Topical treatment of immune checkpoint inhibitor-induced diarrhea, colitis, or enterocolitis using antibodies and their fragments

Abstract

The present invention relates to local therapeutic use of a composition comprising an antibody molecule, a functional fragment or a derivative specifically directed against tumor necrosis factor alpha (TNFα) for treating or preventing adverse events induced by immune checkpoint (ICP) inhibitors.

Description

Field of the Invention

[0001] The present invention relates to the therapeutic use of compositions comprising antibody molecules, functional fragments and derivatives specific for tumor necrosis factor alpha (TNFα) in the local treatment of at least one gastrointestinal adverse event induced by cancer therapy in a patient, preferably induced by one or more immune checkpoint (ICP) inhibitors. In particular, the present invention relates to the therapeutic use of such compositions in the local treatment of diarrhea, colitis and / or enterocolitis induced by immune checkpoint (ICP) inhibitors, especially in patients who have not stopped taking ICP inhibitors as cancer treatment; and the use of such compositions as at least partial prevention or treatment of diarrhea, colitis and / or enterocolitis caused by ICP inhibitors in patients receiving ICP inhibitor treatment. Background Art

[0002] In recent years, the use of inhibitors that target so-called immune checkpoints (ICPs) and thereby activate the immune system against cancer cells has emerged as a promising new cancer treatment strategy.

[0003] ICP inhibitors are immunostimulatory agents that do not block existing immune responses or the initiation of immune responses, and have been shown to be very effective in treating certain types of cancer. In recent years, the ICP members that have received the most attention in ICP inhibitor research are programmed death-1 (PD-1), programmed death-ligand 1 (PD-L1), and cytotoxic T lymphocyte-associated protein 4 (CTLA-4). For example, PD-1, PD-L1, and CTLA-4 inhibitors have shown significant potential in improving the overall survival of patients with malignant melanoma, malignant non-small cell lung cancer, head and neck squamous cell carcinoma, and renal cell carcinoma (Wang et al., Inflamm Bowel Dis, 24(8):1695–1705, July 2018). In addition, ICP inhibitors have also shown encouraging results in phase I and II clinical trials in patients with malignant lymphoma, and the PD-1 inhibitor nivolumab was approved by the US Food and Drug Administration in May 2016 for the treatment of relapsed or refractory classical Hodgkin's lymphoma (Hude et al., Haematologica, 2017 Jan, 102(1):30–42).

[0004] CTLA-4, PD-1, and its ligands are members of the B7-CD28 family of signaling molecules that play an important role in all stages of T cell function and other cellular functions. The PD-1 receptor is expressed on the surface of activated T cells (and B cells) and normally binds to its ligands (PD-L1 and PD-L2) expressed on the surface of antigen-presenting cells (such as dendritic cells or macrophages). This interaction sends a signal to the T cell, essentially shutting it down / inhibiting it.

[0005] Currently available ICP inhibitors target pathways that inhibit cytotoxic T cell activation, proliferation, and function. These include antibodies specific for CTLA-4, PD-1, and PD-L1. For example, monoclonal antibodies targeting PD-1 or PD-L1 can block this binding and increase the immune response against cancer cells. These inhibitors have shown great promise in treating a variety of cancer types.

[0006] Although ICP inhibitors enhance the patient's natural adaptive immune system to improve tumor control, nonspecific immune activation often leads to the development of immune-related adverse events that affect various organ systems, especially the gastrointestinal (GI) tract, such as ICP inhibitor-induced colitis, enteritis, and / or diarrhea. In fact, results from clinical trials show that the most common severe or life-threatening (grade 3 and 4) immune-related adverse events occur in the GI tract (Wang et al., Inflamm Bowel Dis, 24(8):1695–1705, July 2018; Wang DY et al., Fatal Toxic Effects Associated With Immune Checkpoint Inhibitors. A Systematic Review and Meta-analysis. JAMA Oncol. Published online on September 13, 2018. doi:10.1001 / jamaoncol.2018.3923).

[0007] For example, immune-related adverse events involving the GI tract, such as ICP-induced colitis, have been reported in approximately 21% to 44% of patients treated with CTL-4 inhibitors and with a slightly lower frequency in patients treated with PD-1 / PD-L1 inhibitors (Wang et al., Inflamm Bowel Dis, 24(8):1695–1705, July 2018).

[0008] Colitis is a condition characterized by inflammation of the large intestine / colon. Enteritis is defined as a condition characterized by inflammation of the small intestine. Enterocolitis is a condition characterized by inflammation of both the small intestine and the large intestine / colon, i.e., a combination of enteritis and colitis. Symptoms that characterize ICP inhibitor-induced colitis include diarrhea, abdominal pain, nausea, cramping, bloody or sticky stools or changes in bowel habits, fever, abdominal distension, hard stools, and constipation (Brahmer et al., Management of Immune-Related Adverse Events in Patients Treated With Immune Checkpoint Inhibitor Therapy: American Society of Clinical Oncology (ASCO) Practice Guideline , J Clin Oncol. 2018 Jun 10; 36(17): 1714-1768). Symptoms characterizing ICP inhibitor-induced enterocolitis include those listed above for ICP inhibitor-induced colitis.

[0009] As described in the ASCO practice guideline for the management of immune-related adverse events in patients treated with immune checkpoint inhibitor therapy, the severity of ICP inhibitor-induced colitis can be clinically categorized into 4 grades based on the National Cancer Institute's CTCAE for diarrhea, 4th edition. Level 1 The clinical features of this phase 2 trial were diarrhea with <4 stools per day over baseline and a small increase in ostomy output compared to baseline. Level 2 The disease was characterized by diarrhea with 4-6 stools per day over baseline and a moderate increase in ostomy output compared to baseline. Level 3 The clinical features of diarrhoea are ≥7 stools per day over baseline; incontinence; a significant increase in stool bulk; and a significant increase in ostomy production compared with baseline; and require hospitalisation. Level 4 Grade 3 is characterized by diarrhea with >10 bowel movements per day above baseline; symptoms are identical to those of Grade 3, but with life-threatening consequences (such as perforation, intestinal obstruction, and / or fever); and hospitalization is usually required (Brahmer et al., Management of Immune-Related Adverse Events in Patients Treated With Immune Checkpoint Inhibitor Therapy: American Society of Clinical Oncology Practice Guideline, J Clin Oncol. 2018 Jun 10; 36(17): 1714-1768). Equivalent grading may be applied to patients suffering from ICP inhibitor-induced colitis or enterocolitis.

[0010] Treatment of ICP inhibitor-induced colitis and enteritis is typically based on symptom severity and involves the use of systemic corticosteroids to immunosuppress moderate to severe symptoms and, in steroid-refractory cases, systemic biologics (eg, infliximab).

[0011] ASCO guidelines recommend administering corticosteroids for grade 2 toxicity associated with ICP inhibitor-induced colitis, unless the diarrhea is transient, and starting with an initial dose of 1 mg / kg / day prednisone or equivalent. During corticosteroid therapy, ICP inhibitor therapy should be discontinued. When symptoms improve to grade 1 or lower, corticosteroids should be tapered over at least 4 to 6 weeks before resuming ICP inhibitor therapy.

[0012] For grade 3 toxicity, the guidelines recommend the administration of corticosteroids, starting with an initial dose of 1 to 2 mg / kg / day of prednisone or equivalent. If the patient recovers to grade 1 or lower, permanent discontinuation of the CTLA-4 agent should be considered, but PD-1 and PD-L1 agents can be restarted. If symptoms persist for more than 3 to 5 days or recur after improvement, intravenous corticosteroids or non-corticosteroid immunosuppressants (e.g., infliximab) should be considered.

[0013] For grade 4 toxicity, the guidelines recommend intravenous administration of corticosteroids at a dose of 1 to 2 mg / kg / day (methyl) prednisone or equivalent until symptoms improve to grade 1, and then gradually reduce corticosteroids over 4 to 6 weeks. ICP inhibitors should be permanently discontinued. If symptoms are refractory within 2 to 3 days, 5-10 mg / kg infliximab (e.g., 5 mg / kg infliximab every 2 weeks) should be considered. Guidelines for the clinical management of ICP inhibitor toxicity are also provided in Hryniewicki et al., J Emerg Med. 2018 Oct; 55(4): 489-502.

[0014] Therefore, persistent grade 2 ICP inhibitor-induced colitis, and almost any grade 3 / 4 ICP inhibitor-induced colitis, will generally require management with systemic corticosteroids. The use of prophylactic steroids (e.g., oral budesonide) has not been shown to be effective in treating the development of ICP inhibitor-induced diarrhea or colitis. In patients who are refractory to systemic corticosteroids, systemic treatment with non-corticosteroids such as the anti-TNFα specific antibody infliximab is recommended. According to the ASCO guidelines, enteritis can generally be managed with a similar approach as for colitis, including corticosteroids and / or infliximab.

[0015] Corticosteroids have several side effects, especially with prolonged use and at high doses. In particular, patients treated with corticosteroids often have an increased susceptibility to infection. Moreover, in addition to higher infection rates, systemic corticosteroid use also leads to other potential side effects, including osteoporosis; bone fractures; osteonecrosis; increased cardiovascular risk; gastritis; peptic ulcer disease; worsening diabetes; nervousness or restlessness; high blood pressure; insomnia; water retention and swelling; cataracts or glaucoma; muscle weakness; sudden mood swings; easy bruising; and weight gain. Therefore, long-term systemic treatment with corticosteroids should be avoided.

[0016] In addition, systemic treatment with higher doses of corticosteroids (e.g., 1-2 mg / kg / day prednisone or equivalent) requires interruption of treatment with ICP inhibitors. Such interruptions can destroy the anticancer effects of these ICP inhibitors. Therefore, preferably, such interruptions should be avoided.

[0017] Nonsteroidal agents with immunosuppressive effects (e.g., anti-TNFα antibodies) do not have as many adverse side effects as systemic treatment with corticosteroids, although anti-TNFα antibodies (such as infliximab) are also commonly administered systemically. In addition, it has been reported that patients with ICP inhibitor-induced colitis who received systemic corticosteroids for a longer duration had a numerically higher infection rate than those who received systemic anti-TNFα antibodies. Therefore, earlier nonsteroidal immunosuppressive therapy may ensure a more beneficial overall outcome (Wang et al., J ImmunoTher Cancer (2018) 6:37).

[0018] Currently, standard therapy for severe ICP inhibitor-induced colitis or enterocolitis using TNFα-specific antibodies involves regular systemic administration of anti-TNFα antibodies via intravenous infusion. However, the use of systemic infusions of anti-TNFα antibodies in cancer patients also requires interruption of any ICP inhibitor therapy, thereby undermining successful cancer treatment with ICP inhibitors.

[0019] Furthermore, intravenous administration can produce complications, including acute infusion reactions, anaphylaxis, and anaphylactic shock. Furthermore, systemic administration of anti-TNFα antibodies carries a variety of risks associated with systemic suppression of the patient's immune defenses against TNFα, including, for example, infectious complications. Furthermore, the immunogenicity of systemically administered antibodies may lead to their neutralization, resulting in loss of the stimulated response. Finally, melanoma and Merkel cell carcinoma have been reported in patients treated with TNF blocker therapy, including infliximab.

[0020] Therefore, currently, treatment of ICP inhibitor-induced adverse events with moderate to severe symptoms in the GI tract (such as ICP inhibitor-induced colitis, enterocolitis and / or diarrhea) requires interruption of ICP inhibitor therapy and usually systemic administration of steroid immunosuppressants. This is detrimental to the effectiveness of cancer therapy and produces undesirable side effects systemically and, in the case of melanoma, recurrence of the underlying disease.

[0021] There is a need for alternative treatments for adverse events of the GI tract caused by ICP inhibitors (e.g., ICP inhibitor-induced diarrhea, colitis, or enterocolitis) that allow specific targeting of inflamed tissues in the GI tract, particularly in the ileum and large intestine, without the need to interrupt treatment with the ICP inhibitor. The treatment should particularly enable effective targeting of tissues in the GI tract affected by inhibitor-induced adverse events, thereby eliminating the need for systemic use. In addition, there is a need for prophylactic treatments to prevent or delay the onset of adverse effects affecting the GI tract caused by treatment with ICP inhibitors; early treatment is particularly needed to prevent serious complications such as intestinal obstruction and perforation and the need for urgent endoscopy, hospitalization, and surgery. Summary of the Invention

[0022] Surprisingly, the present inventors have discovered that by locally treating immune-related adverse events induced by ICP inhibitors in the GI tract, the need for systemic immunosuppressive therapy is eliminated, which overcomes the need to discontinue ICP inhibitor treatment by allowing concomitant local treatment in the GI tract with immunosuppressive TNFα-specific antibodies or functional fragments thereof and systemic treatment with ICP inhibitors, thereby avoiding the need to disrupt successful cancer treatment with ICP inhibitors. Despite the fact that ICP inhibitors have been known for more than 15 years for the treatment of cancer, it has never been demonstrated that TNFα-specific antibodies can be administered topically to treat or prevent immune-related adverse effects of ICP inhibitor therapy.

[0023] The present invention provides a composition for the local treatment of ICP inhibitor-induced diarrhea, colitis, or enterocolitis in the ileum and / or large intestine. In addition, the present invention provides a composition for the local treatment of diarrhea, colitis, or enterocolitis in the ileum and / or large intestine as a preventive therapy for ICP inhibitor-induced diarrhea, colitis, or enterocolitis. In particular, the present invention provides a composition for the early treatment of ICP inhibitor-induced diarrhea to prevent more serious adverse events affecting the gastrointestinal tract, such as colitis or enteritis.

[0024] The present invention therefore relates to the subject matter defined in items 1 to 69 below:

[0025] [1] A pharmaceutical composition comprising an active agent selected from the group consisting of antibodies specific for tumor necrosis factor alpha (TNFα) and functional fragments and derivatives thereof, for treating or preventing at least one gastrointestinal adverse event induced by cancer therapy in a patient, in particular induced by one or more immune checkpoint (ICP) inhibitors, wherein the treatment or prevention comprises administering the composition locally to the affected part of the gastrointestinal tract of the patient, for example, to the ileum and / or large intestine of the patient;

[0026] or

[0027] A pharmaceutical composition comprising an active agent selected from the group consisting of antibodies specific for tumor necrosis factor alpha (TNFα) and functional fragments and derivatives thereof, for treating or preventing at least one gastrointestinal adverse event induced by cancer therapy in a patient, preferably induced by one or more immune checkpoint (ICP) inhibitors, wherein the treatment or prevention comprises orally administering the composition to the patient.

[0028] [2] A pharmaceutical composition comprising an active agent selected from the group consisting of antibodies specific for tumor necrosis factor alpha (TNFα) and functional fragments and derivatives thereof, for preventing the progression or worsening of at least one gastrointestinal adverse event induced by cancer therapy in a patient, preferably induced by one or more immune checkpoint (ICP) inhibitors, wherein the prevention comprises topically administering the composition to an affected area of ​​the gastrointestinal tract of the patient, for example, to the ileum and / or large intestine of the patient; or

[0029] A pharmaceutical composition comprising an active agent selected from the group consisting of antibodies specific for tumor necrosis factor alpha (TNFα) and functional fragments and derivatives thereof, for preventing the progression or worsening of at least one gastrointestinal adverse event induced by cancer therapy in a patient, preferably induced by one or more immune checkpoint (ICP) inhibitors, wherein the prevention comprises orally administering the composition to the patient.

[0030] [3] The composition according to item 1 or item 2, which allows uninterrupted treatment, preferably systemic treatment, with one or more ICP inhibitors.

[0031] [4] The composition according to any one of items 1 to 3, wherein the patient is receiving treatment with one or more ICP inhibitors, preferably systemic treatment.

[0032] [5] The composition according to item 4, wherein the interruption of treatment with the one or more ICP inhibitors is less than 4 weeks, preferably less than 2 weeks, more preferably 1 week, even more preferably less than 5, 4, 3, 2 days or 1 day, and most preferably wherein there is no interruption of treatment.

[0033] [6] The composition according to any of the preceding items, wherein the patient suffers from, or is at risk of developing, ICP inhibitor-induced colitis.

[0034] [7] The composition according to item 6, wherein the ICP inhibitor-induced colitis is characterized by grade 1 toxicity, and optionally wherein the treatment or prevention prevents progression to higher grade toxicity.

[0035] [8] The composition of claim 6, wherein the ICP inhibitor-induced colitis is characterized by grade 2 toxicity, and optionally wherein the treatment or prevention prevents progression to higher grade toxicity.

[0036] [9] The composition of claim 6, wherein the ICP inhibitor-induced colitis is characterized by grade 3 toxicity, and optionally wherein the treatment or prevention prevents progression to higher grade toxicity.

[0037]

[10] The composition according to claim 6, wherein the ICP inhibitor-induced colitis is characterized by grade 4 toxicity.

[0038]

[11] The composition according to any one of items 7 to 10, wherein the treatment or prevention prevents discontinuation of treatment with one or more ICP inhibitors.

[0039]

[12] The composition according to any of the preceding items, wherein the patient suffers from, or is at risk of developing, ICP inhibitor-induced enterocolitis.

[0040]

[13] The composition of claim 12, wherein the ICP inhibitor-induced enterocolitis is characterized by grade 1 toxicity, and optionally wherein the treatment or prevention prevents progression to higher grade toxicity.

[0041]

[14] The composition of claim 12, wherein the ICP inhibitor-induced enterocolitis is characterized by grade 2 toxicity, and optionally wherein the treatment or prevention prevents progression to higher grade toxicity.

[0042]

[15] The composition of claim 12, wherein the ICP inhibitor-induced enterocolitis is characterized by grade 3 toxicity, and optionally wherein the treatment or prevention prevents progression to higher grade toxicity.

[0043]

[16] The composition according to claim 12, wherein the ICP inhibitor-induced enterocolitis is characterized by grade 4 toxicity.

[0044]

[17] The composition according to any one of items 13 to 16, wherein the treatment or prevention prevents discontinuation of treatment with one or more ICP inhibitors.

[0045]

[18] The composition according to any of the preceding items, wherein the patient suffers from, or is at risk of developing, ICP inhibitor-induced diarrhea.

[0046]

[19] The composition according to item 18, wherein the ICP inhibitor-induced diarrhea is characterized by grade 1 toxicity, and optionally wherein the treatment or prevention prevents progression to higher grade toxicity.

[0047]

[20] The composition according to item 18, wherein the ICP inhibitor-induced diarrhea is characterized by grade 2 toxicity, and optionally wherein the treatment or prevention prevents progression to higher grade toxicity.

[0048]

[21] The composition according to item 18, wherein the ICP inhibitor-induced diarrhea is characterized by grade 3 toxicity, and optionally wherein the treatment or prevention prevents progression to higher grade toxicity.

[0049]

[22] The composition according to item 12, wherein the ICP inhibitor-induced diarrhea is characterized by grade 4 toxicity.

[0050]

[23] The composition according to any one of items 19 to 22, wherein the treatment or prevention prevents discontinuation of treatment with one or more ICP inhibitors.

[0051]

[24] A composition according to any of the preceding items, wherein the composition is used as a first-line treatment for a patient suffering from ICP inhibitor-induced diarrhea, colitis or enterocolitis; and / or wherein the patient is not symptomatically treated with corticosteroids; and / or wherein the patient has not been treated with corticosteroids.

[0052]

[25] The composition according to any one of the preceding items, wherein the composition is used as a second-line treatment for patients with steroid-refractory ICP inhibitor-induced diarrhea, colitis or enterocolitis.

[0053]

[26] A pharmaceutical composition comprising an active agent selected from the group consisting of antibodies specific for tumor necrosis factor alpha (TNFα) and functional fragments and derivatives thereof, for local treatment in the ileum and / or large intestine of a patient as a preventive therapy for ICP inhibitor-induced diarrhea, colitis or enterocolitis.

[0054]

[27] The composition according to item 26, wherein the local treatment prevents the occurrence of grade 3 toxicity, preferably grade 2 toxicity, more preferably grade 1 toxicity, most preferably any symptoms associated with ICP inhibitor-induced adverse events in the patient, preferably ICP inhibitor-induced diarrhea, colitis or enterocolitis in the ileum or colon of the patient.

[0055]

[28] The composition according to item 27, which allows for simultaneous treatment, preferably systemic treatment, with one or more ICP inhibitors.

[0056]

[29] The composition according to items 26 to 28, wherein the patient is concurrently receiving systemic treatment with one or more ICP inhibitors.

[0057]

[30] The composition according to items 26 to 29, wherein the composition is used to prevent or minimize and / or alleviate symptoms associated with ICP inhibitor-induced diarrhea, colitis or enterocolitis.

[0058]

[31] A composition according to any of the preceding items, wherein the ICP inhibitor is capable of targeting an immune checkpoint selected from the group consisting of CTLA-4, PD-1, PD-L1, PDL2, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, CSF-1R, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, ADAR1, CD47, ICOS, TIGIT, and B-7 family ligands.

[0059]

[32] A composition according to any of the preceding items, wherein the one or more ICP inhibitors are selected from the following: an antibody specific for cytotoxic T lymphocyte-associated protein 4 (CTLA-4), an antibody specific for programmed cell death protein 1 (PD-1) and an antibody specific for programmed death ligand 1 (PD-L1).

[0060]

[33] The composition according to item 32, wherein the antibody specific for PD-1 is selected from the following: pembrolizumab and nivolumab; the antibody specific for PD-L1 is selected from the following: atezolizumab, avelumab and durvalumab; or the antibody specific for CTLA-4 is selected from the following: ipilimumab and tremelimumab.

[0061]

[34] The composition according to item 32 or item 33, wherein the one or more ICP inhibitors are a combination of an antibody specific for PD-1 and / or PD-L1 and an antibody specific for CTLA-4, preferably a combination of an antibody specific for PD-1 or PD-L1 and an antibody specific for CTLA-4.

[0062]

[35] The composition according to any of the preceding items, wherein the one or more ICP inhibitors modulate the immune response in the patient.

[0063]

[36] The composition according to any of the preceding items, wherein the one or more ICP inhibitors inhibit tumor cell growth in the patient.

[0064]

[37] The composition according to any of the preceding items, wherein the patient suffers from an infectious disease.

[0065]

[38] The composition according to any of the preceding items, wherein the patient is a cancer patient.

[0066]

[39] The composition according to item 38, wherein the cancer is selected from the group consisting of melanoma; lymphoma, such as classical Hodgkin's lymphoma; glioma; urothelial carcinoma; Merkel cell carcinoma; renal cancer; head and neck squamous cell carcinoma, prostate cancer; breast cancer; colon cancer; and lung cancer.

[0067]

[40] The composition according to item 39, wherein the cancer is selected from the group consisting of bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, anal region cancer, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia. , acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors of childhood, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal pelvic cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal tumors, brain stem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, environmentally induced cancers (including those induced by asbestos), and combinations of said cancers.

[0068]

[41] A composition according to any of the preceding items, wherein the composition provides a therapeutically effective dose of the TNFα-specific antibody or a functional fragment or derivative thereof in the lumen of the patient's ileum or large intestine to prevent, minimize, delay or alleviate symptoms associated with ICP inhibitor-induced diarrhea, colitis or enterocolitis.

[0069]

[42] A composition according to any of the preceding items, wherein the functional antibody fragment or derivative specific for TNFα is a Fab fragment, a F(ab')2 fragment, a Fab' fragment, a scFv, a dsFv, a diabody, a trimer, a tetrabody, an Fc fusion protein, a camel antibody, a VHH, a vorabody, a VNAR or a minibody.

[0070]

[43] The composition according to any one of the preceding items, wherein the antibody specific for TNFα and its functional fragments and derivatives are selected from the following: infliximab, adalimumab, etanercept, certolizumab pegol, golimumab and its functional fragments and derivatives; from anti-TNFα antibodies and their functional fragments and derivatives, which have a light chain variable domain and / or a heavy chain variable domain comprising a complementarity determining region (CDR) according to the originally filed patent application having an amino acid sequence as disclosed in claim 2 of WO 2017 / 158092, claim 2 of WO 2017 / 158097, claim 2 of WO 2017 / 158079 and / or claim 2 of WO 2017 / 158084; from anti-TNFα antibodies and their functional fragments and derivatives, which have a light chain variable domain and / or a heavy chain variable domain comprising a complementarity determining region (CDR) according to the originally filed patent application having an amino acid sequence as disclosed in claim 4 of WO 2017 / 158079, claim 5 of WO 2017 / 158084, claim 6 of WO 2017 / 158085, claim 7 of WO 2017 / 158097, claim 8 of WO 2017 / 158099 and / or claim 9 of WO 2017 / 158084. The heavy chain variable domain amino acid sequence and / or light chain variable domain amino acid sequence of claim 5 and claim 6 of WO 2017 / 158097 and claim 4 of WO 2017 / 158084; and combinations thereof.

[0071]

[44] The composition according to item 43, wherein the anti-TNFα antibody is infliximab, adalimumab, etanercept, certolizumab pegol or golimumab.

[0072]

[45] The composition according to item 43, wherein the anti-TNFα antibody is infliximab.

[0073]

[46] The composition according to item 43, wherein the TNFα antibody is adalimumab.

[0074]

[47] The composition according to any one of items 1 to 43, wherein the functional fragment or derivative is a functional fragment or derivative of fliximab, adalimumab, etanercept, certolizumab pegol or golimumab.

[0075]

[48] ​​The composition according to any one of the preceding items, wherein the amino acid sequence of the antibody specific for TNFα or its functional fragment or derivative comprises

[0076] (i) amino acids 233P, 234V, 235A, and a deletion at amino acid position 236; and optionally amino acid 434A or amino acids 252Y, 254T, and 256E; and optionally amino acids 239D, 330L, and 332E or amino acids 326A, 332E, and 333A; and / or

[0077] (ii) amino acids 380A and 434A, and optionally amino acid 307T; and / or

[0078] (iii) amino acid 434W, and optionally amino acid 428E and / or amino acid 311R,

[0079] The amino acid numbers refer to EU numbers.

[0080]

[49] A composition according to any one of items 1 to 43 and 48, wherein the active agent is an antibody comprising: V L A domain comprising a CDR1 region having the amino acid sequence as shown in SEQ ID NO: 3, a CDR2 region having the amino acid sequence as shown in SEQ ID NO: 4, and a CDR3 region having the amino acid sequence as shown in SEQ ID NO: 5; and V H A domain comprising a CDR1 region having the amino acid sequence shown in SEQ ID NO: 6, a CDR2 region having the amino acid sequence shown in SEQ ID NO: 7, and a CDR3 region having the amino acid sequence shown in SEQ ID NO: 8.

[0081]

[50] The composition according to any one of items 1 to 43 and 48, wherein the active agent is an antibody comprising a V having an amino acid sequence as shown in SEQ ID NO: 10. L domain and a V domain having an amino acid sequence as shown in SEQ ID NO: 9 H domain.

[0082]

[51] A composition according to any one of items 1 to 43, 49 and 50, wherein the active agent is an antibody comprising an Fc region comprising amino acids 236 to 451 of the amino acid sequence shown in SEQ ID NO: 11.

[0083]

[52] A composition according to any one of items 1 to 43, wherein the active agent is an antibody comprising a light chain having the amino acid sequence shown in SEQ ID NO: 1 and a heavy chain having the amino acid sequence shown in SEQ ID NO: 11, 14, 15 or 16.

[0084]

[53] The composition according to any of the preceding items, wherein the treatment comprises oral administration of the composition.

[0085]

[54] The composition according to item 53, wherein the composition is a solid dosage form, which is in the form of a pill, granule, microparticle, nanoparticle, microtablet, sphere, capsule, tablet or multiparticulate drug delivery system, which is coated with a delayed-release coating that prevents the release of the active agent before entering the ileum, ileocolic region or large intestine of the gastrointestinal (GI) tract.

[0086]

[55] The composition according to item 54, wherein the delayed-release coating comprises at least one component selected from the group consisting of a coating material that disintegrates pH-dependently, a coating material that disintegrates time-dependently, a coating material that disintegrates due to enzymatic triggering in the intestinal environment, and combinations thereof.

[0087]

[56] The composition according to item 53 or item 54, wherein

[0088] - The pH-dependent disintegrating coating material is selected from the following: polyvinyl acetate phthalate; cellulose acetate trimellitate; hydroxypropyl methylcellulose phthalate HP-50, HP-55 or HP-55S; cellulose acetate phthalate; hydroxypropyl methylcellulose acetate succinate (HPMCAS); poly (methacrylic acid-ethyl acrylate) 1:1 ( L100-55, L30D-55), poly (methacrylic acid-methyl methacrylate) 1:1 ( L-100, L12.5), poly (methacrylic acid-methyl methacrylate) 1:2 ( S-100, S12,5, FS30D) and combinations thereof;

[0089] - the time-dependently disintegrating coating material is selected from the group consisting of poly(ethyl acrylate-methyl methacrylate) 2:1 (e.g. NM 30D or Eudragit NE 30D); poly(ethyl acrylate-methyl methacrylate-methacrylic acid) 7:3:1 (e.g., RS 30D); ethyl cellulose (e.g., or Aquacoat ECD); poly (ethyl acrylate-methyl methacrylate-methacryloyloxyethyl trimethyl ammonium chloride) 1:2:0.1 (e.g., RS 30D); polyvinyl acetate (e.g. SR 30D); and combinations thereof; and

[0090] - A coating material that disintegrates due to an enzymatic trigger in the intestinal environment selected from the group consisting of hemicellulose, chondroitin sulfate; cyclodextrin; pectin; guar gum; chitosan; inulin; lactulose; raffinose; stachyose; alginate; dextran; xanthan gum; locust bean gum; arabinogalactan; amylose; pullulan; carrageenan; scleroglucan; chitin; curdulan; levan; pullulan; starch; resistant starch; azo compounds degraded by azo bond cleavage bacteria; and combinations thereof.

[0091]

[57] A composition according to any one of items 54 to 56, wherein the delayed-release coating comprises a combination of at least one coating material that disintegrates pH-dependently and at least one coating material that disintegrates due to an enzymatic trigger in the intestinal environment.

[0092]

[58] A composition according to any one of items 54 to 56, wherein the delayed-release coating comprises at least one component selected from the group consisting of polyvinyl acetate phthalate, cellulose acetate trimellitate, hydroxypropyl methylcellulose phthalate HP-50, HP-55 or HP-55S, cellulose acetate phthalate, hydroxypropyl methylcellulose acetate succinate (HPMCAS), poly(methacrylic acid-ethyl acrylate) 1:1, poly(methacrylic acid-methyl methacrylate) 1: 1. poly(methacrylic acid-methyl methacrylate) 1:2, chondroitin sulfate, pectin, guar gum, chitosan, inulin, lactulose, raffinose, stachyose, alginate, dextran, xanthan gum, locust bean gum, arabinogalactan, amylose, cyclodextrin, pullulan, carrageenan, scleroglucan, chitin, curdulan, levan, amylopectin, starch, resistant starch, azo compounds degraded by azo bond cleavage bacteria, and combinations thereof.

[0093]

[59] The composition according to any one of items 53 to 58, wherein, after oral administration of the composition, the antibody or functional fragment thereof begins to be released in the ileum, terminal ileum, ileocolic region, ascending colon, transverse colon or descending colon.

[0094]

[60] A composition according to any one of items 54 to 59, wherein the delayed-release coating comprises at least one pH-sensitive (enteric) polymer, preferably poly(methacrylic acid-methyl methacrylate) 1:2 (e.g. S or L), and at least one polysaccharide selected from the group consisting of hemicellulose, chondroitin sulfate, cyclodextrin, chitosan, dextran, arabinogalactan, amylose, amylose, carrageenan, scleroglucan, chitin, curcumin, levan, amylopectin, starch, resistant starch, azo compounds degraded by azo bond cleavage bacteria, and combinations thereof, preferably combinations of resistant starch.

[0095]

[61] A composition according to any one of items 54 to 59, wherein the delayed-release coating comprises one or more pH-sensitive polymers (e.g., S and / or L) as the sole polymer; or wherein the delayed-release coat consists essentially of at least one pH-sensitive polymer (e.g., S and / or L).

[0096]

[62] A composition according to any one of items 1 to 52, wherein the treatment comprises rectal administration of the composition, and / or wherein the composition is an enema, gel, foam or suppository.

[0097]

[63] The composition according to any of the preceding items, comprising at least one additive selected from the group consisting of a hydrophilic polymer, a filler, a hydrophilic binder, a disintegrant, an anti-adherent, a surfactant, a stabilizer, a protease resistance enhancer, a plasticizer, a coagulant, a lubricant, a buffer and / or an acidifier.

[0098]

[64] The composition according to any one of the above items, which is an electronic drug capsule engineered to deliver the active agent directly to the ileum and / or colon; or it is provided by genetically modified cells expressing TNFα binding protein.

[0099]

[65] The composition according to any of the above items, wherein the local treatment results in retention of the antibody or functional fragment thereof in the gastrointestinal tract wall without substantial systemic release in the rest of the body.

[0100]

[66] The composition according to any of the preceding items, wherein the treatment comprises administering the antibody or functional fragment thereof to the patient once a week, twice a week, once every three days, once every two days, once a day, twice a day or three times a day.

[0101]

[67] The composition according to any of the preceding items, wherein the local treatment is in the ileum, terminal ileum, cecum, ascending colon, transverse colon and / or descending colon of the patient.

[0102]

[68] A composition according to any of the preceding items, wherein the topical treatment of ICP inhibitor-induced diarrhea, colitis or enterocolitis is indicated by one or more symptoms selected from the group consisting of diarrhea less than 4 stools above baseline, diarrhea 4-6 stools above baseline, diarrhea greater than 6 stools above baseline, diarrhea greater than 7 stools above baseline, abdominal pain, nausea, cramping, bloody or sticky stools or change in bowel habits, fever, abdominal distension, constipation and constipation.

[0103]

[69] The composition of any of the preceding items, wherein the topical treatment prevents, minimizes and / or slows or alleviates one or more symptoms associated with ICP inhibitor-induced diarrhea, colitis or enterocolitis selected from the group consisting of diarrhea of ​​less than 4 stools above baseline, diarrhea of ​​4-6 stools above baseline, diarrhea of ​​more than 6 stools above baseline, diarrhea of ​​more than 7 stools above baseline, abdominal pain, nausea, cramping, bloody or sticky stools or change in bowel habits, fever, bloating, hard stools and constipation. BRIEF DESCRIPTION OF THE DRAWINGS

[0104] Figure 1 The percentage change in body weight relative to day 0 is shown, see Example 3. Data are presented as mean ± SE. n = 5-12 animals / group.

[0105] Figure 2 Histological and endoscopic scores of treated animals over time are shown, see Example 3. *: p < 0.05, as determined by one-way ANOVA with Dunnett's multiple comparison post hoc test comparing all groups to the vehicle control group. Data are presented as mean ± SE. n = 5-12 animals / group.

[0106] Figure 3 Histological scoring of the mice at sacrifice (d49) is described in Example 3. **: p < 0.01, ***: p < 0.005, as determined by one-way ANOVA with Dunnett's multiple comparison post hoc test for all groups compared to the vehicle control group. Data are presented as mean ± SE. n = 5-12 animals / group.

[0107] Figure 4 Cytokine concentrations in the colon of treated mice, determined on day 49, are shown, as described in Example 3. *: p < 0.05, as determined by one-way ANOVA with Dunnett's multiple comparison post hoc test comparing all groups to the vehicle control group. Data are presented as mean ± SE. n = 5-12 mice / group.

[0108] Figure 5 The effect of cV1q-huF treatment on histological scores is shown in Example 4. The total histological score is the sum of the individual scores (0-5, respectively) for inflammation, crypt damage, erosion, hyperplasia, and edema in the proximal, mid, and distal colon at day 42. *: p < 0.05, **: p < 0.01, ****: p < 0.0001, as determined by Kruskal-Wallace combined with Dunn's multiple comparison post hoc test for all groups compared to the respective vehicle control groups. Data are presented as mean ± SE. n = 6-12 animals / group. cV1q was administered intraperitoneally as a control.

[0109] Figure 6 Effects on cytokines in the colon are shown in Example 4. Cytokines in colonic tissue were measured by ELISA, and the results were normalized to mg tissue weight. *: p < 0.05, **: p < 0.01, ***: p < 0.005, ****: p < 0.001, as determined by one-way ANOVA with Dunnett's multiple comparison post hoc test comparing all groups to the vehicle control group. Data are presented as mean ± SE. n = 6-12 animals / group. cV1q was administered intraperitoneally as a control.

[0110] Figure 7 The mean plasma drug concentrations of cV1q-huFc in healthy and colitis Tg32-SCID mice following rectal administration are described (see Example 5). Plasma was collected at 1, 2, 4, 8, 24, and 48 h for single-dose (SD) animals and 24, 48, 72, 96, 120, and 144 h after the first dose for multiple-dose (MD) animals. Plasma concentrations of cV1q-huFc were determined by immuno-PCR. Due to blood collection limitations, SD and MD characteristics were based on two subgroups (SD: 4 and 24 h from subgroup 1 and 2, 8, and 48 h from subgroup 2; MD: 24, 72, and 120 h from subgroup 1 and 48, 96, and 144 h from subgroup 2). Data are presented as mean ± SD (n = 5-7).

[0111] Figure 8Median plasma concentrations of cV1q-huFc following intracecal administration to healthy and colitis mice are shown (see Example 6). Plasma samples were collected 1, 4, 8, 12, and 24 h after a single dose in Tg32-SCID mice and 120 h after the first dose in multidose animals. Plasma concentrations of cV1q-huFc were determined by immuno-PCR. Data are presented as mean ± IQR (n = 4). IC: intracecal; IR: intrarectal.

[0112] Figure 9 The median concentration of cV1q-huFc in the proximal and distal colon after intracecal or rectal administration is shown (see Example 6). Animals were sacrificed and proximal and distal colon tissue samples were collected 1, 4, 8, 12, and 24 hours after a single dose to Tg32-SCID mice and 120 hours after the first dose of multidose animals. Tissue samples were rinsed, weighed, and homogenized in 0.3 mL PBS. The cV1q-huFc concentration in the supernatant was determined using a commercially available IgG ELISA. The obtained concentration (pg / mL) was then normalized to tissue weight. Data are presented as mean ± IQR (n = 4). IC: intracecal; IR: intrarectal. *: Median values ​​are BLQs for healthy animals at 1 and 4 hours, for colitis mice treated with 300 μg of cV1q-huFc at 1, 4, and 8 hours in the proximal colon, and for colitis animals treated with 100 μg of the agent at 12 hours in the distal colon.

[0113] Figure 10 The effects of Ab-REW and infliximab on IFNγ production after nivolumab treatment are shown (see Example 7). CD14+ monocytes were isolated from PBMCs and differentiated into monocyte-derived dendritic cells (mo-DCs) over 7 days in cell culture medium supplemented with GM-CSF and IL-4. CD4+ T cells were also isolated from PBMCs. CD4+ T cells were combined with mo-DCs at a T-cell:DC ratio of 5:1 and incubated with nivolumab (10 μg / mL) and serial dilutions of Ab-REW or infliximab (50–0.003 μg / mL) for 5 days. IFNγ concentrations in cell culture supernatants were then determined by ELISA. Data are presented as mean ± SE, with n = 3 technical replicates. 4-PL nonlinear regression was performed where possible. The lower dashed line represents the mean IFNγ secretion from untreated cells. The upper dashed line represents the mean IFNγ secretion in the presence of nivolumab alone (n=6 for all control groups). DETAILED DESCRIPTION

[0114] The present invention is described below with respect to antibodies specific for tumor necrosis factor alpha (TNFα) and its functional fragments and derivatives, which represent the most preferred embodiments. All embodiments described below are also applicable to antibodies and their functional fragments and derivatives directed against other targets (antigens), with necessary modifications in detail. The targets of these antibodies and their functional fragments include, but are not limited to, anti-inflammatory cytokines (such as IL-13) and their receptors; pro-inflammatory cytokines (such as CD20, IL-6, IL-12 and IL-23 (IL-12 / IL-23p40, IL-23p19), IL-17, IL-21) and their receptors; cell adhesion molecules (such as MadCAM-1, ICAM-1); CC chemokine receptors (such as CCR5, CCR9) and their ligands; integrins (such as α4β7, β7, α2β1, αEβ7); toll-like receptors (such as TLR2, TLR9); eotaxin (such as eotaxin-1); tumor necrosis factor receptor superfamily members (such as OX40); matrix metalloproteinases (such as MMP-9); CXC motif chemokines (such as IP-10); and other proteins (such as CD20). Preferred targets are α4β7 integrin and IL-23.

[0115] According to a first aspect of the present invention, the present invention relates to a pharmaceutical composition comprising an active agent selected from the group consisting of antibodies specific for tumor necrosis factor alpha (TNFα) and functional fragments and derivatives thereof, for treating or preventing at least one gastrointestinal adverse event induced by cancer therapy in a patient, preferably induced by one or more immune checkpoint (ICP) inhibitors, wherein the treatment or prevention comprises local administration of the composition to the ileum and / or large intestine of the patient.

[0116] The present invention also relates to a pharmaceutical composition comprising an active agent selected from the following: antibodies specific for tumor necrosis factor alpha (TNFα) and functional fragments and derivatives thereof, which are used for local treatment in the ileum and / or large intestine of patients with immune checkpoint (ICP) inhibitor-induced colitis or enterocolitis or diarrhea. The composition of the present invention allows for uninterrupted systemic treatment with one or more ICP inhibitors. This has the advantage of not undermining successful cancer treatment using ICP inhibitors. In addition, local treatment using antibodies specific for TNFα and functional fragments and derivatives thereof means that there is no need for the use of corticosteroids and systemically administered antibodies specific for TNFα and functional fragments and derivatives thereof, thereby minimizing the undesirable side effects of such treatments.

[0117] In the context of the present invention, the term "antibody" is used synonymously with "immunoglobulin" (Ig), which is defined as a protein belonging to the class IgG, IgM, IgE, IgA or IgD (or any subclass thereof), and includes all conventionally known antibodies and functional fragments and derivatives thereof. In the context of the present invention, a "functional fragment or derivative" of an antibody / immunoglobulin is defined as an antigen-binding fragment or other derivative of a parent antibody that substantially retains the antigen-binding properties of such parent antibody. In a preferred embodiment, the functional fragment or derivative of an antibody comprises a functional Fc portion that at least exhibits effector function. In one embodiment, the antibody is capable of binding to human FcRn.

[0118] The "antigen binding fragment or derivative" of an antibody / immunoglobulin is defined as a fragment (e.g., a variant region of IgG) or derivative that retains an antigen binding region. The "antigen binding region" of an antibody is typically present in one or more hypervariable regions of the antibody, i.e., CDR-1, -2, and / or -3 regions. The "antigen binding fragment" of the present invention includes the domains of F(ab')2 fragments and Fab fragments. The "functional fragments and derivatives" of the present invention include Fab fragments, F(ab')2 fragments, Fab' fragments, scFv, dsFv, diabodies, tribodies, tetrabodies, Fc fusion proteins, vorabodies, camel antibodies, VHH, VNARs, and miniantibodies. F(ab')2 or Fab domains can be engineered to minimize or completely remove the interaction of intermolecular disulfides present between the CH1 and CL domains. The antibody or functional fragment of the present invention can be a part of a bifunctional or multifunctional construct.

[0119] Functional fragments and derivatives of the present invention include, but are not limited to, Fab fragments, F(ab')2 fragments, Fab' fragments, scFv and diabodies.

[0120] Fab fragments can be obtained as purified digestion products after digestion of TNFα-specific antibodies using cysteine ​​proteases (such as papain, EC 3.4.22.2). F(ab')2 fragments can be obtained as purified digestion products after digestion of TNFα-specific antibodies using pepsin (EC 3.4.23.1) or IdeS (immunoglobulin degrading enzyme from Streptococcus pyogenes; EC 3.4.22). Fab' fragments can be obtained from F(ab')2 fragments under mild reducing conditions, wherein two Fab' fragments are produced per F(ab')2 molecule.

[0121] scFv is a single-chain Fv fragment in which the variable light chain (“V L ”) and variable heavy chain (“V H”) domains are linked together by peptide bridges.

[0122] A "diabody" is a dimer composed of two fragments, each having a variable region linked together by a linker or the like (hereinafter referred to as a diabody-forming fragment), and generally contains two V L and two V H The dimeric fragment consists of V L and V H 、V L and V L 、V H and V H Those composed of, preferably V H and V L In the dimer-forming fragment, the linker connecting the variable regions is not particularly limited, but is preferably short enough to avoid non-covalent bonds between the variable regions in the same fragment. The length of such a linker can be appropriately determined by those skilled in the art, but 2-14 amino acids, preferably 3-9 amino acids, and particularly 4-6 amino acids are generally used. In this case, the V encoded on the same fragment L and V H Connected together by a linker short enough to avoid V on the same chain L and V H The invention also provides a method for forming a dimer between the two fragments by non-covalent bonds between the fragments and to avoid forming a single chain variable region fragment, which can form a dimer with another fragment. The dimer can be formed between the two-body forming fragments by covalent or non-covalent bonds or both.

[0123] In addition, the diabody-forming fragments can be linked together by a linker or, for example, to form a single-chain diabody (sc(Fv)2). By linking the diabody-forming fragments together using a long linker of about 15-20 amino acids, non-covalent bonds can be formed between the diabody-binding fragments present on the same chain to form a dimer. Based on the same principle as for preparing diabodies, multimeric antibodies such as trimers or tetramers can also be prepared by linking three or more diabody-forming fragments.

[0124] In one embodiment, the functional antibody fragment or derivative is a Fab fragment, a F(ab')2 fragment, a Fab' fragment, a scFv, a dsFv, a diabody, a triabody, a tetrabody, an Fc fusion protein, a vorabody, a camelid antibody, a VHH, a VNAR, or a minibody. Preferred functional fragments or derivatives used in the present invention are Fab fragments, F(ab')2 fragments, Fab' fragments, scFv, and diabodies.

[0125] The antibodies or functional fragments or derivatives thereof contained in the compositions of the present invention specifically bind to TNFα, but are not otherwise specifically limited. The terms "anti-TNFα antibody," "TNFα antibody," "TNFα-specific antibody," and "antibody specific for TNFα" are used interchangeably herein. In the most general form (and when no reference to a definition is mentioned), "specificity" and "specific binding" refer to the ability of an antibody or functional fragment or derivative to distinguish between human TNFα and unrelated biomolecules, as determined, for example, according to specificity determination methods known in the art. Such methods include, but are not limited to, Western blot and enzyme-linked immunosorbent assay (ELISA) assays. For example, a standard ELISA assay can be performed. Typically, binding specificity is determined by using a group (about 3 to 5) of unrelated biomolecules (e.g., milk powder, BSA, transferrin, etc.) rather than a single reference biomolecule. In one embodiment, specific binding refers to the ability of an antibody or fragment to distinguish between human TNFα and human TNFβ. In a preferred embodiment of the present invention, the TNFα antibody or its functional fragment is a TNFα antibody. In an alternative preferred embodiment of the present invention, the TNFα antibody or its functional fragment is a functional fragment or derivative of a TNFα antibody.

[0126] In a preferred aspect of the present invention, the antibodies of the present invention are non-fucosylated antibodies or antibodies with minimal fucosylation. As used herein, the term "antibody with minimal fucosylation" refers to an antibody in which less than 90% of the N-glycans of the antibody are fucosylated. Methods for determining the percentage of fucosylation are well known in the art. In one embodiment, less than 75%, or less than 50%, or less than 25% of the N-glycans of the antibody are fucosylated. Most preferably, less than 10% of the N-glycans of the antibody are fucosylated. In a specific embodiment, the N-glycans of the antibody of the present invention do not contain any fucose. Preferably, less than 90% of the N-glycans of the antibody are fucosylated at N297 (EU numbering). In another embodiment, less than 75%, or less than 50%, or less than 25% of the N-glycans of the antibody are fucosylated at N297 (EU numbering). Most preferably, less than 10% of the N-glycans of the antibody at N297 (EU numbering) are fucosylated. In another embodiment, the N-glycans of the antibody at position N297 do not contain any fucose.

[0127] Non-fucosylated antibodies are sometimes also referred to as afucosylated antibodies, which can be generated by various methods. For example, the genes for α1,6-fucosyltransferase (FUT8) and GDP-mannose 4,6-dehydrogenase (GMD) in CHO cells can be synergistically knocked down to produce completely afucosylated and ADCC-enhanced monoclonal antibody variants (see, e.g., Imai-Nishiya et al., (2007) BMC Biotechnol. 7, 84). Zinc finger nucleases (ZFNs) (which cleave the FUT8 gene in the region encoding the catalytic core of α1,6-fucosyltransferase and thereby destroy the corresponding enzymatic function in CHO cells) can be used to produce monoclonal antibodies completely lacking core fucose (see, e.g., Malphettes et al., (2010) Biotechnol. Bioeng. 106, 774–783).

[0128] Antibodies with less fucosylation can be prepared by adding a decoy substrate such as 2-deoxy-2-fluoro-2-fucose to the culture medium (see, e.g., Dekker et al., (2016) Sci Rep 6:36964), thereby reducing the fucose introduced into IgG-Fc.

[0129] In another embodiment, the antibodies of the invention have a higher sialic acid content. Sialylation can be increased, for example, by co-transfection of cytidine monophosphate-sialic acid synthase (CMP-SAS), cytidine monophosphate-sialic acid transporter (CMP-SAT), and α2,3-sialyltransferase (see, e.g., Son et al., (2011) Glycobiology 21, 1019-1028).

[0130] Preferably, the antibodies of the present invention have a high affinity for human FcRn at pH 6. The high affinity binding of the antibody to human FcRn at pH 6 is characterized by a K D Preferably, the K for high affinity binding at pH 6 is less than 500 nM. D The value is less than 400 nM, or less than 300 nM, or less than 200 nM. For example, a K value characterizing affinity at pH 6 D Values ​​may range from 1 to 500 nM, or 2 to 400 nM, or 3 to 300 nM, or 4 to 200 nM, or 5 to 100 nM.

[0131] In one embodiment, the affinity of an antibody of the invention for human FcRn at pH 6 is higher than the affinity of infliximab for human FcRn at pH 6.

[0132] The affinity of the antibodies of the present invention for human FcRn is preferably determined by surface plasmon resonance (SPR).

[0133] Several monoclonal antibodies against TNFα are described in the prior art. Meager et al. (Hybridoma, 6, 305-311, 1987) described mouse monoclonal antibodies against recombinant TNFα. Fendly et al. (Hybridoma, 6, 359-370, 1987) described the use of mouse monoclonal antibodies against recombinant TNFα to define neutralizing epitopes on TNF. In addition, recombinant antibodies specific for TNFα, including CDR-grafted antibodies, are disclosed in International Patent Application WO 92 / 11383. U.S. Patent No. 5,919,452 discloses anti-TNFα chimeric antibodies and their use in treating conditions associated with the presence of TNFα. Other anti-TNFα antibodies are disclosed in Stephens et al. (Immunology, 85, 668-674, 1995), GB-A-2 246 570, GB-A-2 297 145, US 8,673,310, US 2014 / 0193400, EP 2 390 267 B1, US 8,293,235, US 8,697,074, WO 2009 / 155723 A2 and WO 2006 / 131013 A2.

[0134] Currently approved anti-TNFα antibodies include (i) infliximab, a chimeric IgG monoclonal antibody (ii) Etanercept, a TNFR2 dimer fusion protein with IgG1 Fc (iii) Adalimumab, a fully human monoclonal antibody (mAb) (iv) Certolizumab PEGylated, a PEGylated Fab fragment and (v) golimumab, a human IgG1 monoclonal antibody Any INN antibody or functional fragment or variant mentioned herein encompasses its biosimilars and biooptimal forms thereof. In one embodiment of the present invention, the anti-TNFα antibody or functional fragment or derivative thereof is selected from infliximab, adalimumab, etanercept, certolizumab pegol, and golimumab, or a functional fragment or derivative thereof. In another embodiment of the present invention, the antibody or functional fragment or derivative thereof is selected from the anti-TNFα antibodies or functional fragments or derivatives disclosed in WO 2017 / 158092, WO 2017 / 158097, WO 2017 / 158084, and WO 2017 / 158079. In another embodiment of the present invention, at least one antibody or functional fragment or derivative thereof is an anti-TNFα antibody or functional fragment or derivative thereof having a light chain variable domain and / or a heavy chain variable domain comprising complementarity determining regions (CDRs) having amino acid sequences as disclosed in PCT applications WO 2017 / 158092, WO 2017 / 158097, WO 2017 / 158084 and WO 2017 / 158079.

[0135] In a preferred embodiment of the present invention, the antibody or its functional fragment or derivative is selected from an anti-TNFα antibody or its functional fragment or derivative having a light chain variable domain and / or a heavy chain variable domain comprising one or more CDRs having an amino acid sequence as disclosed in SEQ ID NOs: 7, 9, 12, 14, 24 and 25 of WO 2017 / 158079, SEQ ID NOs: 7-11 and 6 of WO 2017 / 158097, SEQ ID NOs: 7-12 of WO 2017 / 158092 and SEQ ID NOs: 1-4, 7 and 6 of WO 2017 / 158084, and combinations thereof. In another preferred embodiment of the present invention, the antibody or its functional fragment or derivative is selected from an anti-TNFα antibody or its functional fragment or derivative having a light chain variable domain and a heavy chain variable domain comprising CDRs having an amino acid sequence as disclosed in claim 2 of the originally filed application WO 2017 / 158079, claim 2 of WO 2017 / 158097, claim 2 of WO 2017 / 158092 and / or claim 2 of WO 2017 / 158084. In another preferred embodiment of the present invention, the antibody TNFα or its functional fragment or derivative is selected from the following: an anti-TNFα antibody or its functional fragment or derivative having a heavy chain variable domain amino acid sequence and / or a light chain variable domain amino acid sequence according to claim 4 of WO 2017 / 158079, claims 5 and 6 of WO 2017 / 158097, claims 5 and 6 of WO 2017 / 158092, claim 4 of WO 2017 / 158084, and combinations thereof.

[0136] In a preferred embodiment, the antibody comprises (i) V L A domain comprising a CDR1 region having the amino acid sequence as shown in SEQ ID NO: 3, a CDR2 region having the amino acid sequence as shown in SEQ ID NO: 4, and a CDR3 region having the amino acid sequence as shown in SEQ ID NO: 5; and (ii) V H A structural domain comprising a CDR1 region having an amino acid sequence as shown in SEQ ID NO: 6, a CDR2 region having an amino acid sequence as shown in SEQ ID NO: 7, and a CDR3 region having an amino acid sequence as shown in SEQ ID NO: 8. More preferably, the antibody comprises a V domain having an amino acid sequence as shown in SEQ ID NO: 10. L domain and a V domain having an amino acid sequence as shown in SEQ ID NO: 9 H domain.

[0137] In another preferred embodiment (which may be combined with any of the embodiments in the preceding paragraphs), the antibody comprises an Fc region comprising amino acids 236 to 451 of the amino acid sequence shown in SEQ ID NO:11.

[0138] More preferably, the antibody comprises a light chain having the amino acid sequence shown in SEQ ID NO: 1 and a heavy chain having the amino acid sequence shown in SEQ ID NO: 11, 14, 15 or 16.

[0139] In one embodiment, the antibody comprises a light chain having the amino acid sequence as shown in SEQ ID NO: 1 and a heavy chain having the amino acid sequence as shown in SEQ ID NO: 11. In another embodiment, the antibody comprises a light chain having the amino acid sequence as shown in SEQ ID NO: 1 and a heavy chain having the amino acid sequence as shown in SEQ ID NO: 14. In another embodiment, the antibody comprises a light chain having the amino acid sequence as shown in SEQ ID NO: 1 and a heavy chain having the amino acid sequence as shown in SEQ ID NO: 15. In another embodiment, the antibody comprises a light chain having the amino acid sequence as shown in SEQ ID NO: 1 and a heavy chain having the amino acid sequence as shown in SEQ ID NO: 16.

[0140] In yet another embodiment, the antibody comprises a light chain consisting of the amino acid sequence as shown in SEQ ID NO: 1 and a heavy chain consisting of the amino acid sequence as shown in SEQ ID NO: 11. In yet another embodiment, the antibody comprises a light chain consisting of the amino acid sequence as shown in SEQ ID NO: 1 and a heavy chain consisting of the amino acid sequence as shown in SEQ ID NO: 14. In yet another embodiment, the antibody comprises a light chain consisting of the amino acid sequence as shown in SEQ ID NO: 1 and a heavy chain consisting of the amino acid sequence as shown in SEQ ID NO: 15. In yet another embodiment, the antibody comprises a light chain consisting of the amino acid sequence as shown in SEQ ID NO: 1 and a heavy chain consisting of the amino acid sequence as shown in SEQ ID NO: 16.

[0141] In a further embodiment, the TNFα antibody is infliximab, adalimumab, etanercept, certolizumab pegol or golimumab. In an alternative further embodiment, the functional fragment or derivative of the TNFα antibody is infliximab, adalimumab, etanercept, certolizumab pegol or golimumab.

[0142] Antibodies specific for TNFα or their functional fragments or derivatives may also include one or more modifications, for example, in the form of additions or substitutions of residues that improve stability, specificity or targeting. These may include any such modifications known in the art. In one embodiment of the invention, at least one antibody specific for TNFα or its functional fragments or derivatives is included in any such modification disclosed in US8,871,204, US 6,737,056, US 8,742,074, WO 2002 / 060919, WO2017 / 158426, International Patent Application PCT / EP2018 / 074525 (WO 2019 / 057567 A1), PCT / EP2018 / 074523 (WO 2019 / 057565 A1) or PCT / EP2018 / 074522 (WO 2019 / 057564A1).

[0143] In one specific embodiment, the amino acid sequence of the antibody heavy chain comprises i) amino acids 233P, 234V, 235A, and a deletion at amino acid position 236; and amino acid 434A or amino acids 252Y, 254T and 256E; and optionally amino acids 239D, 330L and 332E or amino acids 326A, 332E and 333A (EU numbering); and / or ii) amino acids 380A and 434A, and optionally amino acid 307T (EU numbering); and / or iii) amino acid 434W, and optionally amino acid 428E and / or amino acid 311R (EU numbering). In another specific embodiment, the amino acid sequence of the antibody heavy chain comprises a mutation at one or more positions selected from the group consisting of: 311, 428, 434, 435 and 438 (EU numbering). Preferably, the amino acid sequence of the antibody heavy chain comprises amino acid 434W, amino acid 428E and amino acid 311R (EU numbering). Most preferably, the antibody comprises an Fc domain comprising amino acids 236 to 451 of the amino acid sequence as shown in SEQ ID NO: 11. Antibodies having the specific amino acids described in this paragraph can be obtained by introducing substitutions into the heavy chain sequence (e.g., introducing the sequence as shown in SEQ ID NO: 2).

[0144] Unless otherwise stated herein, residue numbers referred to in the amino acid sequences of antibodies correspond to the residue numbering of the EU numbering system (also known as the EU index) as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, Md., 1991. (See, e.g., WO 2006 / 073941).

[0145] The pharmaceutical composition of the present invention may comprise as an active agent one type of antibody specific for TNFα or a functional fragment or derivative thereof, or several different antibodies specific for TNFα and / or functional fragments or derivatives of antibodies specific for TNFα. For example, the composition of the present invention may comprise 1, 2, 3, 4 or 5 different antibodies specific for TNFα and functional fragments or derivatives of antibodies specific for TNFα.

[0146] As used herein, the terms "treat," "treating," and "treatment" with respect to ICP inhibitor-induced diarrhea, colitis, or enterocolitis refer to the administration of a TNFα-specific antibody, or a functional fragment or derivative thereof, as part of the present invention to prevent, minimize, or delay the onset of symptoms, complications, or similar biochemical signs of ICP inhibitor-induced diarrhea, colitis, or enterocolitis, or to alleviate or minimize symptoms, complications, or similar biochemical signs of ICP inhibitor-induced diarrhea, colitis, or enterocolitis.

[0147] In the context of the present invention, the term "local treatment" is used to describe a composition for local application, as opposed to systemic application of a composition containing a TNFα antibody as used in commercial products (e.g., by intravenous infusion or subcutaneous injection). However, local treatment in the lumen of the ileum and large intestine is not limited by the manner in which the composition is administered. In this context, the term "administering" relates to the manner and form in which the composition first comes into contact with the patient's body. This means that the composition, in an appropriate form, can be administered orally, rectally, or in any other manner that allows the composition to accumulate at the site of local administration.

[0148] In the present invention, the term "local treatment in the ileum and / or large intestine" refers to a local application as defined above in the lumen of the ileum and / or large intestine, in other words, somewhere inside the combined and continuous interior consisting of the ileum of the small intestine and the large intestine. The "large intestine" is the last part of the gastrointestinal (GI) tract and can be further subdivided into the cecum, colon and rectum. The "colon" can be further subdivided into the ascending colon, transverse colon and descending colon. The "ileum" of the small intestine is the last part of the small intestine and is located at one end connected to the cecum. The "terminal ileum" is the last part of the ileum, which is directly adjacent to the cecum. As used herein, the term "gastrointestinal tract" or "GI" describes the organ system of the human body, which includes all structures that form a continuous passage between the mouth and the anus and is responsible for digesting ingested substances, absorbing nutrients and eliminating feces. In one embodiment of the present invention, the composition of the present invention is used for local treatment in the terminal ileum, and / or large intestine, preferably the colon. In another embodiment of the present invention, the composition of the present invention is used for local treatment in the terminal ileum, cecum, ascending colon, transverse colon and / or descending colon of a patient.

[0149] Local treatment using the pharmaceutical composition of the present invention allows for local application of TNFα antibodies and functional fragments or derivatives thereof in the lumen of the patient's ileum and / or large intestine. Thus, systemic application of TNFα antibodies and functional fragments or derivatives thereof can be avoided. In this way, systemic uptake and distribution of TNFα antibodies and functional fragments or derivatives thereof can be minimized. Thus, TNFα antibodies and functional fragments or derivatives thereof do not significantly resist successful cancer treatment using ICP inhibitors at a systemic level. In addition, the multiple risks associated with systemic suppression of TNFα immune defense function in patients (e.g., including infectious complications and accumulation of anti-TNFα antibody-specific antibodies in the patient's body, resulting in loss of anti-TNFα antibody response) can be kept to a minimum.

[0150] According to a first aspect of the present invention, the composition of the present invention is used to treat a patient experiencing ICP inhibitor-induced adverse events in the ileum and / or colon (ie, ICP inhibitor-induced diarrhea, colitis or enterocolitis).

[0151] As used herein, "ICP inhibitors" are any molecules (e.g., small molecules, proteins, peptides, nucleic acid molecules, or antibodies) that are administered to a patient for the purpose of treating a disease (e.g., cancer or infectious disease) to stimulate the patient's immune system (e.g., by not blocking an existing immune response or by not blocking the initiation of an immune response). ICP inhibitors can target the components of any immune checkpoint known in the art to stimulate the immune system, including but not limited to CTLA-4, PD-1, PD-L1, PDL2, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, CSF-1R, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, ADAR1, CD47 (targeted by CD47 inhibitors), ICOS (targeted by ICOS agonists), TIGIT (targeted by TIGIT inhibitors), and B-7 family ligands. An inhibitor for a single component of an immune checkpoint or a combination of different inhibitors for different components of an immune checkpoint can be used. Immune checkpoints are typically part of or consist of an inhibitory or stimulatory pathway that maintains self-tolerance and contributes to an immune response. ICP inhibitors according to the present invention include molecules that block pathways and molecules that stimulate pathways. For example, TIGIT inhibitors and ICOS agonists are ICP inhibitors in the sense of the present invention. Marin-Acevedo et al., (2018) Journal of Hematology & Oncology 11, article No. 39 summarizes inhibitory and stimulatory pathways and their targets that can be targeted by suitable ICP inhibitors. The contents of Marin-Acevedo et al., the entire contents of which are incorporated herein.

[0152] As used herein, an "adverse event" is an unfavorable and usually unexpected, even undesirable sign (including abnormal laboratory results), symptom, or disease associated with the use of a medical treatment. For example, an adverse event may be associated with activation of the immune system or expansion of immune cells (e.g., T cells) in response to a treatment. A medical treatment may have one or more associated adverse events and each adverse event may have the same or varying severity.

[0153] As used herein, in the context of the present invention, "colitis" refers to an inflammatory condition of the large intestine, and particularly in the colon, which may be associated with symptoms including diarrhea, abdominal pain, nausea, cramping, bloody or sticky stools or changes in bowel habits, fever, abdominal distension, constipation, and edema, congestion, and / or brittle walls of the large intestine. As used herein, in the context of the present invention, "enterocolitis" refers to an inflammatory condition of the large intestine, and particularly in the colon and ileum of the small intestine, preferably the terminal ileum, which may be associated with symptoms including diarrhea, abdominal pain, nausea, cramping, bloody or sticky stools or changes in bowel habits, fever, abdominal distension, constipation, or edema, congestion, and / or brittle walls of the ileum or large intestine.

[0154] As used herein, the terms "ICP inhibitor-induced colitis" and "ICP inhibitor-induced enterocolitis" refer to colitis and enterocolitis, respectively, that have the following characteristics: (1) that first occurs in a patient concurrently with or shortly after (i.e., days or weeks, e.g., 1 to 60 weeks, 1 to 48 weeks, 13 to 60 weeks, or 7 to 26 weeks thereafter) the first administration of one or more ICP inhibitors; (2) that is identified by a physician (e.g., based on criteria set forth in ASCO practice guidelines) as ICP inhibitor-induced colitis or enterocolitis; and (3) that is not identified by a physician as having another etiology (e.g., Clostridium difficile toxin).

[0155] Except where specifically stated, the terms "patient" or "subject" are used interchangeably and refer to mammals (e.g., human patients and non-human primates), as well as experimental animals (e.g., rabbits, rats, and mice, among others). Animals include all vertebrates, e.g., mammals and non-mammals, such as sheep, dogs, cows, chickens, amphibians, and reptiles.

[0156] ICP inhibitor-induced colitis in living patients can be graded from 1 to 4 based on the toxicity that occurs in the large intestine, and particularly the colon, induced by treatment of the patient with one or more ICP inhibitors. The grading for ICP inhibitor-induced colitis is as follows:

[0157] Grade 1 colitis presents asymptomatically with only radiographic or histologic findings.

[0158] Grade 2 colitis is characterized by the presence of sticky or bloody stools and abdominal pain.

[0159] Grade 3 colitis is characterized by severe abdominal pain, fever, change in bowel habits, and peritoneal signs.

[0160] Grade 4 colitis is a life-threatening condition in which there is perforation or hemorrhage or signs of ischemia or necrosis and toxic megacolon develops.

[0161] Grade 5 colitis is death caused by colitis.

[0162] Similarly, ICP inhibitor-induced enterocolitis can be graded from 1 to 4 based on the toxicity that occurs in the small intestine, and particularly the ileum, and in the large intestine, and particularly the colon, resulting from treatment of a patient with one or more ICP inhibitors. The grades of ICP inhibitor-induced enterocolitis are as follows:

[0163] Grade 1 enterocolitis presents asymptomatically with only radiographic or histologic findings.

[0164] Grade 2 enterocolitis is characterized by the presence of mucus or blood in the stool and abdominal pain;

[0165] Grade 3 enterocolitis is characterized by severe or persistent abdominal pain, fever, intestinal obstruction, and abdominal signs.

[0166] Grade 4 enterocolitis is a life-threatening condition in which there is perforation or hemorrhage or signs of ischemia or necrosis and toxic megacolon develops.

[0167] Grade 5 enterocolitis is death caused by colitis.

[0168] ICP inhibitor-induced diarrhea is graded as follows:

[0169] Grade 1 diarrhea is defined as an increase of <4 stools over baseline;

[0170] Grade 2 diarrhea characterized by 4-6 stools more than baseline;

[0171] Grade 3 diarrhea characterized by ≥7 stools over baseline;

[0172] Grade 4 diarrhea is characterized by life-threatening consequences due to diarrhea;

[0173] Grade 5 diarrhea is fatal.

[0174] Diagnostic tests for determining toxicity and grading ICP inhibitor-induced diarrhea, colitis, or enterocolitis can be performed, for example, as described in the ASCO guidelines (Brahmer et al., Management of Immune-Related Adverse Events in Patients Treated With Immune Checkpoint Inhibitor Therapy: American Society of Clinical Oncology Practice Guideline, J Clin Oncol. 2018 Jun 10; 36(17): 1714-1768). According to the ASCO guidelines, the diagnostic workup for grade 2 toxicity should include the following:

[0175] -Blood tests (CBC, comprehensive metabolic panel, thyroid-stimulating hormone [TSH], erythrocyte sedimentation rate [ESR], C-reactive protein [CRP]), stool (culture, Clostridium difficile, parasites, cytomegalovirus [CMV]) or other viral etiologies, egg protein, and parasites) should be obtained.

[0176] - Lactoferrin can be tested for patient stratification to identify those requiring urgent endoscopy, and calprotectin can be provided to track disease activity;

[0177] -In patients at high risk for those infections and in appropriately selected patients, laboratory screening (HIV, hepatitis A and B, and blood quantiferon for TB) should be performed routinely based on evaluation by an infectious disease specialist to prepare the patient for initiation of infliximab;

[0178] - Imaging with computed tomography (CT) scans of the abdomen and pelvis and GI endoscopy with tissue biopsy may be performed, as there is evidence that the presence of ulcers in the colon predicts a course refractory to corticosteroids, which may require early infliximab. Infliximab or other tumor necrosis factor (TNF) blockers should not be delayed while awaiting the results of these screening tests;

[0179] - Endoscopy may be repeated in patients who do not respond to immunosuppressants. Endoscopy should be repeated only when clinically indicated and when resumption of therapy is planned to monitor disease.

[0180] According to the ASCO guidelines, the diagnostic workup for grade 3 to 4 toxicity should include the following:

[0181] -All studies listed for Level 2 (blood, stool, imaging, and tissue biopsy) should be completed promptly;

[0182] - Endoscopy may be repeated in patients who do not respond to immunosuppressants. Endoscopy should be repeated only when clinically indicated and when ICP inhibitors are planned to be resumed to monitor disease.

[0183] According to one embodiment of the present invention, treatment with antibodies or fragments or derivatives thereof can prevent adverse effects (eg, diarrhea, colitis and / or enterocolitis) from progressing to higher levels of toxicity. The following embodiments are preferred.

[0184] Patients are to be treated for a Grade 1 toxic adverse effect (e.g., diarrhea, colitis, and / or enterocolitis) and treatment with the antibody, or fragment or derivative thereof, can prevent the adverse effect from progressing to a higher grade toxicity (e.g., Grade 2 toxicity).

[0185] Patients are to be treated for Grade 2 toxic adverse effects (e.g., diarrhea, colitis, and / or enterocolitis) and treatment with the antibody, or fragment or derivative thereof, can prevent progression of the adverse effect to a higher grade toxicity (e.g., Grade 3 toxicity).

[0186] Patients who are to be treated have a Grade 3 toxic adverse effect (e.g., diarrhea, colitis, and / or enterocolitis) and treatment with the antibody, or fragment or derivative thereof, can prevent progression of the adverse effect to a higher grade toxicity (e.g., Grade 4 toxicity).

[0187] In one aspect, the present invention relates to an active agent selected from the following: antibodies specific for tumor necrosis factor alpha (TNFα) and functional fragments and derivatives thereof, for preventing the progression or worsening of at least one gastrointestinal adverse event induced by cancer therapy in a patient, preferably induced by one or more immune checkpoint (ICP) inhibitors, wherein the prevention comprises local administration of the compound to the ileum and / or large intestine of the patient. Preferably, the local use is oral administration. Preferably, the adverse event is selected from the following: diarrhea, colitis and enterocolitis and combinations thereof, and the progression from grade 1 toxicity to grade 2 toxicity or any higher grade can be prevented. In another embodiment, the adverse event is selected from the following: diarrhea, colitis and enterocolitis and combinations thereof, and the progression from grade 2 toxicity to grade 3 toxicity or any higher grade can be prevented. In another embodiment, the adverse event is selected from the following: diarrhea, colitis and enterocolitis and combinations thereof, and the progression from grade 3 toxicity to grade 4 toxicity or any higher grade can be prevented. The preferred embodiments of this aspect of the invention correspond to the preferred embodiments of the other aspects of the invention described herein with necessary modifications.

[0188] According to another embodiment of the invention, the patient to be treated with the antibody or fragment or derivative thereof suffers from ICP inhibitor-induced diarrhea but does not suffer from colitis or enterocolitis, and said treatment prevents the development or onset of colitis and enterocolitis in said patient.

[0189] According to one embodiment of the first aspect of the present invention, the patient is also receiving treatment with one or more ICP inhibitors. "Also receiving treatment with one or more ICP inhibitors" means that the treatment has not been finally stopped, but is currently ongoing or temporarily suspended. According to a preferred embodiment, the patient is also receiving systemic treatment with one or more ICP inhibitors. Systemic treatment or therapy refers to treatment that reaches and affects substantially all cells of the body through the systemic circulation.

[0190] Prior to administration of the composition of the present invention, treatment with one or more ICP inhibitors may be temporarily suspended for a few weeks or days, or not at all, upon development of toxicity associated with ICP inhibitor-induced diarrhea, colitis, or enterocolitis. According to one embodiment of the present invention, treatment with one or more ICP inhibitors is suspended for less than 4 weeks, preferably less than 2 weeks, more preferably 1 week, even more preferably less than 5, 4, 3, 2 days, or 1 day, and most preferably without discontinuation of treatment.

[0191] There is no specific limitation on the degree of toxicity in patients with ICP inhibitor-induced diarrhea, colitis, or enterocolitis to whom the compositions of the present invention are to be administered. The degree of toxicity of diarrhea, colitis, or enterocolitis caused by an ICP inhibitor can be determined by a physician, for example, using the ASCO operating guidelines. In one embodiment, the patient has ICP inhibitor-induced diarrhea, colitis, or enterocolitis of grade 1 toxicity or greater. In another embodiment, the patient has ICP inhibitor-induced diarrhea, colitis, or enterocolitis of grade 2 toxicity or greater. In yet another embodiment, the patient has ICP inhibitor-induced diarrhea, colitis, or enterocolitis of grade 3 toxicity or greater. In a further embodiment, the patient has ICP inhibitor-induced diarrhea, colitis, or enterocolitis of grade 4 toxicity or greater.

[0192] According to a preferred embodiment, the composition of the present invention is used as a first-line treatment for patients with ICP inhibitor-induced diarrhea, colitis or enterocolitis, to replace steroid immunosuppressants such as systemic corticosteroids. In one embodiment, the patient is not currently being treated and has not yet been treated with corticosteroids.

[0193] In an alternative embodiment, the patient first receives a steroid immunosuppressant (e.g., a corticosteroid such as prednisone, prednisolone, methylprednisolone, dexamethasone or budesonide). The composition of the present invention is then administered to the patient when the diarrhea, colitis or enteritis induced by the ICP inhibitor is refractory to steroid therapy. As used herein, the term "steroid-refractory" and: steroid-refractory diarrhea, colitis or enterocolitis refers to diarrhea, colitis or enterocolitis induced by one or more ICP inhibitors that is not responsive to steroid therapy. Therefore, according to a specific alternative embodiment, the composition of the present invention is used as a second-line treatment in patients with ICP inhibitor-induced steroid-refractory diarrhea, colitis or enterocolitis.

[0194] In a specific embodiment, the pharmaceutical composition of the present invention is administered in a maintenance therapy. That is, the composition is administered to prevent the recurrence or recurrence of an adverse event (diarrhea, colitis, and / or enterocolitis). If the adverse event has been successfully treated, the composition may be continued to prevent the recurrence or recurrence of the adverse event. The maintenance dose of the active agent may be the dose used to treat the adverse event.

[0195] In a second aspect, the present invention further relates to a pharmaceutical composition comprising an active agent selected from the group consisting of antibodies specific for TNFα and functional fragments or derivatives thereof, for use in local treatment in the ileum and / or large intestine of a patient as a prophylactic therapy for diarrhea, colitis or enterocolitis induced by an ICP inhibitor. With respect to the terms "antibodies specific for TNFα and functional fragments or derivatives thereof", "local treatment in the ileum and / or large intestine" and "diarrhea, colitis or enterocolitis induced by an ICP inhibitor", it refers to what has been described in detail above.

[0196] As used in the context of the present invention, the term "prophylactic therapy" refers to the administration of an active agent as a preventive therapy. As used in the context of the present invention, the term "prophylactic therapy against ICP inhibitor-induced diarrhea, colitis or enterocolitis" refers to the administration of a composition of the present invention as a preventive or semi-prophylactic therapy prior to the onset of symptoms associated with ICP inhibitor-induced diarrhea, colitis or enterocolitis.

[0197] The compositions of the present invention used as a local treatment of the ileum and / or large intestine of a patient as a preventative therapy allow for simultaneous systemic treatment with one or more ICP inhibitors. In the context of the present invention, "concurrently" refers to a treatment regimen in which the patient receives simultaneous treatment with one or more ICP inhibitors.

[0198] In a preferred embodiment of the present invention, the patient is simultaneously receiving treatment with one or more ICP inhibitors. In another preferred embodiment, the patient is simultaneously receiving systemic treatment with one or more ICP inhibitors. To this end, one or more ICP inhibitors can be administered as part of a separate composition simultaneously with the composition of the present invention using the same dosage frequency or at different times and / or using different dosage frequencies. In addition, one or more ICP inhibitors can be administered by the same route of administration or by different routes of administration. With respect to the route of administration of the ICP inhibitor, it relates to the description of the ICP inhibitor below.

[0199] For example, the one or more ICP inhibitors can be administered intravenously or by subcutaneous or intramuscular injection, while the compositions of the present invention are administered orally or rectally. Alternatively, both the one or more ICP inhibitors and the compositions of the present invention are administered orally or rectally. Where the one or more ICP inhibitors and the compositions of the present invention are administered by the same route of administration and with the same dosing frequency, the one or more ICP inhibitors can be added to the compositions of the present invention as an additional active agent.

[0200] By administering the compositions of the present invention as a prophylactic therapy, the onset of symptoms associated with ICP inhibitor-induced diarrhea, colitis, or enterocolitis can be delayed and / or minimized, or even prevented. Thus, in one embodiment, as a prophylactic therapy for ICP inhibitor-induced diarrhea, colitis, or enterocolitis, the compositions of the present invention used to locally treat the ileum and / or large intestine of a patient can prevent or minimize and / or mitigate symptoms associated with ICP inhibitor-induced diarrhea, colitis, or enterocolitis. In a preferred embodiment, the patient has ICP inhibitor-induced diarrhea, and the onset of symptoms associated with ICP inhibitor-induced colitis or enterocolitis can be delayed and / or minimized, or even prevented. Thus, as a prophylactic therapy for ICP inhibitor-induced colitis or enterocolitis, the compositions of the present invention used to locally treat the ileum and / or large intestine of a patient can prevent or minimize and / or mitigate symptoms associated with ICP inhibitor-induced colitis or enterocolitis.

[0201] In another embodiment, as a prophylactic treatment for ICP inhibitor-induced diarrhea, colitis, or enterocolitis, the compositions of the present invention for local treatment of the ileum and / or large intestine of a patient can prevent the onset of diarrhea, colitis, or enterocolitis caused by an ICP inhibitor. In another embodiment, as a prophylactic treatment for ICP inhibitor-induced diarrhea, colitis, or enterocolitis, the compositions of the present invention for local treatment of the ileum and / or large intestine of a patient can prevent the onset of one or more symptoms associated with ICP inhibitor-induced diarrhea, colitis, or enterocolitis.

[0202] In an alternative embodiment, as a prophylactic therapy for ICP inhibitor-induced diarrhea, colitis, or enterocolitis, the compositions of the present invention used to locally treat the ileum and / or large intestine of a patient can delay the onset and / or minimize the severity of one or more symptoms associated with ICP inhibitor-induced diarrhea, colitis, or enterocolitis. According to a specific embodiment, the local treatment can prevent the occurrence of grade 3 toxicity, preferably grade 2 toxicity, more preferably grade 1 toxicity, and most preferably any symptom associated with ICP inhibitor-induced diarrhea, colitis, or enterocolitis in the patient.

[0203] Local treatment of the ileum and / or large intestine in patients with ICP inhibitor-induced diarrhea, colitis or enterocolitis or prophylactic treatment of ICP inhibitor-induced diarrhea, colitis or enterocolitis using the compositions of the present invention in place of, for example, standard treatment regimens recommended by ASCO guidelines can reduce the incidence and / or severity of symptoms of ICP inhibitor-induced diarrhea, colitis or enterocolitis associated with the use of standard treatment regimens for ICP inhibitor-induced diarrhea, colitis or enterocolitis.

[0204] In a specific embodiment, treatment according to the present invention is associated with or results in a patient having an overall survival that is greater than that of a patient not treated according to the present invention. Preferably, treatment according to the present invention is associated with or results in a patient having an overall survival that is greater than that of a patient treated with corticosteroids. Treatment according to the present invention is associated with or results in a patient having an overall survival that is greater than that of a patient not treated according to the present invention nor treated with corticosteroids.

[0205] The present invention is beneficial in that it may not require the administration of corticosteroids. Corticosteroids often lead to increased infection rates, which in turn require the administration of antibiotics. The present invention avoids the need for corticosteroids and antibiotics. Therefore, in a preferred embodiment, the composition of the present invention is administered after the first signs of enterocolitis, colitis, and / or diarrhea have been detected.

[0206] The present invention also provides prophylactic uses of the compositions of the present invention. In another embodiment, therefore, the compositions of the present invention are administered before signs of enterocolitis, colitis or diarrhea can be detected or have been detected. In another embodiment, the compositions of the present invention are administered to a patient, preferably a cancer patient, more preferably a cancer patient being treated with at least one ICP inhibitor, wherein the patient has not been diagnosed with enterocolitis, colitis and / or diarrhea. In another embodiment, the compositions of the present invention are administered to a patient, preferably a cancer patient, more preferably a cancer patient being treated with at least one ICP inhibitor, wherein the patient does not suffer from enterocolitis; and / or wherein the patient does not suffer from colitis; and / or wherein the patient does not suffer from diarrhea. The patient may be at risk of developing enterocolitis, colitis and / or diarrhea. The patient may be suspected of developing enterocolitis, colitis and / or diarrhea. The present invention relates to the use of the compositions described herein for preventing the onset or occurrence of enterocolitis, colitis and / or diarrhea in a patient, preferably a cancer patient, more preferably a cancer patient being treated with at least one ICP inhibitor.

[0207] In one embodiment, administration of the active agent improves the effectiveness of treatment with an ICP inhibitor.

[0208] ICP inhibitors can be administered to patients to treat diseases (such as cancer) or to prevent the recurrence of such diseases. Such treatment results in adverse events caused by ICP inhibitors in most cases at a certain site, of which ICP inhibitor-induced adverse events in the GI tract (such as inhibitor-induced diarrhea, colitis or enterocolitis) are the most common adverse events.

[0209] "Cancer" refers to a broad group of diseases characterized by uncontrolled, abnormal, usually endogenous cell growth in the body. Unregulated cell division and growth can form malignant tumors that invade adjacent tissues and can also metastasize to distant parts of the body via lymphocytes or the bloodstream. The terms "cancer," "tumor," and "neoplasm" are used interchangeably herein.

[0210] There are no specific limitations on the cancers that can be treated by ICP inhibitors. Exemplary cancers that can be treated by ICP inhibitors include, but are not limited to, melanoma; lymphomas, such as classical Hodgkin's lymphoma; gliomas; urothelial carcinoma; renal cancer; head and neck squamous cell carcinoma, prostate cancer; breast cancer; colon cancer; and lung cancer. According to one embodiment of the present invention, the one or more ICP inhibitors are used to treat a cancer selected from the group consisting of bone cancer, pancreatic cancer, skin cancer, head and neck cancer, malignant melanoma of the skin or eye, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or Acute leukemias, acute myeloid leukemias, chronic myeloid leukemias, acute lymphoblastic leukemias, chronic lymphocytic leukemias, solid tumors of childhood, lymphocytic lymphomas, bladder cancer, kidney or ureteral cancer, renal pelvic cancer, central nervous system (CNS) neoplasms, primary CNS lymphomas, tumor angiogenesis, spinal tumors, brain stem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermoid carcinomas, squamous cell carcinomas, T-cell lymphomas, environmentally induced cancers (including those induced by asbestos), and combinations of such cancers.

[0211] ICP inhibition (e.g., by inhibiting (blocking) ICP components) can be performed for various ICP components (including, for example, PD-1, PD-L1, CTLA-4, LAG-3, ADAR1, and TIM-3, as well as combinations of such ICP components). ICP components can be, but are not limited to, receptors or ligands on, for example, tumor cells or immune cells (such as T cells, monocytes, microglia, and macrophages). ICP inhibitors do not need to be antibodies, but can also be small molecules or other polymers. If the ICP inhibitor is an antibody, it can be a polyclonal, monoclonal, fragment, single chain, or other antibody variant construct. ICP inhibitors include, for example, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-CTLA4 antibodies, anti-LAG-3 antibodies, and anti-TIM-3 antibodies.

[0212] ICP inhibitors can target any component of ICP known in the art to cause stimulation of the immune system, including but not limited to CTLA-4, PD-1, PD-L1, PDL2, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, CSF-1R, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, ADAR1, TIGIT, CD47, ICOS, and B-7 family ligands. A combination of an inhibitor of a target component of a single ICP and different inhibitors of target components of different ICPs can be used.

[0213] ICP inhibitors can be administered simultaneously with, before, or after the pharmaceutical composition of the present invention. ICP inhibitors can be administered by any suitable method known in the art for specific inhibitors to treat specific cancers or infectious diseases. Typically, ICP inhibitors are administered systemically. Suitable methods known in the art for administering ICP inhibitors include, for example, intravenous, oral, intraperitoneal, sublingual, intrathecal, intracavitary, intramuscular, and subcutaneous administration.

[0214] Preferred routes of administration of one or more ICP inhibitors include intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal or other non-parenteral routes of administration, such as by injection or infusion. As used herein, the term "parenteral administration" refers to modes of administration other than enteral and topical administration, typically by injection, and includes but is not limited to intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion. In a more preferred embodiment of the present invention, one or more ICPs are administered intravenously. Alternatively, one or more ICP inhibitors can be administered by non-parenteral routes, such as topical, epidermal or mucosal routes of administration, for example, intranasal, oral, vaginal, rectal, sublingual or topical.

[0215] Currently available ICP inhibitors include antibodies specific for CTLA-4, PD-1, and PD-L1. According to one embodiment of the present invention, the one or more ICP inhibitors are selected from the following: antibodies specific for CTLA-4, antibodies specific for PD-1, and antibodies specific for PD-L1. Monoclonal antibodies that are specific for and thus target PD-1 or PD-L1 can block the binding between these two ICP components and enhance the immune response against cancer cells. These drugs have shown considerable promise in treating various cancer types.

[0216] Examples of PD-1 specific antibodies currently used as ICP inhibitors in cancer therapy include the monoclonal antibody pembrolizumab and nivolumab These antibodies have been shown to be effective in treating several types of cancer, including melanoma of the skin, non-small cell lung cancer, kidney cancer, bladder cancer, head and neck cancer, and Hodgkin's lymphoma.

[0217] Examples of PD-L1-specific antibodies currently used as ICP inhibitors in cancer therapy include the monoclonal antibody atezolizumab Avelumab and durvalumab These antibodies have been shown to be effective in treating different types of cancer, including bladder cancer, non-small cell lung cancer, and Merkel cell skin cancer (Merkel cell carcinoma).

[0218] CTLA-4 is another B7-CD28 family member that inhibits T cell function. It is constitutively expressed by regulatory T cells, but can also be upregulated by other T cell types, especially CD4+ T cells upon activation. CTLA-4 mediates immunosuppression by indirectly attenuating signal transduction through the co-stimulatory receptor CD28. CTLA-4 signal transduction has been shown to attenuate immune responses against infection and tumor cells (Curran MA et al., Proc Natl Acad Sci US A. 2010 Mar 2; 107 (9): 4275-80). Examples of CTLA-4 specific antibodies currently used or being studied as ICP inhibitors in cancer therapy include the monoclonal antibody ipilimumab. and tremelimumab (Pfizer).

[0219] According to one embodiment of the present invention, the one or more ICP inhibitors are a combination of an antibody specific for PD-1 and / or PD-L1 and an antibody specific for CTLA-4, preferably a combination of an antibody specific for PD-1 or PD-L1 and an antibody specific for CTLA-4. According to another embodiment of the present invention, the antibody specific for PD-1 is selected from the following: pembrolizumab and nivolumab, the antibody specific for PD-L1 is selected from the following: atezolizumab, avelumab and durvalumab, and the antibody specific for CTLA-4 is selected from the following: ipilimumab and tremelimumab. According to another embodiment of the present invention, the patient to be treated with the composition of the present invention suffers from ICP inhibitor-induced diarrhea, colitis or enterocolitis, which is caused by treatment with any one or more of the ICP inhibitors listed above.

[0220] In order to administer antibodies specific for CTLA-4, PD-1 and PD-L1, for example, a suitable dosage range is about 0.0001 to 100 mg / kg, preferably 0.01 to 5 mg / kg patient body weight. An exemplary dosage can be 0.3 mg / kg body weight, 1 mg / kg body weight, 3 mg / kg body weight, 5 mg / kg body weight or 10 mg / kg body weight or within the range of 1-10 mg / kg body weight. The antibody is typically administered multiple times. An exemplary dosage regimen needs to be administered at the following frequencies: once a week, once every two weeks, once every three weeks, once every four weeks, once a month, once every three months, once every three to six months, or once every six months. Intervals may also be irregular, as indicated by measuring blood levels of antibodies to the target antigen in the patient. In some cases, the dose is adjusted to reach a specific plasma antibody concentration, for example, about 1-1000 μg / ml or about 25-300 μg / ml.

[0221] Preferred dosage regimens for antibodies specific for CTLA-4, PD-1 or PD-L1 include, but are not limited to, about 1 to 10 mg / kg body weight administered intravenously, wherein one antibody or combination of antibodies is administered at the following frequencies: (i) once every four weeks for 6 doses, then once every three months; (ii) once every three weeks; (iii) once every two weeks; or (iv) once a week.

[0222] As used herein, "about" means within an acceptable error range for a particular value as determined by one skilled in the art, which depends in part on the method of measurement or determination of the value, i.e., the limitations of the measurement system. For example, "about" can mean within one or more standard deviations, as practiced in the art. Alternatively, "about" can mean a range of up to 20%. Furthermore, particularly with respect to biological systems or processes, the term can mean up to an order of magnitude or up to 5 times a value.

[0223] In some cases, two or more CTLA-4, PD-1, and / or PD-L1-specific antibodies are administered simultaneously, in which case the dosage of each antibody administered may be within the dosage ranges indicated above. If more than one CTLA-4, PD-1, or PD-L1-specific antibody is administered, it may have different binding specificities for CTLA-4, PD-1, or PD-L1, respectively.

[0224] Alternatively, the ICP inhibitor can be administered as a sustained release formulation, in which case less frequent administration is required. Dosage and frequency vary depending on the half-life of the antibody in the patient. In general, human antibodies show the longest half-life, followed by humanized antibodies, chimeric antibodies, and non-human antibodies.

[0225] ADAR1 is an RNA editing enzyme that catalyzes the hydrolytic deamination of adenosine to inosine in double-stranded RNA (UniProt No. P55265). ADAR1 function can be inhibited by antibodies against ADAR1 and functional fragments thereof or by gene silencing techniques (e.g., using siRNA).

[0226] In a patient treated with one or more ICP inhibitors, the one or more ICP inhibitors may modulate the immune response in the patient, and / or they may inhibit the growth of tumor cells in the patient.

[0227] The patient to be treated with the compositions of the present invention may be a cancer patient. In one embodiment, the patient is a cancer patient and is currently undergoing or has been treated with one or more ICP inhibitors, preferably one or more of the ICP inhibitors listed above. Preferably, the cancer patient is currently undergoing treatment with one or more ICP inhibitors. Preferably, the treatment with one or more ICP inhibitors is systemic treatment.

[0228] In certain embodiments, the patient is undergoing cancer vaccine therapy.For example, the patient can receive a combination therapy with an ICP inhibitor and a cancer vaccine.

[0229] As used herein, the term "cancer vaccine" has its usual meaning in the art and refers to a composition capable of inducing active immunity against at least one cancer antigen. Cancer vaccines typically contain a source of cancer-related substances or cells (antigens) that may be autologous or allogeneic for the subject, as well as other components (e.g., adjuvants) to further stimulate and strengthen the immune response against the antigen. Cancer vaccines can result in stimulation of the subject's immune system to produce antibodies against one or more specific antigens, and / or to produce killer T cells to attack cancer cells with these antigens. Cancer vaccines can produce antibodies or only activate certain cells, particularly antigen-presenting cells, T lymphocytes (particularly T-CD8+ cells) and B lymphocytes. Cancer vaccines can be a composition for preventive purposes or for therapeutic purposes or both.

[0230] There are many types of cancer vaccines. Non-limiting examples of cancer vaccines include tumor cell vaccines, antigen vaccines, dendritic cell vaccines, DNA vaccines, and vector-based vaccines.

[0231] Typically, the cancer vaccine of the present invention comprises a tumor-associated antigen or a nucleic acid sequence (e.g., DNA) encoding a tumor-associated antigen. A variety of tumor-associated antigens are well known in the art. Exemplary tumor-associated antigens include, but are not limited to, 5α-reductase, α-fetoprotein, AM-1, APC, April, BAGE, β-catenin, Bell 2, bcr-abl, CA-125, CASP-8 / FLICE, autolysin, CD 19, CD20, CD21, CD23, CD22, CD33, CD35, CD44, CD45, CD46, CD5, CD52, CD55, CD59, CDC27, CDK4, CEA, c-myc, Cox-2, DCC, DcR3, E6 / E7, CGFR, EMBP, Dna78, farnesyltransferase, FGF8b, FGF8a, FLK-1 / KDR, folate receptor, G2 50, GAGE ​​family, Gastrin 17, Gastrin-releasing hormone, GD2 / GD3 / GM2, GnRH, GnTV, GP1, hCG, Heparinase, Her2 / neu, HMTV, Hsp70, hTERT, IGFR1, IL-13R, iNOS, Ki67, KIAA0205, K-ras, H-ras, N-ras, KSA, LKLR-FUT, MAGE family, Lactoglobulin, MAP 17, melanin-A / MART-1, mesothelin, MIC AB, MT-MMP, mucin, NY-ESO-1, osteonectin, P170 / MDR1, p53, p97 / melanin transferrin, PAI-1, PDGF, uPA, PRAME, probasin, progenipoientin, PSA, PSM, RAGE-1, Rb, RCAS1, SART-1, SSX-family, STAT3, STn, TAG-72, TGF-α, TGF-β, thymosin-β-15, TNF-α, TYRP-, TYRP-2, tyrosinase, VEGF, and glutathione-S-transferase.

[0232] In some embodiments, the vaccine is a DNA vaccine. The vector can be engineered to contain specific DNA that can be injected into a subject so that the DNA is taken up by cells. Once the cells take up the DNA, the DNA programs the cells to produce specific antigens, which can then stimulate the desired immune response.

[0233] In some embodiments, the vaccine consists of a recombinant virus encoding or expressing a cancer antigen. In some embodiments, the recombinant virus is a poxvirus, and more specifically an orthopoxvirus, such as, but not limited to, vaccinia virus, modified vaccinia Ankara virus, or MVA-BN, that expresses a tumor antigen.

[0234] In some embodiments, the vaccine composition comprises at least one antigen presenting cell group presenting the antigen selected. Antigen presenting cells (or stimulator cells) typically have class I or class II MHC molecules on their surface, and in one embodiment, are essentially unable to present the antigen selected to class I or class II MHC molecule loads. Preferably, the antigen presenting cell is a dendritic cell. Suitably, the dendritic cell is an autologous dendritic cell pulsed with a target antigen (e.g., a peptide). T cell therapy uses autologous dendritic cells, which are autologous dendritic cells pulsed with peptides from tumor-associated antigens disclosed in Murphy et al., (1996) The Prostate 29,371-380 and Tjua et al., (1997) The Prostate 32,272-278. Therefore, in some embodiments, the vaccine composition contains at least one antigen presenting cell pulsed or loaded with one or more antigenic peptides. Alternatively, the antigen presenting cell comprises an expression construct encoding an antigenic peptide. The polynucleotide may be any suitable polynucleotide and preferably is capable of transducing dendritic cells, resulting in peptide presentation and induction of an immune response.

[0235] In some embodiments, the vaccine composition comprises one or more adjuvants. Adjuvants are substances that non-specifically enhance or improve immune responses (e.g., immune responses to antigens mediated by CD8-positive T cells and helper T cells) and are therefore considered useful in the medicaments of the present invention.

[0236] In one specific embodiment, the patient is a cancer patient having a cancer selected from the group consisting of: melanoma; lymphoma, such as classical Hodgkin's lymphoma; glioma; urothelial carcinoma; renal cancer; squamous cell carcinoma of the head and neck, prostate cancer; breast cancer; colon cancer; and lung cancer; and combinations thereof. In another specific embodiment, the cancer is selected from the group consisting of: bone cancer, pancreatic cancer, skin cancer, head and neck cancer, malignant melanoma of the skin or eye, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, cancer of the endocrine system, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia, acute myeloid leukemia, leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors of childhood, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal pelvic cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal tumors, brain stem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, environmentally induced cancers (including those induced by asbestos), and combinations of such cancers.

[0237] According to the present invention, the present composition can be in any form that allows local treatment in the lumen of the ileum and / or large intestine of the patient when administered. The composition can be a solid dosage form, which is a pill, granule, microparticle, nanoparticle, mini tablet, capsule or tablet, etc. How to prepare a solid dosage form is well known in the art, for example, see "Aulton's Pharmaceutics: The Design and Manufacture of Medicines", Churchill Livingstone title, 4th revised edition, 2013 (ISBN: 978-0-7020-4290-4). Specific examples of methods for preparing solid dosage forms comprising antibodies and functional fragments or derivatives are included in those disclosed in any one of International Patent Applications PCT / EP2018 / 074521, PCT / EP2018 / 074520 and PCT / EP2018 / 074524.

[0238] According to one embodiment, the present composition comprises at least one additive. Suitable additive comprises hydrophilic polymer, filler, hydrophilic binder, disintegrating agent, antiadherent, surfactant, stabilizing agent, protease resistance reinforcing agent, plasticizer, coalescing agent, lubricant, buffer, acidulant and / or compounding agent. Suitable hydrophilic polymer, filler, hydrophilic binder, disintegrating agent, antiadherent, surfactant, plasticizer, coalescing agent, lubricant, buffer and / or acidulant are well known to those skilled in the art.

[0239] In one embodiment of the invention, the composition for local treatment in the ileum and / or large intestine is orally administered. Oral administration in the context of the present invention refers to introducing the composition into the gastrointestinal tract by mouth. In a preferred embodiment of the present invention, the composition is a solid dosage form, preferably a pill, granule, microparticle, nanoparticle, microtablet, sphere, capsule, tablet or multi-particulate drug delivery system form, which is coated with a delayed release coating. As used herein, the term "delayed release" refers to preventing the antibody or its functional fragment or derivative from releasing before the ileum, before the terminal ileum, before the ileocolic region or before the small intestine colon. The ileocolic region is the region where the small intestine merges with the large intestine in the gastrointestinal tract, i.e., the terminal ileum. In another embodiment, the composition is a delayed release solid dosage form, preferably a pill, granule, microparticle, nanoparticle, microtablet, sphere, capsule, tablet or multi-particulate drug delivery system form, which comprises a sustained release coating or a sustained release matrix. In general, as used herein, the term "sustained release" refers to time-dependent release, i.e., the release of the antibody or its functional fragment or derivative over an extended period of time, for example, over at least 6 h, preferably at least 8 h, at least 10 h, at least 12 h, at least 14 h, at least 16 h, at least 18 h or at least 24 h, etc.

[0240] In a preferred embodiment, the composition comprises a sustained release core and a delayed release coating. In this embodiment, the composition typically does not comprise a sustained release coating.

[0241] Preferably, the sustained release core comprises at least one sustained release agent selected from the group consisting of: 7,000,000, nonionic poly(ethylene oxide) polymers, HPMC type 2208 having a viscosity in water at 2 wt.-% at 20° C. of 3 to 100,000 mPa·s, preferably about 2,308 to 9,030 mPa·s, more preferably 2,663-4,970 mPa·s, xanthan gum, guar gum, tragacanth gum, locust bean gum, gum arabic, chitosan, carbomer, ethylcellulose, polyvinyl acetate, glyceryl (di)behenate, glyceryl palmitostearate, polymethacrylates such as poly(ethyl acrylate-methyl methacrylate-methacryloyloxyethyltrimethylammonium chloride) 1:2:0.1, poly(ethyl acrylate-methyl methacrylate-methacryloyloxyethyltrimethylammonium chloride) 1:2:0.2, or poly(ethyl acrylate-methyl methacrylate) 2:1, and combinations thereof.

[0242] In another preferred embodiment, the composition comprises an immediate release core, a sustained release coating and a delayed release coating. Typically, the sustained release coating is between the core and the delayed release coating.

[0243] In yet another embodiment, the composition comprises an immediate release core and a delayed release coating, but no sustained release core.

[0244] After oral administration, the delayed release of solid dosage forms, particularly for the coating material for targeted release in ileum or large intestine is well known in the art. It can be further divided into a coating material that disintegrates higher than a specific pH, a coating material that disintegrates in the gastrointestinal tract after a specific residence time, and a coating material that disintegrates due to the specific enzymatic triggering of the microflora in a specific intestinal region. The coating materials for targeting these three different categories of large intestine have been reviewed in (Polim.Med.2014,44,2,109-118) such as Bansal. For example, these purposes of such coating materials are also described in WO 2007 / 122374 A2, WO 01 / 76562 A1, WO 03 / 068196 A1, WO 2008 / 135090 A1 and GB 2367002 A. In one embodiment of the present invention, the delayed release coating comprises at least one component selected from the group consisting of coating materials that disintegrate pH-dependently, coating materials that disintegrate time-dependently, coating materials that disintegrate due to enzymatic triggering in the large intestine environment, and combinations thereof.

[0245] Preferred coating materials for pH-dependent disintegration are selected from the following: polyvinyl acetate phthalate, cellulose acetate trimellitate, hydroxypropyl methylcellulose phthalate HP-50, HP-55 or HP-55S, cellulose acetate phthalate, hydroxypropyl methylcellulose acetate succinate (HPMCAS), poly (methacrylic acid-ethyl acrylate) 1:1 ( L100-55, L30D-55), poly (methacrylic acid-methyl methacrylate) 1:1 ( L-100, L12.5), poly (methacrylic acid-methyl methacrylate) 1:2 ( S-100, S12,5, FS30D) and combinations thereof. Preferred coating materials for time-dependent disintegration are selected from the following: poly(ethyl acrylate-methyl methacrylate) 2:1 (e.g., NM30D or Eudragit NE 30D), poly(ethyl acrylate-methyl methacrylate-methacrylic acid) 7:3:1 (e.g., RS 30D), ethyl cellulose (e.g. or Aquacoat ECD), poly(ethyl acrylate-methyl methacrylate-methacryloyloxyethyltrimethylammonium chloride) 1:2:0.1 (e.g., RS 30D), polyvinyl acetate (e.g. SR 30D) and combinations thereof. Preferred coating materials that disintegrate due to enzymatic triggering in the intestinal environment are selected from the group consisting of hemicellulose, chondroitin sulfate, cyclodextrin, pectin, guar gum, chitosan, inulin, lactulose, raffinose, stachyose, alginate, dextran, xanthan gum, locust bean gum, arabinogalactan, amylose, pullulan, carrageenan, scleroglucan, chitin, curdulan, levan, pullulan, starch, resistant starch, azo compounds degraded by azo bond cleavage bacteria, and combinations thereof. The delayed-release coating material optionally comprises one or more other excipients.

[0246] In one embodiment of the present invention, the coating material used for the delayed-release coating comprises one, two, three, etc., components selected from the group consisting of the pH-dependent disintegrating coating materials listed above, the time-dependent disintegrating coating materials, the coating materials that disintegrate due to enzymatic triggering in the large intestine environment, and combinations thereof. In another embodiment of the present invention, the delayed-release coating comprises a combination of at least one coating material that disintegrates pH-dependently and at least one coating material that disintegrates due to enzymatic triggering in the large intestine environment.

[0247] For example, the delayed-release coating material can be designed to focus on delivering the composition comprising the antibody or its functional fragment or derivative intact to the large intestine, starting in the cecum and continuing through the ascending, transverse and descending colons, and ending in the sigmoid colon. Alternatively, for example, the delayed-release coating can be designed to start delivering the antibody or its functional fragment or derivative in the ileum and ultimately release in the transverse colon. There are many possibilities and combinations.

[0248] In a different embodiment of the present invention, the composition for local treatment in the ileum and / or large intestine is administered rectally. Rectal administration in the context of the present invention refers to the introduction of the composition into the gastrointestinal tract via the anus. In a preferred embodiment of the present invention, the composition is administered in the form of an enema, gel, foam or suppository.

[0249] Those skilled in the art can readily determine the effective amount of the TNFα-specific antibody or its functional fragment or derivative to be administered as part of the composition for use in the present invention. In general, the effective amount of the TNFα-specific antibody or its functional fragment or derivative according to the present invention is the minimum amount required to produce a therapeutic effect (i.e., to treat diarrhea, colitis, or enterocolitis induced by one or more ICP inhibitors). The precise amount of the TNFα-specific antibody or its functional fragment or variant administered to the patient can vary depending on the state and severity of the condition and the patient's physical condition.

[0250] According to one embodiment of the present invention, the compositions of the present invention provide a therapeutically effective amount of a TNFα-specific antibody or a functional fragment or derivative thereof in the lumen of the patient's ileum and / or large intestine, thereby preventing, minimizing, and / or delaying or alleviating symptoms associated with diarrhea, colitis, or enterocolitis caused by ICP inhibitors. A "therapeutically effective amount" is the amount of at least one TNFα-specific antibody or a functional fragment or derivative thereof required to provide the desired therapeutic effect. The precise amount may vary for different antibodies or functional fragments or derivatives thereof and / or for individual patients, but can be determined by those skilled in the art.

[0251] For example, a therapeutically effective amount can be achieved in the lumen of the ileum and / or large intestine by targeting a composition of the present invention comprising a TNFα antibody or a functional fragment or derivative thereof. The manner and means of targeting the composition of the present invention to the lumen of the ileum and large intestine are not particularly limited and can be achieved by methods known in the art. These methods include taking advantage of inherent processes in the gastrointestinal tract that result in, for example, differences in pH and microflora, and the specific residence time of ingested material in different parts of the gastrointestinal tract. Methods for collecting a certain concentration of a specific protein (including a specific antibody) in the intestinal lumen are known in the art. For example, a sample can be collected from discharged feces or using a flexible tube inserted through the anus. The concentration of the TNFα antibody is then determined using an ELISA or Western blot or other immunochemical technique similar to that described by Nicholls et al. (J Clin Pathol. 1993 Aug; 46(8): 757–760) for measuring fecal TNFα concentration using an antibody specific for the TNFα antibody used in the composition or a functional fragment or derivative thereof.

[0252] One unit dose of the composition of the invention can contain, for example, an amount of active agent ranging from about 0.05 mg to about 1,000 mg, 0.1 mg to about 200 mg, or from about 1 mg to about 100 mg, or from about 10 mg to about 50 mg.

[0253] The composition comprising the TNFα-specific antibody or its functional fragment or derivative according to the present invention can be administered to the patient, for example, once a day, twice a day or three times a day. In a preferred embodiment, the composition is administered once a day.

[0254] SEQ ID NO: Description of amino acid sequences 1 Light chain of humanized anti-TNFα antibody (= light chain of Ab-REW) 2 Heavy chain of humanized anti-TNFα antibody 3 CDR L1 of antibody Ab-REW 4 CDR L2 of antibody Ab-REW 5 CDR L3 of antibody Ab-REW 6 CDR H1 of antibody Ab-REW 7 CDR H2 of antibody Ab-REW 8 CDR H3 of antibody Ab-REW 9 <![CDATA[V of antibody Ab-REW H > 10 <![CDATA[V of antibody Ab-REW L > 11 Heavy chain of antibody Ab-REW 12 Light chain of surrogate antibody cV1q-huFc 13 Heavy chain of surrogate antibody cV1q-huFc 14 Antibody Ab-REW modified heavy chain 15 Heavy chain of antibody Ab-AA 16 Antibody Ab-AA modified heavy chain

[0255] Example

[0256] Example 1 and Example 2

[0257] Materials and Methods in the Examples

[0258] Preparation of Citrate-TRIS Buffer pH 7: Prepare 100 mM sodium citrate solution (2.942 g and make up to 100.0 mL with purified water). Prepare 100 mM citric acid solution (dissolve 3.842 g and dilute to 200.0 ml with purified water). Adjust the pH of the citric acid solution to 3.5 with sodium citrate solution. Prepare 1 M TRIS solution (12.114 g and make up to 100.0 ml with purified water). Adjust the pH of the citrate buffer to pH 7.0 with TRIS solution.

[0259] Preparation of pellets

[0260] Preparation components (pellet core)

[0261]

[0262] *Relative to the dry mixture before addition of adalimumab

[0263] Preparation steps:

[0264] Dry Blending: The excipients required for each batch (batch size: 10 g) were mixed using the mixer attachment (twin-blade mixer) from a Caleva Multilab apparatus at 50 rpm for a predetermined period of approximately 5 minutes.

[0265] Wet Blending: Following the dry blend step, the adalimumab solution containing 0.1% w / v polysorbate 20 was slowly added to the powder blend of excipients with mixing and mixed at 50 rpm for 10 minutes.

[0266] Extrusion: The wet mass was then discharged from the mixer and extruded through a 1 mm diameter and 1 mm depth hole of an extrusion die using a screw extruder at a constant speed (150 rpm) until all the wet mass was extruded.

[0267] Spheroidization: The wet extrudate is then fed into a pelletizer attachment consisting of a slotted plate which, by rotation, breaks the wet extrudate into smaller fragments which then round off (wet spheres) depending on time, speed, and the properties of the individual components of the extrudate. The extrudate is pelletized at 1500 rpm for a predetermined amount of time.

[0268] The wet pellets obtained from the spheronization step were then collected into disposable weigh boats and dried in an oven at 40°C overnight.

[0269] Delayed (enteric) release coating

[0270] Use the enteric coating containing Eudragit L30D-55 to coat the adalimumab pellets (from Example 1 and Example 2). Prepare the coating suspension by blending the required amount of Eudragit L30D-55 dispersion and glyceryl monostearate (GMS) emulsion. Prepare the GMS emulsion by dissolving polysorbate 80 in water and then adding GMS. Then, the mixture is heated to 75 ° C and kept at this temperature and under continuous magnetic stirring for 15 minutes. Then, the cooled GMS emulsion is added to Eudragit L30D-55, followed by triethyl citrate (plasticizer). The suspension is stirred for 30 minutes before coating.

[0271] Subsequently, the coating suspension was sprayed onto the pellets containing adalimumab to a final polymer weight gain of 30%. Coating was performed using a MiniGlatt fluidized bed coater equipped with a microkit (bottom spray) to allow for smaller batches of coating. Eudragit L30D-55 suspension was sprayed under the following conditions: inlet temperature: 40°C, product temperature: 33.0–34.5°C, air flow: 24–28 m 3 / h, and atomizing air pressure: 0.2-0.3 bar.

[0272] Dissolution of adalimumab from pellets

[0273] A certain amount of loaded adalimumab pellets were placed in 5 mL cryovials and 4.0 ml of buffer was added to produce a nominal adalimumab concentration of 1 mg / ml (based on the calculated theoretical adalimumab loading). Citrate-TRIS buffer pH 7 was used as buffer. During the entire duration of the experiment, the sample was agitated. Supernatant samples were collected at predetermined time points, centrifuged, and the total protein content of the supernatant was analyzed. In the case of enteric coated pellets, the pellets were first exposed to 0.1 N HCl for 2 hours under continuous agitation, the fluid was then taken out, and then pH 7.0 citrate-TRIS buffer was added in the buffering phase as described above.

[0274] Total protein content quantification (Bradford): Total protein was quantified by colorimetry using Coomassie Plus assay (Thermo Fisher Scientific) according to the Bradford method. Briefly, 6.6 μl of sample was pipetted to the bottom of a 96-well plate and 200 μl of Coomassie Plus reagent was added, and the mixture was stirred at 500 rpm for 30 s. The sample was then incubated at room temperature for 10 min, and the absorbance at 595 nm was recorded (Tecan microplate reader) and background was subtracted. Quantification was performed using a freshly prepared standard curve.

[0275] result

[0276] Release from adalimumab immediate- and sustained-release pellet cores

[0277] The uncoated pellets of Example 1 disintegrated rapidly in the citrate-TRIS pH 7.0 buffer, resulting in a rapid and almost complete release of adalimumab within 1 hour. On the other hand, adalimumab was continuously released from the uncoated pellets of Example 2 for the duration of the experiment.

[0278]

[0279] Release from Adalimumab Delayed (Enteric-Coated) Pellets Cores

[0280] Two batches of adalimumab pellets were coated with Eudragit L30D-55 dispersion. After 2 hours in 0.1N HCl, the coated pellets were completely acid-resistant (no adalimumab release). After exchanging to citrate-TRIS pH 7.0 buffer (to simulate the pH of the small intestine), adalimumab began to be released (release ≥ 5%).

[0281] Example 3

[0282] The therapeutic efficacy of anti-TNFα antibodies after topical administration was studied in an IBD mouse model. Due to the species specificity of Ab-REW, which does not bind to mouse TNFα, a surrogate antibody (cV1q) consisting of the variable domains of the cV1q antibody (Echtenacher et al., 1990) and the constant region of mouse IgG2a was used. The T cell transfer model was chosen as the gene expression pattern in this model most closely reflects the altered gene expression in IBD (if compared with DSS- and TNBS-induced colitis) (teVelde et al., Inflamm Bowel Dis 2007; 13(3): 325-30). The clinical characteristics of this model are progressive weight loss and soft stools. Histopathologically, inflammation reaches the rectum from the cecum and is accompanied by macrophage infiltration (with a moderate number of activated CD4+ lymphocytes), mucin depletion, and epithelial hyperplasia, resulting in glandular elongation and mucosal thickening. Increased gene expression of TNF-α, IFN-γ, IL-6, CCR1, CCR2, CCR5, CXC chemokine receptor 3 and their ligands was described (Nagaoka and Radi, Front Biosci 2012 Jun 1;4:1295-314).

[0283] Colitis was induced by adoptive transfer of naive CD4+ T cells from WT mice into immunodeficient mice to induce transmural colitis in the recipients. Briefly, T cells (CD44 - / CD62L + ) and purified, and 0.5 x 10 6 cells were injected intraperitoneally into RAG2 - / - T cell engraftment was confirmed on day 20 in mice.

[0284] Treatment was performed using a rat / mouse chimeric monoclonal antibody of the IgG2a isotype (the mouse homologue of human IgG1) specific for mouse TNFα (Echtenacher et al., J Immunol 1990; 145:3762-66).

[0285] Mice were divided into 4 groups (n=5 for naive mice; n=12 for all other groups) as follows:

[0286] Group 1: Naive mice.

[0287] Group 2 received vehicle intrarectally.

[0288] Group 3 received IgG2a intrarectally.

[0289] Group 4 received intraperitoneal IgG2a as a positive control.

[0290] When colitis had been established in the animals and persisted for 28 days, anti-TNFα treatment was started on day 21 after T cell transfer. Animals receiving rectal administration (300 μg / d IgG2a or 200 μl vehicle) were treated once daily, while animals receiving intraperitoneal injections of IgG2a were treated twice weekly. Endpoints were body weight, endoscopic scores (days 14-49), histological scores (day 49), and cytokines in the colon (day 49).

[0291] result:

[0292] Mice in the naive and IgG2a (ip, ir) treated groups maintained or continued to gain weight, whereas mice in the vehicle treated group continued to lose weight ( Figure 1 ).

[0293] Significant reductions in endoscopic scores and reductions in histological scores were observed in the IgG2a treatment groups (ir and ip); see Figure 2 and Figure 3 During endoscopy, colitis was scored on a scale of 0-4 (0: normal; 1: loss of vascularity; 2: loss of vascularity and friability; 3: friability and erosion; 4: ulceration and bleeding). The overall histological score is the sum of the individual scores for inflammation, edema, goblet cell depletion, and epithelial damage (each 0-4).

[0294] Intraperitoneal treatment with IgG2a resulted in the greatest reduction of cytokines in the colon, followed by intrarectal treatment with IgG2a. Figure 4 The values ​​given are the mean of data from the proximal, middle, and distal colon.

[0295] In conclusion, local treatment was effective in a mouse T cell transfer model as shown by reduced endoscopic scores, reduced histological scores, decreased cytokines in the colon, and reduced weight loss.

[0296] Example 4

[0297] Dose-response study of colitis treatment in a colitis mouse model

[0298] In a dose-response study, a therapeutically effective dose of a rectal cV1q-huFc surrogate antibody was evaluated in a T cell transfer IBD mouse model using Tg32-SCID mice (human FcRn transgenic mice). Due to the species specificity of Ab-REW, which does not bind to mouse TNFα, a surrogate antibody (cV1q-huFc) consisting of the variable domains of the cV1q antibody (Echtenacher et al., 1990) and the Ab-REW constant region was used. The light and heavy chain sequences of the surrogate antibody are shown in SEQ ID NOs: 12 and 13, respectively. Colitis was induced by adoptive transfer of naive CD4+ T cells from C57Bl / 6 mice into Tg32-SCID mice. Treatment with cV1q-huFc was started on day 14 after adoptive transfer and continued until day 42. Mice were administered cV1q-huFc intrarectally daily (10, 30, 100, and 300 μg / mouse), intrarectally daily (300 μg / mouse), or intraperitoneally twice weekly at 10 mg / kg (control). Following rectal treatment, dose-dependent improvements in the total histological score (comprised of colonic inflammation (submucosal and muscularis / serosa), crypt damage, erosions, hyperplasia, and edema) were observed in both the proximal and distal colon. Figure 5 In addition, rectal treatment can reduce histological scores. In addition, a decrease in colonic cytokines IL-6, IL-17A, KC, TNFα, and MIP-2 was observed after rectal and cecal administration ( Figure 6 ), which was more pronounced in the case of IL-17A, KC and MIP-2 after intracecal administration. In conclusion, local administration of TNFα resulted in a reduction of inflammation in the colon.

[0299] Example 5

[0300] Systemic exposure after rectal administration

[0301] Studies were conducted in healthy and colitis-affected mice to determine the systemic exposure of cV1q-huFc following rectal administration. Colitis was induced by exposing Tg32-SCID mice to 3% DSS in drinking water from day 0 to day 5. Healthy and diseased mice were rectally administered a single dose (SD) or multiple doses (MD; 0.1 or 0.3 mg) of cV1q-huFc. Due to limitations in blood collection, only three blood samples could be drawn from each mouse (left eye, right eye, and terminal). Therefore, for blood collection, each treatment group (n = 10-14 mice) was divided into two subgroups (n = 5-7 mice). Single-dose animals received cV1q-huFc antibody on day 10 after colitis was established, and blood samples were collected 1, 4, and 24 hours (Subgroup 1) and 2, 8, and 48 hours (Subgroup 2) after dosing. Mice that received multiple doses of cV1q-huFc on days 10-14 received daily rectal administration, and blood samples were collected on day 10 at 24, 72, and 120 h (subgroup 1) and 48, 96, and 144 h (subgroup 2) after the first dose of cV1q-huFc, always just before the next dose. In parallel, two groups (healthy and diseased) received a single intravenous injection of 5 mg / kg cV1q-huFc. Blood samples were collected as in mice that received a single dose of rectal treatment. Immuno-PCR analysis of plasma samples.

[0302] Systemic exposure is very low after rectal administration compared to intravenous administration ( Figure 7 Colitis animals had slightly, but not significantly, higher levels of leukemia than healthy animals. A single rectal dose of cV1q-huFc resulted in a calculated systemic bioavailability of <0.01% when comparing dose-normalized exposure (AUC). After 5 daily doses, no systemic accumulation was observed over time.

[0303] Example 6

[0304] Systemic exposure and tissue concentrations after intracecal administration

[0305] cV1q-huFc tissue concentrations and systemic exposure were measured in healthy mice and mice with colitis after intracecal administration. Male Tg32-SCID mice underwent surgery, where a cannula was placed from the cecum to the back between the shoulder blades, the cannula was exposed, and the tip was fixed in place using sutures, wound clips, and tissue glue. Animals received buprenorphine (for 3 days) and Baytril (for 5 days) after surgery and were allowed to recover for 2-4 weeks before being assigned to the study. Colitis was induced by exposing mice to drinking water containing 3% DSS from day 0 to day 5. Healthy mice and diseased mice were treated with a single dose (n=16 / group) or multiple doses (n=8 / group) of cV1q-huFc. Healthy mice received a dose of 0.3 mg / administration, while two dose levels (0.1 and 0.3 mg / administration) were tested in colitis mice. When dosing the animals, 0.1 mL of cV1q-huFc was applied through the cannula, followed by a 0.05 mL saline flush to prevent any drug substance from being retained in the cannula and reaching the cecum. Four mice were sacrificed at each time point, and colon and blood (plasma) were collected. Single-dose animals received cV1q-huFc antibody on day 10 after establishment of colitis, and animals were sacrificed 1, 4, 8, and 12 h after dosing. Mice receiving multiple doses of cV1q-huFc received daily intracecal administration on days 10-14 and were sacrificed 24 and 120 h after the first dose of cV1q-huFc on day 10. In parallel, two groups with induced colitis received single or multiple rectal administrations at 0.3 mg / application. Blood (plasma) samples were collected as for animals treated intracecally. Plasma samples were analyzed by Immuno-PCR. At sacrifice, the colon was excised, flushed, weighed, and trimmed to 5 cm in length. 1 cm sections from the proximal and distal ends were snap-frozen for ELISA analysis.

[0306] Plasma concentrations measured after intracecal administration were highly variable, as was observed after rectal administration. Systemic exposure (AUC) increased approximately 60-fold (single dose) or 200-fold (multiple doses) after intracecal administration compared to rectal administration, however, overall systemic exposure remained very low ( Figure 8 After 5 daily doses, mild accumulation of cV1q-huFc in plasma was observed in most groups. Similar to plasma concentrations, cV1q-huFc levels in tissues were highly variable across groups and time points. Both intracecal and intrarectal administration routes produced measurable antibody concentrations in the proximal and distal colon ( Figure 9In contrast to the significant differences in systemic exposure observed between rectal and cecal treatments, tissue levels were comparable for both routes of administration. No significant differences were observed between healthy and diseased animals. Repeated daily administration for 5 consecutive days resulted in mild accumulation of cV1q-huFc in the proximal and distal colon in most groups.

[0307] Example 7

[0308] Nivolumab-stimulated human CD4 + The impact of T cells

[0309] Immune checkpoint inhibitors targeting PDF-1 or CTLA-4 are very effective cancer treatments. However, they are associated with some serious immune-related adverse events, such as colitis. Therefore, we investigated the effect of Ab-REW on nivolumab-stimulated human CD4 + Effects of nivolumab on T cells. It was shown that nivolumab removed CD4 + T cells induced higher levels of IFNγ secretion. Co-incubation with nivolumab and Ab-REW or infliximab inhibited IFNγ secretion in a dose-dependent manner, but significant variability was observed between different donor pairs and runs ( Figure 10 ). However, this trend remained consistent across runs and donor pairs, suggesting that this is a true biological phenomenon. Donor pairs capable of secreting higher levels of IFNγ did not appear to have as robust a response as donor pairs showing weaker IFNγ secretion, suggesting that donor variability and the strength of the initial stimulation may affect the effectiveness of anti-TNFα agents in this setting. Interestingly, the IC of Ab-REW 50 The values ​​(range 0.028–0.266 μg / mL) were generally lower than those of infliximab (range 0.068–0.326 μg / mL), suggesting that Ab-REW may be a more potent inhibitor of nivolumab-mediated IFNγ secretion than infliximab. Sequence Listing <110> Tilotes Pharmaceuticals Co., Ltd. <120> Topical treatment of immune checkpoint inhibitor-induced diarrhea, colitis, or enterocolitis using antibodies and their fragments <130> P010-P015 <150> PCT / EP2018 / 084057 <151> 2018-12-07 <160> 16 <170> PatentIn version 3.5 <210> 1 <211> 221 <212> PRT <213> artificial sequence <220> <223> Ab-REW's chain link <400> 1 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Lys Cys Gln Ala Ser Gln Ser Ile Phe Ser Gly 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Gly Leu Gln Pro 65 70 75 80 Ala Asp Phe Ala Thr Tyr Tyr Cys Gln Ser Tyr Tyr Tyr Ser Ser Ser 85 90 95 Ser Ser Asp Gly Ser Tyr Ala Phe Gly Gly Gly Thr Lys Leu Thr Val 100 105 110 Leu Gly Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser 115 120 125 Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn 130 135 140 Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala 145 150 155 160 Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys 165 170 175 Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp 180 185 190 Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu 195 200 205 Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 220 <210> 2 <211> 452 <212> PRT <213> Artificial sequence <220> <223> Heavy chain of humanized anti-TNFα antibody <220> <221> MISC_FEATURE <222> (452)..(452) <223> Xaa is lysine or missing (absent) <400> 2 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Lys Ala Ser Gly Ile Asp Phe Asn Asn Tyr 20 25 30 Gly Ile Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Thr Tyr Ile Tyr Pro Gly Phe Ala Ile Thr Asn Phe Ala Asn Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Pro Val Tyr Ala Thr Ser Ser Gly Tyr Phe Asp Leu Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Xaa 450 <210> 3 <211> 11 <212> PRT <213> artificial sequence <220> <223> Antibody Ab-REW's CDR L1 <400> 3 Gln Ala Ser Gln Ser Ile Phe Ser Gly Leu Ala 1 5 10 <210> 4 <211> 7 <212> PRT <213> artificial sequence <220> <223> CDR L2 of antibody Ab-REW <400> 4 Gly Ala Ser Lys Leu Ala Ser 1 5 <210> 5 <211> 15 <212> PRT <213> Artificial sequence <220> <223> CDR L3 of antibody Ab-REW <400> 5 Gln Ser Tyr Tyr Tyr Ser Ser Ser Ser Ser Asp Gly Ser Tyr Ala 1 5 10 15 <210> 6 <211> 10 <212> PRT <213> Artificial sequence <220> <223> CDR H1 of antibody Ab-REW <400> 6 Gly Ile Asp Phe Asn Asn Tyr Gly Ile Gly 1 5 10 <210> 7 <211> 17 <212> PRT <213> Artificial sequence <220> <223> CDR H2 of antibody Ab-REW <400> 7 Tyr Ile Tyr Pro Gly Phe Ala Ile Thr Asn Phe Ala Asn Ser Val Lys 1 5 10 15 Gly <210> 8 <211> 13 <212> PRT <213> Artificial sequence <220> <223> CDR H3 of antibody Ab-REW <400> 8 Asp Pro Val Tyr Ala Thr Ser Ser Gly Tyr Phe Asp Leu 1 5 10 <210> 9 <211> 122 <212> PRT <213> Synthetic Sequence <220> <223> VH of antibody Ab-REW <400> 9 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Lys Ala Ser Gly Ile Asp Phe Asn Asn Tyr 20 25 30 Gly Ile Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Thr Tyr Ile Tyr Pro Gly Phe Ala Ile Thr Asn Phe Ala Asn Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Pro Val Tyr Ala Thr Ser Ser Gly Tyr Phe Asp Leu Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 10 <211> 114 <212> PRT <213> Artificial sequence <220> <223> VL of antibody Ab-REW <400> 10 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Lys Cys Gln Ala Ser Gln Ser Ile Phe Ser Gly 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Gly Leu Gln Pro 65 70 75 80 Ala Asp Phe Ala Thr Tyr Tyr Cys Gln Ser Tyr Tyr Tyr Ser Ser Ser 85 90 95[[ID=三十九]] Ser Ser Asp Gly Ser Tyr Ala Phe Gly Gly Gly Thr Lys Leu Thr Val 100 105 110 T Leu Gly <210> 11 <211> 451 <212> PRT <213> Artificial sequence <220> It should be noted that there may be some inaccuracies in the translation due to the unclear nature of some of the original text. For example, the "三十九" in the original might be an error, and it's translated as such in case it's a placeholder or an unknown entity in the specific context. If possible, it would be beneficial to have more context or clarify the original text for a more accurate translation.<223> Heavy chain of antibody Ab-REW <400> 11 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Lys Ala Ser Gly Ile Asp Phe Asn Asn Tyr 20 25 30 Gly Ile Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Thr Tyr Ile Tyr Pro Gly Phe Ala Ile Thr Asn Phe Ala Asn Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Pro Val Tyr Ala Thr Ser Ser Gly Tyr Phe Asp Leu Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Arg Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Glu His Glu Ala Leu His Trp His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly 450 <210> 12 <211> 233 <212> PRT <213> artificial sequence <220> <223> Antibody cV1q-huFc's chain link <400> 12 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Asn Ile Gln Leu Thr Gln Ser Pro Ser Leu Leu Ser 20 25 30 Ala Ser Val Gly Asp Arg Val Thr Leu Ser Cys Lys Gly Ser Gln Asn 35 40 45 Ile Asn Asn Phe Leu Ala Trp Tyr Gln Gln Glu Leu Gly Glu Ala Pro 50 55 60 Lys Leu Leu Ile Tyr Asn Thr Asn Ser Leu Gln Thr Gly Ile Pro Ser 65 70 75 80 Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser 85 90 95 Ser Leu Gln Pro Glu Asp Val Ala Thr Tyr Phe Cys Tyr Gln Tyr Asn 100 105 110 Asn Gly Asn Thr Phe Gly Val Gly Thr Lys Leu Glu Leu Lys Arg Thr 115 120 125 Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu 130 135 140 Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro 145 150 155 160 Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly 165 170 175 Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr 180 185 190 Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His 195 200 205 Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val 210 215 220 Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 13 <211> 467 <212> PRT <213> Artificial sequence <220> <223> Heavy chain of antibody cV1q - huFc <400> 13 Met Glu Trp Ser Trp Val Phe Leu Phe Phe Leu Ser Val Thr Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Lys Glu Ser Gly Pro Gly Leu Val Gln<​​​​35 40 45 Thr Ser Tyr Asn Val His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu 50 55 60 Glu Trp Met Gly Arg Met Arg Tyr Asn Gly Asp Thr Ser Tyr Asn Ser 65 70 75 80 Ala Leu Lys Ser Arg Leu Ser Ile Ser Arg Asp Thr Ser Lys Asn Gln 85 90 95 Val Phe Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Gly Thr Tyr 100 105 110 Tyr Cys Thr Arg Asp Arg Phe Ser Trp Ala Ser Tyr Phe Asp Tyr Trp 115 120 125 Gly Gln Gly Val Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 130 135 140 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 145 150 155 160 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 165 170 175 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 180 185 190 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 195 200 205 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 210 215 220 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 225 230 235 240 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 245 250 255 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 260 265 270 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 275 280 285 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 290 295 300 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 305 310 315 320 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Arg Asp Trp Leu Asn 325 330 335 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 340 345 350 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 355 360 365 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 370 375 380 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 385 390 395 400 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 405 410 415 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 420 425 430 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 435 440 445 Glu His Glu Ala Leu His Trp His Tyr Thr Gln Lys Ser Leu Ser Leu 450 455 460 Ser Pro Gly 465 <210> 14 <211> 452 <212> PRT <213> Synthetic Sequence <220> <223> Heavy chain of a variant of antibody Ab-REW <400> 14 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Lys Ala Ser Gly Ile Asp Phe Asn Asn Tyr 20 25 30 Gly Ile Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Thr Tyr Ile Tyr Pro Gly Phe Ala Ile Thr Asn Phe Ala Asn Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Pro Val Tyr Ala Thr Ser Ser Gly Tyr Phe Asp Leu Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Arg Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Glu His Glu Ala Leu His Trp His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Lys 450 <210> 15 <211> 451 <212> PRT <213> Artificial Sequence <220> <223> Heavy chain of antibody Ab-AA <400> 15 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Lys Ala Ser Gly Ile Asp Phe Asn Asn Tyr 20 25 30 Gly Ile Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Thr Tyr Ile Tyr Pro Gly Phe Ala Ile Thr Asn Phe Ala Asn Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Pro Val Tyr Ala Thr Ser Ser Gly Tyr Phe Asp Leu Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Ala Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly 450 <210> 16 <211> 452 <212> PRT <213> Artificial sequence <220> <223> Heavy chain of modified antibody Ab-AA <400> 16 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Lys Ala Ser Gly Ile Asp Phe Asn Asn Tyr 20 25 30 Gly Ile Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Thr Tyr Ile Tyr Pro Gly Phe Ala Ile Thr Asn Phe Ala Asn Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Pro Val Tyr Ala Thr Ser Ser Gly Tyr Phe Asp Leu Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Ala Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Lys 450

Claims

1. A pharmaceutical composition comprising an active agent selected from antibodies specific for tumor necrosis factor alpha (TNFα) and functional fragments and derivatives thereof for the preparation of a medicament for treating or preventing at least one gastrointestinal adverse event induced by cancer therapy and induced by one or more immune checkpoint (ICP) inhibitors in a patient, wherein the anti-TNFα antibody is Ab-REW antibody having CDR L1 of SEQ ID NO: 3, CDR L2 of SEQ ID NO: 4, and CDR L3 of SEQ ID NO: 5, and CDR H1 of SEQ ID NO: 6, CDR H2 of SEQ ID NO: 7, and CDR H3 of SEQ ID NO: 8, wherein the medicament is suitable for local administration to an affected area of ​​the gastrointestinal tract of the patient, and the medicament allows for an interruption of treatment with one or more ICP inhibitors for less than 4 weeks, wherein the patient is a cancer patient, and wherein the functional fragments and derivatives thereof are antibody Fab fragments, F(ab')2 fragments, Fab' fragments, scFvs and dsFvs that retain antigen-binding properties.

2. Use of a pharmaceutical composition according to claim 1, wherein the medicament is suitable for administration to the ileum and / or large intestine of the patient.

3. Use of a pharmaceutical composition according to claim 1, wherein the interruption of treatment with the one or more ICP inhibitors is less than 2 weeks.

4. Use of a pharmaceutical composition according to claim 3, wherein the treatment with the one or more ICP inhibitors is interrupted for less than 1 week.

5. Use of a pharmaceutical composition according to claim 3, wherein the interruption of treatment with the one or more ICP inhibitors is less than 5 days.

6. Use of a pharmaceutical composition according to claim 3, wherein the interruption of treatment with the one or more ICP inhibitors is less than 4 days.

7. Use of a pharmaceutical composition according to claim 3, wherein the interruption of treatment with the one or more ICP inhibitors is less than 3 days.

8. Use of a pharmaceutical composition according to claim 3, wherein the treatment with the one or more ICP inhibitors is interrupted for less than 2 days.

9. Use of a pharmaceutical composition according to claim 3, wherein the interruption of treatment with the one or more ICP inhibitors is less than 1 day.

10. Use of a pharmaceutical composition according to claim 3, wherein the treatment is not interrupted.

11. Use of the pharmaceutical composition according to claim 1, wherein the adverse event is selected from the group consisting of diarrhea induced by an ICP inhibitor, colitis induced by an ICP inhibitor, enterocolitis induced by an ICP inhibitor, and combinations thereof.

12. Use of a pharmaceutical composition according to claim 11, wherein the ICP inhibitor-induced diarrhea, colitis and / or enterocolitis is characterized by grade 1 toxicity or grade 2 toxicity.

13. Use of a pharmaceutical composition according to claim 11, wherein the medicament is suitable for preventing the ICP inhibitor-induced diarrhea, colitis and / or enterocolitis from developing into higher-grade toxicity.

14. Use of a pharmaceutical composition according to claim 1, wherein the medicament is suitable for prophylactic therapy against ICP inhibitor-induced diarrhea, colitis and / or enterocolitis.

15. Use of the pharmaceutical composition according to claim 14, wherein the patient is concurrently undergoing treatment with one or more ICP inhibitors.

16. Use of a pharmaceutical composition according to claim 1, wherein the patient suffers from ICP inhibitor-induced diarrhea but not from ICP inhibitor-induced colitis or ICP inhibitor-induced enterocolitis, and wherein the medicament is for preventing the progression or onset of ICP inhibitor-induced colitis and / or ICP inhibitor-induced enterocolitis in the patient.

17. Use of the pharmaceutical composition according to claim 16, wherein the patient suffers from ICP inhibitor-induced diarrhea of ​​grade 2 toxicity.

18. Use of the pharmaceutical composition according to claim 16, wherein the patient suffers from ICP inhibitor-induced diarrhea of ​​grade 1 toxicity.

19. The use of a pharmaceutical composition according to claim 1, wherein the one or more ICP inhibitors are selected from the group consisting of an antibody specific for cytotoxic T lymphocyte-associated protein 4 (CTLA-4), an antibody specific for programmed cell death protein 1 (PD-1), and an antibody specific for programmed death ligand 1 (PD-L1).

20. The use of a pharmaceutical composition according to claim 19, wherein the antibody specific for PD-1 is selected from the following: pembrolizumab and nivolumab; the antibody specific for PD-L1 is selected from the following: atezolizumab, avelumab, and durvalumab; or the antibody specific for CTLA-4 is selected from the following: ipilimumab and tremelimumab.

21. The use of the pharmaceutical composition according to claim 1, wherein the amino acid sequence of the antibody specific for TNFα and its functional fragment comprises i) amino acids 233P, 234V, 235A, and a deletion at amino acid position 236; and amino acid 434A or amino acids 252Y, 254T, and 256E; and optionally amino acids 239D, 330L, and 332E or amino acids 326A, 332E, and 333A; and / or ii) amino acids 380A and 434A, and optionally amino acid 307T; and / or iii) amino acid 434W, and optionally amino acid 428E and / or amino acid 311R, The amino acid numbers refer to EU numbers.

22. Use of a pharmaceutical composition according to any preceding claim, wherein said topical administration comprises or consists of oral administration of said drug.

23. Use of a pharmaceutical composition according to claim 22, wherein the drug is a delayed release formulation.

24. Use of a pharmaceutical composition according to claim 23, wherein the drug is a solid dosage form in the form of pellets, granules, microparticles, nanoparticles, minitablets, spheres, capsules, tablets or multiparticulate drug delivery systems, which is coated with a delayed release coating that prevents the release of the active agent prior to entering the ileum, ileocolic region or large intestine of the gastrointestinal (GI) tract.

25. The use of a pharmaceutical composition according to claim 24, wherein the delayed-release coating comprises at least one component selected from the group consisting of polyvinyl acetate phthalate, cellulose acetate trimellitate, hydroxypropylmethylcellulose phthalate HP-50, HP-55 or HP-55S, cellulose acetate phthalate, hydroxypropylmethylcellulose acetate succinate (HPMCAS), poly(methacrylic acid-ethyl acrylate) 1:1, poly(methacrylic acid-methyl methacrylate) 1:1 , poly(methacrylic acid-methyl methacrylate) 1:2, chondroitin sulfate, pectin, guar gum, chitosan, inulin, lactulose, raffinose, stachyose, alginate, dextran, xanthan gum, locust bean gum, arabinogalactan, amylose, cyclodextrin, pullulan, carrageenan, scleroglucan, chitin, curdulan, levan, pullulan, starch, resistant starch, azo compounds degraded by azo bond cleavage bacteria, and combinations thereof.

26. Use of a pharmaceutical composition according to claim 22, comprising a sustained release coating or a sustained release matrix.

27. Use of a pharmaceutical composition according to claim 26, wherein the sustained release coating or sustained release matrix comprises a material that disintegrates over time.

28. Use of a pharmaceutical composition according to claim 27, wherein the material that disintegrates over time is selected from the group consisting of: poly(ethyl acrylate-methyl methacrylate) 2:1; poly(ethyl acrylate-methyl methacrylate-methacryloyloxyethyltrimethylammonium chloride) 1:2:0.1; ethylcellulose; poly(ethyl acrylate-methyl methacrylate-methacryloyloxyethyltrimethylammonium chloride) 1:2:0.2; polyvinyl acetate; and combinations thereof.

29. Use of a pharmaceutical composition according to claim 22, wherein the medicament comprises a sustained release core coated with a prolonged release coating.

30. Use of a pharmaceutical composition according to claim 22, wherein the medicament comprises a core, a sustained release coating and a delayed release coating.

31. Use of a pharmaceutical composition according to any one of claims 1 to 21, wherein said topical administration comprises rectal administration of said drug.

32. The use of the pharmaceutical composition according to claim 1, wherein the anti-TNFα antibody comprises a V having an amino acid sequence as shown in SEQ ID NO:

10. L domain and a V domain having an amino acid sequence as shown in SEQ ID NO: 9 H domain.

33. The use of the pharmaceutical composition according to claim 1, wherein the anti-TNFα antibody is an antibody comprising an Fc region comprising amino acids 236 to 451 of the amino acid sequence shown in SEQ ID NO:

11.

34. The use of the pharmaceutical composition according to claim 1, wherein the anti-TNFα antibody comprises a light chain having the amino acid sequence shown in SEQ ID NO: 1 and a heavy chain having the amino acid sequence shown in SEQ ID NO: 11, 14, 15 or 16.

35. Use of a pharmaceutical composition according to claim 31, wherein the medicament is an enema, a gel, a foam or a suppository.

Images