Patent Application
20220340672
The disclosure is directed to treatment and prevention of immune-related diseases and disorders such as diabetes and related conditions and graft-versus host disease, using TNF Receptor Superfamily Member 25 (TNFRSF25) agonistic antibodies.
The application contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Sep. 28, 2020, is named PEL-016PC_ST25.txt and is 14,496 bytes in size.
Tregs are T cells which have a role in regulating or suppressing other cells in the immune system. Stimulation of tumor necrosis factor receptor superfamily, member 25 (TNFRSF25), in vivo facilitates selective proliferation of Tregs in mice and suppression of immunopathology in allergic lung inflammation, allogeneic heart transplantation and HSV-1 mediated ocular inflammation.
Various immune-related diseases and disorders remain major health problems and, among the most pressing medical needs are treatments for diabetes and graft-versus-host disease (GVHD). Diabetes affects a large number of people in the United States and abroad, and many new cases occur each year. Diabetes is linked to a number of health problems, including microvascular complications, such as retinopathy, neuropathy, and nephropathy. Further, in the United States, diabetes is the leading cause of new blindness in working-age adults, new cases of end-stage renal disease, and non-traumatic lower leg amputations. In addition, cardiovascular complications are now the leading cause of diabetes-related morbidity and mortality, particularly among women and the elderly. In adult patients with diabetes, the risk of cardiovascular disease (CVD) is three-to-five fold greater than in the general population.
Islet cell transplantation has been recognized as an emerging, promising therapeutic approach to treatment of diabetes. However, immunosuppression is required to prevent host rejection of donor islet cells, as is common for allografts in human organ transplantation. Moreover, islet cell transplantation can be complicated by allograft rejection and GVHD, a common immune response to donor's cells in the context of a transplant which can lead to, among other issues, serious immune system complications and even death.
Current therapies for diabetes and related diseases suffer from inadequate responses and/or patient adherence.
Therefore, there remains a need for more effective and accessible methods of treating diabetes, allograft rejection and/or GVHD.
Accordingly, the present invention provides new methods and uses for the treatment and/or prevention of diabetes, including, for example, type 1 and type 2 diabetes, and related diseases (e.g., prediabetes and glucose intolerance), comprising administering an effective amount of TNF Receptor Superfamily Member 25 (TNFRSF25) agonistic antibody or antigen binding fragment thereof to a patient in need thereof. The treatment may expand and/or selectively activate a population of Tregs in the patient.
The described methods can be used to treat and/or prevent diabetes and other related conditions in patients suffering other conditions and/or diseases. For example, the patient may be diagnosed with one or more of insulin resistance, prediabetes, impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and acanthosis nigricans. The patient may have cardiovascular disease or metabolic disease, type 1 diabetes or type 2 diabetes, gestational diabetes or steroid-induced diabetes, or other conditions or diseases.
In embodiments, the patient may be undergoing treatment with one or more of insulin or an insulin analog. The insulin analog can be selected from a rapid acting insulin analog (e.g., lispro, aspart or glulisine) or a long acting insulin analog (e.g., glargine or detemir).
In embodiments, a method for treating diabetes and/or glucose intolerance is provided that comprises administering an effective amount of TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof, wherein the patient is not receiving insulin therapy. In such embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment can stimulate glucose uptake in the patient.
In some aspects, the present invention also provides a method for increasing a graft or transplant survival and preventing or delaying a graft or transplant rejection. The graft, interchangeably referred to herein as a transplant, can be a solid organ graft (e.g., pancreas or other organ). In some embodiments, the graft can comprise islet cells which can be obtained from either a donor (islet allo-transplantation) or a recipient (islet auto-transplantation).
In some embodiments, the administration of TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof results in a Treg expansion. The Treg expansion can include expansion of FoxP3+ cells out of CD4+ T cells, and/or expansion of one or both CD4+FOXP3+ T cells and CD4+CD25+FOXP3+ T cells in the patient. In embodiments in which the patient, which can be afflicted by diabetes or similar disease(s), is a transplant recipient (e.g., a recipient of an islet cells graft, pancreatic graft, etc), the increase in the Treg expansion can result in increase in a graft survival and prevention or delay of a graft rejection and GVHD.
In some embodiments, a method for treating or preventing graft-versus-host disease (GVHD) is provided, comprising administering an effective amount of TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof. The patient may be a transplant recipient, and the transplant recipient may receive the transplant comprising islet cells from a transplant donor. In some embodiments, the transplant may comprise donor T cells, donor hematopoietic cells, donor stem cells, or donor bone marrow cells. The method may involve reducing a graft versus host disease (GVHD), which can be acute graft-versus-host-disease (aGVHD) or chronic graft-versus-host-disease (cGVHD).
In some embodiments, a method for treating or preventing GVHD is provided, comprising administering an effective amount of TNF Receptor Superfamily Member 25 (TNFRSF25) agonistic antibody or antigen binding fragment thereof to a patient in need thereof. The patient may be a transplant recipient, and the transplant recipient may receive the transplant comprising islet cells from a transplant donor. The transplant may comprise donor T regulatory cell, donor hematopoietic cells, donor stem cells, or donor bone marrow cells. The method may involve reducing a graft versus host disease (GVHD), which can be acute graft-versus-host-disease (aGVHD) or chronic graft-versus-host-disease (cGVHD).
The administration of the effective amount of TNFRSF25 agonistic antibody or antigen binding fragment can be to a transplant donor. In some embodiments, the administration to the transplant donor can occur prior to transplant. In some embodiments, the administration is to both the transplant donor and transplant recipient. In some embodiments, the method prevents a transplant rejection, e.g., a solid organ transplant rejection, islet cell transplant rejection, or a stem cell transplant rejection.
In some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence is GFTFSNHDLN (SEQ ID NO: 1), the heavy chain CDR2 sequence is YISSASGLISYADAVRG (SEQ ID NO: 2); and the heavy chain CDR3 sequence is DPAYTGLYALDF (SEQ ID NO: 3) or DPPYSGLYALDF (SEQ ID NO: 4); and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence is TLSSELSWYTIV (SEQ ID NO: 5), the light chain CDR2 sequence is LKSDGSHSKGD (SEQ ID NO: 6), and the light chain CDR3 sequence is CGAGYTLAGQYGWV (SEQ ID NO: 7).
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof further comprises variable region framework (FW) sequences juxtaposed between the CDRs according to the formula (FW1)-(CDR1)-(FW2)-(CDR2)-(FW3)-(CDR3)-(FW4), wherein the variable region FW sequences in the heavy chain variable region are heavy chain variable region FW sequences, and wherein the variable region FW sequences in the light chain variable region are light chain variable region FW sequences. The variable region FW sequences may be human.
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof further comprises human heavy chain and light chain constant regions. The constant regions can be selected from the group consisting of human IgG1, IgG2, IgG3, and IgG4. For example, the constant regions can be IgG1 or IgG4.
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises a heavy chain variable region having the amino acid sequence EVQLVESGGGLSQPGNSLQLSCEASGFTFSNHDLNWVRQAPGKGLEWVAYISSASGLISYADAVRGRFTISRDN AKNSLFLQMNNLKSEDTAMYYCARDPPYSGLYALDFWGQGTQVTVSS (SEQ ID NO: 8), or an amino acid sequence of at least about 85 to about 99% identity thereto. In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises a light chain variable region having the amino acid sequence QPVLTQSPSASASLSGSVKLTCTLSSELSSYTIVWYQQRPDKAPKYVMYLKSDGSHSKGDGIPDRFSGSSSGAH RYLSISNVQSEDDATYFCGAGYTLAGQYGWVFGSGTKVTVL (SEQ ID NO: 9), or an amino acid sequence of at least about 85 to about 99% identity thereto.
In
The present invention is based, in part, on the discovery of surprising immune-modulatory effects of anti-TNFRSF25 agonistic antibodies (e.g., PTX-35). For instance, the present inventors have discovered that anti-TNFRSF25 agonistic antibodies (e.g., PTX-35) cause expansion of Tregs in vivo. The described treatments may expand and/or selectively activate a population of Tregs in the patient. This effect can find use in treating or preventing diabetes, prediabetes, and/or glucose intolerance in patients who may or who may not be receiving insulin therapy. Also, the expansion of Tregs in vivo can be used in treating or preventing a graft rejection, increasing graft survival, and treating or preventing a graft-versus-host disease (GVHD).
The present invention makes use of antibodies targeted to particular epitopes within tumor necrosis factor receptor superfamily member 25 (TNFRSF25). TNFRSF25 is a TNF-receptor superfamily member that is preferentially expressed by activated and antigen-experienced T lymphocytes. The structural organization of the TNFRSF25 protein is most homologous to TNF receptor 1 (TNFR1). The extracellular domain of TNFRSF25 includes four cysteine-rich domains, and the cytoplasmic region contains a death domain known to signal apoptosis. Alternative splicing of the TNFRSF25 gene in B and T cells encounters a program change upon T cell activation, which predominantly produces full-length, membrane bound isoforms, and is involved in controlling lymphocyte proliferation induced by T cell activation. TNFRSF25 is activated by its ligand, TNF-like protein 1A (TL1A), also referred to as TNFSF15, which is rapidly upregulated in antigen presenting cells and in some endothelial cells following Toll-Like Receptor or Fc receptor activation. TL1A has co-stimulatory activity for TNFRSF25-expressing T cells through the activation of NF-κB and suppression of apoptosis by up-regulation of c-IAP2. TNFRSF25 signaling increases the sensitivity of T cells to endogenous IL-2, and enhances T cell proliferation.
The inventors have previously demonstrated that TNFRSF25 is a potent T cell costimulator due to its specificity for expansion of memory CD4+ and CD8+ T cells that are known to maintain tolerance to self antigens, help establish tolerance to allogeneic grafts, suppress differentiation of naïve T cells into effector T cells, and suppress activity of already differentiated T cells. Profileration of T cells (e.g., human T cells, murine T cells, or macaque T cells) can be stimulated by administering an amount of an anti-TNFRSF25 antibody.
In some embodiments, the TNFRSF25 agonistic antibody is PTX-35. PTX-35 is defined by its variable heavy and variable light chain sequences.
Islet transplantation is a method of implantation of pancreatic islets for the treatment of type 1 diabetes mellitus, and it is referred to as Edmonton Protocol established in 2000. See Shapiro et al. (2000) Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen. The New England Journal of Medicine. 343 (4): 230-238. Islet transplantation requires simultaneous and constant immunosuppression to avoid allograft rejection. See Bottino et al. (2002) Pancreas and islet cell transplantation. Best Pract Res Clin Gastroenterol. 16(3):457-474. Islet transplantation normalizes glucose levels in patients with type 1 diabetes and alleviates hypoglycemic episodes associated with traditional insulin therapy. However, the associated challenge is that islet transplantation requires lifelong immunosuppression, and ways to develop a long term insulin independence have not been developed.
A delivery of a transplant, such as, e.g., islet cell transplantation, to a patient in need thereof using existing approaches is typically plagued by a loss of the function of the transplant, which can be caused by allogeneic rejection, recurrence of autoimmunity, and immunosuppressive drug toxicity. See Wang et al.
Radiology vol. 266, 3 (2013): 822-30. The progressive decline in graft function is observed in many islet recipients. See Forbes et al. American Journal of Transplantation. 2016: 16 (91): pages 2704-2713 (first published: 28 Mar. 2016). Accordingly, the long-term survival and function of islet grafts presents a significant challenge. The present disclosure therefore provides a method that comprises administering an effective amount of TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof to cause Treg expansion and thereby prolong transplant (such as, without limitation or antigen binding fragment thereof, an islet cell transplant) survival. The transplant survival is prolonged such that the transplant remains functional and is not rejected by the recipient's body for a longer period of time as compared to survival of a transplant in patient which was not administered a TNFRSF25 agonistic antibody or antigen binding fragment thereof. The patient may be diagnosed with one or more of insulin resistance, prediabetes, diabetes (type I or type II), impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and acanthosis nigricans.
Accordingly, the present disclosure provides methods for expanding the population of T regulatory cells, e.g., FoxP3+ cells out of CD4+ T cells, which demonstrates that administration of anti-TNFRSF25 agonistic antibodies (e.g., PTX-35) can lead to immunological tolerance, which can prevent the use of chronic immunosuppression in islet transplantation. In embodiments, the present methods relate to a method of islet transplantation in which the present anti-TNFRSF25 agonistic antibodies (e.g., PTX-35) are administered simultaneously with the islet transplantation and/or as a maintenance therapy subsequent to the islet transplantation.
In some embodiments, the expansion of T regulatory cells comprises increase in FoxP3+ cells out of CD4+ T cells, increase in CD4+FOXP3+, and/or increase in CD4+CD25+FOXP3+ cells. In some embodiments, the increase in FoxP3+ cells out of CD4+ T cells is an increase at which at least about 10% of CD4+ T-cells are FOXP3+Tregs. In some embodiments, the increase in FoxP3+ cells out of CD4+ T cells is an increase at which at least about 11%, or at least about 12%, or at least about 13%, or at least about 14%, or at least about 15%, or at least about 16%, or at least about 17%, or at least about 18%, or at least about 19%, or at least about 20% of CD4+ T-cells are FOXP3+Tregs. In some embodiments, the increase in FoxP3+ cells out of CD4+ T cells is an increase at which at least about 25%, or at least about 30%, or at least about 35%, or at least about 40% of CD4+ T-cells are FOXP3+Tregs.
In some embodiments, administration of an anti-TNFRSF25 agonistic antibody in accordance with the present disclosure results in significant Treg expansion in a subject. In some embodiments, the subject is a diabetic patient. The expansion of the Treg populations comprises expansion of CD4+FOXP3+ and/or CD4+CD25+FOXP3+ cells. In some embodiments, the anti-TNFRSF25 agonistic antibody comprises PTX-35 or an antigen binding fragment thereof. In some embodiments, the anti-TNFRSF25 agonistic antibody comprises PTX-25 or an antigen binding fragment thereof.
In some embodiments, an anti-TNFRSF25 agonistic antibody comprises a suitable anti-TNFRSF25 agonistic antibody. In embodiments, an anti-TNFRSF25 agonistic antibody, also referred to an TNFRSF25 agonist (or “DR3”) refers to a substance that binds to the TNFRSF25 receptor and triggers a response in the cell on which the TNFRSF25 receptor is expressed, similar to a response that would be observed by exposing the cell to a natural TNFRSF25 ligand, e.g., TL1A. An agonist is the opposite of an antagonist in the sense that while an antagonist may also bind to the receptor, it fails to activate the receptor and actually completely or partially blocks it from activation by endogenous or exogenous agonists. A partial agonist activates a receptor but does not cause as much of a physiological change as docs a full agonist. Alternatively, another example of a TNFRSF25 agonist is an antibody that is capable of binding and activating TNFRSF25. An example of an anti-TNFRSF25 antibody is 4C12 (agonist). (Deposited under the Budapest Treaty on Behalf of: University of Miami; Date of Receipt of seeds/strain(s) by the ATCC®: May 5, 2009; ATCC® Patent Deposit Designation: PTA-10000. Identification Reference by Depositor: Hybridoma cell line; 4C12; The deposit was tested Jun. 4, 2009 and on that date, the seeds/strain(s) were viable. International Depository Authority: American Type Culture Collection (ATCC®), Manassas, Va., USA). In some embodiments, an anti-TNFRSF25 agonistic antibody is PTX-35, which is shown by the inventors to have an activity similar to that of 4C12.
In embodiments, an anti-TNFRSF25 agonistic antibody is from a genus of anti-TNFRSF25 agonistic antibodies that bind to the TNFRSF25 receptor and trigger a response in the cell on which the TNFRSF25 receptor is expressed. The genus comprises 4C12, PTX-25, PTX-35 (such as mPTX-35 or a humanized PTX-35), 11H08 anti-TNFRSF25 agonistic antibody (see U.S. Patent Application No. 2012/0014950), anti-DR3 mouse monoclonal antibody F05 (see Wen et al., The Journal of Biological Chemistry, 2003; 278:39251-39258), another anti-DR3 monoclonal antibody (see U.S. Pat. No. 7,357,927; see also Papadakis et al., J Immunol 2004; 172:7002-7007 (“an agonistic anti-DR3 mAb synergize with IL-12/IL-18 to augment IFN-γ production in human peripheral blood T cells and NK cells”), or any other anti-TNFRSF25 agonistic antibody.
In some embodiments, administration of an TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof results in an increase in serum levels of IL-5.
In some embodiments, the present disclosure provides methods for treating or preventing diabetes, prediabetes, and/or glucose intolerance, comprising administering an effective amount of an TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof. In various embodiments, a TNFRSF25 agonistic antibody helps provide a patient with glycemic control, as monitored by, for example, average glucose and/or glycosylated hemoglobin levels. In various embodiments, the anti-diabetic effects of TNFRSF25 agonistic antibody are insulin-independent.
In some embodiments, the present disclosure provides methods for treating or preventing diabetes, prediabetes, and/or glucose intolerance, comprising administering an effective amount of TNFRSF25 agonistic antibody or antigen binding fragment thereof, in combination with an immunosuppressant or anti-inflammatory agent, to a patient in need thereof. In some embodiments, the immunosuppressant or anti-inflammatory agent is an anti-CTLA4 antibody. In some embodiments, the immunosuppressant or anti-inflammatory agent is an IL-1 receptor antagonist. In some embodiments, the IL-1 receptor antagonist is Kineret®. In various embodiments, TNFRSF25 agonistic antibody, in combination with an immunosuppressant or anti-inflammatory agent, helps provide a patient with glycemic control, as monitored by, for example, average glucose and/or glycosylated hemoglobin levels. In various embodiments, the anti-diabetic effects of TNFRSF25 agonistic antibody are insulin-independent.
The patient may be affected by various conditions or diseases. For example, in some embodiments, the patient being treated may be suffering from insulin resistance. As another example, the patient may be diagnosed with one or more of insulin resistance, prediabetes, impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and acanthosis nigricans. In some embodiments, the patient has cardiovascular disease or metabolic disease. As yet another example, the patient can have type 1 diabetes or type 2 diabetes. In some embodiments, the patient has gestational diabetes or steroid-induced diabetes.
The TNFRSF25 agonistic antibody or antigen binding fragment thereof can be administered as a regimen that decreases blood glucose level, stimulates peripheral glucose disposal, and/or inhibits hepatic glucose production. In some embodiments, the patient can have one or more of an average hemoglobin A1c value of more than about 10% and an average glucose of more than about 200 mg/dl (11 mmol/I) at the start of treatment with conventional diabetic therapy. In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof is administered to a patient that has one or more of an average hemoglobin Alc value of more than about 11%, or more than about 12%, or more than about 13%, or more than about 14%, or more than about 15% at the start of treatment with conventional diabetic therapy. In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof is administered to a patient that has an average glucose of more than about 210 mg/dl, or more than about 220 mg/dl, or more than about 230 mg/dl, or more than about 240 mg/dl, or more than about 250 mg/dl at the start of treatment with conventional diabetic therapy. In various embodiments, the conventional diabetic therapy is any one of those described herein, including, for example, insulin therapy and non-insulin diabetes agent therapy. The non-insulin diabetes agents may include, e.g., metformin, sulfonylureas, glipizide and glimepiride, thiazolidinediones, DPP-4 inhibitors, GLP-1 receptor agonists, and SGLT2 inhibitors.
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof administration is effective for providing average glucose of below about 200 mg/dl (11 mmol/I). In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof administration is effective for providing an average glucose of below about 190 mg/dl, or about 180 mg/dl, or about 170 mg/dl, or about 160 mg/dl, or about 150 mg/dl, or about 140 mg/dl, or about 130 mg/dl, or about 120 mg/dl, or about 120 mg/dl, or about 110 mg/dl, or about 100 mg/dl, or about 90 mg/dl, or about 80 mg/dl, or about 70 mg/dl. In some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof administration is effective for providing average glycosylated hemoglobin levels (hemoglobin A1c) values of about 8% or less. For example, in some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof administration is effective for providing an average glycosylated hemoglobin levels (hemoglobin A1c) values of about 8%, or about 7%, or about 6%, or about 5%, or about 4%. In some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof administration is effective for providing average glycosylated hemoglobin levels (hemoglobin A1c) values of less than about 8%, or less than about 7%, or less than about 6%, or less than about 5%, or less than about 4%.
In various embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof administration does not cause a patient to experience an increase of insulin upon TNFRSF25 agonistic antibody or antigen binding fragment thereof administration. Accordingly, in some embodiments, TNFRSF25 agonistic antibody's anti-diabetic effects are insulin-independent.
In various embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof is administered as an adjuvant therapy. For instance, a diabetic patient may receive treatment with insulin or an insulin analog, or any of the agents listed herein (e.g. sulfonylureas, biguanides, meglitinides, thiazolidinediones, DPP-4 inhibitors, SGLT2 Inhibitors, Alpha-glucosidase inhibitors, and bile acid sequestrants) and TNFRSF25 agonistic antibody or antigen binding fragment thereof is administered to supplement these treatments. For example, in some embodiments, the use of TNFRSF25 agonistic antibody or antigen binding fragment thereof as an adjuvant therapy with a long-acting insulin offsets the high frequency of hypo- and hyperglycemic excursions and modest reduction in HbA1c seen with these agents.
In some embodiments, the patient is undergoing treatment with one or more of insulin or an insulin analog. The insulin analog may be selected from a rapid acting or long acting insulin analog. Non-limiting examples of the rapid acting insulin analog comprise lispro, aspart or glulisine. Non-limiting examples of the long acting insulin analog comprise glargine or detemir.
In embodiments of the present disclosure, administration of TNFRSF25 agonistic antibody or antigen binding fragment thereof does not cause one or more of common side effects of standard diabetes care, such as hypoglycemia or hypokalemia.
In various aspects, the present methods provide for the treatment of diabetes with TNFRSF25 agonistic antibody or antigen binding fragment thereof in specific patient populations in need thereof. For example, in various embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof may supplement various agents in a treatment regimen for diabetes, including type 1 or type 2 diabetes, or may supplant various agents in a treatment regimen for diabetes, including type 1 or type 2 diabetes. For example, in some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof is an adjuvant therapy for type 1 or type 2 diabetes.
Type 1 diabetes, once known as juvenile diabetes or insulin-dependent diabetes, is a chronic condition in which the pancreas produces little or no insulin. Treatment is often via intensive insulin regimens, which attempt to mimic the body's normal pattern of insulin secretion, and often involve basal and bolus insulin coverage. For example, one common regimen is the administration of a long-acting insulin (as described herein and including, for example, glargine/detemir) once or twice a day with rapid acting insulin (as described herein and including, for example, aspart, glulisine, lispro) preprandially or postprandially and as needed to correct high blood sugars (as monitored by a glucose meter, for example). Doses administered preprandially or postprandially or as needed to correct high blood sugars may be referred to as bolus administrations. Another common regimen is involves dosing, including continuous dosing, via an insulin pump (or continuous subcutaneous insulin infusion device (CSII)) of, for example a rapid acting insulin (as described herein and including, for example, aspart, glulisine, lispro). In various embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof may replace any of the insulins used in various regimens, including instances in which the insulins are not providing effective therapy in the patient. TNFRSF25 agonistic antibody or antigen binding fragment thereof may cause an increase in patient compliance as it may allow for easier self-dosing relative to various forms of insulin, which must be administered as various doses throughout the day, even in the context of an insulin pump, which requires programming.
Further, TNFRSF25 agonistic antibody or antigen binding fragment thereof can offset common frustration of diabetic patient dosing, such as, for example, the dawn phenomenon. Alternatively, TNFRSF25 agonistic antibody or antigen binding fragment thereof may be used adjuvant to any of the type 1 diabetes treatments described herein to, for example, normalize a patient's regimen and avoid blood sugar “dips” (e.g. hypoglycemia, e.g. blood sugar of below about 70 mg/dL) and “spikes” (e.g. hyperglycemia, e.g. blood sugar of below about 200 mg/dL) that afflict many patients. Accordingly, in some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof may treat or prevent symptoms associated with hypoglycemia, including for example, shakiness, anxiety, nervousness, palpitations, tachycardia, pallor, coldness, clamminess, dilated pupils (mydriasis), hunger, borborygmus, nausea, vomiting, abdominal discomfort, headache, abnormal mentation, impaired judgment, nonspecific dysphoria, paresthesia, negativism, irritability, belligerence, combativeness, rage, personality change, emotional lability, fatigue, weakness, apathy, lethargy, daydreaming, sleep, confusion, amnesia, lightheadedness or dizziness, delirium, staring, “glassy” look, blurred vision, double vision, flashes of light in the field of vision, automatism, difficulty speaking, slurred speech, ataxia, incoordination, focal or general motor deficit, paralysis, hemiparesis, paresthesia, headache, stupor, coma, abnormal breathing, generalized or focal seizures, memory loss, and amnesia. Accordingly, in some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof may treat or prevent symptoms associated with hyperglycemia, including for example, polyphagia, polydipsia, polyuria, blurred vision, fatigue, weight loss, poor wound healing, dry mouth, dry or itchy skin, tingling in feet or heels, erectile dysfunction, recurrent infections, external ear infections (e.g. swimmer's ear), cardiac arrhythmia, stupor, coma, and seizures. In various regimens, a type 1 diabetes may receive additional agents to supplement insulin therapy. In some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof are used in this manner. TNFRSF25 agonistic antibody or antigen binding fragment thereof may provide additional therapeutic benefits in patients that are struggling to manage type 1 diabetes with insulin therapy alone. In some embodiments, patients that are struggling to manage type 1 diabetes with insulin therapy alone have poor glycemic control as described herein.
Patients with type 2 diabetes may be instructed to manage their diabetes with healthy eating and exercise. However, certain non-insulin diabetes agents (e.g. selected from metformin (e.g. GLUCOPHAGE, GLUMETZA); sulfonylureas (e.g. glyburide (e.g. DIABETA, GLYNASE), glipizide (e.g. GLUCOTROL) and glimepiride (e.g. AMARYL)); thiazolidinediones (e.g. rosiglitazone (e.g. AVANDIA) and pioglitazone (e.g. ACTOS)); DPP-4 inhibitors (e.g. sitagliptin (e.g. JANUVIA), saxagliptin (e.g. ONGLYZA) and linagliptin (e.g. TRADJENTA)); GLP-1 receptor agonists (e.g. exenatide (e.g. BYETTA) and liraglutide (e.g. VICTOZA)); and SGLT2 inhibitors (e.g. canagliflozin (e.g. NVOKANA) and dapagliflozin (e.g. FARXIGA))) and/or insulin may be used in treatment. For example, certain patients may be able to manage diabetes with diet and exercise alone (e.g. along with glucose monitoring). However, often this is not the case and therapeutic agents are needed. A first line of treatment may be a non-insulin diabetes agent (e.g. selected from metformin (e.g. GLUCOPHAGE, GLUMETZA); sulfonylureas (e.g. glyburide (e.g. DIABETA, GLYNASE), glipizide (e.g. GLUCOTROL) and glimepiride (e.g. AMARYL)); thiazolidinediones (e.g. rosiglitazone (e.g. AVANDIA) and pioglitazone (e.g. ACTOS)); DPP-4 inhibitors (e.g. sitagliptin (e.g. JANUVIA), saxagliptin (e.g. ONGLYZA) and linagliptin (e.g. TRADJENTA)); GLP-1 receptor agonists (e.g. exenatide (e.g. BYETTA) and liraglutide (e.g. VICTOZA)); and SGLT2 inhibitors (e.g. canagliflozin (e.g. NVOKANA) and dapagliflozin (e.g. FARXIGA)). However, some of these agents provide side effects (e.g., in the case of metformin, abdominal or stomach discomfort, cough or hoarseness, decreased appetite, diarrhea, fast or shallow breathing, fever or chills, general feeling of discomfort, lower back or side pain, muscle pain or cramping, painful or difficult urination, and sleepiness) or negative drug interactions (e.g., in the case of metformin, certain imaging and contrast agents). In some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof is used instead of a non-insulin diabetes agent or in combination with one or more non-insulin diabetes agents (e.g., to lower the dose of the non-insulin diabetes agents and increase their therapeutic windows).
In some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof is used to improve an ineffective treatment regimen. In certain embodiments, use of TNFRSF25 agonistic antibody or antigen binding fragment thereof increases patient compliance and increases the likelihood of effective type 2 diabetes management. In certain embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof replaces a non-insulin diabetes agent in a patient's treatment regimen in a patient whose diabetes is not well-managed by a non-insulin diabetes agent (e.g. those having uncontrolled, cardiovascular complications and/or blood glucose levels). In some embodiments, a patient whose diabetes is not well-managed by a non-insulin diabetes agent has poor glycemic control as described herein.
In some type 2 diabetes patients, diet and exercise and/or non-insulin diabetes agents are insufficient for treatment of diabetes and treatment with insulin therapy is needed. In various embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof may prevent the need to turn to insulin therapy in type 2 diabetes patients or reduce the amount (e.g. frequency of administration) of insulin therapy in type 2 diabetes patients. For example, TNFRSF25 agonistic antibody or antigen binding fragment thereof may be used in certain type 2 diabetes patient populations that are often at risk for needing insulin therapy, including patients afflicted having: acute infections or other serious illnesses, pregnancy, major surgery, congestive heart failure, kidney disease, liver disease, use of other drugs (e.g. prednisone and some psychiatric medications), overeating or excessive weight gain (including obesity), and progressive loss of beta cell function. In some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof may be administered to patients having onset of diabetes prior to age thirty, or a duration over fifteen years to prevent the need for insulin therapy. Further, in some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof is used to treat diabetes in patients at risk for uncontrolled or poorly controlled type 2 diabetes (overweight and/or obese patients, patients with high abdominal fat distribution, inactive patients, patients with a family history. of type 2 diabetes, patients of certain racial groups (e.g., blacks, Hispanics, American Indians and Asian-Americans), older patients (e.g. over the age of about 45), patients previously afflicted with gestational diabetes and/or who have birthed a baby weighing more than about 9 pounds, and patients having polycystic ovary syndrome).
In some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof is used to treat type 2 diabetes patients that have uncontrolled or poorly controlled type 2 diabetes and are facing a nontraumatic lower extremity amputation (LEA). In some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof is used to treat type 2 diabetes patients that have uncontrolled or poorly controlled type 2 diabetes and have some degree of vision loss and/or blindness (by way of non-limiting example, diabetic retinopathy, which may include one or more of non-proliferative diabetic retinopathy (including, for example, treating microanuerysms) and proliferative diabetic retinopathy (including, for example, treating vitreous, clouding vision, detachment of the retina and glaucoma). In some embodiments, the determination of whether a patient is afflicted with or has a high risk for some degree of vision loss and/or blindness comprises diagnostic methods known in the art (e.g. ophthalmoscopy, fluorescein angiography). In some embodiments, TNFRSF25 agonistic antibody or antigen binding fragment thereof is used to treat type 2 diabetes patients that have uncontrolled or poorly controlled type 2 diabetes and have end-stage renal disease (including, for example, end-stage renal disease).
In some aspects, the present disclosure relates a method for treating diabetes and/or glucose intolerance, comprising administering an effective amount of TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof, wherein the patient is not receiving insulin therapy. In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof stimulates glucose uptake in the patient. In some embodiments, the glucose uptake is mediated by glucose transporter type 4 (GLUT4). In some embodiments, the glucose uptake is in muscle or fat cells.
The present invention provides, in some aspects, methods of treatment of diabetes and/or glucose intolerance in certain patient populations, including, for example, patients not receiving insulin therapy (e.g., not receiving one or more of basal, preprandial, and postprandial insulin therapy) for various reasons (e.g., ineffectiveness of insulin therapy, allergies, side effects, and lack of compliance) and/or patients that are not receiving non-insulin diabetes agents (e.g., metformin, sulfonylureas, glipizide and glimepiride, thiazolidinediones, DPP-4 inhibitors, GLP-1 receptor agonists, and SGLT2 inhibitors). For example, in some embodiments, the patient is not receiving one or more of basal, preprandial, and postprandial insulin therapy. In some embodiments, the patient is not receiving preprandial or postprandial insulin therapy but is receiving basal insulin therapy. In some embodiments, the patient has not received insulin therapy in up to about 1 hour, or up to about 2 hours, or up to about 3 hours, or up to about 4 hours, or up to about 5 hours, or up to about 6 hours, or up to about 7 hours, or up to about 8 hours, or up to about 12 hours or up to about 16 hours or up to about 20 hours, or up to about 24 hours, up to about 2 days, up to about 3 days, up to about 4 days, up to about 5 days, up to about 6 days, up to about 7 days.
In various embodiments, the patient has experienced one or more instances of lipodystrophy that is caused by injection (e.g., injection of insulin). In some embodiments, the patient is afflicted with or is at risk of having hypokalemia. In some embodiments, the patient is afflicted with or is at risk of having an insulin allergy or allergy to a an agent, such as zinc, commonly used to formulate insulin (e.g., a patient having or who has previously had an immediate hypersensitive reaction upon insulin injection (e.g., injection site swelling, redness and/or itching, local tender subcutaneous nodules which develop about 0.5 to about 6 hours after an insulin injection, inflammation of the lymph glands, or a serum sickness reaction and arthralagia).
In some embodiments, the patient is receiving one or more non-insulin diabetes agents selected from metformin (e.g., GLUCOPHAGE, GLUMETZA); sulfonylureas (e.g., glyburide (e.g., DIABETA, GLYNASE), glipizide (e.g., GLUCOTROL) and glimepiride (e.g., AMARYL)); thiazolidinediones (e.g., rosiglitazone (e.g., AVANDIA) and pioglitazone (e.g., ACTOS)); DPP-4 inhibitors (e.g., sitagliptin (e.g. JANUVIA), saxagliptin (e.g., ONGLYZA) and linagliptin (e.g., TRADJENTA)); GLP-1 receptor agonists (e.g., exenatide (e.g. BYETTA) and liraglutide (e.g., VICTOZA)); and SGLT2 inhibitors (e.g., canagliflozin (e.g. NVOKANA) and dapagliflozin (e.g., FARXIGA)).
In some embodiments, the patient is not receiving one or more non-insulin diabetes agents selected from metformin (e.g., GLUCOPHAGE, GLUMETZA); Sulfonylureas (e.g., glyburide (e.g., DIABETA, GLYNASE), glipizide (e.g., GLUCOTROL) and glimepiride (e.g., AMARYL)); thiazolidinediones (e.g., rosiglitazone (e.g., AVANDIA) and pioglitazone (e.g., ACTOS)); DPP-4 inhibitors (e.g., sitagliptin (e.g., JANUVIA), saxagliptin (e.g., ONGLYZA) and linagliptin (e.g., TRADJENTA)); GLP-1 receptor agonists (e.g., exenatide (e.g., BYETTA) and liraglutide (e.g., VICTOZA)); and SGLT2 inhibitors (e.g., canagliflozin (e.g., NVOKANA) and dapagliflozin (e.g., FARXIGA)).
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof is an adjuvant therapy to an insulin therapy and/or a non-insulin diabetes agent. In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof is an insulin replacement therapy, wherein the insulin analog can be selected from a rapid acting insulin analog (e.g., lispro, aspart or glulisine) or a long acting insulin analog (e.g., glargine or detemir). The patient may have type 1 diabetes or type 2 diabetes, gestational diabetes or steroid-induced diabetes. In some embodiments, the treatment of diabetes and/or glucose intolerance in a patients not receiving insulin therapy comprises one or more of a decrease of the blood glucose level, stimulation of peripheral glucose disposal, and inhibition of hepatic glucose production. The TNFRSF25 agonistic antibody or antigen binding fragment thereof administration can be effective for providing glycemic control. The patient may suffer from insulin resistance. In some embodiments, the patient can be diagnosed with one or more of insulin resistance, prediabetes, impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and acanthosis nigricans. The patient may have cardiovascular disease or metabolic disease.
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof is administered as a regimen that decreases blood glucose level, stimulates peripheral glucose disposal, and/or inhibits hepatic glucose production. In some embodiments, the patient has one or more of an average hemoglobin A1c value of more than about 10% and an average glucose of more than about 200 mg/dl (11 mmol/1) at the start of treatment with conventional diabetic therapy, which can be insulin therapy and/or non-insulin diabetes agent therapy. In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof administration is effective for providing average glucose of below about 200 mg/dl (11 mmol/1) and/or providing average glycosylated hemoglobin levels (hemoglobin A1c) values of about 8% or less. In some embodiments, the patient does not experience an increase of insulin production upon TNFRSF25 agonistic antibody or antigen binding fragment administration and/or the TNFRSF25 agonistic antibody or antigen binding fragment administration does not cause one or more of hypoglycemia and hypokalemia.
In some aspects, the present disclosure provides a method for increasing a graft survival. The method can comprise administering an effective amount of TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof. In some embodiments, patient is a transplant recipient. The transplant may comprise regulatory T cells (Tregs) from a transplant donor. In some embodiments, the transplant comprises donor hematopoietic cells, donor stem cells, or donor bone marrow cells.
For example, in some embodiments, the method increases a survival of a graft such as a solid organ transplant rejection. In some embodiments, the method reduces the likelihood of solid organ transplant rejection.
The solid organ can be selected from lung, kidney, heart, liver, pancreas, thymus, gastrointestinal tract, cornea, eye, and composite allografts. Composite allograft transplantation can be, in embodiments, intestinal/multivisceral transplantation (which may or may not include liver) and its variants.
In some embodiments, Treg expansion, caused by administration of TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof, is correlated with graft survival, such as with islet transplant survival. In some embodiments, Treg expansion correlated significantly with a fold increase in one or both CD4+FOXP3+ and CD4+CD25+FOXP3+ cells. In embodiments, patient is a diabetes patient. In some embodiments, the patient, which may be afflicted by a diabetes or by another insulin-related condition or disease, is a recipient of a transplant, such as an islet cells transplant, pancreatic transplant, or another transplant.
In some aspects, the present disclosure provides a method for increasing graft survival, comprising administering an effective amount of TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof. In some embodiments, the patient is a transplant recipient. The transplant may comprise islet cells transplant or a solid organ (e.g., pancreas) from a transplant donor. In some embodiments, the patient may be afflicted by a diabetes or by another insulin-related condition or disease. In some embodiments, the increase in the graft survival comprises an increase in at least in at least one month, or at least about 2 months, or at least about 3 months, or at least about 4 months, or at least about 5 months, or at least about 6 months, or at least about 7 months, or at least about 8 months, or at least about 9 months, or at least about 10 months, or at least about 11 months, or at least about 12 months, or at least about a year.
In embodiments, the present methods provide for an increase in a graft survival of greater than about one month, or greater than about 2 months, or greater than about 3 months, or greater than about 4 months, or a greater than about 5 months, or greater than about 6 months, or greater than about 7 months, or greater than about 8 months, or greater than about 9 months, or greater than about 10 months, or greater than about 11 months, or greater than about 12 months, or greater than about year.
In some aspects, the present disclosure provides a method for treating or preventing a graft rejection, comprising administering an effective amount of TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof. In some embodiments, the patient is a transplant recipient. The transplant may comprise islet cells or a solid organ (e.g., pancreas) from a transplant donor. In some embodiments, the patient may be afflicted by a diabetes or by another insulin-related condition or disease.
The present disclosure provides, in some aspects, a method for treating or preventing graft-versus-host disease (GVHD), comprising administering an effective amount of TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof. In some embodiments, the patient is a transplant recipient.
The transplant may comprise islet cells or a solid organ (e.g., pancreas) from a transplant donor. In some embodiments, the transplant comprises donor hematopoietic cells, donor stem cells, or donor bone marrow cells.
In embodiments, the effectiveness of the present methods is assayed by beta cell function. See Forbes et al. American Journal of Transplantation. 2016: 16 (91): pages 2704-2713 (first published: 28 Mar. 2016). In some embodiments, beta cell function is assessed using a single fasting blood sample factoring in the dose of insulin required per day and the patient's body weight. In particular, a BETA-2 score is determined, which is a validated composite score of beta cell function that incorporates the continuous variables glucose, C-peptide, HbA1c and insulin dose and that correlates strongly with other validated measures of graft function. See Forbes et al. American Journal of Transplantation. 2016: 16 (91): pages 2704-2713 (first published: 28 Mar. 2016). The BETA-2 score allows tracking islet engraftment over time, and permits early detection of graft dysfunction. See id. In embodiments, the present methods provide an improvement in BETA-2 score or a comparable measure. It should be appreciated that other approaches can be used additionally or alternatively to access a graft function.
In some embodiments, the treatment or prevention of GVHD using TNFRSF25 agonistic antibody or antigen binding fragment results in reduction of a graft versus host disease. In some embodiments, the graft versus host disease is acute graft-versus-host-disease (aGVHD). In some embodiments, the graft versus host disease is chronic graft-versus-host-disease (cGVHD). In various embodiments, the administration is also to the transplant donor. In some embodiments, the administration to the transplant donor occurs prior to transplant. In some embodiments, the administration to the transplant recipient occurs after the transplant. In some embodiments, administration is to both the transplant donor and transplant recipient.
GVHD is the deterioration of cells or tissues that are transplanted from a donor to a recipient due to the recognition by the immune system of the recipient that the cells or tissues are foreign. Thus, because Class I MHC are on more cells of the body, it is most desirable to transplant cells and tissues from people that have highest matching Class I MHC profiles followed by the highest matching Class MHC profiles. Thus, in most transplant recipients, GVHD is due to activation of the immune system to mismatched Class MHC molecules and other polymorphic proteins (minor histocompatibility antigens).
In some embodiments, the present methods relate to acute and chronic forms of GVHD. The acute or fulminant form of the disease (aGVHD) is normally observed within the first 100 days post-transplant, and is a major challenge to the effectiveness of transplants owing to the associated morbidity and mortality. The chronic form of graft-versus-host-disease (cGVHD) normally occurs after 100 days. The appearance of moderate to severe cases of cGVHD adversely influences long-term survival. After bone marrow transplantation, T cells present in the graft, either as contaminants or intentionally introduced into the host, attack the tissues of the transplant recipient after perceiving host tissues as antigenically foreign. The T cells produce an excess of cytokines, including TNF alpha and interferon-gamma (IFNγ). A wide range of host antigens can initiate graft-versus-host-disease, among them the human leukocyte antigens (HLAs). However, graft-versus-host disease can occur even when HLA-identical siblings are the donors. Classically, acute graft-versus-host-disease is characterized by selective damage to the liver, skin and mucosa, and the gastrointestinal tract. Additional studies show that that other graft-versus-host-disease target organs include the immune system (such as the bone marrow and the thymus) itself, and the lungs in the form of idiopathic pneumonitis. Chronic graft-versus-host-disease also attacks the above organs, but over its long-term course can also cause damage to the connective tissue and exocrine glands.
In some embodiments, the present methods relate to treating or preventing acute GVHD. In embodiments, the present methods treat a patient who has one or more risk factors of acute GVHD, such as HLA “mismatch,” or unrelated donor, older patient age, female donor to male recipient, intensity of the conditioning regimen or total body irradiation during conditioning regimen, and donor lymphocyte infusion. In embodiments, the present methods treat or prevent symptoms of acute GVHD, such as skin rash, gastrointestinal (GI) tract disorders, and liver symptoms.
In some embodiments, the present methods relate to treating or preventing chronic GVHD. In embodiments, the present methods treat a patient who has one or more risk factors of chronic GVHD, such as HLA mismatch or unrelated donor, older patient age, older donor age, female donor for male recipient and number of children the female donor has had, stem cell source, stem cells retrieved from peripheral blood have a higher risk of causing chronic GVHD than stem cells retrieved from bone marrow, stem cells retrieved from cord blood have the lowest risk of causing chronic GVHD, and prior acute GVHD. In embodiments, the present methods treat or prevent symptoms of chronic GVHD, such as symptoms of the eyes, mouth, skin, nails, scalp and body hair, gastrointestinal (GI) tract, lungs, liver, muscles and joints, and genitals and sex organs.
In some embodiments, the present methods relate to GVHD as defined by one of more of the Billingham Criteria: 1) administration of an immunocompetent graft, with viable and functional immune cells; 2) the recipient is immunologically histoincompatible; and 3) the recipient is immunocompromised and therefore cannot destroy or inactivate the transplanted cells.
In some embodiments, the present methods relate to treating or preventing GVHD in a patient/transplant recipient who is undergoing a GVHD treatment. In embodiments, the present methods relate to treating or preventing GVHD in using the present fusion proteins in combination therapies with a GVHD treatment. In embodiments, the present fusion proteins reduce or ameliorate one or more side effects of a GVHD treatment. Illustrative GVHD treatments are immunosuppression agents, such as corticosteroids (such as methylprednisolone or prednisone) and other immunosuppressive drugs. An illustrative GVHD treatment, in some embodiments, is prednisone. Other illustrative GVHD treatments include ibrutinib (e.g. IMBRUVICA), mycophenolate mofetil, mTOR inhibitors, such as sirolimus (rapamycin), everolimus, calcineurin inhibitors, such as tacrolimus or cyclosporine, ciclosporin, monoclonal antibodies such as infliximab (e.g. REMICADE), tocilizumab (e.g. ACTEMRA), alemtuzumab (e.g. CAMPATH), basiliximab (e.g. SIMULECT), daclizumab (e.g. ZINBRYTA), and denileukin diftitox (e.g. ONTAK), antithymocyte globulin (ATG), anti-lymphocyte globulin (ALG), pentostatin (e.g. NIPENT), ruxolitinib (e.g. JAKAFI), and photopheresis.
In some embodiments, the present methods pertain to patients who fail to respond to steroid therapy are labeled “steroid-refractory”. In embodiments, the present methods pertain to patients who fail one or more lines of systemic GVHD therapy.
In embodiments, administration of an TNFRSF25 agonistic antibody or antigen binding fragment thereof to a patient in need thereof results in an increase in serum levels of IL-5.
In various embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment causes a sustained increase in Treg cells in the transplant donor and/or transplant recipient. In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment does not cause substantial Treg suppression in the transplant donor and/or transplant recipient. In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof does not cause substantial Treg anergy in the transplant donor and/or transplant recipient.
In various embodiments of the present disclosure, the method for treating or preventing GVHD prevents a transplant rejection. For example, in some embodiments, the method prevents a solid organ transplant rejection. In some embodiments, the method reduces the likelihood of solid organ transplant rejection. The solid organ can be selected from lung, kidney, heart, liver, pancreas, thymus, gastrointestinal tract, cornea, eye, and composite allografts. Composite allograft transplantation can be, e.g., intestinal/multivisceral transplantation (which may or may not include liver) and its variants.
The TNFRSF25 agonistic antibody or antigen binding fragment thereof can cause a sustained increase in Treg cells in the solid organ transplant recipient. The sustained increase in Treg cells can comprise a substantially similar level of Treg cells in the solid organ transplant recipient after the first and last administrations of the TNFRSF25 agonistic antibody or antigen binding fragment. For example, if the administration of the TNFRSF25 agonistic antibody or antigen binding fragment occurs 3 times, the increase in Treg cells remains at a substantially similar level in the solid organ transplant recipient after the first and third administrations of the TNFRSF25 agonistic antibody or antigen binding fragment.
In some embodiments, the method prevents a rejection of a transplant that is islet cells transplant. In some embodiments, the method reduces the likelihood of islet cells transplant rejection. Composite allograft transplantation can be, e.g., islet cells transplant rejection and a solid organ transplant such as, pancreas.
The TNFRSF25 agonistic antibody or antigen binding fragment thereof can cause a sustained increase in Treg cells in the islet cells transplant recipient. The sustained increase in Treg cells can comprise a substantially similar level of Treg cells in the islet cells transplant recipient after the first and last administrations of the TNFRSF25 agonistic antibody or antigen binding fragment. For example, if the administration of the TNFRSF25 agonistic antibody or antigen binding fragment occurs 3 times, the increase in Treg cells remains at a substantially similar level in the islet cells transplant recipient after the first and third administrations of the TNFRSF25 agonistic antibody or antigen binding fragment.
In some embodiments in accordance with the present disclosure, the TNFRSF25 agonistic antibody or antigen binding fragment thereof can be administered in various doses. For example, the administration can occur at least 7 times, at least 10 times, at least 14 times, about 3-7 times, about 3-14 times, about 3-21 times, about 3 times, about 7 times, about 10 times, or about 14 times. In some embodiments, the administration occurs daily, which can be, for example, twice daily. In some embodiments, the administration occurs daily for 3-7 days, daily for 7-14 days, daily for 7-21 days, daily for at least 7 days, daily for at least 10 days, or daily for at least 21 days. The administration in any of the above doses can occur before the transplant or concurrently with the transplant. In embodiments in which the transplant is a solid organ transplant, the administration of the TNFRSF25 agonistic antibody or antigen binding fragment thereof in any of the above doses can occur after the solid organ transplant, or before and after the solid organ transplant.
A combination therapy in which the TNFRSF25 agonistic antibody or antigen binding fragment thereof described herein can be administered sequentially or simultaneously with one or more anti-rejection drugs (and in which the anti-rejection drugs can be administered sequentially or simultaneously) can include administration of interleukin-2 (IL-2) (e.g., a low dose of IL-2), which can also be administered sequentially or simultaneously with the TNFRSF25 agonistic antibody or antigen binding fragment thereof and/or with one or more anti-rejection drugs.
Accordingly, in some embodiments, the method of administering TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises comprising administering IL-2. In some embodiments, the IL-2 is a low dose of IL-2. In some embodiments, the low dose of IL-2 is less than 1 million units per square meter per day. In some embodiments, the low dose of IL-2 is an amount in the range of about 30,000 to about 300,000 units per square meter per day. In some embodiments, the low dose of IL-2 is about 300,000 units per square meter per day. In some embodiments, the low dose of IL-2 is about 30,000 units per square meter per day.
In some embodiments, the administration of low dose IL-2 can be sequential with the administration of the TNFRSF25 agonistic antibody or antigen binding fragment thereof. In some embodiments, the administration of low dose IL-2 is concurrent with the administration of the TNFRSF25 agonistic antibody or antigen binding fragment thereof.
In various embodiments, the present invention relates to the generation or modulation of Tregs. A “T regulatory cell” or “Treg cell” refers to a cell that can modulate a T cell response. Treg cells express the transcription factor Foxp3, which is not upregulated upon T cell activation and discriminates Tregs from activated effector cells. Tregs are identified by the cell surface markers CD25, CTLA4, and GITR. Several Treg subsets have been identified that have the ability to inhibit autoimmune, immune, and chronic inflammatory responses and to maintain immune tolerance in tumor-bearing hosts. These subsets include interleukin 10- (IL-10-) secreting T regulatory type 1 (Tr1) cells, transforming growth factor-β- (TGF-β-) secreting T helper type 3 (Th3) cells, and “natural” CD4+/CD25+Tregs (Trn) (Fehervari and Sakaguchi. J. Clin. Invest. 2004, 114:1209-1217; Chen et al. Science. 1994, 265: 1237-1240; Groux et al. Nature. 1997, 389: 737-742).
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises a TNFRSF25 agonistic antibody or antigen binding fragment, such as PTX-35, which is described in PCT/US2017/036817 (WO2017214547), which is incorporated herein by reference in its entirety.
In any of the methods of treatment or prevention of a disease or condition in accordance with the present disclosure, the TNFRSF25 agonistic antibody or antigen binding fragment thereof can comprise (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence is GFTFSNHDLN (SEQ ID NO: 1), the heavy chain CDR2 sequence is YISSASGLISYADAVRG (SEQ ID NO: 2); and the heavy chain CDR3 sequence is DPAYTGLYALDF (SEQ ID NO: 3) or DPPYSGLYALDF (SEQ ID NO: 4); and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence is TLSSELSWYTIV (SEQ ID NO: 5), the light chain CDR2 sequence is LKSDGSHSKGD (SEQ ID NO: 6), and the light chain CDR3 sequence is CGAGYTLAGQYGWV (SEQ ID NO: 7). In some embodiments, heavy chain CDR1, CDR2, and CDR3 sequences and/or light chain CDR1, CDR2, and CDR3 sequences each include at least one, or at least 2, or at least 3, or at least 4, or at least 5 mutations such as amino acid substitutions. In some embodiments, heavy chain CDR1, CDR2, and CDR3 sequences and/or light chain CDR1, CDR2, and CDR3 sequences each include more than 5 mutations.
In some embodiments, an TNFRSF25 agonistic antibody includes a set of six CDRs that include no more than two, or no more than three, or no more than four, or no more than five, or no more than six total amino acid substitutions in the set of six CDRs having the amino acid sequences set forth in SEQ ID NOS: 1, 2, 3 or 4, 5, 6, and 7. In some embodiments, an TNFRSF25 agonistic antibody includes a set of six CDRs that include at least one, or at least two, or at least three, or at least four, or at least five total amino acid substitutions in the set of six CDRs having the amino acid sequences set forth in SEQ ID NOS: 1, 2, 3 or 4, 5, 6, and 7. In some embodiments, an TNFRSF25 agonistic antibody includes a set of six CDRs that include one, or two, or three, or four, or five, or more than five total amino acid substitutions in the set of six CDRs having the amino acid sequences set forth in SEQ ID NOS: 1, 2, 3 or 4, 5, 6, and 7.
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof further comprises variable region framework (FW) sequences juxtaposed between the CDRs according to the formula (FW1)-(CDR1)-(FW2)-(CDR2)-(FW3)-(CDR3)-(FW4), wherein the variable region FW sequences in the heavy chain variable region are heavy chain variable region FW sequences, and wherein the variable region FW sequences in the light chain variable region are light chain variable region FW sequences. In some embodiments, the variable region FW sequences are human. In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof further comprises human heavy chain and light chain constant regions. The constant regions can be selected from the group consisting of human IgG1, IgG2, IgG3, and IgG4. For instance, in some embodiments, the constant regions are IgG1. In some embodiments, the constant regions are IgG4.
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises a heavy chain variable region having the amino acid sequence EVQLVESGGGLSQPGNSLQLSCEASGFTFSNHDLNWVRQAPGKGLEWVAYISSASGLISYADAVRGRFTISRDN AKNSLFLQMNNLKSEDTAMYYCARDPPYSGLYALDFWGQGTQVTVSS (SEQ ID NO: 8), or an amino acid sequence of at least about 85% to about 99% identity thereto. In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises a light chain variable region having the amino acid sequence QPVLTQSPSASASLSGSVKLTCTLSSELSSYTIVWYQQRPDKAPKYVMYLKSDGSHSKGDGIPDRFSGSSSGAH RYLSISNVQSEDDATYFCGAGYTLAGQYGWVFGSGTKVTVL (SEQ ID NO: 9), or an amino acid sequence of at least about 85% to about 99% identity thereto.
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises a heavy chain variable region having the amino acid sequence EVQLVESGGGLSQPGNSLQLSCEASGFTFSNHDLNWVRQAPGKGLEWVAYISSASGLISYADAVRGRFTISRDN AKNSLFLQMNNLKSEDTAMYYCARDPPYSGLYALDFWGQGTQVTVSS (SEQ ID NO: 8), or an amino acid sequence of at least about 85%, or at least about 86%, or at least about 87%, or at least about 88%, or at least about 89%, or at least about 90%, or at least about 91%, or at least about 92%, or at least about 93%, or at least about 94%, or at least about 95%, or at least about 96%, or at least about 97%, or at least about 98%, or at least about 99% identity thereto.
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises a light chain variable region having the amino acid sequence QPVLTQSPSASASLSGSVKLTCTLSSELSSYTIVWYQQRPDKAPKYVMYLKSDGSHSKGDGIPDRFSGSSSGAH RYLSISNVQSEDDATYFCGAGYTLAGQYGWVFGSGTKVTVL (SEQ ID NO: 9), or an amino acid sequence of at least about 85%, or at least about 86%, or at least about 87%, or at least about 88%, or at least about 89%, or at least about 90%, or at least about 91%, or at least about 92%, or at least about 93%, or at least about 94%, or at least about 95%, or at least about 96%, or at least about 97%, or at least about 98%, or at least about 99% identity thereto.
In some embodiments, a heavy chain variable region has an amino acid sequence of SEQ ID NO: 8, or an antigen binding fragment thereof, but with one to 24 sequence modifications, as well as polypeptides having at least about 80% (e.g., about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99%) amino acid sequence identity to SEQ ID NO: 8, or an antigen binding fragment thereof. In some embodiments, a heavy chain variable region polypeptide can contain 24 or less (e.g., 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, ten, nine, eight, seven, six, five, four, three, two, or one) amino acid substitution as compared to SEQ ID NO: 8, or an antigen binding fragment thereof.
In some embodiment, a light chain variable region has an amino acid sequence of SEQ ID NO: 9, or an antigen binding fragment thereof, but with one to 23 sequence modifications, as well as polypeptides having at least about 80% (e.g., about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99%) amino acid sequence identity to SEQ ID NO: 9, or an antigen binding fragment thereof. In some embodiments, a light chain variable region polypeptide can contain 23 or less (e.g., 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, ten, nine, eight, seven, six, five, four, three, two, or one) amino acid substitutions as compared to SEQ ID NO: 9, or an antigen binding fragment thereof.
In some embodiments, an TNFRSF25 agonistic antibody or antigen binding fragment thereof is PTX-25 antibody or antigen binding fragment thereof. The PTX-25 antibody is described, for example, in PCT/US2015/061082 (WO2016081455), which is incorporated herein by reference in its entirety. In some embodiments, an TNFRSF25 agonistic antibody comprises (i) a heavy chain variable region sequence comprising the amino acid sequence EVQLVESGGGLSQPGNSLQLSCEAS GFTFSNHDLNWVRQAPGKGLEWVAYISSASGLISYADAVRGRFTISRDNAKNSLFLQMNNLKSEDTAMYYCARD PPYSGLYALDFWGQGTQVTVSS (SEQ ID NO: 10) or the amino acid sequence of SEQ ID NO: 10 with no more than 12 total amino acid substitutions (e.g., no more than 12, or no more than 11, or no more than ten, or no more than nine, or no more than eight, or no more than seven, or no more than six, or no more than five, or no more than four, or no more than three, or no more than two total amino acid substitutions); and (ii) a light chain variable region sequence comprising the amino acid sequence QPVLTQSPSASASLSGSVKLTCTLSSEL SSYTIVWYQQRPDKAPKYVMYLKSDGSHSKGDGIPDRFSGSSSGAHRYLSISNVQSEDDATYFCGAGYTLAGQ YGWVFGSGTKVTVL (SEQ ID NO:11) or the amino acid sequence of SEQ ID NO: 11 with no more than 11 total amino acid substitutions (e.g., no more than 11, or no more than ten, or no more than nine, or no more than eight, or no more than seven, or no more than six, or no more than five, or no more than four, or no more than three, or no more than two total amino acid substitutions, or no more than one total amino acid substitution). An amino acid substitution refers to the replacement of one amino acid residue with another amino acid in a peptide sequence.
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises a heavy chain variable region having the amino acid sequence EVQLVESGGGLSQPGNSLQLSCEAS GFTFSNHDLNWVRQAPGKGLEWVAYISSASGLISYADAVRGRFTISRDNAKNSLFLQMNNLKSEDTAMYYCARD PPYSGLYALDFWGQGTQVTVSS (SEQ ID NO: 10) or an amino acid sequence of at least about 85 to about 99% (e.g., an amino acid sequence of at least about 85%, or at least about 86%, or at least about 87%, or at least about 88%, or at least about 89%, or at least about 90%, or at least about 91%, or at least about 92%, or at least about 93%, or at least about 94%, or at least about 95%, or at least about 96%, or at least about 97%, or at least about 98%, or at least about 99%) identity thereto.
In some embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises a light chain variable region having the amino acid sequence QPVLTQSPSASASLSGSVKLTCTLSSELSSYTIVWYQQRPDKAPKYVMYLKSDGSHSKGDGIPDRFSGSSSGAH RYLSISNVQSEDDATYFCGAGYTLAGQYGWVFGSGTKVTVL (SEQ ID NO: 11), or an amino acid sequence of at least about 85 to about 99% (e.g., an amino acid sequence of at least about 85%, or at least about 86%, or at least about 87%, or at least about 88%, or at least about 89%, or at least about 90%, or at least about 91%, or at least about 92%, or at least about 93%, or at least about 94%, or at least about 95%, or at least about 96%, or at least about 97%, or at least about 98%, or at least about 99%) identity thereto.
In some embodiments, an TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises an isolated heavy chain variable region polypeptide that binds specifically to TNFRSF25, where the polypeptide includes heavy chain CDR1, CDR2, and CDR3 sequences, where the CDR1 sequence is GFTFSNHDLN (SEQ ID NO: 12), the CDR2 sequence is YISSASGLISYADAVRG (SEQ ID NO: 13); and (c) the CDR3 sequence is DPPYSGLYALDF (SEQ ID NO: 14). The isolated heavy chain variable region polypeptide can further include variable region heavy chain framework (FW) sequences juxtaposed between the heavy chain CDRs according to the formula (FW1)-(CDR1)-(FW2)-(CDR2)-(FW3)-(CDR3)-(FW4). The heavy chain framework sequences can be human. In some embodiments, the isolated heavy chain variable region polypeptide can be combination with a light chain variable region polypeptide. In some embodiments, the light chain variable region polypeptide comprises light chain CDR1, CDR2, and CDR3 sequences, wherein the CDR1 sequence is TLSSELSSYTIV (SEQ ID NO: 15), the CDR2 sequence is LKSDGSHSKGD (SEQ ID NO: 16), and the CDR3 sequence is GAGYTLAGQYGWV (SEQ ID NO: 17). The TNFRSF25 agonistic antibody can be a humanized monoclonal antibody that specifically binds to TNFRSF25.
In some embodiments, an TNFRSF25 agonistic antibody includes a set of six CDRs that include no more than two, or no more than three, or no more than four, or no more than five, or no more than six total amino acid substitutions in the set of six CDRs having the amino acid sequences set forth in SEQ ID NOS: 12, 13, 14, 15, 16, and 17. In some embodiments, an TNFRSF25 agonistic antibody includes a set of six CDRs that include at least one, or at least two, or at least three, or at least four, or at least five, or at least 6 total amino acid substitutions in the set of six CDRs having the amino acid sequences set forth in SEQ ID NOS: 12, 13, 14, 15, 16, and 17. In some embodiments, an TNFRSF25 agonistic antibody includes a set of six CDRs that include one, or two, or three, or four, or five, or more than five total amino acid substitutions in the set of six CDRs having the amino acid sequences set forth in SEQ ID NOS: 12, 13, 14, 15, 16, and 17.
In embodiments, the TNFRSF25 agonistic antibody or antigen binding fragment thereof comprises a heavy chain comprising an amino acid of SEQ ID NO: 18, 19, 20, or 21, and a light chain comprising an amino acid of SEQ ID NO: 22 or 23, as follows:
In embodiments, an TNFRSF25 agonistic antibody or antigen binding fragment thereof is any of the antibodies or antibody fragments (or combinations thereof) described in PCT/US2015/061082 (WO2016081455).
In embodiments, variable region light chain framework (FW) sequences can be juxtaposed between the light chain CDRs according to the formula (FW1)-(CDR1)-(FW2)-(CDR2)-(FW3)-(CDR3)-(FW4). The light chain framework sequences can be human. The antibody or antigen binding fragment can further include a human constant region (e.g., a constant region selected from the group consisting of human IgG1, IgG2, IgG3, and IgG4), or a murine constant region (e.g., a constant region selected from the group consisting of murine IgG1, IgG2A, IgG2B, and IgG3).
In some embodiments, an TNFRSF25 agonistic antibody or antigen binding fragment thereof is as described in PCT/US2010/044218 (WO2011017303), which is incorporated herein by reference in its entirety.
Various embodiments of the present disclosure make use of TNFRSF25 agonistic antibodies or antigen binding fragment thereof. In various embodiments, the antibody is an antibody (e.g., human, hamster, feline, mouse, cartilaginous fish, or camelid antibodies), and any derivative or conjugate thereof, that specifically binds to TNFRSF25. Non-limiting examples of antibodies include monoclonal antibodies, polyclonal antibodies, humanized antibodies, multi-specific antibodies (e.g., bi-specific antibodies), single-chain antibodies (e.g., single-domain antibodies, camelid antibodies, and cartilaginous fish antibodies), chimeric antibodies, feline antibodies, and felinized antibodies. Monoclonal antibodies are homogeneous populations of antibodies to a particular epitope of an antigen. Polyclonal antibodies are heterogeneous populations of antibody molecules that are contained in the sera of the immunized animals.
An isolated polypeptide can yield a single major band on a non-reducing polyacrylamide gel. An isolated polypeptide can be at least about 75% pure (e.g., at least 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% pure). Isolated polypeptides can be obtained by, for example, extraction from a natural source, by chemical synthesis, or by recombinant production in a host cell or transgenic plant, and can be purified using, for example, affinity chromatography, immunoprecipitation, size exclusion chromatography, and ion exchange chromatography. The extent of purification can be measured using any appropriate method, including, without limitation, column chromatography, polyacrylamide gel electrophoresis, or high-performance liquid chromatography.
In embodiments, an antigen binding fragment that specifically binds to TNFRSF25 is provided. Such antigen binding fragment, in embodiments, is any portion of a full-length antibody that contains at least one variable domain (e.g., a variable domain of a mammalian (e.g., feline, human, hamster, or mouse) heavy or light chain immunoglobulin, a camelid variable antigen binding domain (VHH), or a cartilaginous fish immunoglobulin new antigen receptor (Ig-NAR) domain) that is capable of specifically binding to an antigen. Non-limiting examples of antibody fragments include Fab, Fab′, F(ab′)2, and Fv fragments, diabodies, linear antibodies, and multi-specific antibodies formed from antibody fragments. Additional antibody fragments containing at least one camelid VHH domain or at least one cartilaginous fish Ig-NAR domain include mini-bodies, micro-antibodies, subnano-antibodies, and nano-antibodies, and any of the other forms of antibodies described, for example, in U.S. Publication No. 2010/0092470.
An antibody can be of the IgA-, IgD-, IgE, IgG- or IgM-type, including IgG- or IgM-types such as, without limitation, IgG1-, IgG2-, IgG3-, IgG4-, IgM1- and IgM2-types. For example, in some cases, the antibody is of the IgG1-, IgG2- or IgG4-type.
In some embodiments, antibodies as provided herein can be fully human or humanized antibodies. In embodiments, the human antibody is an antibody that is encoded by a nucleic acid (e.g., a rearranged human immunoglobulin heavy or light chain locus) present in the genome of a human. In some embodiments, a human antibody can be produced in a human cell culture (e.g., feline hybridoma cells). In some embodiments, a human antibody can be produced in a non-human cell (e.g., a mouse or hamster cell line). In some embodiments, a human antibody can be produced in a bacterial or yeast cell.
Human antibodies can avoid certain problems associated with xenogeneic antibodies, such as antibodies that possess murine or rat variable and/or constant regions. For example, because the effector portion is human, it can interact better with other parts of the human immune system, e.g., to destroy target cells more efficiently by complement-dependent cytotoxicity or antibody-dependent cellular cytotoxicity. In addition, the human immune system should not recognize the antibody as foreign. Further, half-life in human circulation will be similar to naturally occurring human antibodies, allowing smaller and less frequent doses to be given. Methods for preparing human antibodies are known in the art.
In embodiments, the antibody is a humanized antibody, e.g., an antibody that contains minimal sequence derived from non-human (e.g., mouse, hamster, rat, rabbit, or goat) immunoglobulin. Humanized antibodies generally are chimeric or mutant monoclonal antibodies from mouse, rat, hamster, rabbit or other species, bearing human constant and/or variable region domains or specific changes. In non-limiting examples, humanized antibodies are human antibodies (recipient antibody) in which hypervariable region (HVR) residues of the recipient antibody are replaced by HVR residues from a non-human species (donor) antibody, such as a mouse, rat, rabbit, or goat antibody having the desired specificity, affinity, and capacity. In some embodiments, Fv framework residues of the human immunoglobulin can be replaced by corresponding non-human residues. In some embodiments, humanized antibodies can contain residues that are not found in the recipient antibody or in the donor antibody. Such modifications can be made to refine antibody performance, for example.
In some embodiments, a humanized antibody can contain substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops (CDRs) correspond to those of a non-human immunoglobulin, while all or substantially all of the framework regions are those of a human immunoglobulin sequence. A humanized antibody also can contain at least a portion of an immunoglobulin constant (Fc) region, typically that of a human immunoglobulin.
In some embodiments, a humanized antibody or antigen binding fragment as provided herein can have reduced or minimal effector function (e.g., as compared to corresponding non-humanized antibody), such that it does not stimulate effector cell action to the same extent that a corresponding non-humanized antibody would.
Techniques for generating humanized antibodies are well known to those of skill in the art. In some embodiments, controlled rearrangement of antibody domains joined through protein disulfide bonds to form new, artificial protein molecules or “chimeric” antibodies can be utilized (Konieczny et al., Haematologia (Budap.) 14:95, 1981). Recombinant DNA technology can be used to construct gene fusions between DNA sequences encoding mouse antibody variable light and heavy chain domains and human antibody light and heavy chain constant domains (Morrison et al., Proc Natl Acad Sci USA 81:6851, 1984). For example, DNA sequences encoding antigen binding portions or CDRs of murine monoclonal antibodies can be grafted by molecular means into DNA sequences encoding frameworks of human antibody heavy and light chains (Jones et al., Nature 321:522, 1986; and Riechmann et al., Nature 332:323, 1988). Expressed recombinant products are called “reshaped” or humanized antibodies, and contain the framework of a human antibody light or heavy chain and antigen recognition portions, CDRs, of a murine monoclonal antibody.
Other methods for designing heavy and light chains and for producing humanized antibodies are described in, for example, U.S. Pat. Nos. 5,530,101; 5,565,332; 5,585,089; 5,639,641; 5,693,761; 5,693,762; and 5,733,743. Yet additional methods for humanizing antibodies are described in U.S. Pat. Nos. 4,816,567; 4,935,496; 5,502,167; 5,558,864; 5,693,493; 5,698,417; 5,705,154; 5,750,078; and 5,770,403, for example.
In embodiments, the antibody is a single-chain antibody, e.g. a single polypeptide that contains at least one variable binding domain (e.g., a variable domain of a mammalian heavy or light chain immunoglobulin, a camelid VHH, or a cartilaginous fish (e.g., shark) Ig-NAR domain) that is capable of specifically binding to an antigen. Non-limiting examples of single-chain antibodies include single-domain antibodies.
In embodiments, the antibody is a single-domain antibody, e.g. a polypeptide that contains one camelid VHH or at least one cartilaginous fish Ig-NAR domain that is capable of specifically binding to an antigen. Non-limiting examples of single-domain antibodies are described, for example, in U.S. Publication No. 2010/0092470.
In embodiments, the antibody specifically binds to a particular antigen, e.g., TNFRSF25, when it binds to that antigen in a sample, and does not recognize and bind, or recognizes and binds to a lesser extent, other molecules in the sample. In some embodiments, an antibody or an antigen binding fragment thereof can selectively bind to an epitope with an affinity (Kd) equal to or less than, for example, about 1×10−6 M (e.g., equal to or less than about 1×10−9 M, equal to or less than about 1×10−10 M, equal to or less than about 1×10−11 M, or equal to or less than about 1×10−12 M) in phosphate buffered saline. The ability of an antibody or antigen binding fragment to specifically bind a protein epitope can be determined using any of the methods known in the art or those methods described herein (e.g., by Biacore/Surface Plasmon Resonance). This can include, for example, binding to TNFRSF25 on live cells as a method to stimulate caspase activation in live transformed cells, binding to an immobilized target substrate including human TNFRSF25 fusion proteins as detected using an ELISA method, binding to TNFRSF25 on live cells as detected by flow cytometry, or binding to an immobilized substrate by surface plasmon resonance (including ProteOn).
Antibodies having specific binding affinity for TNFRSF25 can be produced using standard methods. For example, a TNFRSF25 polypeptide can be recombinantly produced, purified from a biological sample (e.g., a heterologous expression system), or chemically synthesized, and used to immunize host animals, including rabbits, chickens, mice, guinea pigs, or rats. Various adjuvants that can be used to increase the immunological response depend on the host species and include Freund's adjuvant (complete and incomplete), mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet hemocyanin and dinitrophenol. Monoclonal antibodies can be prepared using a TNFRSF25 polypeptide and standard hybridoma technology. In particular, monoclonal antibodies can be obtained by any technique that provides for the production of antibody molecules by continuous cell lines in culture such as described by Kohler et al. (Nature 256:495, 1975), the human B-cell hybridoma technique of Kosbor et al. (Immunology Today, 4:72, 1983) or Cote et al. (Proc. Natl. Acad. Sci. USA, 80:2026, 1983), and the EBV-hybridoma technique described by Cole et al. (Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96, 1983). Such antibodies can be of any immunoglobulin class including IgG, IgM, IgE, IgA, IgD, and any subclass thereof. The hybridoma producing the monoclonal antibodies can be cultivated in vitro and in vivo.
In some embodiments, amino acid substitutions can be made by selecting conservative substitutions that do not differ significantly in their effect on maintaining (a) the structure of the peptide backbone in the area of the substitution, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain. For example, naturally occurring residues can be divided into groups based on side-chain properties: (1) hydrophobic amino acids (norleucine, methionine, alanine, valine, leucine, and isoleucine); (2) neutral hydrophilic amino acids (cysteine, serine, and threonine); (3) acidic amino acids (aspartic acid and glutamic acid); (4) basic amino acids (asparagine, glutamine, histidine, lysine, and arginine); (5) amino acids that influence chain orientation (glycine and proline); and (6) aromatic amino acids (tryptophan, tyrosine, and phenylalanine). Substitutions made within these groups can be considered conservative substitutions. Non-limiting examples of conservative substitutions include, without limitation, substitution of valine for alanine, lysine for arginine, glutamine for asparagine, glutamic acid for aspartic acid, serine for cysteine, asparagine for glutamine, aspartic acid for glutamic acid, proline for glycine, arginine for histidine, leucine for isoleucine, isoleucine for leucine, arginine for lysine, leucine for methionine, leucine for phenylalanine, glycine for proline, threonine for serine, serine for threonine, tyrosine for tryptophan, phenylalanine for tyrosine, and/or leucine for valine. In some embodiments, an amino acid substitution can be non-conservative, such that a member of one of the amino acid classes described above is exchanged for a member of another class.
Pharmaceutical Compositions
In addition, the disclosure also provides pharmaceutical compositions for the present methods of treating diabetes and related disorders, and GVHD (acute or chronic), which include an antibody or antigen binding fragment, as described herein, in combination with a pharmaceutically acceptable carrier. A “pharmaceutically acceptable carrier” (also referred to as an “excipient” or a “carrier”) is a pharmaceutically acceptable solvent, suspending agent, stabilizing agent, or any other pharmacologically inert vehicle for delivering one or more therapeutic compounds to a subject (e.g., a mammal, such as a human, non-human primate, dog, cat, sheep, pig, horse, cow, mouse, rat, or rabbit), which is nontoxic to the cell or subject being exposed thereto at the dosages and concentrations employed. Pharmaceutically acceptable carriers can be liquid or solid, and can be selected with the planned manner of administration in mind so as to provide for the desired bulk, consistency, and other pertinent transport and chemical properties, when combined with one or more of therapeutic compounds and any other components of a given pharmaceutical composition. Typical pharmaceutically acceptable carriers that do not deleteriously react with amino acids include, by way of example and not limitation: water, saline solution, binding agents (e.g., polyvinylpyrrolidone or hydroxypropyl methylcellulose), fillers (e.g., lactose and other sugars, gelatin, or calcium sulfate), lubricants (e.g., starch, polyethylene glycol, or sodium acetate), disintegrates (e.g., starch or sodium starch glycolate), and wetting agents (e.g., sodium lauryl sulfate). Pharmaceutically acceptable carriers also include aqueous pH buffered solutions or liposomes (small vesicles composed of various types of lipids, phospholipids and/or surfactants which are useful for delivery of a drug to a mammal). Further examples of pharmaceutically acceptable carriers include buffers such as phosphate, citrate, and other organic acids, antioxidants such as ascorbic acid, low molecular weight (less than about 10 residues) polypeptides, proteins such as serum albumin, gelatin, or immunoglobulins, hydrophilic polymers such as polyvinylpyrrolidone, amino acids such as glycine, glutamine, asparagine, arginine or lysine, monosaccharides, disaccharides, and other carbohydrates including glucose, mannose or dextrins, chelating agents such as EDTA, sugar alcohols such as mannitol or sorbitol, salt-forming counterions such as sodium, and/or nonionic surfactants such as TWEEN™, polyethylene glycol (PEG), and PLURONICS™.
Pharmaceutical compositions can be formulated by mixing one or more active agents with one or more physiologically acceptable carriers, diluents, and/or adjuvants, and optionally other agents that are usually incorporated into formulations to provide improved transfer, delivery, tolerance, and the like. A pharmaceutical composition can be formulated, e.g., in lyophilized formulations, aqueous solutions, dispersions, or solid preparations, such as tablets, dragées or capsules. A multitude of appropriate formulations can be found in the formulary known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences (18th ed, Mack Publishing Company, Easton, Pa. (1990)), particularly Chapter 87 by Block, Lawrence, therein. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic) containing vesicles (such as LIPOFECTIN™), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, emulsions carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. Any of the foregoing mixtures may be appropriate in treatments and therapies as described herein, provided that the active agent in the formulation is not inactivated by the formulation and the formulation is physiologically compatible and tolerable with the route of administration. See, also, Baldrick, Regul Toxicol Pharmacol 32:210-218, 2000; Wang, Int J Pharm 203:1-60, 2000; Charman, J Pharm Sci 89:967-978, 2000; and Powell et al. PDA J Pharm Sci Technol 52:238-311, 1998), and the citations therein for additional information related to formulations, excipients and carriers well known to pharmaceutical chemists.
Pharmaceutical compositions include, without limitation, solutions, emulsions, aqueous suspensions, and liposome-containing formulations. These compositions can be generated from a variety of components that include, for example, preformed liquids, self-emulsifying solids and self-emulsifying semisolids. Emulsions are often biphasic systems comprising of two immiscible liquid phases intimately mixed and dispersed with each other; in general, emulsions are either of the water-in-oil (w/o) or oil-in-water (o/w) variety. Emulsion formulations have been widely used for oral delivery of therapeutics due to their ease of formulation and efficacy of solubilization, absorption, and bioavailability.
Compositions and formulations can contain sterile aqueous solutions, which also can contain buffers, diluents and other suitable additives (e.g., penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers). Compositions additionally can contain other adjunct components conventionally found in pharmaceutical compositions. Thus, the compositions also can include compatible, pharmaceutically active materials such as, for example, antipruritics, astringents, local anesthetics or anti-inflammatory agents, or additional materials useful in physically formulating various dosage forms of the compositions provided herein, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers. Furthermore, the composition can be mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings, and aromatic substances. When added, however, such materials should not unduly interfere with the biological activities of the polypeptide components within the compositions provided herein. The formulations can be sterilized if desired.
In some embodiments, a composition containing an antibody or antigen binding fragment as used herein can be in the form of a solution or powder with or without a diluent to make an injectable suspension. The composition may contain additional ingredients including, without limitation, pharmaceutically acceptable vehicles, such as saline, water, lactic acid, mannitol, or combinations thereof, for example.
In one aspect, the disclosure provides a method of making an anti-TNFRSF25 antibody or antigen binding fragment thereof, comprising: a) providing the host cell as described herein; b) culturing said host cell under conditions wherein said antibody is expressed; and c) recovering said antibody from the host cell.
Any appropriate method can be used to administer an antibody or antigen binding fragment as described herein to a mammal. Administration can be, for example, parenteral (e.g., by subcutaneous, intrathecal, intraventricular, intramuscular, or intraperitoneal injection, or by intravenous drip). Administration can be rapid (e.g., by injection) or can occur over a period of time (e.g., by slow infusion or administration of slow release formulations). In some embodiments, administration can be topical (e.g., transdermal, sublingual, ophthalmic, or intranasal), pulmonary (e.g., by inhalation or insufflation of powders or aerosols), or oral. In addition, a composition containing an antibody or antigen binding fragment as described herein can be administered prior to, after, or in lieu of surgical resection of a tumor.
A composition containing an anti-TNFRSF25 antibody or antigen binding fragment can be administered to a mammal in any appropriate amount, at any appropriate frequency, and for any appropriate duration effective to achieve a desired outcome. For example, an anti-TNFRSF25 antibody or antigen binding fragment can be administered to a subject in an amount effective to stimulate proliferation of T cells in vitro or in vivo (e.g., human, murine, hamster, or macaque T cells, including CD8+ T cells and/or CD4+FoxP3+ regulatory T cells), to stimulate apoptosis of tumor cells that express TNFRSF25, to reduce tumor size, or to increase progression-free survival of a cancer patient. In some embodiments, an anti-TNFRSF25 antibody or antigen binding fragment can be administered at a dosage of about 0.1 mg/kg to about 10 mg/kg (e.g., about 0.1 mg/kg to about 1 mg/kg, about 1 mg/kg to about 5 mg/kg, or about 5 mg/kg to about 10 mg/kg), and can be administered once every one to three weeks (e.g., every week, every 10 days, every two weeks, or every three weeks).
In some cases, a composition containing an anti-TNFRSF25 antibody or antigen binding fragment as described herein can be administered to a subject in an amount effective to increase proliferation of T cells (e.g., by at least about 10 percent, about 20 percent, about 25 percent, about 50 percent, about 60 percent, about 70 percent, about 75 percent, about 80 percent, about 90 percent, about 100 percent, or more than 100 percent), as compared to the “baseline” level of T cell proliferation in the subject prior to administration of the composition, or as compared to the level of T cell proliferation in a control subject or population of subjects to whom the composition was not administered. The T cells can be, for example, CD4+FoxP3+ T cells, regulatory T cells. Any suitable method can be used to determine whether or not the level of T cell proliferation is increased in the subject. Such methods can include, without limitation, flow cytometry analysis of antigen specific T cells (e.g., flow cytometry analysis of the proportion of antigen specific CD4+FoxP3+ T cells as a fraction of the total CD4+ T cell pool), analysis of cell proliferation markers (e.g., expression of Ki67) in CD4+ T cells, increased counts of CD4+ T cells, or increased proportions of individual TCR sequences of a particular clone of CD4+ T cells.
Any aspect or embodiment described herein can be combined with any other aspect or embodiment as disclosed herein.
In order that the invention disclosed herein may be more efficiently understood, examples are provided below. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting the invention in any manner.
Streptozotocin (STZ, 2-deoxy-2-(3-(methyl-3-nitrosoureido)-D-glucopyranose)) is a naturally occurring alkylating antineoplastic agent that selectively destroys insulin-producing beta cells of the pancreas in mammals. STZ is used to induce experimental diabetes in rodents. As shown in
In the study of
Results shown in
In the present study, the effect of 4C12 on regulatory T cells (Tregs) expansion and the role of 4C12 in islet transplantation in vivo were assessed, using a murine model.
Administration of 4C12 Leads to Pronounced Treg Expansion
Groups were compared using paired t tests to assess within group differences and unpaired t tests to assess between group differences.
In the present study, flow cytometry analysis was conducted at different time points to characterize the kinetics of the Treg response to 4C12 administration. Thus,
Administration of 4C12 Prolongs Graft Survival
Treg Expansion is Correlated with Graft Survival
Further, in the present study, it was shown that Treg expansion is correlated with graft survival, as was shown by experiments in which the fold increase in Tregs (percentage at day 4/day 0) was correlated with graft survival. A significant correlation of Treg expansion with the fold increase in CD4+FOXP3+ and CD4+CD25+FOXP3+ cells was observed.
Administration of 4C12 Leads to Increased Serum Levels of IL-5
Serum levels of several Treg-related cytokines and biomarkers were measured.
Systemic Treg Expansion is not Correlated with Intra-Graft Treg Infiltration
An acute graft rejection study was conducted in which an islet graft was obtained from nephrectomy of mice at day 7 post-transplantation. Immunohistochemistry analysis of the islet graft was then performed.
Tregs Numbers in the Control and 4C12 Groups
The results of these experiments show that administration of the 4C12 antibody results in an increase in production and expansion of T regulatory cells, which delays allograft rejection and leads to an improvement in allograft survival.
In the present study, the effect of mPTX-35 on regulatory T cells expansion and the role of mPTX-35 in islet transplantation in vivo were assessed, using a murine model.
Administration of mPTX-35 Leads to Increased Treg Expansion
In this study, the effect of mPTX-35 on Treg expansion was assessed. In the experiments, mPTX-35 was administered four days before transplantation, concomitantly with STZ. This led to a significant expansion of the Treg populations, including both CD4+FoxP3+ populations and CD4+CD25+FoxP3+(double positive populations). Thus,
Further, in the present study, a flow cytometry analysis was conducted at different time points to characterize the kinetics of the Treg response.
Interestingly, as illustrated in
Administration of mPTX-35 Prolongs Graft Survival
The results of these experiments show that administration of the mPTX-35 antibody results in an increase in production and expansion of T regulatory cells that delays graft rejection and leads to an improvement in allograft survival. Also, the effect of mPTX-35 antibody on expansion of Tregs and delaying graft rejection and increasing graft survival and, surprisingly, is substantially the same as the effect of the 4C12 antibody on expansion of Tregs and delaying graft rejection and increasing graft survival.
It is to be understood that while the disclosure has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the disclosure, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.
All patents and publications referenced herein are hereby incorporated by reference in their entireties. The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. As used herein, all headings are simply for organization and are not intended to limit the disclosure in anyway.
This application claims the priority and benefit of U.S. Provisional Patent Application No. 62/906,438, filed Sep. 26, 2019, the disclosure of which is hereby incorporated by reference in its entirety.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US20/53085 | 9/28/2020 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 62906438 | Sep 2019 | US |
