylactically effective amount of teplizumab, otelixizumab or foralumab, determining, by flow cytometry, the frequency of TIGIT+KLRG1+CD8+ T cells in peripheral blood mononuclear cells of the non-diabetic subject, wherein an increase in the frequency indicates responsiveness to an anti-CD3 antibody, such as teplizumab, otelixizumab or foralumab, thereby being shown and being related to a method comprising In some embodiments, the method may further comprise determining that the non-diabetic subject is (1) substantially free of antibodies to zinc transporter 8 (ZnT8), (2) HLA-DR4+,

[0020] and / or (3) not HLA-DR3+ It will be apparent to those skilled in the art that the aspects and / or embodiments described herein can be combined with each other and

[0021] It will be apparent to those skilled in the art that the aspects and / or embodiments described herein can be combined with each other BRIEF DESCRIPTION OF THE DRAWINGS

[0022]

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Figure 7A-B

Mode for Carrying Out the Invention

[0023] In some embodiments, the present disclosure provides the surprising finding that non-diabetic subjects who respond to treatment with an anti-CD3 antibody, such as teplizumab, do not have antibodies against ZnT8. In certain embodiments, such non-diabetic subjects are HLA-DR4+ and not HLA-DR3+. Unexpectedly, in such non-diabetic subjects who respond to treatment with an anti-CD3 antibody, an increase in the frequency (or relative amount) of TIGIT+KLRG1+CD8+ T cells in peripheral blood mononuclear cells (e.g., by flow cytometry) is demonstrated after administration of teplizumab (e.g., 1 month later, 2 months later, 3 months later, or more or less before or after that). In some embodiments, it prevents or delays the onset of clinical type 1 diabetes (T1D). In certain embodiments, such non-diabetic subjects are HLA-DR4+ and not HLA-DR3+. Unexpectedly, in such non-diabetic subjects who respond to treatment with an anti-CD3 antibody, an increase in the frequency (or relative amount) of TIGIT+KLRG1+CD8+ T cells in peripheral blood mononuclear cells (e.g., by flow cytometry) is demonstrated after administration of teplizumab (e.g., 1 month later, 2 months later, 3 months later, or more or less before or after that). In some embodiments, non-diabetic subjects who respond to treatment with an anti-CD3 antibody, such as teplizumab, do not have antibodies against ZnT8. In certain embodiments, such non-diabetic subjects are HLA-DR4+ and not HLA-DR3+. Unexpectedly, in such non-diabetic subjects who respond to treatment with an anti-CD3 antibody, an increase in the frequency (or relative amount) of TIGIT+KLRG1+CD8+ T cells in peripheral blood mononuclear cells (e.g., by flow cytometry) is demonstrated after administration of teplizumab (e.g., 1 month later, 2 months later, 3 months later, or more or less before or after that). In some embodiments, non-diabetic subjects who respond to treatment with an anti-CD3 antibody, such as teplizumab, do not have antibodies against ZnT8. In certain embodiments, such non-diabetic subjects are HLA-DR4+ and not HLA-DR3+. Unexpectedly, in such non-diabetic subjects who respond to treatment with an anti-CD3 antibody, an increase in the frequency (or relative amount) of TIGIT+KLRG1+CD8+ T cells in peripheral blood mononuclear cells (e.g., by flow cytometry) is demonstrated after administration of teplizumab (e.g., 1 month later, 2 months later, 3 months later, or more or less before or after that).

[0024] In some embodiments, it prevents or delays the onset of clinical type 1 diabetes (T1D). A method comprising the steps of preparing a non-diabetic subject at risk of T1D, and a non-diabetic subject being , (1) substantially free of antibodies against zinc transporter 8 (ZnT8), ([ 2) being HLA-DR4+, and / or (3) not being HLA-DR3+ determining, and administering to the non-diabetic subject a prophylactically effective amount of an anti-CD3 antibody, such as teplizumab is provided herein.

[0025] In certain embodiments, a method of predicting responsiveness to an anti-CD3 antibody, such as teplizumab, in preventing or delaying the onset of T1D is provided. The method comprises the steps of preparing a non-diabetic subject at risk of T1D, administering to the non-diabetic subject a prophylactically effective amount of an anti-CD3 antibody, such as teplizumab, and determining by flow cytometry the frequency of TIGIT +KLRG1+CD8+ T cells in the peripheral blood mononuclear cells of the non-diabetic subject , wherein an increase in the frequency indicates responsiveness to the anti-CD3 antibody, such as teplizumab and may comprise the step.

[0026] [Definitions] Certain terms are defined herein below. Additional definitions are provided throughout this application.

[0027] As used herein, the articles "a" and "an" refer to one or more than one of the grammatical object of the article, e.g., at least one. The use of the word "a" or "an" when used in conjunction with the term "comprising", herein means "one" when used in conjunction with the term "comprising", herein means "one" may mean "one or more", "at least one", and is also consistent with the meaning of "one or more than one". The meaning of "at least one" is also consistent with that of "one or more than one".

[0028] As used herein, "about" and "approximately" generally mean the degree of error tolerated with respect to the measured quantity, taking into account the nature or precision of the measurement. Exemplary degrees of error are within 20 percent (%) of the range of a given value, generally within 10%, more generally within 5%. The term "substantially" means more than 50%, preferably more than 80%, and most preferably more than 90% or 95%.

[0029] As used herein, the term "comprising" or "comprises" is used with respect to the compositions, methods, and their respective components (which may be plural) present in a given embodiment, and is open to the inclusion of unspecified elements.

[0030] As used herein, the term "consisting essentially of" refers to the elements required for a given embodiment. This term allows the presence of additional elements that do not substantially affect the basic and novel or functional characteristics (which may be plural) of that embodiment of the present disclosure.

[0031] The term "consisting of" refers to the compositions, methods, and their respective components described herein, as described in the description of that embodiment. No element is excluded.

[0032] As used herein, the term "antibody" is used in the broadest sense and is not limited to, but includes monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and various antibody structures including antibody fragments so long as they exhibit the desired antigen-binding activity.

[0033] "Antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody and binds to an antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab’, Fab’-SH, F(ab’)2; diabody( diabody); linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments.

[0034] As used herein, the term "preventive agent" refers to a CD3-binding molecule such as teprilizumab that can be used for the prevention, treatment, management, or amelioration of one or more symptoms of T1D.

[0035] As used herein, the term "onset" of a disease associated with type 1 diabetes refers to a patient meeting the criteria established by the American Diabetes Association for the diagnosis of type 1 diabetes (see Mayfield et al., 2006, Am. Fam. Physician 5 8:1355-1362).

[0036] As used herein, the terms "prevent", "preventing", and "prevention" refer to a preventive agent or prevention of the onset of one or more symptoms of T1D in a subject due to administration of a therapeutic agent refers to.

[0037] As used herein, "protocol" includes a dosing schedule and dosing regimen. The protocol herein is the method of use and includes a prevention protocol and a treatment protocol. "Dosing regimen" or "course of treatment" may include administering a therapeutic or prophylactic agent in several doses over a period of 1 to 20 days.

[0038] As used herein, the terms "subject" and "patient" are used interchangeably. As used herein, the terms "subject" and "subjects" refer to animals, preferably mammals including non - primates (e.g., cows, pigs, horses, goats, dogs, rats, and mice) and primates (e.g., monkeys or humans), more preferably humans.

[0039] As used herein, the term "prophylactically effective amount" refers to an amount of teplizumab sufficient to effect a delay or prevention of the occurrence, recurrence, or onset of one or more symptoms of T1D. In some embodiments, the prophylactically effective amount preferably refers to an amount of teplizumab that delays the onset of T1D in a subject by at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%.

[0040] Various aspects of the present disclosure are described in further detail below. Additional definitions are detailed throughout this specification. throughout this specification.

[0041] <Anti-CD3 Antibodies and Pharmaceutical Compositions> The terms "anti-CD3 antibody" and "antibody that binds to CD3" refer to an antibody or antibody fragment that can bind with sufficient affinity to surface antigen cluster 3 (CD3), and thus, the antibody is useful as a prophylactic, diagnostic, and / or therapeutic agent in the targeting of CD3. In one embodiment, the degree of binding of the anti-CD3 antibody to an irrelevant protein other than CD3 is, for example, less than about 10% of the binding of the antibody to CD3 as measured by, for example, radioimmunoassay (RIA). In certain embodiments, the dissociation constant (Kd) of the antibody that binds to CD3 is <1 μM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0 .01 nM, or <0.001 nM (e.g., 10 M or less, e.g., 10 M to 10 -8 M, e.g., 10 -8 M to 10 -13 M, e.g., 10 -9 M to 10 -13 M). In certain embodiments, the anti-CD3 antibody binds to an epitope of CD3 that is conserved among CD3s from different species.

[0042] In one embodiment, the anti-CD3 antibody can be ChAglyCD3 (otelixizumab). Otelixizumab is a humanized Fc non-binding anti-CD3 that was first evaluated in a Phase 2 trial by the Belgian Diabetes Registry (BDR), then developed by Tolerx, and then Tolerx, in collaboration with GSK, conducted a Phase 3 D The EFEND New-Onset T1D Trials (NCT00678886, NCT01123083, N CT00763451) were conducted. Otelixizumab was administered IV by infusion over 8 days . For example, all of which are hereby incorporated by reference in their entirety, Wiczling et al . J Clin Pharmacol. 50(5) (May 2010) 494 - 50 6; Keymeulen et al., N Engl J Med. 2005; 352 :2598 - 608; Keymeulen et al., Diabetologia. 2010; 53:614 - 23; Hagopian et al., Diabetes. 2013; 62:3901 - 8; Aronson et al., Diabetes C are. 2014; 37:2746 - 54; Ambery et al., Diabet Med. 2014; 31:399 - 402; Bolt et al., Eur. J. I mmunol. lYY3. 23: 403 - 411; Vlasakakis et al ., Br J Clin Pharmacol(2019)85 704 - 714; Gu glielmi et al, Expert Opinion on Biologic al Therapy, 16:6, 841 - 846; Keymeulen et al. , N Engl J Med 2005; 352:2598 - 608; Keymeule n et al., BLOOD 2010, VOL 115, No.6; Sprange rs et al., Immunotherapy(2011)3(11), 1303 - 1316; Daifotis et al., Clinical Immunology (2013)149, 268 - 278. See also

[0043] In another embodiment, the anti-CD3 antibody can be visilizumab (also referred to as HuM291; Nuvion). Visilizumab is a humanized anti-CD3 monoclonal antibody characterized by a mutant IgG2 isotype, lack of binding to Fcγ receptors, and the ability to selectively induce apoptosis in activated T cells. Visilizumab has been evaluated in patients with graft-versus-host disease (NCT00720629; NCT00032279) as well as ulcerative colitis (NCT00267306) and Crohn's disease (NCT00267709). See, for example, Sandborn et al., Gut 59(11)(Nov 2010)1485-1492, which is incorporated herein by reference in its entirety. In another embodiment, the anti-CD3 antibody can be visilizumab (also referred to as HuM291; Nuvion). Visilizumab is a humanized anti-CD3 monoclonal antibody characterized by a mutant IgG2 isotype, lack of binding to Fcγ receptors, and the ability to selectively induce apoptosis in activated T cells. Visilizumab has been evaluated in patients with graft-versus-host disease (NCT00720629; NCT00032279) as well as ulcerative colitis (NCT00267306) and Crohn's disease (NCT00267709). See, for example, Sandborn et al., Gut 59(11)(Nov 2010)1485-1492, which is incorporated herein by reference in its entirety. In another embodiment, the anti-CD3 antibody can be visilizumab (also referred to as HuM291; Nuvion). Visilizumab is a humanized anti-CD3 monoclonal antibody characterized by a mutant IgG2 isotype, lack of binding to Fcγ receptors, and the ability to selectively induce apoptosis in activated T cells. Visilizumab has been evaluated in patients with graft-versus-host disease (NCT00720629; NCT00032279) as well as ulcerative colitis (NCT00267306) and Crohn's disease (NCT00267709). See, for example, Sandborn et al., Gut 59(11)(Nov 2010)1485-1492, which is incorporated herein by reference in its entirety. In another embodiment, the anti-CD3 antibody can be visilizumab (also referred to as HuM291; Nuvion). Visilizumab is a humanized anti-CD3 monoclonal antibody characterized by a mutant IgG2 isotype, lack of binding to Fcγ receptors, and the ability to selectively induce apoptosis in activated T cells. Visilizumab has been evaluated in patients with graft-versus-host disease (NCT00720629; NCT00032279) as well as ulcerative colitis (NCT00267306) and Crohn's disease (NCT00267709). See, for example, Sandborn et al., Gut 59(11)(Nov 2010)1485-1492, which is incorporated herein by reference in its entirety. In another embodiment, the anti-CD3 antibody can be visilizumab (also referred to as HuM291; Nuvion). Visilizumab is a humanized anti-CD3 monoclonal antibody characterized by a mutant IgG2 isotype, lack of binding to Fcγ receptors, and the ability to selectively induce apoptosis in activated T cells. Visilizumab has been evaluated in patients with graft-versus-host disease (NCT00720629; NCT00032279) as well as ulcerative colitis (NCT00267306) and Crohn's disease (NCT00267709). See, for example, Sandborn et al., Gut 59(11)(Nov 2010)1485-1492, which is incorporated herein by reference in its entirety. In another embodiment, the anti-CD3 antibody can be visilizumab (also referred to as HuM291; Nuvion). Visilizumab is a humanized anti-CD3 monoclonal antibody characterized by a mutant IgG2 isotype, lack of binding to Fcγ receptors, and the ability to selectively induce apoptosis in activated T cells. Visilizumab has been evaluated in patients with graft-versus-host disease (NCT00720629; NCT00032279) as well as ulcerative colitis (NCT00267306) and Crohn's disease (NCT00267709). See, for example, Sandborn et al., Gut 59(11)(Nov 2010)1485-1492, which is incorporated herein by reference in its entirety. In another embodiment, the anti-CD3 antibody can be visilizumab (also referred to as HuM291; Nuvion). Visilizumab is a humanized anti-CD3 monoclonal antibody characterized by a mutant IgG2 isotype, lack of binding to Fcγ receptors, and the ability to selectively induce apoptosis in activated T cells. Visilizumab has been evaluated in patients with graft-versus-host disease (NCT00720629; NCT00032279) as well as ulcerative colitis (NCT00267306) and Crohn's disease (NCT00267709). See, for example, Sandborn et al., Gut 59(11)(Nov 2010)1485-1492, which is incorporated herein by reference in its entirety. In another embodiment, the anti-CD3 antibody can be visilizumab (also referred to as HuM291; Nuvion). Visilizumab is a humanized anti-CD3 monoclonal antibody characterized by a mutant IgG2 isotype, lack of binding to Fcγ receptors, and the ability to selectively induce apoptosis in activated T cells. Visilizumab has been evaluated in patients with graft-versus-host disease (NCT00720629; NCT00032279) as well as ulcerative colitis (NCT00267306) and Crohn's disease (NCT00267709). See, for example, Sandborn et al., Gut 59(11)(Nov 2010)1485-1492, which is incorporated herein by reference in its entirety.

[0044] In another embodiment, the anti-CD3 antibody can be faralumab, a fully human anti-CD3 monoclonal antibody being developed by Tiziana Life Sciences, PLC for NASH and T2D (NCT03291249). See, for example, Ogura et al., Clin Immunol. 2017;183:240-246; Ishikawa et al., Diabetes. 2007;56(8):2103-9; Wu et al., J Immunol. 2010;185(6):3401-7, which are incorporated herein by reference in their entireties. In another embodiment, the anti-CD3 antibody can be faralumab, a fully human anti-CD3 monoclonal antibody being developed by Tiziana Life Sciences, PLC for NASH and T2D (NCT03291249). See, for example, Ogura et al., Clin Immunol. 2017;183:240-246; Ishikawa et al., Diabetes. 2007;56(8):2103-9; Wu et al., J Immunol. 2010;185(6):3401-7, which are incorporated herein by reference in their entireties. In another embodiment, the anti-CD3 antibody can be faralumab, a fully human anti-CD3 monoclonal antibody being developed by Tiziana Life Sciences, PLC for NASH and T2D (NCT03291249). See, for example, Ogura et al., Clin Immunol. 2017;183:240-246; Ishikawa et al., Diabetes. 2007;56(8):2103-9; Wu et al., J Immunol. 2010;185(6):3401-7, which are incorporated herein by reference in their entireties. In another embodiment, the anti-CD3 antibody can be faralumab, a fully human anti-CD3 monoclonal antibody being developed by Tiziana Life Sciences, PLC for NASH and T2D (NCT03291249). See, for example, Ogura et al., Clin Immunol. 2017;183:240-246; Ishikawa et al., Diabetes. 2007;56(8):2103-9; Wu et al., J Immunol. 2010;185(6):3401-7, which are incorporated herein by reference in their entireties. In another embodiment, the anti-CD3 antibody can be faralumab, a fully human anti-CD3 monoclonal antibody being developed by Tiziana Life Sciences, PLC for NASH and T2D (NCT03291249). See, for example, Ogura et al., Clin Immunol. 2017;183:240-246; Ishikawa et al., Diabetes. 2007;56(8):2103-9; Wu et al., J Immunol. 2010;185(6):3401-7, which are incorporated herein by reference in their entireties. In another embodiment, the anti-CD3 antibody can be faralumab, a fully human anti-CD3 monoclonal antibody being developed by Tiziana Life Sciences, PLC for NASH and T2D (NCT03291249). See, for example, Ogura et al., Clin Immunol. 2017;183:240-246; Ishikawa et al., Diabetes. 2007;56(8):2103-9; Wu et al., J Immunol. 2010;185(6):3401-7, which are incorporated herein by reference in their entireties. In another embodiment, the anti-CD3 antibody can be faralumab, a fully human anti-CD3 monoclonal antibody being developed by Tiziana Life Sciences, PLC for NASH and T2D (NCT03291249). See, for example, Ogura et al., Clin Immunol. 2017;183:240-246; Ishikawa et al., Diabetes. 2007;56(8):2103-9; Wu et al., J Immunol. 2010;185(6):3401-7, which are incorporated herein by reference in their entireties.

[0045] In another embodiment, the anti-CD3 antibody can be teprilizumab, which is also known as hOKT3γl(Ala-Ala) (containing alanine at positions 234 and 235) and mediates the functional alteration of T lymphocytes that mediate the destruction of insulin-producing β cells in pancreatic islets. In another embodiment, the anti-CD3 antibody can be teprilizumab, which is also known as hOKT3γl(Ala-Ala) (containing alanine at positions 234 and 235) and mediates the functional alteration of T lymphocytes that mediate the destruction of insulin-producing β cells in pancreatic islets. In another embodiment, the anti-CD3 antibody can be teprilizumab, which is also known as hOKT3γl(Ala-Ala) (containing alanine at positions 234 and 235) and mediates the functional alteration of T lymphocytes that mediate the destruction of insulin-producing β cells in pancreatic islets. An anti-CD3 antibody that has been engineered to be further made. Teplizumab binds to the epitope of the CD3s chain expressed on mature T cells, thereby changing their function. The sequence and composition of teplizumab are incorporated herein by reference in their entirety from U.S. Patent Nos. 6,491,916; 8,663,634; and 9,056, . The complete sequences of the light and heavy chains are described below. The bolded portions are Complementary determining regions. Teplizumab light chain (SEQ ID NO: 1): JPEG0007696871000001.jpg1296 Teplizumab heavy chain (SEQ ID NO: 2): JPEG0007696871000002.jpg2496

[0046] In some embodiments, pharmaceutical compositions are provided herein. Such compositions contain a prophylactically effective amount of an anti-CD3 antibody and a pharmaceutically acceptable carrier. In certain embodiments, the term "pharmaceutically acceptable" means approved by a federal or state government regulatory authority for use in animals, more particularly in humans, or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeias. The term "carrier" refers to a diluent, adjuvant (e.g., Freund's adjuvant (complete and incomplete)), excipient, or vehicle with which the therapeutic agent is co-administered. Such pharmaceutical carriers can be sterile liquids such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, mineral oil, sesame oil, etc. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Aqueous saline solutions as well as aqueous dextrose and glycerol solutions The liquid can also be used as a liquid carrier, particularly as an injection solution. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, cereal flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol, etc. (for example, Handbook of Pharmaceutical Excipients, Arthur H. Kibbe (ed., 2000, the entire is incorporated herein by reference), see Am. Pharmaceutical Association, Washington, D.C).

[0047] If desired, the composition may also contain trace amounts of wetting agents or emulsifiers, or pH buffering agents. These compositions may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained release formulations, etc. Oral formulations may contain standard carriers such as, for example, pharmaceutical grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutical carriers are described in “Remington’s Pharmaceutical Sci ences” by E.W. Martin. Such compositions contain a prophylactically effective amount or therapeutically effective amount of a prophylactic or therapeutic agent, preferably in a purified form, together with an appropriate amount of carrier to provide a form suitable for proper administration to a patient. The formulation should be suitable for the mode of administration. In a preferred embodiment, the pharmaceutical composition is sterile and is for a subject, preferably an animal subject. It is a form suitable for administration to an elephant, more preferably a mammalian subject, and most preferably a human subject. .

[0048] In certain embodiments, it may be desirable to locally administer the pharmaceutical composition to the area in need of treatment, which can be achieved, for example, but not limited to, by local injection or by using an implant, and the implant is a membrane such as a silicone membrane, or a porous material, a non-porous material, or a gelatinous material including fibers. Preferably, when administering the anti-CD3 antibody, care must be taken to use a material that is not absorbed by the anti-CD3 antibody. For example, but not limited to, by local injection or by using an implant, and the implant is a membrane such as a silicone membrane, or a porous material, a non-porous material, or a gelatinous material including fibers. It can be achieved by injection or by using an implant, and the implant is a membrane such as a silicone membrane, or a porous material, a non-porous material, or a gelatinous material including fibers. It can be achieved by injection or by using an implant, and the implant is a membrane such as a silicone membrane, or a porous material, a non-porous material, or a gelatinous material including fibers. It can be achieved by injection or by using an implant, and the implant is a membrane such as a silicone membrane, or a porous material, a non-porous material, or a gelatinous material including fibers. Preferably, when administering the anti-CD3 antibody, care must be taken to use a material that is not absorbed by the anti-CD3 antibody. It can be achieved by injection or by using an implant, and the implant is a membrane such as a silicone membrane, or a porous material, a non-porous material, or a gelatinous material including fibers. Preferably, when administering the anti-CD3 antibody, care must be taken to use a material that is not absorbed by the anti-CD3 antibody.

[0049] In another embodiment, the composition can be delivered as vesicles, particularly liposomes (Langer, Science 249:1527-1533 (1990); Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; generally see the same book). Langer, Science 249:1527-1533 (1990); Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; generally see the same book). Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; generally see the same book). Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; generally see the same book). Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; generally see the same book). Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; generally see the same book). Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; generally see the same book).

[0050] In yet another embodiment, the composition can be delivered in a controlled release or sustained release system. In one embodiment, a pump can be used to achieve controlled release or sustained release (Langer, supra; Sefton, (1987), CRC Crit. Ref. Biomed. Eng. 14:20; Buchwald et al., 19 In yet another embodiment, the composition can be delivered in a controlled release or sustained release system. In one embodiment, a pump can be used to achieve controlled release or sustained release (Langer, supra; Sefton, (1987), CRC Crit. Ref. Biomed. Eng. 14:20; Buchwald et al., 19 In yet another embodiment, the composition can be delivered in a controlled release or sustained release system. In one embodiment, a pump can be used to achieve controlled release or sustained release (Langer, supra; Sefton, (1987), CRC Crit. Ref. Biomed. Eng. 14:20; Buchwald et al., 19 .Ref.Biomed.Eng.14:20;Buchwald et al.,19 80, Surgery 88:507; Saudek et al., 1989, N.E ngl. J. Med. 321:574). In another embodiment, a polymeric material is used to achieve controlled or sustained release of the antibody or fragment thereof of the present invention (e.g., Medical Applications of Controll ed Release, Langer and Wise (eds.), CRC Pre s., Boca Raton, Fla. (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley , New York (1984); Ranger and Peppas, (1983) , J., Macromol. Sci. Rev. Macromol. Chem. 23:61 ; see also, Levy et al., 1985, Science 228 :190; During et al., 1989, Ann. Neurol. 25:35 1; Howard et al., 1989, J. Neurosurg. 71:105); U.S. Patent No. 5,679,377; U.S. Patent No. 5,916,597; U.S. Patent No. 5, 912,015; U.S. Patent No. 5,989,463; U.S. Patent No. 5,128,326 ; PCT Publication No. WO99 / 15154; and PCT Publication No. WO99 / 20253 ). Examples of polymers used in sustained release formulations include, but are not limited to poly(2-hydroxyethyl methacrylate), poly(methyl methacrylate), poly( acrylic acid), poly(ethylene-co-vinyl acetate), poly(methacrylic acid), polyglyceryl Poly(vinyl chloride), polyanhydride, poly(N-vinyl pyrrolidone), poly(vinyl alkenyl) Poly(ethylene glycol), polyacrylamide, polylactic acid (PLA), Poly(lactide-co-glycolide) (PLGA), and polyorthoesters. In a preferred embodiment, the polymers used in the sustained release formulation are inert and do not leach. It is free of possible impurities, stable during storage, sterile, and biodegradable. In another embodiment, a controlled release or sustained release system is placed in proximity to the therapeutic target, i.e., the lungs. and thus only a fraction of the systemic dose is required (e.g., Goodson, in Medical Applications of Controlled Re See Lease, Supra, Vol. 2, pp. 115-138 (1984). stomach).

[0051] Controlled release systems are reviewed by Langer (1990, Science 249: 1527-1533). Sustained release formulations comprising one or more antibodies or fragments thereof of the invention may be made. See, for example, U.S. Pat. No. 4,526, each of which is incorporated herein by reference in its entirety. ,938; PCT Publication No. WO91 / 05548; PCT Publication No. WO96 / 20698 No.;Ning et al.,1996,Radiotherapy & Oncolo gy 39:179-189;Song et al.,1995,PDA Journal al of Pharmaceutical Science & Technolog y 50:372-397; Cleek et al., 1997, Pro. Int’l . Symp. Control. Rel. Bioact. Mater. 24:853-85 4; and Lam et al., 1997, Proc. Int’l. Symp. Con trol Rel. Bioact. Mater. 24:759-760, see.

[0052] The pharmaceutical composition can be formulated to be compatible with its intended route of administration. The examples of routes of administration include, but are not limited to, parenteral administration, such as intravenous administration, intradermal administration, subcutaneous administration, oral administration, intranasal administration (e.g., inhalation), transdermal (topical) administration, transmucosal administration, and rectal administration. In certain embodiments, the composition is formulated into a pharmaceutical composition suitable for intravenous, subcutaneous, intramuscular, oral, intranasal, or topical administration to humans according to conventional procedures. In a preferred embodiment, the pharmaceutical composition is formulated for subcutaneous administration to humans according to conventional procedures. Generally, compositions for intravenous administration are solutions in sterile isotonic aqueous buffers. If necessary, the composition may also contain solubilizing agents and local anesthetics such as lidocaine to relieve the pain at the injection site.

[0053] The composition can be formulated for parenteral administration by injection, such as by bolus injection or continuous infusion. Injectable formulations can be in unit dosage form, for example, in ampoules or in multi-dose containers, and can be provided with added preservatives. The composition can take the form of a suspension, solution or emulsion in an oily or aqueous vehicle, and can contain formulatory agents such as suspending agents, stabilizers It is possible. Alternatively, the active ingredient may be in powder form for constitution using a suitable vehicle, for example, sterile pyrogen-free water before use.

[0054] In certain embodiments, the present disclosure provides a dosage form (e.g., related to a pump or other device for such delivery) that enables continuous administration of an anti-CD3 antibody over a period of hours or days, e.g., 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 1 6 hours, 20 hours, 24 hours, 30 hours, 36 hours, 4 days, 5 days, 7 days, 10 days or 14 days. In other specific embodiments, the present invention provides a dosage form that enables continuous increase in dosage, e.g., from 51 μg / m / day to 82 2 6 μg / m / day and administration over a period of 24 hours, 30 hours, 36 hours, 4 days, 5 days, 7 2 days, 10 days or 14 days.

[0055] The composition can be formulated in neutral or salt form. Pharmaceutically acceptable salts include salts formed using anions such as those derived from hydrochloric acid, phosphoric acid, acetic acid, oxalic acid, tartaric acid, etc., and salts formed using cations such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxide, isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, procaine, etc.

[0056] Generally, the components of the compositions disclosed herein can be formulated as unit dosage forms separately or in combination. ​​​​​​In any of these, for example, as a dry lyophilized powder or a water-free concentrate, the active agent is supplied in a sealed container such as an ampoule or sachet indicating the quantity of the agent. When the composition is administered by infusion, it can be dispensed using an infusion bottle containing sterile pharmaceutical grade water or saline . When the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the components can be mixed before administration.

[0057] In particular, the present disclosure provides an anti-CD3 antibody or a pharmaceutical composition thereof that can be packaged in a sealed container such as an ampoule or sachet indicating the quantity of the agent. In one embodiment, the anti-CD3 antibody or a pharmaceutical composition thereof is supplied as a dry, sterile lyophilized powder or a water-free concentrate in a sealed container, and this can be reconstituted to a concentration suitable for administration to a subject using, for example, water or saline . The anti-CD3 antibody or a pharmaceutical composition thereof is provided as a dry, sterile lyophilized powder in a sealed container in a unit dose of at least 5 mg, more preferably at least 10 m g, at least 15 mg, at least 25 mg, at least 35 mg, at least 45 m g, at least 50 mg, at least 75 mg, or at least 100 mg. The lyophilized prophylactic agent or pharmaceutical composition herein should be stored between 2°C and 8°C in its original container, and the prophylactic or therapeutic agent or pharmaceutical composition of the present invention should be administered within 1 week, preferably within 5 days, within 72 hours, within 48 hours, within 24 hours, within 12 hours, within 6 hours, within 5 hours , within 3 hours, or within 1 hour after reconstitution. In an alternative embodiment, the pharmaceutical composition should be stored between 2°C and 8°C in its original container, and the prophylactic or therapeutic agent or pharmaceutical composition of the present invention should be administered within 1 week, preferably within 5 days, within 72 hours, within 48 hours, within 24 hours, within 12 hours, within 6 hours, within 5 hours , within 3 hours, or within 1 hour after reconstitution. should be administered within 1 week, preferably within 5 days, within 72 hours, within 48 hours, within 24 hours, within 12 hours, within 6 hours, within 5 hours The article provides a method of treating a subject having or at risk of having a disease or condition associated with T1D, the method comprising administering to the subject a composition of the invention in an amount effective to prevent or ameliorate one or more symptoms associated with T1D. The liquid form of the composition is preferably supplied in a sealed container at at least 0.25 mg / ml, more preferably at least 0.5 mg / ml, at least 1 mg / ml, at least 2.5 mg / ml, at least 5 mg / ml, at least 8 mg / ml, at least 10 mg / ml, at least 15 m g / kg, at least 25 mg / ml, at least 50 mg / ml, at least 75 mg / ml or at least 100 mg / ml. The liquid form should be stored in its original container between 2°C and 8°C.

[0058] In certain embodiments, the disclosure provides a composition of the invention packaged in a sealed container such as an ampoule or sachet indicating the quantity of the anti-CD3 antibody.

[0059] The composition may, if desired, be provided in a pack or dispenser device containing one or more unit dosage forms containing the active ingredient. The pack may include, for example , a metal or plastic foil, such as, for example, a blister pack.

[0060] The amount of the composition of the invention effective to prevent or ameliorate one or more symptoms associated with T1D can be determined by standard clinical techniques. The exact dosage to be used in the formulation depends also on the route of administration, and the severity of the condition and should be decided according to the judgment of the practitioner and the circumstances of each patient. The effective dosage can be estimated from the dosage-response curve derived from in vitro or animal model test systems.

[0061] <Methods and Uses> In certain embodiments, the present disclosure provides anti-human CD3 antibodies, such as teprotumumab, to individuals who have a predisposition to developing type 1 diabetes or have preclinical stage type 1 diabetes but do not meet the diagnostic criteria established by the American Diabetes Association or the Immunology of Diabetes Society, in order to prevent or delay the onset of type 1 diabetes and / or prevent or delay the need for insulin administration in such patients. In certain embodiments, high-risk factors for identifying individuals with a predisposition include having a first- or second-degree relative diagnosed with type 1 diabetes, abnormal fasting glucose levels (e.g., at least one determination of glucose levels between 100-125 mg / dL after an 8-hour fast), impaired glucose tolerance in response to a 75g OGTT (e.g., at least one determination of glucose levels between 140-199 mg / dL at 2 hours after a 75g OGTT), in Caucasians, having an HLA type of DR3, DR4, or DR7; in African Americans, having an HLA type of DR3 or DR4; in Japanese individuals, having an HLA type of DR3, DR4, or DR9; exposure to viruses (e.g., Coxsackie B virus, enterovirus, adenovirus, rubella, cytomegalovirus, Epstein-Barr virus), a positive diagnosis of at least one other autoimmune disorder (e.g., thyroid disease, celiac disease) according to criteria recognized in the art, and / or detection of autoantibodies in serum or other tissues, particularly ICA and type 1 diabetes-related autoantibodies. In certain embodiments, subjects identified as having a predisposition to developing type 1 diabetes are ​​​​​​​​​​​​​​​​​ at least one of the risk factors described herein and / or known in the art having. The present disclosure also encompasses the identification of a subject having a predisposition to develop type 1 diabetes, where the subject exhibits two or more, three or more, four or more, or more than five combinations of the risk factors disclosed herein or known in the art.

[0062] Serum autoantibodies associated with type 1 diabetes or a predisposition to develop type 1 diabetes are pancreatic islet cell auto antibodies (e.g., anti-ICA512 autoantibodies), glutamic acid decarboxylase autoantibodies ( e.g., anti-GAD65 autoantibodies), IA2 antibodies, ZnT8 antibodies, and / or anti-insulin autoantibodies. Thus, in certain examples according to this embodiment, the invention provides for treating an individual with a detectable autoantibody (e.g., anti-IA2, anti-ICA512, anti-GAD, or anti-insulin autoantibody) associated with a predisposition to develop type 1 diabetes or early stage type 1 diabetes, where the individual has not been diagnosed with type 1 diabetes and / or is a first-degree or second-degree relative of a type 1 diabetic patient. In certain embodiments the presence of the autoantibody is detected by ELISA, electrochemiluminescence (ECL), radioassay ( e.g., see Yu et al., 1996, J.Clin.Endocrinol.Met ab.81:4264-4267), agglutination PCR (Tsai et al , ACS Central Science 2016 2(3), 139-147), or by any other method for immunospecifically detecting antibodies described herein or known to those of skill in the art.

[0063] ​​​Before, during, and after treatment, β-cell function can be evaluated by the methods described herein or by any method known to those skilled in the art. For example, the Diabetes Control and Complications Trial (DCCT) Research Group established monitoring of percentage glycosylated hemoglobin (HA1 and HA1c) as a standard for evaluating blood glucose control (DCCT, (1993), N. Engl. J. Med. 329:977-986). Alternatively, daily insulin requirements, C-peptide levels / responses, hypoglycemic episodes, and / or characterization of the FPIR can be used as markers of β-cell function or to establish a therapeutic index (see Keymeulen et al., 2005, N. Engl. J. Med. 352:2598-2608; Herold et al., 2005, Diabetes 54:1763-1769; U.S. Patent Application Publication No. 2004 / 0038867 A1; and Greenbaum et al., 2001, Diabetes 50:470-476, respectively). For example, the FPIR is calculated as the sum of insulin values at 1 and 3 minutes after an IGTT performed according to the Islet Cell Antibody Register User’s Study protocol (see, for example, Bingley et al., 1996, Diabetes 45:1720-1728 and McCulloch et al., 1993, Diabetes Care 16:911-915). or by any method known to those skilled in the art. For example, the Diabetes s Control and Complications Trial (DCCT) Research Group established monitoring of percentage glycosylated hemoglobin (HA1 and HA1c) as a standard for evaluating blood glucose control (DCCT, (1993), N. Engl. J. Med. 329:977-986). Alternatively, or, the daily insulin requirements, C-peptide levels / responses, hypoglycemic episodes and / or characterization of the FPIR can be used as markers of β-cell function or to establish a therapeutic index (see Keymeulen et al., 2005, N. Engl. J. Med. 352:2598-2608; Herold et al., 2005, Diabetes 54:1763-1769; U.S. Patent Application Publication No. 2004 / 0038867 A1; and Greenbaum et al., 2001, Diabetes 50:470-476, respectively). For example, the FPIR is calculated as the sum of insulin values at 1 and 3 minutes after an IGTT performed according to the Islet Cell Antibody Register User’s Study protocol (see, for example, Bingley et al., 1996, Diabetes 45:1720-1728 and McCulloch et al., 1993, Diabetes Care 16:911-915). (see, for example, Bingley et al., 1996, Diabetes 45:1720-1728 and McCulloch et al., 1993, Diabetes Care 16:911-915). Islet Cell Antibody Register User’s Study protocol (see, for example, Bingley et al., 1996, Diabetes 45:1720-1728 and McCulloch et al., 1993, Diabetes Care 16:911-915). (see, for example, Bingley et al., 1996, Diabetes 45:1720-1728 and McCulloch et al., 1993, Diabetes Care 16:911-915). s 45:1720-1728 and McCulloch et al., 1993, Diabetes Care 16:911-915). (see, for example, Bingley et al., 1996, Diabetes 45:1720-1728 and McCulloch et al., 1993, Diabetes Care 16:911-915).

[0064] In some embodiments, an individual having a predisposition to develop T1D can be a non-diabetic subject who is a blood relative of a T1D patient. In certain embodiments, the non-diabetic subject can have two or more diabetes-related autoantibodies selected from islet cell antibodies (ICA), insulin autoantibodies (IAA), and antibodies against glutamic acid decarboxylase (GAD), tyrosine phosphatase (IA-2 / ICA512), or ZnT8.

[0065] In various embodiments, the non-diabetic subject has an abnormality in glucose tolerance in an oral glucose tolerance test (OGTT). An abnormality in glucose tolerance in the OGTT is defined as a fasting glucose level of 110 - 125 mg / dL, or a plasma 2-hour value of 140 mg / dL or higher and less than 200 mg / dL, or an intermediate glucose value at 30 minutes, 60 minutes, or 90 minutes during the OGTT that is greater than 200 mg / dL.

[0066] In some embodiments, a non-diabetic subject who responds to an anti-CD3 antibody, such as teplizumab, does not have an antibody against ZnT8. In certain embodiments, such a non-diabetic subject is HLA-DR4+ and not HLA-DR3+. In some embodiments, in such non-diabetic subjects who respond to an anti-CD3 antibody, such as teplizumab, an increase in the frequency (or relative amount) of TIGIT+KLRG1+CD8+ T cells in peripheral blood mononuclear cells is demonstrated (e.g., by flow cytometry) after administration (e.g., 1 month, 2 months, 3 months, or later or earlier than that).

[0067] In various embodiments, a prophylactically effective amount is 10 - 1000 micrograms per square meter (μg / m 2) of an anti-CD3 antibody, such as teplizumab. 2 ​​​​​​​​​​​) includes a 10 - 14 day cool down by subcutaneous (SC) injection or intravenous (IV) infusion. In one embodiment, the prophylactically effective amount is an anti - CD3 antibody, such as teplizumab, administered at 0 - 3 days at 51 μg / m , 103 μg / m , 2 , 207 μg / m 2 , , 413 μg / m 2 by IV infusion, and includes a 14 - day cool down with a single dose of 826 μg / m 2 by IV infusion on each day from 4 - 13 days. In certain embodiments , the prophylactically effective amount delays the median time to clinical diagnosis of T1D by at least 50%, at least 2 80%, or at least 90%, or by at least 12 months, at least 18 months, , at least 24 months, at least 36 months, at least 48 months, or at least 6 0 months, or longer.

[0068] In certain embodiments, the dosing cool down with an anti - CD3 antibody, such as teplizumab, can be repeated at intervals of 2 months, 4 months, 6 months, 8 months, 9 months, 10 months, 12 months, 15 months, 18 months , 24 months, 30 months, or 36 months. In certain embodiments , the effectiveness of treatment with an anti - CD3 antibody, such as teplizumab, is determined as described herein or as known in the art 2 months after, 4 months after, 6 months after, 9 months after, 12 months after, 15 months after, 18 months after, 24 months after, 30 months after, or 36 months after the previous treatment.

[0069]

[0069] In another embodiment, to prevent, treat, or ameliorate one or more symptoms of T1D in a subject, approximately 0.5 - 50 μg / kg, approximately 0.5 - 40 μg / kg ​, approximately 0.5 to 30 μg / kg, approximately 0.5 to 20 μg / kg, approximately 0.5 to 15 μg / kg, approximately 0.5 to 10 μg / kg, approximately 0.5 to 5 μg / kg, approximately 1 to 5 μg / kg, approximately 1 to 10 μg / kg, approximately 20 to 40 μg / kg, approximately 20 to 30 μg / kg, approximately 22 to 28 μg / kg or approximately 25 to 26 μg / kg of anti-C D3 antibody, such as teplizumab, in one or more unit doses. In another embodiment the subject is administered one or more unit doses of from about 200 μg / kg, 178 μg / kg, 180 μg / kg, 128 μg / k g, 100 μg / kg, 95 μg / kg, 90 μg / kg, 85 μg / kg, 80 μg / kg, 75 μg / kg, 70 μg / kg, 65 μg / kg, 60 μg / kg, 55 μg / kg, 50 μg / kg, 45 μg / kg, 40 μg / kg, 35 μg / kg, 30 μg / kg, 26 μg / kg, 25 μg / kg, 20 μg / kg, 15 μg / kg, 13 μg / kg, 10 μg / kg, 6.5 μg / kg, 5 μg / kg, 3.2 μg / kg, 3 μg / kg, 2.5 μg / kg, 2 μg / kg, 1.6 μg / kg, 1.5 μg / kg, 1 μg / kg, 0.5 μg / kg, 0.25 μg / kg, 0.1 μg / kg, or 0.05 μ g / kg of anti-CD3 antibody, such as teplizumab, in one or more unit doses are administered to the subject. In certain embodiments, the subject is administered one or more doses of from about 5 to 1200 μg / m

[0070] , preferably 51 to 826 2 μg / m of anti-CD3 antibody, such as teplizumab. 2 In certain embodiments, the subject is administered one or more doses of from about 5 to 1200 μg / m 。In another embodiment, to a subject, to prevent, treat, slow the progression of, delay the onset of, or reverse one or more symptoms of T1D, 1200 μg / m μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m 2 μg / m μg / m 2 μg / m 2 μg / m, or 5μg / m 2 of an anti-CD3 antibody, such as teplizumab administer one or more unit doses that are

[0071] In another embodiment, to a subject, administer a treatment regimen comprising a prophylactically effective amount of one or more doses of an anti-CD3 antibody, such as teplizumab, wherein the treatment cycle is for 2 days, 3 days days Administered over 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days or 14 days. In one embodiment, the treatment regimen is prophylactically effective doses are administered daily, every other day, every two days, every three days, or every four days including. In certain embodiments, the treatment regimen is a prophylactically effective dose, 14 doses, 13 doses, 13 doses, 12 doses, 11 doses, 10 doses, 9 doses, or 8 doses are administered until, on Monday, Tuesday, Wednesday, Thursday of a given week, and not on Friday, Saturday and Sunday of the same week a prophylactically effective dose is administered. In certain embodiments the dose administered is the same on each day of the regimen.

[0072] In certain embodiments, a subject is administered a treatment regimen comprising one or more doses of a prophylactically effective amount of an anti-CD3 antibody, such as teprilizumab, where the prophylactically effective amount is 200 μg / kg / day, 175 μg / kg / day, 150 μg / kg / day, 125 μg / kg / day, 1 00 μg / kg / day, 95 μg / kg / day, 90 μg / kg / day, 85 μg / kg / day, 80 μg / kg / day, 75 μg / kg / day, 70 μg / kg / day, 65 μg / kg / day, 60 μg / kg / day, 55 μg / kg / day, 50 μg / kg / day, 45 μg / kg / day, 40 μg / kg / day, 35 μg / kg / day, 30 μg / kg / day, 26 μg / kg / day, 25 μg / kg / day, 20 μg / kg / day, 15 μg / kg / day, 13 μg / kg / day, 10 μg / kg / day, 6.5 μg / kg / day, 5 μg / kg / day, 3.2 μg / kg / day 10 μg / kg / day, 6.5 μg / kg / day, 5 μg / kg / day, 3.2 μg / kg / day 3 μg / kg / day, 2.5 μg / kg / day, 2 μg / kg / day, 1.6 μg / kg / day , 1.5 μg / kg / day, 1 μg / kg / day, 0.5 μg / kg / day, 0.25 μg / k g / day, 0.1 μg / kg / day, or 0.05 μg / kg / day; and / or the prophylactically effective amount is 1200 μg / m 2 / day, 1150 μg / m 2 / day, 1100 μg / m 2 / day, 1050 μg / m 2 / day, 1000 μg / m 2 / day, 950 μg / m 2 / day, 900 μg / m 2 / day, 850 μg / m 2 / day, 800 μg / m 2 / day, 750 μg / m 2 / day, 700 μg / m 2 / day, 650 μg / m 2 / day, 600 μg / m 2 / day, 550 μg / m 2 / day, 500 μg / m 2 / day, 450 μg / m 2 / day, 400 μg / m 2 / day , 350 μg / m 2 / day, 300 μg / m 2 / day, 250 μg / m 2 / day, 200 μg / m 2 / day, 150 μg / m 2 / day, 100 μg / m 2 / day, 50 μg / m 2 / day, 40 μ g / m 2 / day, 30 μg / m 2 / day, 20 μg / m 2 / day, 15 μg / m 2 / day, 10 μ g / m 2 / day, or 5 μg / m 2 / day.

[0073] In another embodiment, preventing, treating, or To improve, 1200 μg / m 2 Hereinafter, 1150 μg / m 2 Hereinafter, 1100 μg / m 2 Hereinafter, 1050 μg / m 2 Hereinafter, 1000 μg / m 2 Hereinafter, 950 μg / m 2 Hereinafter, 900 μg / m 2 Hereinafter, 850 μg / m 2 Hereinafter, 800 μg / m 2 Hereinafter, 750 μg / m 2 Hereinafter, 700 μg / m 2 Hereinafter, 650 μg / m 2 Hereinafter, 600 μg / m 2 Hereinafter, 550 μg / m 2 Hereinafter, 500 μg / m 2 Hereinafter, 450 μg / m 2 Hereinafter, 400 μg / m 2 Hereinafter , 350 μg / m 2 Hereinafter, 300 μg / m 2 Hereinafter, 250 μg / m 2 Hereinafter, 200 μg / m 2 Hereinafter, 150 μg / m 2 Hereinafter, 100 μg / m 2 Hereinafter, 50 μg / m 2 Hereinafter, 40 μ g / m 2 Hereinafter, 30 μg / m 2 Hereinafter, 20 μg / m 2 Hereinafter, 15 μg / m 2 Hereinafter, 10 μ g / m 2 Hereinafter, or 5 μg / m 2 Hereinafter, an intravenous dose of an anti-CD3 antibody such as teprilizumab for about 24 hours, about 22 hours, about 20 hours, about 18 hours, about 16 hours, about 14 hours, about 12 hours, about 10 hours, about 8 hours, about 6 hours, about 4 hours, about 2 hours, about 1.5 hours, about 1 hour, about 50 minutes, about 40 minutes, about 30 minutes, about 20 minutes, about 10 minutes, about 5 minutes, about 2 minutes, about 1 minute 1 hour, about 50 minutes, about 40 minutes, about 30 minutes, about 20 minutes, about 10 minutes, about 5 minutes, about 2 minutes, about 1 minute Administer over about 30 seconds or about 10 seconds. The total dose over the duration of the regimen is, in total, 9000 μg / m 2 less than, 8000 μg / m 2 less than, 7000 μg / m 2 less than, 6000 μg / m 2 It is preferably less than, 5000 μg / m 2 less than, 4000 μg / m 2 less than, 3000 μg / m 2 less than, 2000 μg / m 2 less than, or 1000 μ g / m 2 It may be less than. In certain embodiments, the total dose administered in the regimen is 10 0 μg / m 2 ~200 μg / m 2 , 100 μg / m 2 ~500 μg / m 2 , 100 μg / m 2 ~1000 μg / m 2 or, 500 μg / m 2 ~1000 μg / m 2 is.

[0074] In preferred embodiments, the dose of the therapeutic regimen is increased gradually over the first quarter, the first half or the first two-thirds (e.g., over 10 days, 12 days, 14 days, 16 days, 18 days or 20 days at 1 dose per day, the first 2 days, 3 days, 4 days, 5 days, or 6 days of a 12-day, 14-day, 16-day, 18-day or 20-day regimen) until a prophylactically effective daily amount of an anti-CD3 antibody, such as teprotumumab, is achieved. In certain embodiments, a subject is administered a therapeutic regimen comprising one or more doses of a prophylactically effective amount of an anti-CD3 antibody, such as teprotumumab, where the prophylactically effective amount is, for example, increased by 0.01 μg / kg per day as the treatment progresses, 0 ​0.02 μg / kg each, 0.04 μg / kg each, 0.05 μg / kg each, 0.06 μ g / kg each, 0.08 μg / kg each, 0.1 μg / kg each, 0.2 μg / kg each 、0.25 μg / kg each, 0.5 μg / kg each, 0.75 μg / kg each, 1 μg / kg each, 1.5 μg / kg each, 2 μg / kg each, 4 μg / kg each, 5 μg / kg each, 10 μg / kg each, 15 μg / kg each, 20 μg / kg each, 25 μg / kg each, 30 μg / kg each, 35 μg / kg each, 40 μg / kg each, 45 μg / kg each, 50 μg / kg each, 55 μg / kg each, 60 μg / kg each, 65 μg / kg each, 70 μg / kg each, 75 μg / kg each, 80 μg / kg each, 85 μg / kg each, 90 μg / kg each, 95 μg / kg each, 100 μg / kg each, or 12 5 μg / kg each; or, for example, 1 μg / m 2 each day, 5 μg / m 2 each , 10 μg / m 2 each, 15 μg / m 2 each, 20 μg / m 2 each, 30 μg / m 2 each , 40 μg / m 2 each, 50 μg / m 2 each, 60 μg / m 2 each, 70 μg / m 2 each , 80 μg / m 2 each, 90 μg / m 2 each, 100 μg / m 2 each, 150 μg / m 2 each, 200 μg / m 2 each, 250 μg / m 2 each, 300 μg / m 2 each, 350 μg / m 2 each, 400 μg / m 2 each, 450 μg / m2 each, 500 μg / m 2 each , 550 μg / m 2 each, 600 μg / m 2 each, or 650 μg / m 2 each, and increase it. In certain embodiments, a subject is administered a treatment regimen comprising one or more doses of a prophylactically effective amount of an anti-CD3 antibody, such as teplyzumab, where the prophylactically effective amount is increased 1.25-fold, 1.5-fold , 2-fold, 2.25-fold, 2.5-fold, or 5-fold until the one-day prophylactically effective amount of the anti-CD 3 antibody, such as teplyzumab, is achieved.

[0075] In certain embodiments, to prevent, treat, or ameliorate one or more symptoms of T1D in a subject, 200 μg / kg or less, preferably 175 μg / kg or less, 1 50 μg / kg or less, 125 μg / kg or less, 100 μg / kg or less, 95 μg / kg or less , 90 μg / kg or less, 85 μg / kg or less, 80 μg / kg or less, 75 μg / kg or less , 70 μg / kg or less, 65 μg / kg or less, 60 μg / kg or less, 55 μg / kg or less , 50 μg / kg or less, 45 μg / kg or less, 40 μg / kg or less, 35 μg / kg or less , 30 μg / kg or less, 25 μg / kg or less, 20 μg / kg or less, 15 μg / kg or less , 10 μg / kg or less, 5 μg / kg or less, 2.5 μg / kg or less, 2 μg / kg or less , 1.5 μg / kg or less, 1 μg / kg or less, 0.5 μg / kg or less, or 0.5 μg / kg or less of an anti-CD3 antibody, such as teplyzumab, otelixizumab, or foralumab is administered intramuscularly in one or more doses.

[0076] In another embodiment, to prevent, treat, or or to prevent, treat, or improve, administer one or more doses subcutaneously of an anti-CD3 antibody at 200 μg / kg or less, preferably 175 μg / kg or less, 15 0 μg / kg or less, 125 μg / kg or less, 100 μg / kg or less, 95 μg / kg or less , 90 μg / kg or less, 85 μg / kg or less, 80 μg / kg or less, 75 μg / kg or less , 70 μg / kg or less, 65 μg / kg or less, 60 μg / kg or less, 55 μg / kg or less , 50 μg / kg or less, 45 μg / kg or less, 40 μg / kg or less, 35 μg / kg or less , 30 μg / kg or less, 25 μg / kg or less, 20 μg / kg or less, 15 μg / kg or less , 10 μg / kg or less, 5 μg / kg or less, 2.5 μg / kg or less, 2 μg / kg or less, 1.5 μg / kg or less, 1 μg / kg or less, 0.5 μg / kg or less, or 0.5 μg / kg or less, such as teplizumab, otelixizumab, or faralimab. In another embodiment, to prevent, treat, or improve one or more symptoms of T1D in a subject, administer one or more doses subcutaneously of an anti-CD3 antibody at 100 μg / kg or less, preferably 95 μg / kg or less, 90 μ

[0077] g / kg or less, 85 μg / kg or less, 80 μg / kg or less, 75 μg / kg or less, 70 μ g / kg or less, 65 μg / kg or less, 60 μg / kg or less, 55 μg / kg or less, 50 μ g / kg or less, 45 μg / kg or less, 40 μg / kg or less, 35 μg / kg or less, 30 μ g / kg or less, 25 μg / kg or less, 20 μg / kg or less, 15 μg / kg or less, 10 μ g / kg or less, 5 μg / kg or less, 2.5 μg / kg or less, 2 μg / kg or less, 1.5 μ g / kg or less, 1 μg / kg or less, 0.5 μg / kg or less, or 0.5 μg / kg or less ​​One anti-CD3 antibody, such as teplizumab, otelixizumab or faralimab Administer one or more doses intravenously. In another embodiment, one or more symptoms of T1D To prevent, treat, or ameliorate, 100 μg / kg or less, 95 μg / k g or less, 90 μg / kg or less, 85 μg / kg or less, 80 μg / kg or less, 75 μg / k g or less, 70 μg / kg or less, 65 μg / kg or less, 60 μg / kg or less, 55 μg / k g or less, 50 μg / kg or less, 45 μg / kg or less, 40 μg / kg or less, 35 μg / k g or less, 30 μg / kg or less, 25 μg / kg or less, 20 μg / kg or less, 15 μg / k g or less, 10 μg / kg or less, 5 μg / kg or less, 2.5 μg / kg or less, 2 μg / kg or less, 1.5 μg / kg or less, 1 μg / kg or less, 0.5 μg / kg or less, or 0.5 μg / kg or less of an anti-CD3 antibody, such as teplizumab, otelixizumab or faral mab, as an intravenous dose over about 6 hours, about 4 hours, about 2 hours, about 1.5 hours, about 1 hour, about 50 minutes, about 40 minutes, about 30 minutes, about 20 minutes, about 10 minutes, about 5 minutes, about 2 minutes, about 1 minute, about 30 seconds or about 10 seconds.

[0078] In another embodiment, to a subject to prevent, treat, or ameliorate one or more symptoms of T1D, 100 μg / kg or less, preferably 95 μg / kg or less, 90 μ g / kg or less, 85 μg / kg or less, 80 μg / kg or less, 75 μg / kg or less, 70 μ g / kg or less, 65 μg / kg or less, 60 μg / kg or less, 55 μg / kg or less, 50 μ g / kg or less, 45 μg / kg or less, 40 μg / kg or less, 35 μg / kg or less, 30 μ g / kg or less, 25 μg / kg or less, 20 μg / kg or less, 15 μg / kg or less, 10 μ less than g / kg, less than 5 μg / kg, less than 2.5 μg / kg, less than 2 μg / kg, less than 1.5 μ g / kg, less than 1 μg / kg, less than 0.5 μg / kg, or less than 0.5 μg / kg of an anti-CD3 antibody, such as teplizumab, otrexup, or foralumab, one or a plurality of doses is administered orally. In another embodiment, one or more symptoms of T1D are prevented, treated, or ameliorated with an oral dose of an anti-CD3 antibody of 100 μg / kg or less, 95 μg / kg or less, 90 μg / kg or less, 85 μg / kg or less, 80 μg / kg or less, 75 μg / kg or less, 70 μg / kg or less, 65 μg / kg or less, 60 μg / kg or less, 55 μg / kg or less, 50 μg / kg or less, 45 μg / kg or less, 40 μg / kg or less, 35 μg / kg or less, 30 μg / kg or less, 25 μg / kg or less, 20 μg / kg or less, 15 μg / kg or less, 10 μg / kg or less, 5 μg / kg or less, 2.5 μg / kg or less, 2 μg / kg or less than 1.5 μg / kg, 1 μg / kg or less, 0.5 μg / kg or less, or 0.5 μ g / kg or less of an anti-CD3 antibody, such as teplizumab, otrexup, or foraluma b, which is administered over a period of about 6 hours, about 4 hours, about 2 hours, about 1.5 hours, about 1 hour, about 50 minutes, about 40 minutes, about 30 minutes, about 20 minutes, about 10 minutes, about 5 minutes, about 2 minutes, about 1 minute, about 30 seconds, or about 10 seconds.

[0079] In certain embodiments where escalating doses are administered over the first few days of the dosing regimen, the dose on day 1 of the regimen is 5 - 100 μg / m / day, preferably 51 μg / m 2 / day, and by day 3, 4, 5, 6, or 7, the daily dose as described immediately above 2 is and up to the 1-day dose described immediately above by day 3, 4, 5, 6, or 7 of the regimen gradually increase up to a certain amount. For example, for a subject, approximately 51 μg / m 2 / day of dosage is administered on the first day and approximately 103 μg / m 2 / day is administered on the second day, and approximately 207 μg / m 2 / day is administered on the third day, and approximately 413 μg / m 2 / day is administered on the fourth day, and for the subsequent several days of the regimen( for example, days 5 - 14), 826 μg / m 2 / day is administered. In another embodiment, for the subject , approximately 227 μg / m 2 / day of dosage is administered on the first day, and approximately 459 μg / m 2 / day is administered on the second day, and approximately 919 μg / m 2 / day is administered on the third day and for the subsequent several days . In another embodiment, for the subject, approximately 284 μg / m 2 / day of dosage is administered on the first day, and on the second day, approximately 574 μg / m 2 / day is administered, and on the third day and for the subsequent several days, approximately 11 48 μg / m 2 / day is administered.

[0080] In other embodiments, the initial dose is 1 / 4, 1 / 2, the same amount as, or but in divided doses at intervals of 6 hours, 8 hours, 10 hours, or 12 hours of the last - day dose of the regimen. For example, a dose of 13 μg / kg / day is administered in 4 divided doses of 3 - 4 μg / kg at 6 - hour intervals to reduce the level of cytokine release caused by the administration of the antibody. In certain embodiments, to reduce the likelihood of cytokine release and other adverse effects, the first 1 dose, 2 doses, 3 doses, or 4 doses or all doses of the regimen are administered more slowly by intravenous administration. For example, a dose of 51 μg / m 2 / day is administered over about 5 minutes, about 15 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, and administered over about 22 hours. In certain embodiments, the dose is administered by slow infusion, for example, over a period of 20 to 24 hours. In certain embodiments the dose is administered by a pump, preferably increasing the concentration of the antibody being administered as the infusion proceeds.

[0081] In other embodiments, a specified fraction of the dose corresponding to the above 2 51 μg / m 2 / day to 826 μg / m / day regimen is administered as a gradually increasing dose. In certain embodiments, the fraction is 1 / 10, 1 / 4, 1 / 3, 1 / 2, 2 / 3, or 3 / 4 of the daily dose of the above regimen. Thus, when the fraction is 1 / 10, the daily dose is 5.1 μg / m 2 on day 1, 10.3 μg / m 2 on day 2, 20.7 μg / m 2 on day 3, 41.3 μg / m 2 on day 4, and 82.6 μg / m 2 on days 5 to 14. When the fraction is 1 / 4, the dose is 12.75 μg / m 2 on day 1, 25.25 μg / m 2 on day 2, 51 μg / m 2 on day 3, 103 μg / m 2 on day 4, and 207 μg / m 2 on days 5 to 14. When the fraction is 1 / 3, the dose is 17 μg / m 2 on day 1, 34.3 μg / m 2 on day 2, 69 μg / m 2 on day 3, 137.6 μg / m 2, and on the 5th to 14th days is 275.3 μ g / m 2 becomes. When the fraction is 1 / 2, the dosage is 25.5 μg / m on the first day 2 , the second day is 51 μg / m 2 , the third day is 103 μg / m 2 , the fourth day is 207 μg / m 2 , and on the 5th 2 ~14th days is 413 μg / m / m 2 , the second day is 69 μg / m 2 , the third day is 137.6 μg / m 2 , the fourth day is 275.3 μg / m 2 , and on the 5th to 14th days is 550.1 μg / m 2 becomes. When the fraction is 3 / 4, the dosage is 38.3 μg / m on the first day 2 , the second day is 77.3 μg / m 2 , the third day is 155. 3 μg / m 2 , the fourth day is 309.8 μg / m 2 , and on the 5th to 14th days is 620 μg / m 2 becomes. In other embodiments, the regimen is the same as one of the above regimens, but covers only the 1st to 4th days, the 1st to 5th days, or the 1st to 6th days. For example, in a particular embodiment , the dosage is 17 μg / m on the first day 2 , the second day is 34.3 μg / m 2 , the third day is 69 μg / m 2 , the fourth day is 137.6 μg / m 2 , and on the 5th and 6th days is 275.3 μg / m 2 becomes.

[0082] In a particular embodiment, an anti-CD3 antibody, such as teplizumab, otrexup, or f Rather than administering oral mab at a daily dose over several days, it is administered by infusion in a continuous manner over 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 15 hours, 18 hours, 20 hours, 24 hours, 30 hours or 36 hours. The infusion may be constant. For example, the first 1 hour, 2 hours, 3 hours, 5 hours, 6 hours, or 8 hours of the infusion may start at a low dose and then increase to a higher dose. Over the course of the infusion, the patient receives a dose equal to the amount administered in the above 5-20 day regimen. For example, about 150 μg / m, 200 μg / m, 250 μg / m, 500 μg / m, 750 μg / m, 1000 μg / m, 1500 μg / m, 2000 μg / m, 3000 μg / m, 4000 μg / m, 5000 μg / m, 6000 μg / m, 7000 μg / m, 8000 μg / m, or 9000 μg / m dose. In particular, the infusion speed and duration are designed so that the level of free anti-CD3 antibody, such as teprilizumab, otrexizumab or foralumab, in the subject after administration is minimized. In certain embodiments, the level of free anti-CD3 antibody, such as teprilizumab, should not exceed 200 ng per ml of free antibody. Further, the infusion is designed so that at least 50%, 60%, 70%, 80%, 90%, 95% or 100% of the T cell receptor coating and modulation of the joints are achieved. Among them, 10 hours, 12 hours, 15 hours, 18 hours, 20 hours, 24 hours, 30 hours or 3 6 hours of continuous infusion. The infusion may be constant, for example , the first 1 hour, 2 hours, 3 hours, 5 hours, 6 hours, or 8 hours of the infusion starts at a low dose and then can increase to a higher dose. Over the course of the infusion, the patient receives a dose equal to the amount administered in the above 5-20 day regimen. For example, about 150 μg / m 2 200 μg / m 2 250 μg / m 2 500 μg / m 2 750 μg / m 2 1000 μg / m 2 1500 μg / m 2 2000 μg / m 2 3000 μg / m 2 4000 μg / m 2 5000 μg / m 2 6000 μg / m 2 7000 μg / m 2 8000 μg / m 2 or 9000 μg / m 2 dose. In particular, the infusion speed and duration are designed so that the level of free anti-CD3 antibody, such as teplizumab , otrexizumab or foralumab, in the subject after administration is minimized. In a particular embodiment, the level of free anti-CD3 antibody, such as teplizumab, should not exceed 200 ng per ml of free antibody. Further, the infusion is adjusted so that at least 50%, 6 0%, 70%, 80%, 90%, 95% or 100% of the T cell receptor coating and modulation of the joints are achieved.

[0083] In other embodiments, to treat, prevent, or slow or delay the onset or worsening of, or reverse, one or more symptoms of type 1 diabetes, an anti-CD3 antibody, such as teplizumab, otelixizumab, or faralumab, is administered over a long period of time. For example, in certain embodiments, an anti-CD3 antibody, such as teplizumab, is administered at a low dose, as an alternative to, or to enhance or maintain the effect after, the above-described dosing regimen of 6 to 14 days, once a month, twice a month, three times a month, once a week, or more frequently. Such low doses can be any dose from 1 μg / m to 100 μg / m , for example, approximately 5 μg / m , 10 μg / m 2 , 15 μg / m 2 , 20 μg / m 2 , 25 μg / m 2 2 , 30 μg / m 2 2 , 35 μg / m 2 2

[0084] , 40 μg / m 2 2 , 45 μg / m 2 , or 50 μg / m

[0084] In other embodiments, the subject can be redosed at some point after the administration of the dosing regimen of an anti-CD3 antibody, such as teplizumab, otelixizumab, or faralumab, for example, based on one or more physiological parameters, and of course, redosing can also be done. The administration of such redosing and / or the evaluation of the need for such redosing can be performed 2 months, 4 months, 6 months, 8 months, 9 months, 1 year, 15 months, etc. after the administration of the dosing regimen. It can be performed after 6 months, 18 months, 2 years, 30 months or 3 years, and also every 6 months and may include performing an unrestricted treatment cool every 9 months, every year, every 15 months, every 18 months, every 2 years, every 30 months or every 3 years.

Example

[0085] [Summary] Background: Type 1 diabetes (T1D), which leads to the destruction of insulin-producing β-cells and dependence on exogenous insulin for survival, is a chronic autoimmune disease. Although some interventions have shown success in attenuating the loss of insulin production in patients with clinical disease, no intervention has been able to affect the progression of the disease in individuals at high risk of developing the disease. Although some interventions have shown success in attenuating the loss of insulin production in patients with clinical disease, no intervention has been able to affect the progression of the disease in individuals at high risk of developing the disease. to date.

[0086] Method: A randomized, placebo-controlled, double-blind trial of teprilizumab (FcR non-binding anti-CD3 mAb) was conducted in non-diabetic blood relatives of T1D patients at high risk of clinical disease. Patients were randomized to a single 14-day cool of drug or placebo and followed for disease progression using an oral glucose tolerance test at approximately 6-month intervals. randomized to a single 14-day cool of drug or placebo and followed for disease progression using an oral glucose tolerance test at approximately 6-month intervals. using an oral glucose tolerance test at approximately 6-month intervals.

[0087] Results: A total of 76 subjects were randomly assigned, 44 to the teprilizumab arm and 32 to the placebo arm. 72% of the participants were pediatric. Treatment with teprilizumab delayed the median time to clinical diagnosis of T1D from 24.4 months to 48.4 months (Cox proportional hazards, p = 0.006), and the annual incidence of diabetes decreased from 9.8% to 4.1%. Treatment with the drug resulted in TIGIT+KLRG1+CD ​The percentage of 8+ T cells was increased in participants without antibodies to ZnT8 (p=0.01). , participants who were HLA-DR4+ (p=0.006), and not HLA-DR3+. participants (p=0.05) were most likely to respond to teplizumab.

[0088] Conclusion: Teplizumab delays the diagnosis of T1D in high-risk individuals. Subgroups of individuals may be more likely to respond to the treatment and benefit.

[0089] [method] <Test participants> Participants in the TrialNet Natural History Study 14 Through The study was conducted between July 2011 and November 2018 in the United States, Canada, and the United States. The study was conducted at sites in Germany and Germany. Institutional Review Board (IRB) approval was obtained at each participating site. Written informed consent was obtained from the patient, the patient's parent, or both prior to entering the study. I got the point.

[0090] Eligible participants were nondiabetic relatives of patients with type 1 diabetes and aged 8 years or older at the time of randomization. Participants were at high risk of developing clinical diabetes. had two or more diabetes-associated autoantibodies on two specimens collected within six months prior to damming; Additionally, eligible participants were required to undergo an oral glucose tolerance test (OGTT) within 52 days of enrollment. twice a day, fasting glucose levels between 110 and 125 mg / dL, or 140 mg / dL 2-hour plasma value of >200 mg / dL or 30-hour plasma value during OGTT of >200 mg / dL Impaired glucose tolerance, defined as intermediate glucose values ​​at 10, 60, or 90 minutes after Had. The protocol was amended in 2014 to allow enrollment of participants under 18 years of age with a single abnormal OGTT. This was because the rate of T1D progression was similar with and without a confirmatory OGTT in this age group. In these 8 subjects (5 in the teprilizumab arm and 3 in the placebo arm), a second pre-treatment OGTT was performed on the first day of study drug administration. Individuals with other significant medical history, abnormal clinical chemistry values or blood counts were excluded.

[0091] <Patient Identification> Subjects were identified in the TrialNet Pathway to Prevention (PTP) trial. See Figure 5. The PTP trial enrolls first-degree relatives of T1D patients aged 1 - 45 years, and second- or third-degree relatives up to 20 years of age, and assesses for diabetes autoantibodies to micronutrient insulin (mIAA), glutamic acid decarboxylase-65 (GAD), and insulinoma-associated antigen-2 (IA-2, or ICA512). If at least one of the other antibodies tested was positive, pancreatic islet cell (ICA) and zinc transporter 8 (ZnT8) autoantibodies were measured.

[0092] <Study Design and Intervention> Participants were randomized to receive either teprilizumab or placebo, with equal allocation to each group. Randomization was stratified at each TrialNet study site by age (over 18 years or under 18 years) and glucose between pre-randomization OGTT status. Assignment to treatment was double masked.

[0093] Participants came to the clinical research center as outpatients and received drug administration as previously described​​​​​​​​​ Drug regimen 9,10 Received a 14-day cool-down using teprilizumab or placebo. Specifically, participants assigned to receive the active investigational drug received teprilizumab via a one-day IV infusion of 51 micrograms per square meter (μg / m ), 103 μg / m ), 207 μg / m ), and 413 2 μg / m 2 ), respectively, on Days 0–3 of the study, and a single daily dose of 826 μg / m 2 on each day from Days 4–13 of the study, according to a specific schedule. Participants randomized to the placebo arm received a 14-day cool-down of corresponding IV saline. Participants received ibuprofen and diphenhydramine prior to infusion for the first 5 days and then further dosing of ibuprofen, diphenhydramine, and / or acetaminophen as needed to relieve symptoms. Discontinuation criteria for the investigational drug infusion as defined in the protocol were followed. μg / m 2 throughout the entire study, all subjects had a formal interview with the study staff regarding adverse events and symptoms of diabetes. 2 schedule. Participants randomized to the placebo arm received a 14-day cool-down of corresponding IV saline. Participants received ibuprofen and diphenhydramine prior to infusion for the first 5 days and then further dosing of ibuprofen, diphenhydramine, and / or acetaminophen as needed to relieve symptoms. Discontinuation criteria for the investigational drug infusion as defined in the protocol were followed. saline according to a corresponding IV schedule. Participants received ibuprofen and diphenhydramine prior to infusion for the first 5 days and then further dosing of ibuprofen, diphenhydramine, and / or acetaminophen as needed to relieve symptoms. Discontinuation criteria for the investigational drug infusion as defined in the protocol were followed. saline according to a corresponding IV schedule. Participants received ibuprofen and diphenhydramine prior to infusion for the first 5 days and then further dosing of ibuprofen, diphenhydramine, and / or acetaminophen as needed to relieve symptoms. Discontinuation criteria for the investigational drug infusion as defined in the protocol were followed. throughout the entire study, all subjects had a formal interview with the study staff regarding adverse events and symptoms of diabetes. throughout the entire study, all subjects had a formal interview with the study staff regarding adverse events and symptoms of diabetes. throughout the entire study, all subjects had a formal interview with the study staff regarding adverse events and symptoms of diabetes.

[0094] Throughout the entire study, all subjects had a formal interview with the study staff regarding adverse events and symptoms of diabetes. Throughout the entire study, all subjects had a formal interview with the study staff regarding adverse events and symptoms of diabetes.

[0095] <Endpoints and Assessments> The primary outcome was the time elapsed from randomization to diagnosis of diabetes using the criteria defined by the American Diabetes Association. 15 using the criteria defined by the American Diabetes Association. The primary outcome was the time elapsed from randomization to diagnosis of diabetes using the criteria defined by the American Diabetes Association.

[0096] Scheduled OGTT tests were performed 3 and 6 months after infusion and then every 6 months thereafter. Random glucose screening was performed every 3 months, and random glucose screening was performed every 3 months, and random glucose If the fasting blood glucose level is over 200 mg / dl and accompanied by diabetes symptoms, an OGTT test was performed. It was necessary to sequentially confirm the diabetes OGTT test, and the diagnosis date was identified as the second diagnostic examination time. The outcome was examined without knowing the treatment assignment.

[0097] Blood samples were analyzed using the methods described in the experimental methods, centered around the TrialNet core laboratory. Flow cytometry was used to analyze the CD8+ T cell subset in peripheral blood (Figure 4).

[0098] <Experimental Methods> C-peptide was measured from frozen plasma by enzyme immunoassay (Tosoh Bioscience, South San Francisco, CA) at two sites. HbA 1c was measured using ion exchange high performance liquid chromatography (Variant I I, Bio-Rad Diagnostics, Hercules, CA). For each assay, the reliability coefficient from split duplicate samples exceeded 0.99. mIAA, GAD-65Ab, ICA-512Ab, ZnT8A were measured using radioimmunoassay at the Barbara Davis Diabetes Center, Anschultz CO, and ICA was measured using indirect immunofluorescence at the University of Florida at Gainesville. C-peptide, glucose, and HbA were measured at the Northwest Research Laboratory, Seattle, WA. C-peptide was measured from frozen plasma by enzyme immunoassay (Tosoh Bioscience, Sou ptide, glucose, and HbA 1c were measured at the Northwest Research Laboratory, Seattle, WA. C-peptide was measured from frozen plasma by enzyme immunoassay (Tosoh Bioscience, Sou th San Francisco, CA) at two sites. Measured by (San Francisco, CA), HbA 1c by ion exchange high Performance liquid chromatography (Variant II, Bio-Rad Diagnost ics, Hercules, CA). For each assay, the confidence coefficient from split duplicate samples exceeded 0.99. EBV and CMV virus loads in whole blood were measured using the method previously described at the University of Colorado. 1 Used for measurement.

[0099] <Flow cytometry> Peripheral blood mononuclear cells (PBMCs) were processed and stored in the NIDDK repository. PB MC cryovials were sent to the Benaroya Research Institute for analysis by flow cytometry using the antibody panel shown in Figure 4. For T cell phenotyping of PBMCs was performed as previously described using LSR-Fortessa (BD Biosci ences) with FACS Diva software and analyzed using FlowJo software version 9.5 (Tree Star, Ashland , OR). The frequencies of CD8+ T cells that are TIGIT+KLRG+CD57-, TIGIT-KLRG 1-CD57-, or CD4+CD127 Foxp3+ (CD4+ Treg) were determined as previously described. lo Yes The frequencies of CD8+ T cells were determined as previously described. 2 Quadrants were set based on staining controls. Installed.

[0100] <Test supervision> This study was conducted by the National Institutes of Hea funded by the National Institutes of Health (NIH) (grant no. DK062418) and the Juvenile Diabetes Rese arch Foundation and developed and conducted by Type 1 Diabet es TrialNet.

[0101] Except for CBC and white blood cell fractions and routine chemistry tests analyzed at the infusion sites, the trial coordination, clinical laboratory tests, and data management were centralized. An independent med ical monitor (masked with respect to treatment assignment) reviewed all of the safety data obtained.

[0102] <Statistical analysis> The cumulative incidence of diabetes onset over time from randomization within each group was estimated from the Kaplan-Meier estimates of the “diabetes-free” survival function. The differences between treatment groups in the cumulative incidence functions at 6-month intervals were estimated by hypothesis testing using the hazard ratio (HR) and likelihood ratio tests; 16 both were based on the Cox proportional hazards (PH) model. The critical value of the test statistic for the primary hypothesis was determined by a group sequential procedure. 17 Because the expected enrollment rate was slower than anticipated, the original protocol (n = 144 subjects) was revised (one-sided) to detect a 60% (previously 50%) reduction in the hazard rate with 80% power at an alpha level of 0.025 (HR = 0.4). For this protocol, the trial

[0103] aim was set to enroll at least 71 subjects and follow them until 40 subjects were diagnosed with T1D. 18

[0104] Data on safety and efficacy were evaluated twice a year by an independent Data Safety Monitoring Board (D SMB). Interim analyses were performed when 50% of the predicted number of T1D cases had been observed, at which point formal comparisons were submitted to the DSMB and the Lan DeMets stopping rules were applied . Data were analyzed according to the treatment intent principle. The significance tests reported herein were one-sided tests using a significance threshold of 0.025 according to the design 19 , but two-sided tests for treatment interaction tests. If not otherwise indicated, 95% confidence intervals were reported. Flow cytometry data were analyzed by repeated measures ANOVA. Statistical analysis was performed using either TIBCO Spotfire S+8.2 Workbech or SAS 9 .4 . [Results]

[0105] Patients: Of the 112 subjects screened for eligibility, 76 were enrolled: 44 were randomly assigned to teprilizumab and 32 to placebo (Figure 1). As a result of the randomization process, an unequal proportion occurred in the study groups, which was probably due to an unequal distribution between arms as a result of randomization at study sites with a small number (<3) of enrolled subjects. All participants had at least two or more autoantibodies, and 71% of participants had three or more. The treatment arms were generally well balanced (Table 1 ). The majority of subjects (55, 72%) were pediatric, and about half were siblings of T1D patients. Of the subjects under 18 years of age, 47 had confirmed glucose metabolism abnormalities OGTT before randomization. Of the subjects randomized after a single glucose metabolism abnormalities OGTT, 2 had "diabetes" . ) . . Had OGTT, and 6 had normal pre-treatment OGTT: These 8 individuals had pre-registration blood glucose Were registered based on abnormal OGTT.

[0106] Ninety-three percent ( 41 / 44) and 87.5% (28 / 32) of the subjects randomized to the teprilizumab group and the placebo group, respectively, completed 14 days of drug therapy. The Total dose of teprilizumab administered was 9 (IQR: 9.01 - 9.37) μg / m 2 Was 1 4 . Three subjects treated with drug and four subjects treated with placebo did not complete treatment due to laboratory abnormality (n = 4), inability to establish an intravenous access (n = 2), or rash (n = 1). The median follow-up was 745 days (range 74 - 2683 days). The follow-up duration exceeded 3 years in 75% of the subjects. T1D was diagnosed in 42 (55%) of the participants.

[0107]

[0107] 、 or when adjusted for a predefined covariate, anti-GAD65 antibody, remained statistically different. It remained statistically different.

[0108] The overall rate of worsening to T1D was highest in the first year of the study compared to the second year (n = 10, 24%) , third year (n = 6, 14%), or fourth year (n = 5, 12%) (n = 17, 41%). The effect of treatment with teprilizumab was also highest in the first year of the study (Figure 1B). The hazard ratio was lowest in the first 36 months after study enrollment and then remained relatively constant and statistically significant (p < 0.01).

[0109] Administration and safety of treatment: Generally, treatment with teprilizumab was well tolerated. Adverse events designated as possibly related, probably related, or definitely related to the study drug are shown in Table 2. Lymphocyte counts decreased to a nadir on day 5 at 72.3% (IQR 82.1, 68.4%) (p < 0.0001) but then recovered rapidly. Of the 15 cases (34.1%) of grade 3 events in the teprilizumab group, 15 were associated with lymphopenia in the first 30 days after study drug administration. There were no cases of lymphopenia after day 30 in either treatment arm (Figure 1C). As noted above, a rash that resolved spontaneously occurred in 36% of subjects who received drug treatment. 11 . The infection rate was similar in the two treatment arms.

[0110] At baseline, 30 subjects (39%) (16 treated with teprilizumab and 14 treated with placebo) had antibodies to the EBV virus. After treatment with the study drug, quantifiable EBV virus was detected in all 7 participants in the teprilizumab group between weeks 3 and 6. There was a viral load. Among the participants with a detectable viral load, 1 participant had symptoms of pharyngitis, rhinorrhea, and cough on day 38 The EBV viral load decreased from day 43 until it fell below the level of quantification between days 134 (mean 74 days). At enrollment, 17 participants (10 teprilizumab and 7 placebo) had antibodies against the CMV virus . One teprilizumab subject who was CMV seropositive had a detectable level of C MV virus on day 20, which became undetectable by day 42 .

[0111] Response biomarkers: Regarding changes in CD8+ T cells such as the markers TIM- IGIT, KLRG1, and other expressions, the inventors have previously described this 12,13 . To determine whether clinical outcomes are associated with these changes in CD8+ T cells , the frequencies of CD8+KLRG1+TIM- IGIT+CD57-T cells in the two treatment arms were compared. Treatment with teprilizumab increased the frequencies of these T cells at 3 and 6 months compared to baseline (p = 0.009, 0.007 respectively), and the levels at 3 mos and 6 mos were higher in participants treated with teprilizumab than in those treated with placebo ( p = 0.02, 0.04 respectively). (Figure 3A, Figure 6). No changes in these cells were identified in subjects treated with placebo . Not all T cell subsets were affected by teprilizumab: there were no significant changes in CD4+ Treg or CD8+ KLRG1-TIM- IGIT-CD57- cells in either group (Figure 7A, 7 8,20 B). .

[0112] To determine whether the demographic characteristics of participants were associated with clinical response, in a pre-specified analysis, the effects of teprilizumab were analyzed in subgroups of participants based on age, HLA type, pre-treatment C-peptide, and glucose during OGTT, and autoantibodies (Figure 3B). Participants without anti-ZnT8 antibodies showed a greater response to teprilizumab compared to participants with anti-ZnT8 antibodies (p = 0.004) (Figure 3C). The presence or absence of other autoantibodies was not associated with clinical response. 49% and 65% of the teprilizumab-treated subjects were HLA-DR3 and HLA-DR4, respectively. The presence of HLA-DR4 and the absence of HLA-DR3 were associated with a more robust response to teprilizumab (p = 0.004 and 0.01, respectively, two-sided test) (Figures 3D, E).

[0113] [Discussion] In this Phase II trial, a single course of teprilizumab was found to significantly slow the progression to T1D in nondiabetic relatives with abnormal glucose tolerance during OGTT at the time of study entry. The median delay in diabetes diagnosis was 2 years. Also, at the end of the trial, the frequency of diabetes-free individuals was twice as high in drug-treated subjects (57%) as in placebo-treated subjects (28%). Safety findings in children and adults were favorable, and the predicted adverse events were rash and transient lymphopenia. According to the inventors' findings, this is the first treatment to delay or prevent the onset of T1D. Delaying the onset of clinical T1D, which is associated with the challenge of daily management, is clinically important. Furthermore, the younger the age at diagnosis, the worse the outcome 2,4 ​​​​​​​​​​​​​​​。While being large-scale and well-designed, it failed in prevention Previous tests that failed did not utilize immunotherapy against immune cells, and the discovery of the present inventors supports the view that T1D is a chronic T cell-mediated disease 21,22 。 Furthermore, from the finding that this treatment affects pre-diagnostic disease exacerbation and post-diagnostic loss of β-cell function it is suggested that there is a continuum of the autoimmune process, and attempts to use immunomodulation before the onset of clinical disease are validated 。 9~11,23~25 。

[0114] The effect of the drug was maximal in the first 3 years after administration. Forty-one percent of the individuals in whom diabetes developed developed in the first year after randomization, and the HR was lowest at that time in the individuals exposed to teplizumab The relatively rapid rate of exacerbation to diabetes in the placebo group reflects the very high risk of these individuals 5 。Indeed, the decision of the present inventors to enroll these subjects without clinical disease reflects the inevitability of exacerbation when two or more autoantibodies and glucose abnormalities are found, which is consistent with the report of the present inventors that the rate of β-cell death is high in these individuals 。Furthermore, the rapid onset of clinical T1 26 D may reflect the fact that there are more pediatric participants among the participants with a rapid rate of exacerbation (72.4 %) 。 27,28 。

[0115] Based on the characteristics of the subjects at the time of trial registration, differences were seen in the response to teplizumab. One of the T1 D-related MHC alleles, HLA-DR3, is absent, but another T1D-related MH C allele, HLA-DR4, is present, and the absence of anti-ZnT8 antibodies The individual with the highest likelihood of responding is identified. MHC may regulate responsiveness to teprotumumab through its influence on the T cell repertoire, perhaps by altering the state of T cell activation and susceptibility to drug effects. Anti-ZnT8 antibodies may also be markers for individuals with a more severe immune response, or other characteristics that make T cells more susceptible to the effects of teprotumumab. It is uncertain whether treatment is effective in individuals with the disease at an earlier stage. Further immunological and metabolic studies may identify characteristics that define the individuals most likely to benefit from this treatment. The transient effect of drug treatment on lymphocyte counts most likely reflects release from peripheral blood rather than cell depletion. From our flow cytometry tests, it is suggested that changes in the phenotype of CD8+ T cells are markers of clinical response. These effects are associated with a non-responsive or "exhausted" phenotype, but CD8+ T cells are not rendered inactive as active responses to EBV and CMV were seen in individuals with increasing viral loads. The functional effects of teprotumumab on T cells may be affected by the binding activity of T cells to antigens. T cells with high binding activity, such as virus antigen-reactive cells, may not be affected, but T cells with low binding activity, such as autoreactive T cells, may become inactive. Further tests using antigen-reactive T cells are needed to support this hypothesis. There are some limitations to consider in this clinical trial. The cohort is relatively small, and the... ... ...

[0116] ... ... 29,30 ... ... ... ... ... 31,32 ... ... ... ... ...

[0117] ... The rate of exacerbation to T1D in the placebo group was rapid. The subjects were blood relatives of T1D patients and, therefore, it is unknown whether these findings are generally applicable to non-blood relatives in whom the risk of T1D has been found. From recent reports, it is suggested that in genetically high risk individuals, the rate of diabetes is similar in non-blood relatives and blood relatives 33 . Furthermore , while reflecting the known disease incidence, the population was overwhelmingly non-Hispanic Caucasian . It is desirable for the median of the disease onset delay to increase. In this trial, only one cycle of the drug was administered, and from the analysis of the present inventors regarding HR, repeated dosing may be necessary to capture more individuals with active disease and to extend the treatment effect as suggested 9,25 . It is necessary to explore the identification of the treatment target population and the number of drug cycles .

[0118] In summary, this is the first trial showing the delay or prevention of T1D. Since the age at onset and the duration of diabetes are important determinants of metabolic management and complications, and also from the daily management burden, it has clinical significance whenever not diabetic. The selection of individuals with the highest likelihood of response, repeated dosing, or combinations of other agents with a mechanism of action complementary to teprilizumab may make it possible to prevent clinical disease over the long term

[0119] JPEG0007696871000003.jpg237170JPEG0007696871000004.jpg255170JPEG0007696871000005.jpg60170

[0120] JPEG0007696871000006.jpg148170

[0121] Modifications and variations of the methods and compositions described in this disclosure will be apparent to those skilled in the art without departing from the scope and spirit of this disclosure. This disclosure is described in relation to specific embodiments, but it should be understood that the claimed disclosure should not be unduly limited to such specific embodiments. Indeed, various modifications of the described ways of carrying out this disclosure are intended to fall within the scope of the disclosure as represented by the following claims and will be understood by those skilled in the relevant art to which this disclosure pertains.

[0122] [Incorporation by Reference] All patents and publications mentioned in this specification are hereby incorporated by reference in their entirety as if each individual patent and publication were specifically and individually indicated to be incorporated by reference.

[0123] [References] 1. Menke A, Orchard TJ, Imperatore G, Bullard KM, Mayer-Davis E, Cowie CC. T he prevalence of type 1 diabetes in the United States. Epidemiology 2013;24:773 -4. 2. Miller KM, Foster NC, Beck RW, et al. Current state of type 1 diabetes tr eatment in the U.S.: updated data from the T1D Exchange clinic registry. Diabe tes Care 2015;38:971-8. 3. Livingstone SJ, Levin D, Looker HC, et al. Estimated life expectancy in a Scottish cohort with type 1 diabetes, 2008 - 2010. JAMA 2015;313:37 - 44. 4. Rawshani A, Sattar N, Franzen S, et al. Excess mortality and cardiovascul ar disease in young adults with type 1 diabetes in relation to age at onset: a nationwide, register - based cohort study. Lancet 2018;392:477 - 86. 5. Insel RA, Dunne JL, Atkinson MA, et al. Staging presymptomatic type 1 dia betes: a scientific statement of JDRF, the Endocrine Society, and the American Diabetes Association. Diabetes Care 2015;38:1964 - 74. 6. Atkinson MA, Roep BO, Posgai A, Wheeler DCS, Peakman M. The challenge of modulating beta - cell autoimmunity in type 1 diabetes. Lancet Diabetes Endocrino l 2019;7:52 - 64. 7. Keymeulen B, Vandemeulebroucke E, Ziegler AG, et al. Insulin needs after C D3-antibody therapy in new-onset type 1 diabetes. N Engl J Med 2005;352:2598-608 . 8. Perdigoto AL, Preston-Hurlburt P, Clark P, et al. Treatment of Type 1 diab etes with teplizumab: clinical and immunological follow-up after 7 years from d iagnosis. Diabetologia in press. 9. Herold KC, Gitelman SE, Ehlers MR, et al. Teplizumab (anti-CD3 mAb) treatm ent preserves C-peptide responses in patients with new-onset type 1 diabetes in a randomized controlled trial: Metabolic and immunologic features at baseline i dentify a subgroup of responders. Diabetes 2013. 10. Hagopian W, Ferry RJ, Jr., Sherry N, et al. Teplizumab preserves C-peptid e in recent-onset type 1 diabetes: two-year results from the randomized, placeb o-controlled Protege trial. Diabetes 2013;62:3901-8. 11. Herold KC, Hagopian W, Auger JA, et al. Anti-CD3 monoclonal antibody in n ew-onset type 1 diabetes mellitus. N Engl J Med 2002;346:1692-8. 12. Tooley JE, Vudattu N, Choi J, et al. Changes in T-cell subsets identify r esponders to FcR non-binding anti-CD3 mAb (teplizumab) in patients with Type 1 d iabetes. Eur J Immunol 2015. 13. Long SA, Thorpe J, DeBerg HA, et al. Partial exhaustion of CD8 T cells an d clinical response to teplizumab in new-onset type 1 diabetes. Sci Immunol 2016 ;1. 14. Bingley PJ, Wherrett DK, Shultz A, Rafkin LE, Atkinson MA, Greenbaum CJ. Type 1 Diabetes TrialNet: A Multifaceted Approach to Bringing Disease-Modifying Therapy to Clinical Use in Type 1 Diabetes. Diabetes Care 2018;41:653-61. 15. American Diabetes A. 2. Classification and Diagnosis of Diabetes: Standa rds of Medical Care in Diabetes-2019. Diabetes Care 2019;42:S13-S28. 16. Therneau T, Grambsch P. Modeling survival data: extending the Cox Model. New York: Springer-Verlag; 2000. 17. Cox D. Regression model and life tables. J R Stat Soc Ser C Appl Stat 197 2;34B:187-220. 18. Schoenfeld DA. Sample-size formula for the proportional-hazards regressi on model. Biometrics 1983;39:499-503. 19. K.K. L, DeMets D. Discrete sequential boundaries for clinical trials. B iometrika 1983;70:659-63. 20. Herold KC, Burton JB, Francois F, Poumian-Ruiz E, Glandt M, Bluestone JA. Activation of human T cells by FcR nonbinding anti-CD3 mAb, hOKT3gamma1(Ala-Al a). J Clin Invest 2003;111:409-18. 21. Effects of insulin in relatives of patients with type 1 diabetes mellitus . N Engl J Med 2002;346:1685-91. 22. Gale EA, Bingley PJ, Emmett CL, Collier T. European Nicotinamide Diabetes Intervention Trial (ENDIT): a randomised controlled trial of intervention befo re the onset of type 1 diabetes. Lancet 2004;363:925-31. 23. Herold KC, Gitelman SE, Masharani U, et al. A Single Course of Anti-CD3 Monoclonal Antibody hOKT3{gamma}1(Ala-Ala) Results in Improvement in C-Peptide R esponses and Clinical Parameters for at Least 2 Years after Onset of Type 1 Diab etes. Diabetes 2005;54:1763-9. 24. Perdigoto AL, Preston-Hurlburt P, Clark P, et al. Treatment of type 1 dia betes with teplizumab: clinical and immunological follow-up after 7 years from diagnosis. Diabetologia 2018. 25. Sherry N, Hagopian W, Ludvigsson J, et al. Teplizumab for treatment of ty pe 1 diabetes (Protege study): 1-year results from a randomised, placebo-contro lled trial. Lancet 2011. 26. Herold KC, Usmani-Brown S, Ghazi T, et al. beta Cell death and dysfunctio n during type 1 diabetes development in at-risk individuals. J Clin Invest 2015; 125:1163-73. 27. Greenbaum CJ, Beam CA, Boulware D, et al. Fall in C-peptide During First 2 Years From Diagnosis: Evidence of at Least Two Distinct Phases From Composite TrialNet Data. Diabetes 2012. 28. Wherrett DK, Chiang JL, Delamater AM, et al. Defining pathways for develo pment of disease-modifying therapies in children with type 1 diabetes: a consen sus report. Diabetes Care 2015;38:1975-85. 29. Esplugues E, Huber S, Gagliani N, et al. Control of TH17 cells occurs in the small intestine. Nature 2011;475:514-8. 30. Waldron-Lynch F, Henegariu O, Deng S, et al. Teplizumab induces human gut -tropic regulatory cells in humanized mice and patients. Sci Transl Med 2012;4: 118ra12. 31. Wherry EJ. T cell exhaustion. Nature Immunology 2011;12:492. 32. Wherry EJ, Ha SJ, Kaech SM, et al. Molecular signature of CD8+ T cell exh austion during chronic viral infection. Immunity 2007;27:670-84. 33. Hippich M, Beyerlein A, Hagopian WA, et al. Genetic Contribution to the D ivergence in Type 1 Diabetes Risk Between Children From the General Population a nd Children From Affected Families. Diabetes 2019. 34. Mantel N. Evaluation of survival data and two new rank order statistics a rising in its consideration. 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Claims

1. A pharmaceutical composition comprising teprotumumab for delaying the onset of stage 3 type 1 diabetes (T1D) in subjects with stage 2 T1D and increasing the median time to clinical diagnosis of stage 3 T1D by about twofold, wherein the pharmaceutical composition is administered intravenously to the subject once daily over a 14-day cool-down period, the total dose of teprotumumab during the 14-day cool-down period is from about 9034 μg / m² to about 14634 μg / m², the subject is 8 years of age or older and has (i) two or more diabetes-related autoantibodies selected from islet cell antibodies (ICA), insulin autoantibodies (IAA), antibodies against glutamic acid decarboxylase (GAD), antibodies against tyrosine phosphatase (IA-2 / ICA512), and antibodies against zinc transporter 8 (ZnT8), and (ii) abnormal blood glucose levels in an oral glucose tolerance test (OGTT), pharmaceutical composition.

2. the total dose of teprotumumab during the 14-day cool-down period is about 13486 μg / m ２ The pharmaceutical composition according to claim 1.

3. The pharmaceutical composition according to claim 1, wherein the subject is a blood relative of a T1D patient.

4. The pharmaceutical composition according to claim 1, wherein the subject has abnormal glucose tolerance in an OGTT.

5. The pharmaceutical composition according to claim 4, wherein the abnormal glucose tolerance in the OGTT is a fasting glucose level of 110-125 mg / dL.

6. The pharmaceutical composition according to claim 4, wherein the abnormal glucose tolerance in the OGTT is a 2-hour value of 140 mg / dL or more and less than 200 mg / dL.

7. The pharmaceutical composition according to claim 4, wherein the abnormal glucose tolerance in the OGTT is a glucose value above 200 mg / dL at 30, 60, or 90 minutes.

8. About 5 to about 1200 μg / m of teprotumumab per day is administered to the subject ２ The pharmaceutical composition according to any one of claims 1 to 7, which is administered by

9. The pharmaceutical composition according to any one of claims 1 to 7, wherein teprilizumab is administered to the subject at a dose of about 10 to about 1000 μg / m2 per day.

10. The pharmaceutical composition according to any one of claims 1 to 7, wherein the administration of the pharmaceutical composition delays the median time to clinical diagnosis of stage 3 T1D by about 24 months to about 48 months.

11. The pharmaceutical composition according to any one of claims 1 to 7, wherein an increase in the relative amount of TIGIT+KLRG1+CD8+ T cells in the peripheral blood mononuclear cells of the subject after administration of the pharmaceutical composition indicates responsiveness to teprilizumab.

12. The pharmaceutical composition according to claim 11, wherein the relative amount of the TIGIT+KLRG1+CD8+ T cells is determined by flow cytometry.

13. The pharmaceutical composition according to claim 1, wherein teprilizumab is administered to the subject at a dose of about 51 μg / m2 on day 0, about 103 μg / m2 on day 1, about 207 μg / m2 on day 2, about 413 μg / m2 on day 3, and 826 μg / m2 on each of days 4 to 13.

14. The pharmaceutical composition according to claim 1, wherein the subject does not have an antibody against ZnT8.

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