Use of anti-CTLA-4 antibodies for treatment of adenoid cystic cancer

Abstract

The present invention relates to the use of an anti-CTLA-4 antibody for the treatment of adenoid cystic carcinoma.

Description

[0001] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] This invention was made with government support in part under Grant No. R44CA250824 awarded by the National Cancer Institute of the National Institutes of Health (NIH). The government has certain rights in this invention. Field of the Invention

[0003] The present invention relates to the use of anti-CTLA-4 antibodies for the treatment of adenoid cystic carcinoma (ACC), including as the sole immunotherapy or as a monotherapy.

[0004] Reference sequence list

[0005] Applicants hereby refer to the sequence listing contained in the file "111005_0604_01PC00_Sequence_Listing.xml" (25kB; created on September 14, 2023), the contents of which are incorporated herein by reference. Background of the Invention

[0007] ACC is a rare cancer with an annual incidence of about 4 cases per million and a 16-year limited duration prevalence of about 33 cases per million. In the United States, there are about 1,400 new cases each year, and about 11,000 patients suffer from ACC. ACC usually originates from the major and minor salivary glands, but can also arise from the breast, prostate, lacrimal glands and other secretory gland tissues. The latter is managed according to the same principles as the salivary gland primary. ACC is typically initially treated with curative surgery and / or radiation, but relapses in most cases. About 50% of these cases have metastases, usually in the lungs. Almost all recurrent and / or metastatic (R / M) ACC are fatal because there is currently no available curative therapy and only a limited number of effective treatments. No therapy is approved by the FDA for ACC patients. Outside of clinical trials, based on limited Phase II data showing moderate clinical efficacy, chemotherapy or multi-target tyrosine kinase inhibitors are used for palliative purposes in R / MACC patients. Therefore, there is a need in the art for effective treatment of ACC. Summary of the invention

[0008] Provided herein are methods of treating adenoid cystic carcinoma (ACC) in a subject in need thereof, which may include administering an anti-CTLA-4 antibody to the subject. Also provided herein are anti-CTLA-4 antibodies for use in treating ACC, compositions comprising anti-CTLA-4 antibodies for treating ACC, and uses of anti-CTLA-4 antibodies in the preparation of a medicament for treating ACC. ACC may be recurrent, metastatic, or both. The anti-CTLA-4 antibody may be the sole immunotherapeutic agent administered to a subject or intended for use in treating ACC. The anti-CTLA-4 antibody may be administered to a subject or intended for use as a monotherapy.

[0009] The anti-CTLA-4 antibody can be Ipilimumab / Yervoy, XTX101, Botensilimab, Zaliferelimab, ADG116, HBM4003, APL-509, BA3071, BMS-986249, Tikab, quavonlimab (MK-1308), or a combination thereof. The anti-CTLA-4 antibody can comprise: (a) a light chain variable region comprising a complementary determining region (CDR) 1 comprising the amino acid sequence shown in SEQ ID NO: 1; a CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs: 2-4; and a CDR3 comprising the amino acid sequence shown in SEQ ID NO: 5; and (b) a heavy chain variable region comprising a CDR1 comprising the amino acid sequence shown in SEQ ID NO: 6; a CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs: 7-9; and a CDR3 comprising the amino acid sequence shown in SEQ ID NO: 10. The anti-CTLA-4 antibody may comprise a light chain variable region comprising a CDR2 comprising the sequence shown in SEQ ID NO:3 and a heavy chain variable region comprising a CDR2 comprising the sequence shown in SEQ ID NO:9. The anti-CTLA-4 antibody may comprise a light chain variable region comprising the sequence shown in SEQ ID NO:12 and a heavy chain variable region comprising the sequence shown in SEQ ID NO:16. The anti-CTLA-4 antibody may comprise a light chain comprising the sequence shown in SEQ ID NO:23 and a heavy chain comprising the sequence shown in SEQ ID NO:21. The anti-CTLA-4 antibody may be ONC-392. The anti-CTLA-4 antibody may have been diluted with a 5% dextrose solution to a final concentration of about 1.0 to about 2.5 mg / mL. Dilution may be from a formulation containing 5 mg / mL anti-CTLA-4 antibody, 20 mM histidine buffer, 8.8% (w / v) trehalose dihydrate, and 0.06% (w / v) PS80 at pH 6.0.

[0010] Also provided herein are compositions comprising anti-CTLA-4 antibodies. The antibody may comprise a light chain variable region comprising a CDR1 comprising the amino acid sequence shown in SEQ ID NO: 1; a CDR2 comprising the amino acid sequence shown in SEQ ID NO: 3; and a CDR3 comprising the amino acid sequence shown in SEQ ID NO: 5. The antibody may further comprise a heavy chain variable region comprising a CDR1 comprising the amino acid sequence shown in SEQ ID NO: 6; a CDR2 comprising the amino acid sequence shown in SEQ ID NO: 9; and a CDR3 comprising the amino acid sequence shown in SEQ ID NO: 10. The antibody may have been diluted with a 5% dextrose solution to a final concentration of about 1.0 to about 2.5 mg / mL. The dilution may be from a formulation containing 5 mg / mL anti-CTLA-4 antibody, 20 mM histidine buffer, 8.8% (w / v) trehalose dihydrate, and 0.06% (w / v) PS80 at pH 6.0. The antibody may comprise a light chain variable region comprising the sequence shown in SEQ ID NO: 12 and a heavy chain variable region comprising the sequence shown in SEQ ID NO: 16. The antibody may comprise a light chain comprising the sequence shown in SEQ ID NO: 23 and a heavy chain comprising the sequence shown in SEQ ID NO: 21. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 A diagram of Part A of the ONC-392 monotherapy clinical trial is shown.

[0012] Figure 2 A diagram of Part B of the Phase I clinical trial of ONC-392 plus pembrolizumab is shown.

[0013] Figure 3 A diagram of Part B of the Phase IB extension of the trial of ONC-392 plus pembrolizumab is shown.

[0014] Figure 4A -B shows the study design and preliminary results of the clinical trial of ONC-392. Figure 4A The dose-finding study design is shown. Figure 4B Preliminary results are shown. *Not confirmed as of October 13, 2021. #IHC of surgical tissue showed abundant CD4+ and CD8+ T cell infiltration within the tumor.

[0015] Figure 5 Shown is the design of a clinical trial testing the efficacy of the anti-CTLA-4 antibody ONC-392 for the treatment of recurrent / metastatic (R / M) ACC.

[0016] Fig. 6A -C shows a magnetic resonance imaging (MRI) image demonstrating the efficacy of anti-CTLA-4 antibody ONC-392 against ACC. Fig. 6A The target lesion at baseline (December 9, 2021) is shown, which was a left anterior temporal soft tissue tumor mass. Figure 6B Target lesions after two cycles of treatment with ONC-392 (on February 24, 2022) are shown. Figure 6C Target lesions after 3 cycles of ONC-392 treatment (on March 24, 2022) are shown. DETAILED DESCRIPTION OF THE INVENTION

[0018] The anti-CTLA-4 antibodies described herein can be used to treat cancer. In particular, surprisingly, the inventors found that anti-CTLA-4 antibodies - even provided as the sole immunotherapeutic agent - are effective against adenoid cystic carcinoma, a rare cancer that is difficult to treat. When provided as the sole immunotherapeutic agent, no anti-CTLA-4 monoclonal antibody has been shown to be effective in treating ACC. ACC can be considered a cold tumor due to its low mutation / neoantigen load. However, the prevalent MYB-NFIB gene fusion can lead to new antigens for T cells, and intratumoral infiltration was observed in 42% of clinical samples of primary and metastatic lesions. Interestingly, none of the tumors analyzed had PD-L1 expression on tumor cells. Transcripts associated with regulatory T cells were most abundant in tumors with immune infiltration compared to tumors without immune infiltration. This, combined with the lack of PD-L1 on tumor cells, suggests that more intensive immunotherapy aimed at eliminating regulatory T cells can overcome immune escape in ACC. Since regulatory T cells express higher levels of CTLA-4 than other immune cell types, the inventors recognized that one of the most effective ways to target regulatory T cells may be to use anti-CTLA-4 antibodies with Fc portion-capable antibody-dependent cell-mediated cytotoxicity (ADCC) or phagocytosis (ADCP).

[0019] Although the approved anti-CTLA-4 antibody ipilimumab is considered ADCC / ADCP competent due to its IgG1 Fc, its ADCC activity on Tregs has been shown to be suboptimal, primarily due to its downregulation of CTLA-4 molecules through lysosomal degradation. In addition, since clinically used anti-CTLA-4 antibodies cause immunotherapy-related adverse events at a high rate, the dose and duration of treatment are severely shortened in the clinic. Preclinical and clinical studies have shown that anti-CTLA-4 antibodies such as ONC-392 have a significantly improved therapeutic index due to their ability to avoid CTLA-4 downregulation.

[0020] Described in more detail below, ONC-392 is a humanized monoclonal antibody generated by transplanting the CDR region from a mouse monoclonal antibody onto a human IgG1 antibody framework. In order to promote antibody-dependent cellular toxicity (ADCC) for tumor rejection, ONC-392 has a human IgG1 Fc region, which has strong ADCC activity. In addition, in order to enhance ADCC activity, the Fc domain of ONC-392 is modified to improve the immunotherapy effect of the antibody. Specifically, three mutations (S298A, E333A, and K334A) are introduced into CH to increase ADCC activity. Another three mutations (M252Y, S254T, and T256E) are introduced to increase the in vivo half-life of the antibody. The ONC-392 drug is a sterile liquid for intravenous (IV) administration, which is supplied in a 20mL glass vial and sealed with a 20mm rubber stopper flip cap. Drug products were formulated in 20 mM histidine buffer, 8.8% (w / v) trehalose dihydrate, 0.06% (w / v) PS80 at pH 6.0 to a protein concentration of 5.0 mg / mL.

[0021] 1. Definition

[0022] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0023] For the recitation of numerical ranges herein, each intervening number with equal precision therebetween is expressly contemplated. For example, for the range of 6-9, in addition to 6 and 9, the numbers 7 and 8 are also contemplated, and for the range of 6.0-7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly contemplated.

[0024] 2. Anti-CTLA-4 Antibodies

[0025] Anti-CTLA-4 antibodies are provided herein. Anti-CTLA-4 antibodies can be one or more of Ipilimumab / Yervoy, XTX101, Botensilimab, Zaliferelimab, ADG116, HBM4003, APL-509, BA3071, BMS-986249, Tikab, quavonlimab (MK-1308). Anti-CTLA-4 antibodies can also be described in U.S. Patent No. 10,618,960, the contents of which are incorporated herein by reference.

[0026] An anti-CTLA-4 antibody may comprise a light chain variable region comprising a complementarity determining region (CDR) 1 comprising the amino acid sequence of RASENIYSNLA (SEQ ID NO: 1); a CDR2 comprising the amino acid sequence of AATNLQS (SEQ ID NO: 2) (LC1), AATNLQD (SEQ ID NO: 3) (LC2), or AATSLQS (SEQ ID NO: 4) (LC3); and a CDR3 comprising the amino acid sequence of QHLWGTPYT (SEQ ID NO: 5).

[0027] The light chain variable region comprising one of LC1-LC3 may also respectively comprise one of the following sequences:

[0028] LC1

[0029]

[0030] LC2

[0031]

[0032] LC3

[0033]

[0034] More specifically, a light chain comprising one of LC1-LC3 may respectively comprise one of the following amino acid sequences:

[0035] LC1

[0036]

[0037] LC2

[0038]

[0039]

[0040] LC3

[0041]

[0042] An anti-CTLA-4 antibody may comprise a heavy chain variable region comprising a CDR1 comprising the amino acid sequence of GFSLTSYGLS (SEQ ID NO:6); a CDR2 comprising the amino acid sequence of YIWYDGNTNFHPSLKSR (SEQ ID NO:7) (HC1), YIWYDGNTNFHSSLKSR (SEQ ID NO:8) (HC2), or YIWYDGNTNFHSPLKSR (SEQ ID NO:9) (HC3); and a CDR3 comprising the amino acid sequence of TEGHYYGSNYGYYALDY (SEQ ID NO:10).

[0043] The heavy chain variable region comprising one of HC1-HC3 may respectively comprise one of the following amino acid sequences:

[0044] HC1

[0045]

[0046] HC2

[0047]

[0048] HC3

[0049]

[0050] The anti-CTLA-4 antibody may comprise a heavy chain constant region from a human Ig protein, which may be IgG, IgE, IgM, IgD, IgA, IgY, IgG1, IgG2, IgG3, IgG4, IgA1 or IgA2. In one example, the constant region is an Fc region from a human IgG1 protein. In one example, the heavy chain constant region comprises the amino acid sequence:

[0051]

[0052] The heavy chain constant region may also comprise one or more mutations. Relative to the sequence shown in SEQ ID NO: 17, one or more mutations may be selected from M135Y, S137T, T139E, S181A, E216A and K217A and combinations thereof. In one example, the heavy chain constant region of the antibody comprises all six mutations. The mutant heavy chain constant region may comprise the amino acid sequence:

[0053]

[0054] Even more specifically, the heavy chains of the anti-CTLA-4 antibodies comprising the heavy chain variable regions HC1-HC3 may each comprise one of the following amino acid sequences:

[0055] HC1

[0056]

[0057]

[0058] HC2

[0059]

[0060] HC3

[0061]

[0062] A C-terminal lysine (K) may be additionally included in the amino acid sequence of the heavy chain shown in SEQ ID NOs: 19-21, which may increase expression levels. The terminal lysine may be naturally cleaved during production of the anti-CTLA-4 antibody or upon administration of the antibody.

[0063] PP4637 (LC2 / HC3): In one example, an anti-CTLA-4 antibody comprises a light chain variable region comprising a CDR1 comprising the sequence shown in SEQ ID NO: 1, a CDR2 comprising the sequence shown in SEQ ID NO: 3, and a CDR3 comprising the sequence shown in SEQ ID NO: 5. The heavy chain variable region comprises a CDR1 comprising the sequence shown in SEQ ID NO: 6, a CDR2 comprising the sequence shown in SEQ ID NO: 9, and a CDR3 comprising the sequence shown in SEQ ID NO: 10. In particular, the light chain variable region may comprise the sequence shown in SEQ ID NO: 12, and the heavy chain variable region may comprise the sequence shown in SEQ ID NO: 16. More particularly, the light chain variable region may comprise the sequence shown in SEQ ID NO: 23, and the heavy chain variable region may comprise the sequence shown in SEQ ID NO: 21. The antibody may be referred to as ONC-392.

[0064] PP4631 (LC2 / HC1): In another example, an anti-CTLA-4 antibody comprises a light chain variable region comprising a CDR1 comprising the sequence shown in SEQ ID NO: 1, a CDR2 comprising the sequence shown in SEQ ID NO: 3, and a CDR3 comprising the sequence shown in SEQ ID NO: 5. The heavy chain variable region comprises a CDR1 comprising the sequence shown in SEQ ID NO: 6, a CDR2 comprising the sequence shown in SEQ ID NO: 7, and a CDR3 comprising the sequence shown in SEQ ID NO: 10. In particular, the light chain variable region may comprise the sequence shown in SEQ ID NO: 13, and the heavy chain variable region may comprise the sequence shown in SEQ ID NO: 14. More particularly, the light chain variable region may comprise the sequence shown in SEQ ID NO: 23, and the heavy chain variable region may comprise the sequence shown in SEQ ID NO: 19.

[0065] PP4638 (LC3 / HC3): In a further example, the anti-CTLA-4 antibody comprises a light chain variable region comprising a CDR1 comprising the sequence shown in SEQ ID NO: 1, a CDR2 comprising the sequence shown in SEQ ID NO: 4, and a CDR3 comprising the sequence shown in SEQ ID NO: 5. The heavy chain variable region comprises a CDR1 comprising the sequence shown in SEQ ID NO: 6, a CDR2 comprising the sequence shown in SEQ ID NO: 9, and a CDR3 comprising the sequence shown in SEQ ID NO: 10. In particular, the light chain variable region may comprise the sequence shown in SEQ ID NO: 12, and the heavy chain variable region may comprise the sequence shown in SEQ ID NO: 16. More particularly, the light chain variable region may comprise the sequence shown in SEQ ID NO: 24, and the heavy chain variable region may comprise the sequence shown in SEQ ID NO: 21.

[0066] 3. Dosage regimen

[0067] The anti-CTLA-4 antibody may be administered to a subject, which may be a human. Administration may be to treat ACC as further described herein. The anti-CTLA-4 antibody may be administered systemically, which may be administered via injection or intravenously (IV). The antibody may be administered as a monotherapy or as a combination therapy. The dosing regimen may include administering one or more doses of the anti-CTLA-4 antibody. Independently, each dose may be about 0.1 mg / kg, 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 5 mg / kg, 6 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 50 mg / kg or 100 mg / kg, or an amount within the range of two of these amounts. The dosing regimen may include periodic administration, wherein one of the aforementioned doses is administered to the subject. In each dosing cycle, the dose may be different from the previous dose. Administration may involve increasing doses. In one example, the anti-CTLA-4 antibody is administered approximately every 1, 2, 3, 4, 5 or 6 weeks. In particular, the antibody is administered about once every 3 weeks.When describing the time period of a dosing cycle, "about" can mean ± 1, 2 or 3 days.

[0068] In particular, the dosage of the anti-CTLA-4 antibody can be about 1, 3, 6 or 10 mg / kg, or an amount within the range of two of these amounts. The dosing regimen can also include two doses of 10 mg / kg, followed by an extended administration of 1-6 mg / kg (ie, each subsequent dose is 1-6 mg / kg). Extended administration can include administration of a dose of 3 mg / kg or 6 mg / kg. In one example, each administration is about once every 3 weeks. In one example, the anti-CTLA-4 antibody is administered about once every 4 weeks. Administration can occur within a time period of about 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51 or 52 weeks or within the range thereof.

[0069] The anti-CTLA-4 antibody can be administered in combination with a second therapeutic agent, or it can be administered alone or mixed with a second therapeutic agent. The therapeutic agent can be an anticancer agent. In one example, the anticancer agent is administered on the same day as the anti-CTLA-4 antibody. In particular, the anticancer agent can be an anti-PD-1 or anti-PD-L1 antibody. In a particular example, the anticancer agent is pembrolizumab (KEYTRUDA). In one example, pembrolizumab is administered once every 21 days at 200 mg / cycle. In a further example, the second therapeutic agent is administered on the same day as the anti-CTLA-4 antibody.

[0070] 4. Preparation

[0071] The anti-CTLA-4 antibody can be formulated or provided in a dosage as described herein. In one example, the formulation comprises 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 15 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 50 mg / mL or 100 mg / mL of an anti-CTLA-4 antibody, or an amount within a range thereof. In one example, the amount is 5 mg / mL. The formulation can include 5, 10, 15, 20, 25, 30, 35 or 40 mM histidine buffer, or an amount within a range of two of these amounts. In one example, the amount is 20 mM. The formulation may also include 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9 or 10.0% (w / v) α, α-trehalose dihydrate, or an amount within the range of two of these amounts. In one example, the amount is 8.8%. The formulation may include 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 or 0.10 (w / v) polysorbate 80, or an amount within the range of two of these amounts. In one example, the amount is 0.06%. The pH of the formulation can be 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4 or 6.5, or a pH within a range thereof. Equivalent ingredients of histidine buffer, α, α-trehalose dihydrate, and polysorbate 80 for formulating antibodies are known in the art and may also be used as substitutes.

[0072] The formulation may comprise 5-100 mg / mL anti-CTLA-4 antibody, 20 mM histidine buffer, 8.8% (w / v) α, α-trehalose dihydrate, and 0.01-0.06% (w / v) polysorbate 80. The pH of the formulation may be 6.0. In another example, the formulation may comprise 5 mg / mL anti-CTLA-4 antibody, 20 mM histidine buffer, 8.8% (w / v) α, α-trehalose dihydrate, and 0.06% (w / v) polysorbate at pH 6.0.

[0073] 5. Cancer treatment

[0074] The compositions and dosing regimens described herein can be used to treat cancer. The cancer can be a solid tumor. The cancer can be one of progressive locally advanced and metastatic cancers. The cancer can be stage IV cancer. The subject may show failure or intolerance to standard care guidelines, which can be the National Comprehensive Cancer Network Oncology Clinical Practice Guidelines (NCCN Guidelines). The cancer can be refractory or resistant to anti-PD-1 / PD-L1 treatment. Resistance can be primary resistance or acquired resistance to disease progression after immunotherapy. Primary PD-1 resistance can be defined as disease progression within 24 weeks of initiation of anti-PD-(L)1 therapy. Acquired PD-1 resistance can be defined as disease control (CR, PR, or SD) for 24 weeks or longer after initiation of anti-PD-(L)1 therapy, and subsequent progression after 24 weeks. The cancer can be immunotherapy-naïve. And can be PD-L1 positive, for example by having a PD-L1 tumor proportion score ≥ 1%. The cancer can be non-small cell lung cancer. In another example, the cancer is ovarian cancer, cervical cancer, gastroesophageal cancer, lung cancer or ovarian cancer.

[0075] The subject may have metastatic disease or locally advanced disease that is not amenable to local therapy. The subject may also not have received established standard medical anticancer therapy, which may not be pembrolizumab for a given tumor type, or may be intolerant to such therapy.

[0076] Cancer may be a neoplasm or tumor caused by abnormal, uncontrolled growth of cells. Cancer may be a leukemia or lymphoma. Cancer may involve cells that have the potential to metastasize to distant sites.

[0077] The cancer may be one of the following: carcinoma, such as bladder cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer, stomach cancer, cervical cancer, thyroid cancer or skin cancer; squamous cell carcinoma; hematopoietic tumors of the lymphoid lineage, such as leukemia, acute lymphoblastic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Burkitt's lymphoma; hematopoietic tumors of the myeloid lineage, such as acute and chronic myeloid leukemia or promyelocytic leukemia; interstitial cell carcinoma, such as myeloid leukemia, acute lymphoblastic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Burkitt's lymphoma; Tumors of mesenchymal origin, such as fibrosarcoma or rhabdomyosarcoma; tumors such as melanoma, seminoma, teratoma, neuroblastoma or glioma; central and peripheral nervous system tumors, such as astrocytoma, neuroblastoma, glioma and schwannoma; tumors of mesenchymal origin, such as fibrosarcoma, rhabdomyosarcoma or osteosarcoma; or tumors such as melanoma, xeroderma pigmentosum, keratoacanthoma, seminoma, follicular thyroid carcinoma or teratoma.

[0078] Cancer may be caused by abnormal apoptosis. Cancer may be follicular lymphoma, cancer with one or more p53 mutations, hormone-dependent tumors of the breast, prostate or ovary, precancerous lesions such as familial adenomatous polyposis or myelodysplastic syndrome. Cancer may be a malignancy or abnormal proliferative changes (such as metaplasia or dysplasia) or a hyperproliferative disorder, and may be in the ovary, bladder, breast, colon, lung, skin, pancreas or uterus. In particular, cancer may also be a sarcoma, melanoma or leukemia.

[0079] The cancer can also be non-small cell lung cancer, advanced solid tumors, metastatic melanoma, metastatic head and neck cancer, metastatic renal cell carcinoma, metastatic colorectal cancer, sarcoma, metastatic prostate cancer, ovarian cancer, small cell lung cancer, metastatic breast cancer, pancreatic cancer, gastric cancer, esophageal cancer, gastroesophageal junction adenocarcinoma, cervical cancer, adenoid cystic carcinoma (ACC), salivary gland cancer, or urothelial carcinoma.

[0080] In particular, the cancer may be ACC. ACC may be recurrent, metastatic, or both. For the treatment of ACC, the anti-CTLA-4 antibody may be administered once every about 3 or 4 weeks, in particular once every about 4 weeks. In one example, the anti-CTLA-4 antibody is the only immunotherapeutic agent administered to the subject. In another example, the anti-CTLA-4 antibody is administered as a monotherapy. In a further example, a second anticancer therapeutic agent is administered. The second anticancer therapeutic agent may be pembrolizumab, which may be administered as described herein.

[0081] The present invention has several aspects, which are illustrated by the following non-limiting examples.

[0082] Example 1

[0083] Safety and efficacy of anti-CTLA-4 antibody therapy

[0084] This example demonstrates the safety and efficacy of anti-CTLA-4 antibody ONC-392 (P4637) for the treatment of cancer, particularly advanced solid tumors and non-small cell lung cancer (NSCLC). The trial was conducted in two parts: A and B. In part A, patients with histologically or cytologically confirmed solid tumors were enrolled according to standard care guidelines (such as NCCN guidelines), who had progressive locally advanced or metastatic disease after failure or intolerance of previous standard medical anticancer treatment.

[0085] In Part B, which evaluated dose-finding and expansion cohorts, patients with advanced / metastatic non-small cell lung cancer (NSCLC) who were immunotherapy-naive and PD-L1-positive (PD-L1 tumor proportion score (TPS) ≥1%) or refractory / resistant to anti-PD-1 / PD-L1 therapy (regardless of PD-L1 status) were enrolled.

[0086] Study Design Summary

[0087] A Phase IA / IB open-label, dose-escalation study was conducted in participants with advanced / metastatic solid tumors and NSCLC, with ONC-392 as a single agent and in combination with pembrolizumab (anti-PD-1, marketed by Merck & Co. Marketed) combination for intravenous (IV) administration.

[0088] The study consists of two related parts:

[0089] Part A is a dose-finding rapid titration, Phase IA trial of ONC-392 as a single agent in patients with advanced disease of various histologies. The objective of this trial is to determine the recommended Phase II dose of ONC-392 monotherapy (RP2D-M).

[0090] Part B is a Phase IA / IB trial of ONC-392 in combination with a standard dose of 200 mg pembrolizumab in patients with NSCLC. The trial consists of dose finding, dose escalation or de-escalation, with the Phase IA portion designed to determine the recommended Phase II dose of ONC-392 in combination with a standard dose of pembrolizumab (RP2D-C), and then progresses to two parallel, single-arm, Phase IB expansion cohorts to test safety and preliminary efficacy in two groups of patients with NSCLC:

[0091] Stage IV NSCLC with PD-L1 positive (PD-L1 TPS ≥ 1%) who have not been treated with anti-PD(L)1 immunotherapy;

[0092] Stage IV NSCLC refractory / resistant to anti-PD(L)1 immunotherapy, regardless of PD-L1 status.

[0093] In the Phase IA portion, both anti-PD(L)1 immunotherapy-naïve and refractory / resistant disease were enrolled.

[0094] Experimental Design and Sample Size

[0095] To be eligible for the study, patients had to be 18 years of age or older, had to have metastatic disease or locally advanced disease that was not amenable to local therapy, and had to have failed established standard medical anticancer therapy other than pembrolizumab for the given tumor type, had been intolerant to such therapy, or, in the opinion of the investigator, had been deemed unsuitable for a specific form of standard therapy for medical reasons. Patients had to have had an Eastern Cooperative Oncology Group (ECOG) performance status of ≤2.

[0096] Part A ONC-392 monotherapy will enroll a minimum of 10 patients and a maximum of 30 subjects.

[0097] The Phase IA dose-finding phase of ONC-392 and pembrolizumab combination therapy in Part B will enroll up to 18 subjects.

[0098] Two cohorts of 18 patients each participated in the Phase IB dose-finding phase of the ONC-392 and pembrolizumab combination therapy in Part B.

[0099] Cohort 1: Part B Phase IB anti-PD(L)-1 treatment-naïve and PD-L1 positive (PD-L1 TPS ≥ 1%) cohort plans to enroll eighteen (18) subjects.

[0100] Cohort 2: Part B Phase IB anti-PD(L)-1 refractory / resistant cohort is planned to enroll eighteen (18) subjects.

[0101] A total of 84 subjects were planned to be enrolled in the study.

[0102] Part A: ONC-392 Single Agent

[0103] Part A of the Phase IA trial tested up to five predetermined dose levels: 0.1 mg / kg, 0.3 mg / kg, 1 mg / kg, 3 mg / kg and 10 mg / kg of ONC-392 as a monotherapy every 21 days (Q3W) by IV infusion. The trial adopted an accelerated titration design. Intra-patient dose escalation was tested in the first patient who received 0.1 mg / kg, 0.3 mg / kg, 1.0 mg / kg without any AEs. The patient's dose was escalated to 3.0 mg / kg and received 3 cycles at this dose without any AEs. The second patient started at 0.3 mg / kg without any AEs. Then, enrollment was converted to a 3+3 design at 3.0 mg / kg and 10.0 mg / kg levels in the following regimen.

[0104] Part B: Combination of ONC-392 and pembrolizumab in NSCLC

[0105] Part B was designed as a Phase IA dose-escalation / de-escalation study, followed by a Phase IB expansion part at RP2D-C, for the combination of ONC-392 with pembrolizumab in two cohorts of patients with NSCLC.

[0106] The dose of pembrolizumab was fixed at 200 mg / cycle, administered once every 21 days (Q3W).

[0107] Part B Phase IA Study

[0108] The Phase IA study was initiated with 200 mg pembrolizumab at a dose one level below the ONC-392 RP2D-M dose, initially enrolling 6 patients. The ONC-392 dose was adjusted based on:

[0109] (1) If 1 / 6 patients develop dose-limiting toxicity (DLT), a dose one level below RP2D-M will be declared RP2D-C.

[0110] or

[0111] (2) If 0 / 6 patients develop DLT, enroll an additional 6 patients at the RP2D-M dose level of ONC-392. If ≤ 1 / 6 additional patients develop DLT, the RP2D-M will be declared PR2D-C.

[0112] or

[0113] (3) When 2 DLTs occur before 6 patients are enrolled, the ONC-392 dose will be reduced to the next dose level until ≤ 1 / 6 patients treated at that dose develop DLT. This dose level is designated RP2D-C. If dose level 1 (0.1 mg / kg) is too toxic using the above rules, further exploration of the drug combination should be stopped.

[0114] Part B Phase IB study

[0115] The Part B, Phase IB expansion cohort is designed for patients with advanced NSCLC and includes an immunotherapy-naive cohort and a refractory / resistant cohort. Six patients treated with RP2D-C in Part B, Phase IA, are evaluable for efficacy. One of the purposes of the expansion cohort is to obtain a more comprehensive safety profile of ONC-392 in combination with pembrolizumab in RP2D-C. To ensure the safety of patients enrolled in both expansion cohorts, a Pocock-type boundary is used to allow early stopping due to excessive toxicity at any given time. The trial is stopped at any time point if the incidence of DLT is significantly higher than θ=20%.

[0116] In patients who were first treated with anti-PD(L)1 immunotherapy, Patients with advanced NSCLC who are PD-L1 positive (PD-L1 TPS ≥ 1% or other indicated for pembrolizumab) will be enrolled in the study. 18 subjects will be enrolled in the Phase IB expansion cohort.

[0117] In the anti-PD(L)1 refractory / resistant population, Patients with advanced NSCLC who have disease progression or are intolerant to anti-PD(L)1-containing therapy (monotherapy, combination therapy, or immunotherapy in combination with chemotherapy) are enrolled in this study after 4 or more cycles. Prior CTLA-4 therapy is allowed. A history of irAE with recovery is allowed. 18 subjects will be enrolled in the Phase IB expansion cohort.

[0118] Response rates for the PD-(L)1 therapy-naïve and refractory / resistant cohorts will be determined separately 6 months after first treatment.

[0119] Dosage / dosage form, route and administration schedule

[0120] For dose escalation in monotherapy, ONC-392 is administered as a minimum 30-minute IV infusion. Five dose levels of ONC-392 will be evaluated: 0.1 mg / kg, 0.3 mg / kg, 1 mg / kg, 3 mg / kg and 10 mg / kg. The dosing interval is 21 days. ONC-392 is given on a Q3W schedule. Intra-patient dose escalation to 3 mg / kg is allowed. If the patient confirms a complete response, or if the patient tolerates treatment and confirms disease progression, treatment should continue for an additional 4 cycles. If the patient's disease state is considered to be "stable disease" or "partial response", treatment will continue for up to 1 year. Treatment will be stopped due to unacceptable toxicity, or the patient voluntarily withdraws or 1 year (whichever occurs first).

[0121] In the combination of ONC-392 and pembrolizumab, ONC-392 will be administered first as a 30±15 minute IV infusion. Pembrolizumab will then be administered as a 30±15 minute IV infusion at a fixed 200 mg / dose. ONC-392 and pembrolizumab will not be mixed in administration.

[0122] ONC-392 and pembrolizumab are given on a Q3W schedule. If the patient has a confirmed complete response, or confirmed disease progression if treatment is likely to be tolerated, treatment will continue for an additional 4 cycles. If the patient's disease status is considered "stable disease" or "partial response," treatment should continue for up to 1 year. Treatment was discontinued due to unacceptable toxicity or voluntary withdrawal of the patient.

[0123] Administration of ONC-392 as a single agent or in combination with pembrolizumab requires hourly monitoring of vital signs during dose escalation from the start of the IV infusion until 4 hours after the end of the infusion. When patients are dosed at the RP2D, monitor vital signs before and after the infusion (+ / - 30 minutes).

[0124] Table 1 Objectives and endpoints

[0125]

[0126] Figure 1 A diagram of the ONC-392 monotherapy trial design is shown. Figure 2 A diagram of Part B of the Phase I clinical trial of ONC-392 plus pembrolizumab is shown. Figure 3 A diagram of Part B of the Phase IB extension of the ONC-392 plus pembrolizumab trial is shown. The table below shows the study eligibility criteria.

[0127] Table 2 Study Eligibility

[0128]

[0129] Safety Results from Part A

[0130] The dosing schedule for Part A of the clinical trial is as follows: Figure 4A The demographics of the patients evaluated in Part A of the ONC-392 trial are shown in the following table.

[0131] Table 3

[0132]

[0133]

[0134] Dosing and preliminary results of the trial are as follows Figure 4B Results showed that ONC-392 was generally well tolerated at both doses. Grade 3 irAEs of pancreatitis and colitis were manageable and reversible. The PR2D for monotherapy was 10 mg / kg, q3w.

[0135] Results showed that ONC-392 was well tolerated. The longest dosing was 3 mg / kg for up to 9 cycles. No DLT or grade 3 / 4 AEs occurred during the DLT observation period at any dose. The maximum tolerated dose was not reached. The recommended phase 2 dose for monotherapy was determined to be 10 mg / kg. The following grade 3 / 4 AEs occurred in 3 patients after 3 or 4 cycles of treatment with 10 mg / kg ONC-392: colitis / hypokalemia (2) and pancreatitis (1). Two of the three patients achieved an unconfirmed complete response, and one patient had stable disease and reduced tumor burden. Other drug-related AEs were grade 1 / 2, and adverse reactions occurring in more than two patients included infusion-related reactions, itching, fatigue, and elevated TSH.

[0136] Clinical Results

[0137] In addition, beneficial activity was observed in 6 / 10 patients. Two of the six patients treated with 10 mg / kg ONC-392 showed a complete response, two of the six patients treated with 10 mg / kg ONC-392 had stable disease, with significant reductions in tumor burden or biomarkers of enhanced T cell activation in the tumor, and two of the four patients treated with 3 mg / kg had stable disease (SD) for more than 7 months. At the first tumor assessment, stable disease was observed in 7 of 10 patients and partial response was observed in 1 of 10 patients. In addition, clinical improvement was observed in three PD-(L)1 refractory / resistant patients with NSCLC (one complete response; one with disease control at more than 24 weeks, eligible for surgery; and one with stable disease at 8 weeks after continued treatment).

[0138] Safety and Efficacy Conclusions

[0139] ONC-392 was generally safe and well tolerated. Treatment-related AEs were manageable. The maximum tolerated dose was not reached at the 10 mg / kg dose. ONC-392 also demonstrated therapeutic antitumor activity. As the first pH-sensitive monoclonal antibody that retains CTLA-4 recycling and avoids lysosomal degradation, ONC-392 may fundamentally change the risk / benefit ratio of CTLA-4 targeting by conferring improved efficacy and reduced toxicity.

[0140] Example 2

[0141] Clinical trials on the efficacy of anti-CTLA-4 antibodies against ACC

[0142] Study Design Summary

[0143] The study is part of a Phase IA / IB / II open-label, dose-escalation and dose-expansion study in which ONC 392 is administered intravenously (IV) as a single agent and in combination with pembrolizumab (anti-PD-1, marketed by Merck & Co., Inc. as a single agent) in patients with advanced / metastatic solid tumors. Part D is designed to study patients with ACC treated with ONC-392 monotherapy. Figure 5 A schematic diagram of the study design is shown in .

[0144] Patients had recurrent and / or metastatic adenoid cystic carcinoma (R / MACC) originating from the salivary glands or other sites and were not suitable for curative surgery or radiotherapy. Prior chemotherapy, targeted therapy, and immunotherapy were allowed. The primary endpoint was the objective response rate (ORR) assessed by blinded independent central review (BICR) based on the solid tumor response evaluation criteria version 1.1 (RECIST 1.1) after patients received ONC-392 treatment as a monotherapy or as a combination therapy with pembrolizumab. The primary safety endpoint was the incidence of treatment-emergent adverse events (TEAEs). Secondary efficacy parameters included objective incidence rate (ORR), duration of occurrence (DoR), best overall occurrence (BoR), and disease control rate (DCR). Other efficacy parameters included progression-free survival (PFS) assessed by the investigator based on RECIST 1.1 and iRECIST; overall survival (OS) after administration of ONC-392; and ORR, DoR, BoR, DCR assessed by BICR based on iRECIST. Additionally, PK parameters were measured to investigate any exposure-response relationships.

[0145] Dosage / dosage form, route and administration schedule

[0146] For dose escalation in monotherapy, five dose levels of ONC-392 were evaluated: 0.1 mg / kg, 0.3 mg / kg, 1.0 mg / kg, 3.0 mg / kg, and 10 mg / kg. For the dose levels of 0.1, 0.3, and 1.0 mg / kg, ONC-392 was administered as an IV infusion over a minimum of 30 minutes, and for the dose level of 3.0 mg / kg, it was administered over a minimum of 60 minutes. At the 10 mg / kg dose level, an infusion time of at least 90 minutes was required for the first dose, and at least 60 minutes was required for subsequent doses. ONC-392 dosing intervals were 21 days (once every 3 weeks [Q3W]). Intra-patient dose escalation was allowed up to 3 mg / kg.

[0147] For the combination of ONC-392 and pembrolizumab, ONC-392 was first administered as an IV infusion over a minimum of 60 minutes, but the first dose of ONC-392 10 mg / kg was administered over a minimum of 90 minutes. For the 6.0 mg / kg ONC-392 dose level, the IV infusion lasted for 60 minutes. Pembrolizumab was then administered IV at a fixed 200 mg / dose over a minimum of 30 minutes. There was a gap of at least 30 minutes between the end of the ONC-392 infusion and the start of the pembrolizumab infusion. ONC-392 and pembrolizumab were not mixed during administration. Both ONC-392 and pembrolizumab were administered Q3W.

[0148] If the patient tolerates treatment, study treatment (both monotherapy and combination therapy) may continue for an additional 4 cycles (optional) after the patient has confirmed progressive disease (PD) according to Immune Response Evaluation Criteria in Solid Tumors (iRECIST).

[0149] Study treatment (both monotherapy and combination therapy) was discontinued due to unacceptable toxicity, voluntary patient withdrawal, or at 1 year (13 or 17 cycles), whichever occurred first (see Section 5.7 for options after 1 year).

[0150] The dosing in Part D (ACC) of the study is ONC-392, 10 mg / kg, IV infusion, Q4W. The treatment period is up to 13 cycles (approximately 1 year). Dose adjustments are allowed.

[0151] Patient selection, enrollment, discontinuation, and withdrawal

[0152] Patient inclusion criteria

[0153] Patients were required to be at least 18 years of age at the time of informed consent. Patients could be male or female (if pregnancy testing was negative). Patients had to have an ECOG performance status of ≤1. Patients had to have a histological or cytological diagnosis of a solid tumor and had progressive metastatic disease or progressive locally advanced disease.

[0154] For Part D, patient inclusion criteria were:

[0155] a. Histologically or cytologically confirmed adenoid cystic carcinoma (ACC) with local recurrence or metastasis, not amenable to curative surgery or radiotherapy. ACC from salivary or non-salivary gland primary sites is permitted.

[0156] b. Have measurable target lesions as determined by RECIST 1.1.

[0157] c. New or progressive lesions on radiographic studies performed within 12 months prior to study entry (disease progression was allowed at any interval) and / or new / worsening disease-related symptoms within 12 months prior to study entry. Note: This assessment is performed by the treating investigator. Photographs or radiographic images with ruler reference are required to document disease progression.

[0158] d. Previous chemotherapy, targeted therapy, or immunotherapy is allowed.

[0159] e. Life expectancy ≥ 12 weeks.

[0160] Measurable disease as determined by RECIST 1.1 was determined as follows:

[0161] a. Mass: must be accurately measured in at least 1 dimension (the longest diameter to be recorded) and the minimum dimension is:

[0162] 1. 10mm by computer tomography (CT) scan (CT scan slide thickness must be <5mm),

[0163] or

[0164] 2. 20mm on chest X-ray (if the margins are clear and surrounded by inflated lungs).

[0165] b. With or without malignant lymph nodes: Short axis ≥ 15 mm during CT scan assessment (CT scan slice thickness must be < 5 mm). Measurement is a two-dimensional measurement in the axial plane. The short axis is perpendicular to the long diameter.

[0166] Patients must also have adequate organ function as indicated by specific laboratory values. Patients voluntarily agreed to participate by giving written informed consent. If female patients were of childbearing potential (WOCBP) and sexually active, they agreed to use adequate and effective birth control from the start of the first dose of study drug until 90 days after the last dose of study therapy. Male patients (if sexually active) agreed to use adequate and effective contraception from the start of the first dose of study drug until 90 days after the last dose of study therapy. Patients agreed to allow the research team access to archived diagnostic tissue (re-cut slides or tumor biopsies).

[0167] Patients were excluded based on the following criteria:

[0168] 1) Patients with AEs due to cancer treatment that have not recovered to NCI CTCAE grade 1 or higher, except for chemotherapy-related peripheral neuropathy (motor or sensory) or endocrine-related AEs, which are allowed to recover to ≤ grade 2. The washout period for cancer treatment drugs (such as chemotherapy, radiotherapy, or targeted therapy) is 21 days. The washout period for treatment regimens containing monoclonal antibodies is 28 days. Best supportive care such as thyroxine, insulin, steroid replacement therapy, blood transfusions, and non-cancer disease therapies are allowed.

[0169] 2) Patients who are currently participating in any other clinical trial testing an investigational drug or using an investigational device, or are concurrently receiving other approved systemic therapies.

[0170] 3) Patients who have received chronic systemic steroid therapy (dose >10 mg / day prednisone or equivalent) or any other form of immunosuppressive medication within 7 days prior to ONC-392 treatment. Topical steroids are permitted.

[0171] 4) Active brain metastases or leptomeningeal metastases. Patients are eligible if brain metastases are adequately treated and the patient is neurologically stable (except for residual signs or symptoms related to central nervous system (CNS) treatment).

[0172] 5) Patients with active infection requiring systemic IV antibiotics within 14 days prior to administration of ONC-392 or combined ONC 392 and pembrolizumab. Regular treatment for urinary tract infection (UTI) and / or topical treatments are permitted.

[0173] 6) Patients with a history or current evidence of any condition, therapy, or laboratory abnormality that could confound study results, interfere with patient participation throughout the study, or make study participation not in the best interest of the patient.

[0174] 7) Patients with known psychiatric or substance abuse disorders that interfere with cooperation with trial requirements.

[0175] 8) Patients who are pregnant or breastfeeding.

[0176] ONC-392 IV Infusion

[0177] All patients were premedicated during their first treatment with ONC-392 to prevent infusion reactions. Premedication was recorded in the concomitant medications.

[0178] ONC-392 IV infusion was administered over a minimum of 30 minutes for the 0.1, 0.3, and 1.0 mg / kg dose levels, over a minimum of 60 minutes for the 3.0 mg / kg and 6.0 mg / kg dose levels, over a minimum of 90 minutes for the first dose of 10.0 mg / kg, and over a minimum of 60 minutes for subsequent doses of 10 mg / kg. For combination therapy, ONC-392 was administered first, and there was an interval of at least 30 minutes before a 200 mg fixed dose of IV pembrolizumab was administered over at least 30 minutes.

[0179] The concentration of ONC-392 drug product is 5 mg / mL. Dilute the drug product with 5% dextrose solution to a final concentration between 1.0 and 2.5 mg / mL. The IV infusion line has a 0.2 μm inline filter. After the infusion, flush the line with 25 mL of normal saline according to local institutional guidelines.

[0180] Composition of the drug

[0181] ONC-392 drug product is a sterile liquid for IV administration, supplied in 20 mL glass vials, sealed with 20 mm rubber stopper flip caps. The drug product is formulated in 20 mM histidine buffer, 8.8% (w / v) trehalose dihydrate, 0.06% (w / v) PS80 at pH 6.0 to a protein concentration of 5.0 mg / mL. For an extractable volume of 16 mL (80 mg), the target fill volume is 17.340 mL / bottle. Long-term storage conditions are 2°C to 8°C, protected from light.

[0182] Example 3

[0183] Anti-CTLA-4 antibody is effective against ACC

[0184] This example demonstrates the efficacy of anti-CTLA-4 antibodies (including ONC-392) for the treatment of ACC. ONC-392 produced a clinical response in a patient with ACC, a 48-year-old female with the following clinical diagnosis and medical history: Left nasal ACC in January 2018. Mass in the posterior maxilla, biopsied on January 29, 2018. Pathological diagnosis: ACC, grade 2, cribriform pattern, involving bones. Immunohistochemistry showed that the cells were positive for AE1 / AE2; and negative for P63, TTF1, ER, PR, GATA3, GCDFP15, and CDX2. The cells were strongly positive for MYB and CAM5.2, and negative for p16 and p40.

[0185] The patient received proton / neutron irradiation at the University of Washington in 2019. The tumor recurred in the left maxilla, and the patient underwent salvage surgery for the recurrence in January 2020, including left maxillectomy / fibula free flap reconstruction / orbital floor reconstruction. The cancer recurred in March 2021, and the patient underwent a left revision maxillectomy, left orbital exenteration, anterior skull base and infratemporal fossa malignancy resection-extradural, left neck exploration with scar release and vascular preparation, followed by left radial forearm free flap (with skin island), left thigh split-thickness skin graft, and free soft tissue transfer reconstruction of midface and orbital defects. Anterior skull base dura mater incision with spinal fluid leak was observed on April 23, 2021. The patient received cisplatin therapy with concurrent chemoradiation from June 8, 2021 to July 22, 2021. The cancer grew progressively during treatment and was diagnosed as lung metastasis with multiple bilateral nodules in December 2021.

[0186] The patient was assigned to the group using ONC-392 drug product 10.0 mg / kg ONC-392, Q4W after participating in the ONC-392 clinical trial described in Example 2. Screening MRI (December 9, 2021) on the orbit, face, and neck showed a significant increase in the volume of the left half-face mass. The lesion was centered in the orbit and supraorbital area and extended to the frontal sinus, residual left maxillary sinus, and left anterior temporal soft tissue. Its maximum lateral and AP dimensions were approximately 7.7x4.1 cm. The patient received the first treatment cycle of 10 mg / kg ONC-392 on January 6, 2022, and the second treatment cycle on February 3, 2022.

[0187] Fig. 6A The target lesion from an MRI scan of the patient performed on December 9, 2021 is shown. The scan showed a tumor mass on the left side of the face and forehead of the patient with ACC. After two doses of ONC-392, the patient showed significant aggressive tumor growth, with a large tumor hemorrhage event occurring two days prior to the 8-week MRI assessment that began on February 24, 2022. Figure 6B The target lesion was reported to be an enlarged expansive mass centered in the left orbital region measuring 10.0x7.4x8.9 cm, which was significantly larger than before. The mass appeared to invade the frontal sinus as well as the left nasal cavity into the left sphenoid sinus. The mass also crossed the midline anteriorly toward the right medial canthal soft tissue, which was progressing compared to before. There was no evidence of invasion into the right intraconal space. The left frontal calvaria had been invaded through the endocardium, and the lesion appeared to be adjacent to the left frontal lobe. Chest CT also showed increased spacing of non-target lesions in the lung metastases.

[0188] The patient had no other treatment options and continued ONC-392 treatment. After the third dose of ONC-392 on March 3, 2022, the facial mass showed rapid regression and was largely resolved within three weeks. An MRI scan of the lesion on March 24, 2022, is shown in Figure 1. Figure 6C As shown. Since the last examination on February 24, 2022, there has been a significant interval decrease in the size of the expansive mass centered in the left orbital region, corresponding to a biopsy confirming a recurrent malignancy. The overall size is difficult to measure due to involvement of many surrounding structures and the diffuse appearance of the mass. Overall, there was a reduction in volume, with the mass estimated to be at least approximately 75%. No radiological pathological lymphadenopathy was found. Based on MRI evaluation, the main lesions in the head showed a 78% reduction in tumor burden from baseline. Chest CT showed interval shortening of metastatic lung lesions. The overall assessment on March 24, 2022 indicated a partial response (PR) and the patient continued treatment. The results from this patient suggest that anti-CTLA-4 antibodies can be used as the sole immunotherapy agent to treat cancer, including ACC.

Claims

1. A method of treating adenoid cystic carcinoma (ACC) in a subject in need thereof, the method comprising administering an anti-CTLA-4 antibody to the subject.

2. The method of claim 1, wherein the ACC is recurrent, metastatic, or both.

3. The method of claim 1 or 2, wherein the anti-CTLA-4 antibody is the only immunotherapeutic agent administered to the subject.

4. The method of claim 3, wherein the anti-CTLA-4 antibody is administered as a monotherapy.

5. The method of any one of claims 1-4, wherein the anti-CTLA-4 antibody is selected from the group consisting of Ipilimumab / Yervoy, XTX101, Botensilimab, Zaliferelimab, ADG116, HBM4003, APL-509, BA3071, BMS-986249, Tikab, and quavonlimab (MK-1308), and combinations thereof.

6. The method of any one of claims 1-4, wherein the anti-CTLA-4 antibody comprises: (a) a light chain variable region comprising a complementarity determining region (CDR) 1 comprising the amino acid sequence shown in SEQ ID NO: 1; a CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs: 2-4; and a CDR3 comprising the amino acid sequence shown in SEQ ID NO: 5; and (b) a heavy chain variable region comprising a CDR1 comprising the amino acid sequence shown in SEQ ID NO:6; a CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs:7-9; and a CDR3 comprising the amino acid sequence shown in SEQ ID NO:

10.

7. The method of claim 6, wherein the anti-CTLA-4 antibody comprises a heavy chain comprising an Fc region of a human Ig protein.

8. The method of claim 7, wherein the human Ig protein is human IgG1. 9 . The method of claim 8 , wherein the Fc region of the human IgG1 protein comprises the sequence shown in SEQ ID NO: 17 or 18.

10. The method of claim 6, wherein the anti-CTLA-4 antibody comprises a light chain variable region comprising a CDR2 comprising the sequence shown in SEQ ID NO:3 and a heavy chain variable region comprising a CDR2 comprising the sequence shown in SEQ ID NO:

9.

11. The method of claim 10, wherein the anti-CTLA-4 antibody comprises a light chain variable region comprising the sequence shown in SEQ ID NO: 12 and a heavy chain variable region comprising the sequence shown in SEQ ID NO:

16.

12. The method of claim 11, wherein the anti-CTLA-4 antibody comprises a light chain comprising the sequence shown in SEQ ID NO:23 and a heavy chain comprising the sequence shown in SEQ ID NO:

21.

13. A method of treating adenoid cystic carcinoma in a subject in need thereof, the method comprising administering an anti-CTLA-4 antibody to the subject, wherein the anti-CTLA-4 antibody comprises a light chain comprising the sequence shown in SEQ ID NO:23 and a heavy chain comprising the sequence shown in SEQ ID NO:

21.

14. The method of claim 13, wherein the anti-CTLA-4 antibody has been diluted with 5% dextrose solution from a formulation at pH 6.0 containing 5 mg / mL anti-CTLA-4 antibody, 20 mM histidine buffer, 8.8% (w / v) trehalose dihydrate, and 0.06% (w / v) PS80 to a final concentration of about 1.0 to about 2.5 mg / mL.

15. The method of claim 13 or 14, wherein the anti-CTLA-4 antibody is the only immunotherapeutic agent administered to the subject.

16. The method of claim 15, wherein the anti-CTLA-4 antibody is administered as a monotherapy.

17. A composition comprising an anti-CTLA-4 antibody, wherein the anti-CTLA-4 antibody comprises: (a) a light chain variable region comprising a complementarity determining region (CDR) 1 comprising the amino acid sequence shown in SEQ ID NO: 1; a CDR2 comprising the amino acid sequence shown in SEQ ID NO: 3; and a CDR3 comprising the amino acid sequence shown in SEQ ID NO: 5; and (b) a heavy chain variable region comprising a CDR1 comprising the amino acid sequence shown in SEQ ID NO:6; a CDR2 comprising the amino acid sequence shown in SEQ ID NO:9; and a CDR3 comprising the amino acid sequence shown in SEQ ID NO:10, The anti-CTLA-4 antibody has been diluted with 5% dextrose solution from a pH 6.0 formulation containing 5 mg / mL anti-CTLA-4 antibody, 20 mM histidine buffer, 8.8% (w / v) trehalose dihydrate, and 0.06% (w / v) PS80 to a final concentration of about 1.0 to about 2.5 mg / mL.

18. The composition of claim 17, wherein the light chain variable region comprises the sequence shown in SEQ ID NO: 12, and the heavy chain variable region comprises the sequence shown in SEQ ID NO: 16.

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