AMHR2-ED cancer vaccine formulation

Abstract

Provided herein are compositions, systems, kits, and methods of using compositions comprising at least a portion of an anti-Mullerian hormone receptor type II extracellular domain (AMHR-ED) and an adjuvant comprising i) squalene oil, ii) a non-ionic surfactant (e.g., Tween 80), iii) an emulsifier (e.g., sorbitan trioleate), and iv) a buffer (e.g., citrate buffer). In certain embodiments, such compositions are administered to a female subject to treat or prevent ovarian or endometrial cancer (e.g., by inducing expression of anti-AMHR2-ED IgG antibodies by the subject in vivo).

Description

Detailed Description of the Invention

[0001] [Technical field] Related Applications This application claims priority to U.S. Provisional Patent Application No. 63 / 310,731, filed February 16, 2022, which is incorporated by reference in its entirety. Sequence Listing The text of the computer readable sequence listing entitled "40282-601_SEQUENCE_LISTING", generated on February 15, 2023, and having a file size of 10,200 bytes, is submitted herewith and is incorporated by reference in its entirety into this specification. Technical Field Provided herein are compositions, systems, kits, and methods of using the compositions, comprising at least a portion of an anti-Mullerian hormone receptor type II extracellular domain (AMHR-ED) and an adjuvant comprising: i) squalene oil; ii) a non-ionic surfactant (e.g., Tween 80); iii) an emulsifier (e.g., sorbitan trioleate); and iv) a buffer (e.g., citrate buffer). In certain embodiments, such compositions are administered to a woman to treat or prevent ovarian or endometrial cancer (e.g., by inducing the expression of anti-AMHR2-ED IgG antibodies by the subject in vivo).

[0002] [Background technology] Epithelial ovarian cancer (EOC) is the most lethal of all malignant gynecological diseases in postmenopausal women, accounting for more than 75% of all cases [1, 2]. The inconclusive set of symptoms associated with EOC onset and the lack of effective biomarkers for early detection often lead to delayed diagnosis at advanced disease stages, which, following current standard treatment, often result in high disease recurrence rates and poor prognosis [1-7]. Thus, there remains an unmet need for more effective methods to control this disease. Induction of ovarian tumor immunity by vaccination is a promising approach, which is supported by the increased overall survival observed in patients whose ovarian tumors are infiltrated by T cells [8].

[0003] Anti-Müllerian hormone receptor type II (AMHR2) has been shown to be expressed in approximately 92% of primary EOC, 78% of borderline malignant tumors, 77-86% of non-EOC ovarian tumors, and 56% of malignant ascites from stage III-IV ovarian cancer [9-12]. AMHR2 is a serine / threonine kinase receptor that is homologous to type II receptors of the transforming growth factor beta (TGFβ) superfamily

[13] . Anti-Müllerian hormone (AMH) is the cognate ligand of AMHR2, and upon binding to the extracellular domain of AMHR2 (AMHR2-ED), it signals to cause cell cycle arrest and programmed cell death, resulting in regression of the Müllerian duct during male fetal development, as well as the control of egg cell development, ovarian reserve, and fertility in adult women [14, 15]. In adult women, the longest AMHR2 transcript encodes a 573 amino acid protein expressed exclusively in the ovary, consisting of a 403 amino acid cytoplasmic domain with kinase activity (AMHR2-CD) and a 26 amino acid hydrophobic transmembrane domain, both of which exhibit extraovarian expression.

[16] This long ovary-specific transcript also encodes a 127 amino acid ligand-binding extracellular domain (AMHR2-ED) that is expressed exclusively in human premenopausal ovaries

[16] , and its expression decreases to non-immunogenic levels in the postmenopausal ovary.

[17]

[0004] Summary of the Invention Provided herein are compositions, systems, kits, and methods of using the compositions, comprising at least a portion of an anti-Mullerian hormone receptor type II extracellular domain (AMHR-ED) and an adjuvant comprising: i) squalene oil; ii) a non-ionic surfactant (e.g., Tween 80); iii) an emulsifier (e.g., sorbitan trioleate); and iv) a buffer (e.g., citrate buffer). In certain embodiments, such compositions are administered to a woman to treat or prevent ovarian or endometrial cancer (e.g., by inducing the expression of anti-AMHR2-ED IgG antibodies by the subject in vivo).

[0005] In some embodiments, provided herein is a composition comprising: a) a polypeptide comprising at least a portion of an anti-Mullerian hormone receptor type II extracellular domain (AMHR-ED); and b) an adjuvant comprising: i) squalene oil; ii) a non-ionic surfactant; iii) an emulsifier; and iv) a buffer. In some embodiments, the polypeptide is configured to activate CD4+ T cells when administered to a subject (e.g., a human subject, such as a human female subject). In certain embodiments, the polypeptide is configured to induce the production of AMHR2-ED-specific IgG in vivo when administered to a subject (e.g., a human subject, such as a human female subject).

[0006] In other embodiments, a system or kit is provided herein that includes: a) a polypeptide comprising at least a portion of an anti-Mullerian hormone receptor type II extracellular domain (AMHR-ED) (e.g., of human origin); and b) an adjuvant comprising i) squalene oil, ii) a non-ionic surfactant, iii) an emulsifier, and iv) a buffer. In some embodiments, the polypeptide is configured to activate CD4+ T cells when administered to a subject (e.g., a human subject, such as a human female subject). In certain embodiments, the polypeptide is configured to induce the production of AMHR2-ED-specific IgG in vivo when administered to a subject (e.g., a human subject, such as a human female subject).

[0007] In some embodiments, provided herein are methods of treating ovarian or endometrial cancer tumors in a female subject, comprising administering to the female subject (e.g., a human female) a composition as described above and herein. In other embodiments, provided herein are methods of preventing ovarian and / or endometrial cancer in a subject (e.g., a human female subject), comprising administering to the subject a composition as described above and herein. In some embodiments, the subject does not have detectable ovarian and / or endometrial cancer.

[0008] In certain embodiments, the ovarian cancer tumor is a primary ovarian cancer tumor. In some embodiments, the ovarian cancer tumor is a metastatic tumor. In certain embodiments, the ovarian cancer tumor is an epithelial ovarian cancer (EOC) tumor.

[0009] In some embodiments, administration of the composition induces an immune response against ovarian and / or endometrial cancer tumors in the subject, hi some embodiments, administration of the composition induces the expression of anti-AMHR2-ED IgG antibodies by the subject.

[0010] In some embodiments, the ovarian and / or endometrial cancer tumor expresses AMHR2-ED. In some embodiments, the method further comprises determining whether the ovarian and / or endometrial cancer tumor expresses AMHR2-ED. In further embodiments, the method further comprises determining whether administration of the composition induces an immune response in the subject against the ovarian or endometrial cancer tumor. In other embodiments, the method further comprises repeating administration of the composition to the subject (e.g., one or more times, two or more times, three or more times, etc.). In certain embodiments, the adjuvant comprises ADDAVAX™ (InvivoGen) or MF59 (Novartis, Siona), which are commercially available.

[0011] In some embodiments, the methods provided herein further comprise administering to the subject an additional anti-cancer agent. In certain embodiments, the additional anti-cancer agent is selected from the group consisting of paclitaxel, cisplatin, topotecan, gemcitabine, bleomycin, etoposide, carboplatin, docetaxel, doxorubicin, topotecan, cyclophosphamide, trabectedin, olaparib, tamoxifen, letrozole, bevacizumab, anti-CTLA4 antibody, anti-PD-1 antibody, and anti-PD-L1 antibody. In some embodiments, the additional agent is an anti-cancer agent. In some embodiments, the anti-cancer agent is paclitaxel, cisplatin, topotecan, gemcitabine, bleomycin, etoposide, carboplatin, docetaxel, doxorubicin, topotecan, cyclophosphamide, trabectedin, olaparib, tamoxifen, letrozole, or bevacizumab. In some embodiments, the anti-cancer agent is an immune checkpoint inhibitor. In some embodiments, the immune checkpoint inhibitor is an inhibitor of CTLA4, such as an anti-CTLA4 antibody (e.g., ipilimumab (BMS), tremelimumab (AstraZeneca), and / or KAHR-102 (Kahr Medical)). In some embodiments, the immune checkpoint inhibitor is an inhibitor of PD-1, such as an anti-PD-1 antibody (e.g., nivolumab (BMS), pembrolizumab / lambrolizumab (Merck), pidilizumab (Curetech), AMP-224 (GSK), AMP-514 (AstraZeneca), STI-A1110 (Sorrento), and / or TSR-042 (Tesaro). In some embodiments, the immune checkpoint inhibitor is an inhibitor of PD-L1 and / or anti-PD-L2 antibody (e.g., RG-7446 (Roche), BMS-936559 (BMS), MEDI-4736 (AstraZeneca), MSB-0020718C (Merck), AUR-012 (Pierre Fabre and inhibitors of PD-L1 and / or PD-L2, such as STI-A1010 (Sorrento), STI-A1010 (Med), STI-A1010 (Sorrento). In some embodiments, the subject has had surgery to remove at least a portion of the ovarian and / or endometrial cancer tumor.

[0012] In some embodiments, the emulsifier comprises sorbitan trioleate. In further embodiments, the non-ionic surfactant comprises polysorbate 80, polyglycerol alkyl ether, glucosyl dialkyl ether, crown ether, ester-linked surfactant, polyoxyethylene alkyl ether, Brij, Span (sorbitan ester), and Tween (polysorbate). In further embodiments, the buffer comprises citrate buffer or phosphate buffered saline. In other embodiments, the pH of the composition is 5.5 to 6.9.

[0013] In certain embodiments, the composition is in the form of an emulsion. In certain embodiments, the emulsion is a nanoemulsion of a first component and a second component, the first component comprising an emulsifier and squalene oil, and the second component comprising a non-ionic surfactant and a buffer. In other embodiments, the emulsifier is present in the first component at about 0.4-0.6% weight / volume (e.g., 0.4%, 0.5%, or 0.6%). In further embodiments, the squalene oil is present in the first component at about 4-6% weight / volume (e.g., 0.4%, 0.5%, or 0.6%). In some embodiments, the non-ionic surfactant is present in the second component at about 0.4-0.6% weight / volume (e.g., 0.4%, 0.5%, or 0.6%). In further embodiments, the buffer is present in the second component at about 4-6% weight / volume (e.g., 4%, 5%, or 6%). In certain embodiments, the nanoemulsion is composed of particles having an average size of about 150-170 nm.

[0014] In further embodiments, the adjuvant further comprises at least one of the following: saponin, cholesterol, phospholipids, phosphatidylcholine, and a second buffer solution. In some embodiments, the second buffer solution comprises phosphate buffered saline.

[0015] In some embodiments, the composition comprises an oil phase and an aqueous phase. In further embodiments, squalene is present at 4.0-5.5% (e.g., 4...4.3...4.6...4.9...5.2...5.5%) of the oil phase. In other embodiments, the nonionic surfactant is present at about 0.4-0.6% (e.g., 0.4%, 0.5%, or 0.6%) of the aqueous phase. In further embodiments, the emulsifier is present at about 0.4-0.6% (e.g., 0.4%, 0.5%, or 0.6%) of the aqueous phase.

[0016] In some embodiments, the adjuvant further comprises α-tocopherol. In certain embodiments, the adjuvant further comprises monophosphoryl lipid A. In certain embodiments, the adjuvant further comprises an immunomodulatory agent. In further embodiments, the immunomodulatory agent comprises synthetic trehalose dicorynomycolate.

[0017] In certain embodiments, the polypeptide comprises the amino acid sequence of SEQ ID NO: 7, 8, 9, or 10. In some embodiments, the polypeptide comprises the complete human AMHR-ED or nearly complete human AMHR2-ED. In some embodiments, the polypeptide comprises at least 20 contiguous amino acids from SEQ ID NO: 7. In further embodiments, the polypeptide comprises at least 40 contiguous amino acids from SEQ ID NO: 7. In certain embodiments, the polypeptide comprises at least 60 contiguous amino acids from SEQ ID NO: 7. In further embodiments, the polypeptide comprises at least 100 contiguous amino acids from SEQ ID NO: 7. In some embodiments, the polypeptide comprises at least 115 contiguous amino acids from SEQ ID NO: 7. In further embodiments, the polypeptide comprises at least 122 contiguous amino acids from SEQ ID NO: 7. In other embodiments, the polypeptide has an amino acid sequence that comprises 100 contiguous amino acids that are at least 80% identical to the amino acid sequence of the extracellular domain of AMHR2.

[0018] BRIEF DESCRIPTION OF THE DRAWINGS Figure 1: AMHR2 gene expression in mouse EOC. qRT-PCR analysis of ID8 and MOVCAR EOC cell lines shows that both cell lines express the AMHR2-ED and AMHR2-CD domains, but expression of each AMHR2 domain in MOVCAR cells was consistently approximately twice the expression level measured in ID8 cells. Error bars indicate ±SD.

[0019] FIG. 2. Formulation of AMHR2-ED vaccine with ADDAVAX™ induces high IgG responses. ELISA analysis shows that the use of ADDAVAX™ as an adjuvant when immunizing C57BL / 6 female mice resulted in (A) antigen-specific IgG responses to the AMHR2-ED immunogen, and (B) antigen-specific IgG isotype responses similar to those obtained using CFA as the adjuvant. Serum was analyzed 8 weeks after immunization using a dilution of 1:10,000. Error bars indicate ±SD.

[0020] FIG. 3 shows that AMHR2-ED vaccination with ADDAVAX™ as an adjuvant is highly effective in inhibiting the growth of mouse EOC. Prophylactic AMHR2-ED vaccination with ADDAVAX™ as an adjuvant resulted in significant inhibition of (A) MOVCAR growth and significant increase in (B) overall survival in TgMISIIR-Tag(low) female mice compared to vaccination with CFA alone or vaccination against AMHR2-ED in CFA. AMHR2-ED vaccination in ADDAVAX™ was also effective in inhibiting the growth of (C) MOVCAR and (D) ID8 tumors when vaccination was administered when tumors were palpable. Error bars = ± SD, asterisks indicate significance.

[0021] FIG. 4: AMHR2-ED vaccination with ADDAVAX™ adjuvant results in T cell infiltration of murine EOC. AMHR2-ED vaccination in ADDAVAX™ resulted in infiltration of CD3+ T cells (arrow, bottom row, left), CD4+ T cells (arrow, bottom row, center), and CD8+ T cells (arrow, bottom row, right) compared to corresponding EOC tissues from mice vaccinated with ADDAVAX™ alone (arrow, top row). Tissues are representative of several examined from several different mice. Tissue magnification = 20x.

[0022] FIG. 5 shows exemplary sequences. Panel A shows an exemplary human AMHR2-ED amino acid sequence (SEQ ID NO: 7). Panel B shows an exemplary N-terminally truncated human AMHR2-ED amino acid sequence (SEQ ID NO: 8). Panel C shows an exemplary C-terminally truncated human AMHR2-ED amino acid sequence (SEQ ID NO: 9). Panel D shows an exemplary N- and C-terminally truncated human AMHR2-ED amino acid sequence (SEQ ID NO: 10). definition For convenience, certain terms employed in the specification, examples, and appended claims are collected here.

[0023] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0024] As used herein, the term "administering" means providing a pharmaceutical agent or composition to a subject (e.g., intravenously), and includes, but is not limited to, administration by a medical professional and self-administration.

[0025] The phrase "pharmaceutical acceptable carrier" as used herein means a pharmaceutical acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, that is involved in carrying or transporting a subject compound from one organ or part of the body to another organ or part of the body. In certain embodiments, the compositions herein contain a pharmaceutical acceptable carrier.

[0026] As used herein, the term "subject" means a human or non-human animal selected for treatment or therapy.

[0027] As used herein, the phrases "therapeutically effective amount" and "effective amount" mean an amount of an agent effective to produce a desired therapeutic effect in at least a subpopulation of cells in a subject, at a reasonable benefit / risk ratio applicable to any medical treatment.

[0028] "Treating" a disease in a subject or a subject having a disease refers to subjecting the subject to medical therapy, e.g., administration of a vaccine, such that at least one symptom of the disease is reduced or prevented from worsening. [Mode for carrying out the invention] Provided herein are compositions, systems, kits, and methods of using the compositions, comprising at least a portion of an anti-Mullerian hormone receptor type II extracellular domain (AMHR-ED) and an adjuvant comprising i) squalene oil, ii) a non-ionic surfactant (e.g., Tween 80), iii) an emulsifier (e.g., sorbitan trioleate), and iv) a buffer (e.g., citrate buffer). In certain embodiments, such compositions are administered to a female subject (e.g., a human) to treat or prevent ovarian or endometrial cancer (e.g., by inducing the expression of anti-AMHR2-ED IgG antibodies by the subject in vivo).

[0029] In certain embodiments, the adjuvant used herein is an adjuvant that is chemically identical and / or highly similar to the adjuvant marketed as ADDAVAX™ (InvivoGen) and / or ADDAVAX™. ADDAVAX™ is a nanoemulsion of two components: i) sorbitan trioleate (0.5% w / v) in squalene oil (5% v / v) and ii) Tween 80 (0.5% w / v) in sodium citrate buffer (10 mM, pH 6.5), where the nanoemulsion is generated using a microfluidizer and filtered through a 0.22 μm filter to remove large droplets and sterilize the final product. The particle size is approximately 160 nm.

[0030] In certain embodiments, the adjuvant used herein is MF59 (Novartis, Siona) and / or an adjuvant that is chemically identical and / or closely similar to the adjuvant commercially available as MF59. MF59 is an approximately 160 nm particle emulsion that contains i) squalene: 9.75 mg, ii) polysorbate 80: 1.175 mg, iii) sorbitan trioleate: 1.175 mg, iv) sodium citrate: 0.66 mg, and v) citric acid: 0.04 mg.

[0031] In certain embodiments, the adjuvant used herein is Abisco-100 (known as Matrix-M when made to GMP standards) and / or an adjuvant that is chemically identical and / or very similar to the commercially available adjuvant Abisco-100. Abisco-100 has the following chemical content: purified saponin obtained from a crude extract of the plant, Quillaja saponaria Molina, cholesterol from lanolin, and phosphatidylcholine (phospholipids) from fresh egg yolk, in a suspension of nano-sized (40 nm) cage-like particles made of the above components in PBS. Matrix M (or Abisco-100) is composed of a mixture of Matrix A and Matrix C in a ratio of 80:20 to 95:5, preferably 85:15. Matrix A provides T cell induction and has low toxicity, while Matrix C induces antibodies and has some toxicity. Matrix C contains the C fraction of QS separation, which corresponds to QS21. Fraction A (in matrix A) corresponds to QS7.

[0032] In some embodiments, the adjuvant used herein is AS03 adjuvant and / or an adjuvant that is chemically identical and / or very similar to the adjuvant marketed as AS03 adjuvant. AS03 adjuvant is an emulsion adjuvant composed of an oil phase (10.69 mg squalene, 11.86 mg DL-α-tocopherol) and an aqueous phase (4.86 mg polysorbate 80), each at a 0.5 mL adult dose. ASO3 is with PBS.

[0033] In certain embodiments, the adjuvant used herein is Ribi adjuvant and / or an adjuvant that is chemically identical and / or very similar to the adjuvant marketed as Ribi adjuvant. Ribi adjuvant is an oil-in-water emulsion containing 2% squalene-Tween 80-water, 0.5 mg monophosphoryl lipid A (a low-toxicity derivative of the lipid A region of lipopolysaccharide (LPS)), and 0.5 mg synthetic trehalose dicorynomycolate (an immunomodulator). This adjuvant is in PBS.

[0034] In certain embodiments, the compositions provided herein comprise an AMHR2-ED polypeptide and / or an immunologically active fragment of an AMHR2-ED polypeptide. Polypeptides with substantial sequence similarity may elicit the same or a very similar immune response in a host animal. Thus, in some embodiments, derivatives, equivalents, variants, fragments, or mutants of an AMHR2-ED protein (e.g., as set forth in SEQ ID NO: 7) or a fragment thereof may also be suitable for the methods, compositions, and kits provided herein.

[0035] In some embodiments, variants or derivatives of AMHR2-ED polypeptides (e.g., SEQ ID NOs: 7-10) are provided herein. Modified polypeptides may have an amino acid sequence that is modified, for example, by conservative substitution, but still elicit an immune response that reacts with the unmodified protein antigen and are considered functional equivalents. As used herein, the term "conservative substitution" refers to the replacement of an amino acid residue with another biologically similar residue. It is well known in the art that amino acids within the same conservative group can typically be substituted for each other without substantially affecting the function of the protein. According to certain embodiments, a derivative, equivalent, variant, or mutant of the ligand-binding domain of an AMHR2-ED polypeptide is a polypeptide that is at least 85% homologous to the sequence of an AMHR2-ED protein or a fragment thereof. In some embodiments, the homology is at least 90%, at least 95%, or at least 98% (e.g., compared to SEQ ID NOs: 7-10).

[0036] In certain aspects, provided herein are pharmaceutical compositions (e.g., vaccine compositions) comprising an AMHR2-ED polypeptide described herein and an adjuvant described herein. In some embodiments, the composition comprises a pharma- ceutically acceptable carrier. In some embodiments, the composition comprises a combination of multiple, e.g., two or more, AMHR2-ED polypeptides or nucleic acids described herein.

[0037] The pharmaceutical compositions disclosed herein may be specially formulated for administration in solid or liquid form, including those adapted for: (1) oral administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., buccal, sublingual, and those intended for systemic absorption, boluses, powders, granules, pastes for application to the tongue; or (2) parenteral administration, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection, e.g., as a sterile solution or suspension, or sustained release formulation.

[0038] Methods of preparing these formulations or compositions include the step of bringing into association an AMHR2-ED polypeptide and adjuvant described herein with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the agents described herein with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0039] Pharmaceutical compositions suitable for parenteral administration include the AMHR2-ED polypeptide and adjuvants described herein in combination with one or more pharma- ceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately prior to use. The pharmaceutical compositions may contain sugars, alcohols, antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0040] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0041] Regardless of the route of administration selected, the agents and / or pharmaceutical compositions provided herein, which may be used in a suitable hydrated form, are formulated into pharma- ceutical acceptable dosage forms by conventional methods known to those of skill in the art.

[0042] In some embodiments, the described pharmaceutical compositions, when administered to a subject, are capable of eliciting an immune response against cells expressing AMHR2-ED. Such pharmaceutical compositions may be useful as vaccine compositions for the prophylactic and / or therapeutic treatment of ovarian or endometrial cancer.

[0043] In certain embodiments, the compositions provided herein also include one or more other agents, such as, but not limited to, a chemotherapeutic agent, an immunotherapeutic agent, an immunomodulatory agent, and / or an anti-angiogenic agent. In some embodiments, the one or more other agents may be naturally occurring or synthetic chemotherapeutic agents, for example, as described in "Cancer Chemotherapeutic Agents", American Chemical Society, 1995, WOFoye Ed.

[0044] In one embodiment, the chemotherapeutic agent is a small molecule receptor antagonist such as vatalanib, SU11248 or AZD-6474, an EGFR or HER2 antagonist such as gefitinib, erlotinib, CI-1033 or herceptin, an antibody such as bevacizumab, cetuximab, rituximab, a DNA alkylating agent such as cisplatin, oxaliplatin or carboplatin, an anthracycline such as doxorubicin or epirubicin, 5-FU, pemetrexed, gemcitabine or capecitabine. and the like, camptothecins such as irinotecan or topotecan, anticancer drugs such as paclitaxel or docetaxel, epipodophyllotoxins such as etoposide or teniposide, proteasome inhibitors such as bortezomib, or anti-inflammatory drugs such as celecoxib or rofecoxib (optionally in the form of a pharma- ceutically acceptable salt, in the form of a hydrate and / or solvate, and optionally in the form of an individual optical isomer, a mixture of the individual enantiomers or a racemate thereof).

[0045] In another embodiment, the chemotherapeutic agent is a small molecule VEGF receptor antagonist such as vatalanib (PTK-787 / ZK222584), SU-5416, SU-6668, SU-11248, SU-14813, AZD-6474, AZD-2171, CP-547632, CEP-7055, AG-013736, IM-842 or GW-786034, a dual EGFR / HER2 antagonist such as gefitinib, erlotinib, CI-1033 or GW-2016, Iressa (ZD-1839), Tarceva (OSI-774), PKI-1 EGFR antagonists such as EKB-569, HKI-272 or Herceptin, mitogen-activated protein kinase antagonists such as BAY-43-9006 or BAY-57-9006, 4-[(3-chloro-4-fluorophenyl)amino]-6-{[4-(N,N-dimethylamino)-1-oxo-2-but-1-yl]amino}-7-((S)-tetrahydrofuran-3-yloxy)quinazoline or 4-[(3-chloro-4-fluoro-phenyl)amino]-6-{[4-(homomorpholin-4-yl)- quinazoline derivatives such as 1-oxo-2-buten-1-yl]amino}-7-[(S)-(tetrahydrofuran-3-yl)oxy]-quinazoline or a pharma- ceutically acceptable salt thereof; protein kinase receptor antagonists not classified under synthetic small molecules such as atrasentan, rituximab, cetuximab, Avastin™ (bevacizumab), IMC-1C11, Erbitux (C-225), DC-101, EMD-72000, vitaxin, and imatinib; protein tyrosine kinases that are fusion proteins such as VEGFtrap; enzyme inhibitors, melphalan, cyclophosphamide, oxazaphosphorine, cisplatin, carboplatin, oxaliplatin, satraplatin, tetraplatin, iproplatin, mitomycin, streptozocin, carmustine (BCNU), lomustine (CCNU), busulfan, ifosfamide, streptozocin, thiotepa, alkylating agents such as chlorambucil or platinum compounds, nitrogen mustards such as mechlorethamine, ethylenimine compounds, alkylsulfonates, daunorubicin, doxorubicin (adriamycin),Liposomal doxorubicin (Doxil), epirubicin, idarubicin, mitoxantrone, amsacrine, dactinomycin, distamycin or its derivatives, netropsin, pibenzimol, mitomycin, CC-1065, duocarmycin, mithramycin, chromomycin, olivomycin, phthalanilides such as propamidine or stilbamidine, anthramycin, aziridine, nitrosoureas or their derivatives, cytarabine, 5-fluorouracil (5-FU), pemetrexed, tegafur / uracil pyrimidine or purine analogs or antagonists, such as uracil mustard, fludarabine, gemcitabine, capecitabine, mercaptopurine, cladribine, thioguanine, methotrexate, pentostatin, hydroxyurea, or folic acid, or inhibitors of nucleoside diphosphate reductase, phleomycin, bleomycin or a derivative or salt thereof, CHPP, BZPP, MTPP, BAPP, ribromycin, acridine or a derivative thereof, rifamycin, actinomycin, adramycin, irinomycin, camptothecins such as camptothecan (camptosar) or topotecan, amsacrine or its analogs, tricyclic carboxamides, histone deacetylase inhibitors such as SAHA, MD-275, trichostatin A, CBHA, LAQ824, or valproic acid, plant-derived anticancer drugs such as paclitaxel (taxol), docetaxel, or taxotere, vinca alkaloids such as navelbine, vinblastine, vincristine, vindesine, or vinorelbine, tropolone alkaloids such as colchicine or its derivatives, macrolides such as roids, maytansine, ansamitocins or rhizoxins; mitotic inhibitor peptides such as phomopsin or dolastatins; epipodophyllotoxin or podophyllotoxin derivatives such as etoposide or teniposide; antimitotic carbamate derivatives such as steganacin, combretastatin or amfetinil; proteasome inhibitors such as procarbazine, bortezomib; enzymes such as asparaginase, pegylated asparaginase (pegaspargase), or thymidine-phosphorylase inhibitors;Gestagens or estrogens such as estramustine (T-66) or megestrol; antiandrogens such as flutamide, casodex, anandrone or cyproterone acetate; aromatase inhibitors such as aminoglutethimide, anastrozole, formestane or letrozole; GNrH analogues such as leuprorelin, buserelin, goserelin or triptorelin; antiestrogens such as tamoxifen or its citrate, droloxifene, trioxyphene, raloxifene or zindoxifene. 17.β-estradiol derivatives such as ICI164,384 or ICI182,780; LH-RH antagonists such as aminoglutethimide, formestane, fadrozole, finasteride, ketoconazole, leuprolide; steroids such as prednisone, prednisolone, methylprednisolone, dexamethasone, budenoside, fluocortolone or triamcinolone; interferons such as interferon-beta; interleukins such as IL-10 or IL-12; anti-TNFα drugs such as etanercept. Antibodies, thalidomide, its R- and S-enantiomers and derivatives, or immunomodulators such as revimid (CC-5013), leukotriene antagonists, mitomycin C, BMY-42355, aziridoquinones such as AZQ or EO-9, misonidazole, 2-nitroimidazoles such as NLP-1 or NLA-1, nitroacridines, nitroquinolines, nitropyrazoloacridines, RSU-1069 or RB-6145, "dual function" nitroaromatics such as CB-1954, nitroamines such as nitroamines, etc. N-oxides of nitrogen mustard, metal complexes of nitrogen mustard, anti-CD3 or anti-CD25 antibodies, tolerance inducers, minodronic acid or its derivatives (YM-529, Ono-5920, YH-529), biphosphonates or their derivatives such as zoledronic acid monohydrate, ibandronate sodium hydrate or clodronate disodium, nitroimidazoles such as metronidazole, misonidazole, benznidazole or nimorazole, nitroaryl compounds such as RSU-1069,Nitroxyls or N-oxides such as SR-4233, halogenated pyrimidine analogues such as bromodeoxyuridine, iododeoxyuridine, thiophosphates such as WR-2721, porfimers, photoactivatable drugs such as Photofrin, benzoporphyrin derivatives, pheophorbide derivatives, merocyanine 540 (MC-540) or tin etioporpurin, ant-templates or antisense RNA or DNA such as oblimersen, acetylsalicylic acid, mesalazine, ibuprofen, naproxen , flurbiprofen, fenoprofen, fenbufen, ketoprofen, indoprofen, pirprofen, carprofen, oxaprozin, pranoprofen, miroprofen, tioxaprofen, suprofen, aluminoprofen, tiaprofenic acid, fluprofen, indomethacin, sulindac, tolmetin, zomepirac, nabumetone, diclofenac, fenclofenac, alclofenac, bromfenac, ibufenac, aceclofenac, acemetacin, fentiazac, clidanac, etodolac, oxypinac ( nonsteroidal inflammatory drugs such as oxpinac, mefenamic acid, meclofenamic acid, flufenamic acid, niflumic acid, tolfenamic acid, diflunisal, flufenisal, piroxicam, tenoxicam, romoxicam, nimesulide, meloxicam, celecoxib, rofecoxib, or pharma- ceutically acceptable salts of nonsteroidal inflammatory drugs; cytotoxic antibiotics; antibodies that target cancer cell surface molecules such as apolizumab or 1D09C3; inhibitors of metalloproteinases such as TIMP-1 or TIMP-2; Inhibitors of oncogenes, rare earth complexes such as heterocyclic complexes of lanthanides, photochemotherapeutic agents such as PUVA, inhibitors of the transcription factor complex ESX / DRIP130 / Sur-2, heat shock protein HSP90 modulators, HER-2 expression inhibitors such as geldanamycin and its derivatives 17-allylaminogeldanamycin or 17-AAG, or IM-842, tetrathiomolybdate, squalamine, combrestatin A4, TNP-470, marimastat, neovastat, bicalutamide, abarelix, oregovomab, mitsumomab,The therapeutic agent is selected from the group consisting of TLK-286, alemtuzumab, ibritumomab, temozolomide, denileukin diftitox, aldesleukin, dacarbazine, floxuridine, plicamycin, mitotane, pipobroman, plicamycin, tamoxifen, and testolactone. Preferred compounds include small molecule VEGF receptor antagonists such as vatalanib (PTK-787 / ZK222584), SU-5416, SU-6668, SU-11248, SU-14813, AZD-6474, EGFR / HER2 antagonists such as CI-1033 or GW-2016, Iressa (gefitinib, ZD-1839), Tarceva (erlotinib, OSI-774), PKI-166, EKB-569, HKI-272 or Herceptin. EGFR antagonists such as rituximab, mitogen-activated protein kinase antagonists such as BAY-43-9006 or BAY-57-9006, atrasentan, rituximab, cetuximab, Avastin™ (bevacizumab), IMC-1C11, Erbitux (C-225), DC-101, EMD-72000, vitaxin, imatinib, melphalan, cyclophosphamide, cisplatin, carboplatin, oxaliplatin, satraplatin, alkylating agents or platinum compounds such as daunorubicin, doxorubicin (adriamycin), liposomal doxorubicin (doxil), epirubicin, idarubicin, pyrimidine or purine analogs or antagonists such as cytarabine, 5-fluorouracil (5-FU), pemetrexed, tegafur / uracil, gemcitabine, capecitabine, mercaptopurine, methotrexate, or inhibitors of nucleoside diphosphate reductase, paclitaxel anticancer drugs such as (Taxol) or docetaxel, vinca alkaloids such as navelbine, vinblastine, vincristine, vindesine or vinorelbine, mitotic inhibitor peptides such as dolastatins, epipodophyllotoxin or podophyllotoxin derivatives such as etoposide or teniposide, nonsteroidal inflammatory drugs such as meloxicam, celecoxib, rofecoxib, antibodies targeting cancer cell surface molecules such as apolizumab or ID09C3,or the heat shock protein HSP90 modulator, geldanamycin and its derivative 17-alkylaminogeldanamycin or 17-AAG.

[0046] In another embodiment, the chemotherapeutic agent is a compound that interacts with or binds to tubulin, a synthetic small molecule VEGF receptor antagonist, a small molecule growth factor receptor antagonist, an inhibitor of the EGF receptor and / or the VEGF receptor and / or the integrin receptor or any other protein tyrosine kinase receptor not classified under synthetic small molecules, an inhibitor directed to the EGF receptor and / or the VEGF receptor and / or the integrin receptor or any other protein tyrosine kinase receptor that is a fusion protein, a compound that interacts with nucleic acid and is classified as an alkylating agent or a platinum compound, a compound that interacts with nucleic acid and includes DNA minor groove binding compounds classified as anthracyclines, as DNA intercalators or as DNA crosslinkers, antimetabolites, naturally occurring, semi-synthetic or synthetic bleomycin type antibiotics, inhibitors of DNA transcriptase, in particular topoisomerases. The agent is selected from the group consisting of topoisomerase I or topoisomerase II inhibitors, chromatin modifying agents, mitosis inhibitors, antimitotic agents, cell cycle inhibitors, proteasome inhibitors, enzymes, hormones, hormone antagonists, hormone inhibitors, inhibitors of steroid biosynthesis, steroids, cytokines, hypoxia selective cytotoxins, cytokines, inhibitors of lymphokines, antibodies against cytokines, oral and parenteral tolerance inducers, supportive agents, chemo-radiosensitizers and protectants, photochemically activated drugs, synthetic polynucleotides or oligonucleotides (optionally modified or conjugated), nonsteroidal anti-inflammatory drugs, cytotoxic antibiotics, antibodies targeting surface molecules of cancer cells, antibodies targeting growth factors or their receptors, inhibitors of metalloproteinases, metals, inhibitors of oncogenes, inhibitors of gene transcription or RNA translation or protein expression, complexes of rare earth elements, and photochemotherapeutic agents.

[0047] In other embodiments, the chemotherapeutic agent is a vinca alkaloid such as paclitaxel (taxol), docetaxel, navelbine, vinblastine, vincristine, vindesine, or vinorelbine, an alkylating agent such as melphalan, cyclophosphamide, oxazaphosphorine, cisplatin, carboplatin, oxaliplatin, satraplatin, tetraplatin, iproplatin, mitomycin, streptozocin, carmustine (BCNU), lomustine (CCNU), busulfan, ifosfamide, streptozocin, thiotepa, or chlorambucil. agents or platinum compounds, nitrogen mustards such as mechlorethamine, thalidomide, its R- and S-enantiomers and derivatives, or immunomodulators such as Revimid (CC-5013), ethylenimine compounds, alkylsulfonates, daunorubicin, doxorubicin (adriamycin), liposomal doxorubicin (doxil), epirubicin, idarubicin, mitoxantrone, amsacrine, dactinomycin, distamycin or its derivatives, netropsin, pigenzimol, mitomycin, CC-1065, duocalmycin pyrimidine or purine analogues or antagonists, such as cytarabine, 5-fluorouracil (5-FU), uracil mustard, fludarabine, gemcitabine, capecitabine, mercaptopurine, cladribine, thioguanine, methotrexate, pentostatin, hydroxyurea, or folic acid, or inhibitors of nucleoside diphosphate reductase, acridine or derivatives thereof, camptothecins such as rifamycin, actinomycin, adramycin, irinotecan (camptosar) or topotecan, amsacrine or its analogues, tricyclic carboxamides, histone deacetylase inhibitors such as SAHA, MD-275, trichostatin A, CBHA, LAQ824, or valproic acid, proteasome inhibitors such as bortezomib, vatalanib (PTK-787 / ZK222584), SU-5416, SU-6668, SU-11248, SU-14813, AZD-6474, AZD-2171,small molecule VEGF receptor antagonists such as CP-547632, CEP-7055, AG-013736, IM-842 or GW-786034; mitogen-activated protein kinase antagonists such as BAY-43-9006 or BAY-57-9006; dual EGFR / HER2 antagonists such as gefitinib, erlotinib, CI-1033 or GW-2016; Iressa (ZD-1839), Tarceva (OS I-774), PKI-166, EKB-569, HKI-272 or Herceptin, EGFR antagonists such as 4-[(3-chloro-4-fluorophenyl)amino]-6-{[-4-(N,N-dimethylamino)-1-oxo-2-but-en-1-yl]amino}-7-((S)-tetrahydrofuran-3-yloxy)-quinazoline or 4-[(3-chloro-4-fluoro-phenyl)amino]-6-{[4-( quinazoline derivatives such as {homomorpholin-4-yl}-1-oxo-2-buten-1-yl}amino-7-[(S)-(tetrahydrofuran-3-yl)oxy]-quinazoline, or a pharma- ceutically acceptable salt thereof; inhibitors of the transcription factor complex ESX / DRIP130 / Sur-2; heat shock protein HSP90 modulators; inhibitors of HER-2 expression such as geldanamycin and its derivatives 17-alkylaminogeldanamycin or 17-AAG; protein kinase receptor antagonists not classified as synthetic small molecules such as atrasentan, rituximab, cetuximab, Avastin™ (bevacizumab), IMC-1C11, Erbitux (C-225), DC-101, EMD-72000, vitaxin, imatinib; and antibodies targeting surface molecules of cancer cells such as apolizumab or 1D09C3. ,

[0048] In some embodiments, the anti-cancer agent is an immune checkpoint inhibitor. In some embodiments, the immune checkpoint inhibitor is an inhibitor of CTLA4, such as an anti-CTLA4 antibody (e.g., ipilimumab (BMS), tremelimumab (AstraZeneca), and / or KAHR-102 (Kahr Medical)). In some embodiments, the immune checkpoint inhibitor is an inhibitor of PD-1, such as an anti-PD-1 antibody (e.g., nivolumab (BMS), pembrolizumab / lambrolizumab (Merck), pidilizumab (Curetech), AMP-224 (GSK), AMP-514 (AstraZeneca), STI-A1110 (Sorrento), and / or TSR-042 (Tesaro). In some embodiments, the immune checkpoint inhibitor is an inhibitor of PD-L1 and / or anti-PD-L2 antibody (e.g., RG-7446 (Roche), BMS-936559 (BMS), MEDI-4736 (AstraZeneca), MSB-0020718C (Merck), AUR-012 (Pierre Fabre and PD-L1 and / or PD-L2 inhibitors such as STI-A1010 (Sorrento), STI-A1010 (Med).

[0049] In certain aspects, provided herein are methods for treating or preventing ovarian or endometrial cancer, and / or for inducing an immune response against ovarian or endometrial cancer tumors expressing AMHR2-ED. In certain embodiments, the methods comprise administering to a subject a pharmaceutical composition as described herein. In some embodiments, the ovarian or endometrial cancer tumor is a primary tumor. In some embodiments, the ovarian or endometrial cancer tumor is a metastatic tumor. In some embodiments, the ovarian cancer tumor is an epithelial ovarian cancer (EOC) tumor. In some embodiments, the ovarian cancer tumor expresses AMHR2-ED.

[0050] The methods described herein may be used to treat any subject in need of treatment. As used herein, "subject in need thereof" includes any subject with ovarian or endometrial cancer, any subject who has had ovarian or endometrial cancer, and / or any subject who is predisposed to ovarian or endometrial cancer. For example, in some embodiments, the subject has an ovarian or endometrial cancer tumor (e.g., an ovarian cancer tumor expressing AMHR2-ED). In some embodiments, the subject has undergone surgery to remove at least a portion of an ovarian or endometrial cancer tumor. In some embodiments, the subject is predisposed to ovarian cancer because they have a BRCA1 or BRCA2 gene mutation in their genome that predisposes the subject to ovarian cancer. In some embodiments, the subject is predisposed to ovarian cancer because they have a TP53 gene mutation, a CHEK2 gene mutation, a RAD51 gene mutation, a BRIP1 gene mutation, and / or a PALB2 gene mutation. In some embodiments, the subject has a family history of ovarian cancer.

[0051] The pharmaceutical compositions disclosed herein may be delivered by any suitable route of administration, including oral and parenteral. In certain embodiments, the pharmaceutical compositions are delivered systemically (e.g., by oral or parenteral administration). In some embodiments, the pharmaceutical compositions are delivered intravenously. In some embodiments, the pharmaceutical compositions are delivered intramuscularly. In some embodiments, the pharmaceutical compositions are delivered subcutaneously.

[0052] The dosage of the subject agent can be determined by referring to the plasma concentration of the agent. For example, the maximum plasma concentration (Cmax) and the area under the plasma concentration-time curve from time 0 to infinity (AUC(0-4)) can be used. Dosages include those that result in the above values ​​of Cmax and AUC(0-4), as well as other dosages that result in larger or smaller values ​​of these parameters.

[0053] Actual dosage levels of the active ingredients in pharmaceutical compositions may be varied to obtain an amount of the active ingredient effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without being toxic to the patient.

[0054] The selected dosage level will depend on a variety of factors, including the activity of the particular agent used, the route of administration, the time of administration, the rate of excretion or metabolism of the particular compound used, the duration of treatment, other drugs, compounds, and / or materials used in combination with the particular compound used, the age, sex, weight, condition, general health, and previous medical history of the patient being treated, and similar factors well known in the medical arts.

[0055] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian can prescribe and / or administer doses of the agents used in the pharmaceutical composition at levels lower than those required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved.

[0056] Generally, a suitable daily dose of an agent described herein will be that amount of the agent that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above.

[0057] In one aspect, provided herein is a method for inducing an immune response in a subject against cells expressing AMHR2-ED (e.g., ovarian cancer tumor cells). The method comprises administering to the subject a pharmaceutical composition as described herein, wherein the pharma- ceutically acceptable composition, when administered to the subject, induces an immune response against cells expressing AMHR2-ED. In general, the immune response can include a humoral immune response, a cell-mediated immune response, or both.

[0058] The humoral response can be determined by standard immunoassays for antibody levels in serum samples from subjects receiving the pharmaceutical composition. The cellular immune response is a response involving T cells and can be determined in vitro or in vivo. For example, the general cellular immune response can be determined as T cell proliferation activity in cells (e.g., peripheral blood leukocytes (PBL)) sampled from a subject at a suitable time after administration of a pharma- ceutical acceptable composition. For example, PBMCs are incubated with a stimulating agent for a suitable period of time, and then [ 3 [H]thymidine incorporation can be measured. Proliferating T cell subsets can be determined using flow cytometry.

[0059] In certain aspects, the methods provided herein include administration to both human and non-human mammals. Veterinary applications are also contemplated. In some embodiments, the subject may be any living female organism in which an immune response can be elicited. Examples of subjects include, but are not limited to, humans, livestock, dogs, cats, mice, rats, and transgenic species thereof.

[0060] In certain embodiments, the subject has a history of ovarian or endometrial cancer and has been administered another treatment modality. The other therapies may include, for example, surgical resection, radiation therapy, chemotherapy, and other modalities of immunotherapy where the subject does not present clinically measurable tumors as a result of other therapies. However, the subject may be one who has been determined to be at risk for cancer recurrence or progression, either near the original tumor site or by metastasis. Such subjects may be further classified into high-risk and low-risk subjects. Subdivision can be performed based on characteristics observed before or after initial treatment. These characteristics are known in the clinical art and are well defined for different cancers. Typical characteristics of the high-risk subgroup are those where the tumor has invaded adjacent tissues or shows lymph node involvement. Thus, for example, the pharmaceutical compositions described herein can be administered to the subject to induce an anti-cancer response, primarily as a preventative measure against recurrence.

[0061] In some embodiments, the pharmaceutical composition can be administered at any suitable time.For example, the administration can be performed before or during the conventional treatment of the subject with ovarian cancer or endometrial cancer tumor, and can be continued after the tumor is no longer clinically detectable.Administration can also be continued in subjects who show signs of recurrence.

[0062] In some embodiments, the pharmaceutical composition can be administered in a therapeutically or prophylactically effective amount. Administration of the pharmaceutical composition to a subject can be carried out using known procedures at dosages and for periods of time sufficient to achieve the desired effect.

[0063] In some embodiments, the pharmaceutical composition can be administered to any suitable site in the subject, for example, a site that is distal or proximal to the primary tumor. The route of administration can be parenteral, intramuscular, subcutaneous, intradermal, intraperitoneal, intranasal, intravenous (including via an indwelling catheter), via afferent lymphatics, or by any other route appropriate given the neoplastic disease being treated and the condition of the subject. Preferably, the dose is administered in an amount and for a period of time effective to produce the desired response, whether to induce an immune response or to induce prophylactic or therapeutic treatment of the neoplastic disease and / or symptoms associated therewith.

[0064] The pharma- ceutically acceptable compositions may be administered after, prior to, or concomitantly with other therapies, including therapies that also elicit an immune response in the subject, For example, the subject may have been previously or concomitantly treated with chemotherapy, radiation therapy, and other forms of immunotherapy, and such other therapies are preferably provided in a manner that does not interfere with the immunogenicity of the compositions described herein.

[0065] Administration can be performed by a medical provider (e.g., physician, veterinarian) at an appropriate time and may depend on the subject's clinical condition, the purpose of administration, and / or other therapies also contemplated or administered. In some embodiments, an initial dose can be administered and the subject monitored for immunological and / or clinical response. Suitable means of immunological monitoring include using the patient's peripheral blood lymphocytes (PBL) as responders and neoplastic cells as stimulators. Immunological response can also be determined by delayed inflammatory response at the administration site. One or more doses following the initial dose can be given as needed, typically on a monthly, bimonthly, or preferably weekly basis until the desired effect is achieved. Additional booster or maintenance doses can then be given as needed, especially when immunological or clinical benefits appear to have subsided. [Example] Example 1 Ovarian cancer vaccine formulation using ADDAVAX™ as an adjuvant induces significant inhibition of murine epithelial ovarian cancer growth Epithelial ovarian cancer (EOC) is the most lethal of all human gynecologic malignancies. A single vaccination of female C57BL / 6 mice with the extracellular domain of anti-Müllerian hormone receptor II (AMHR2-ED) in an emulsion containing complete Freund's adjuvant (CFA) has already been shown to provide significant prophylaxis and treatment against murine EOC. Tumor immunity was found to be mediated by AMHR2-ED-specific IgG, which markedly induced EOC cell death via a caspase-3-dependent apoptotic signaling cascade. However, despite the well-established utility of CFA as a "gold standard" adjuvant in proof-of-principle animal studies, its use in human clinical vaccines has been precluded by its tendency to induce non-resolving granulomas, often accompanied by painful abscess formation. Given that ADDAVAX™ adjuvant has been used safely and successfully in generating high titer antibodies in several human clinical trials, we investigated whether vaccination of female C57BL / 6 mice with AMHR2-ED in ADDAVAX™ would generate effective tumor immunity. We found that formulation of AMHR2-ED in ADDAVAX™ induced high serum titers of IgG and highly significant tumor immunity similar to those obtained using CFA as an adjuvant. Our results indicate that vaccination against AMHR2-ED with ADDAVAX™ should provide a novel and effective method of controlling human EOC. 2. Materials and Methods 2.1. Generation of recombinant mouse AMHR2-ED. The entire 125 amino acid DNA coding sequence of mature native AMHR2-ED lacking the signal peptide was used to generate an expression construct as previously reported

[17] (NCBI reference sequence: NM_144547.2, Uniprot Q8K592). To optimize protein folding and improve overall yield, substitutions of the native codon sequence were made (Dapcel, Cleveland, OH), and optimized DNA was synthesized de novo to include an N-terminal methionine, followed immediately by a FLAG tag, and a C-terminal 6×His tag. The 6×His-tagged AMHR2-ED was purified under denaturing conditions using nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography (Qiagen, Valencia, CA). Prior to in vitro use, the 6×His-tagged AMHR2-ED was further purified by reverse-phase high-performance liquid chromatography (HPLC). Endotoxin levels were determined to be less than 0.05 endotoxin units (<5 pg) per mg of recombinant protein, as previously reported

[17] .

[0066] 2.2. Mouse and murine EOC cell lines. TgMISIIR-Tag(DR26) and TgMISIIR-Tag(low) transgenic mice were kindly provided by Dr. Denise C. Connolly (Developmental Therapeutics Program, Fox Chase Cancer Center, Philadelphia, PA). Female TgMISIIR-Tag(DR26) transgenic mice develop bilateral spontaneous EOC due to expression of SV40 large T antigen (Tag) under the control of the AMHR2 promoter

[27] . Although TgMISIIR-Tag(low) transgenic female mice do not develop spontaneous EOC due to erroneous transgene insertion, these mice are immunologically tolerant to SV40-TAg and effectively function as histocompatible recipients of transplantable mouse ovarian carcinoma (MOVCAR) cells derived from ascites of TgMISIIR-Tag(DR26) mice

[28] . MOVCAR cells were cultured in DMEM (Media Preparation Core, Cleveland Clinic, Cleveland, OH) supplemented with 5% fetal bovine serum (HyClone, Logan, UT), 5% HEPES buffer (Sigma-Aldrich, St. Louis, MO), 2 mM L-glutamine (Thermo Fisher Scientific, Waltham, MA), and 1% penicillin / streptomycin (Invitrogen, Carlsbad, CA). A colony of transgenic mice was established and maintained by mating male transgenic mice with wild-type syngeneic C57BL / 6 females (Jackson Laboratory, Bar Harbor, ME). MOVCAR cells were used to develop EOC by subcutaneous injection into histocompatible TgMISIIR-Tag(low) transgenic female mice.ID8 mouse ovarian surface epithelial cells (MOSECs) were obtained commercially (Sigma-Aldrich) and cultured in DMEM (Media Pressure Core) containing 4% fetal bovine serum (HyClone), 2 mM L-glutamine (Thermo Fisher Scientific), 1% penicillin / streptomycin (Invitrogen), and insulin-transferrin-sodium selenate medium additive (Sigma-Aldrich). Subcutaneous injection of ID8 cells into C57BL / 6 female mice induces EOC

[29] .

[0067] 2.3. Transplantable ovarian tumors and treatment. MOVCAR cells (4 × 10 6 cells) and ID8 cells (5 × 10 6 The cells were suspended in PBS and inoculated subcutaneously into the left dorsal flank of female TgMISIIR-Tag(low) and female C57BL / 6 mice in a total volume of 100 μL, respectively. For the CFA-based formulation, mice were immunized with a single subcutaneous injection into the right flank of 200 μL of an emulsion containing 200 μg of Mycobacterium tuberculosis H37Ra in 100 μL of sterile double-distilled deionized water (DDDH2O) and 100 μL of CFA (Difco, Detroit, MI). ADDAVAX™ emulsion was purchased from Invivogen (San Diego, CA, catalog no. vac-adx-10) and 100 μL was suspended in 100 μL of sterile DDDH2O containing 100 μg of recombinant mouse AMHR2-ED, and a final emulsion volume of 200 μL was injected into the ventral flank of female TgMISIIR-tag(low) or female C57BL / 6 mice. For prophylaxis, mice were vaccinated 15 days prior to tumor inoculation. For therapeutic intervention, mice were vaccinated with AMHR2-ED when tumors were palpable. All experiments were initiated when mice were 6-7 weeks of age. Tumor growth was assessed periodically using calipers, and the endpoint of all experiments involving implantable tumors was defined by a tumor measurement of 17 mm in any direction.

[0068] 2.4. Immunohistochemistry. Briefly, CD3+ T cells were immunostained in 5 μm sections of formalin-fixed paraffin-embedded ID8 tumor tissue using a 1:100 dilution of a primary CD3-specific rabbit monoclonal IgG antibody (Abcam, Cambridge, UK; Cat. No. ab16669), followed by a horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG secondary antibody (Abcam; Cat. No. ab214880). CD4+ and CD8+ T cells were immunostained in 5 μm sections of ovarian tumors by incubation with a 1:1000 dilution of primary CD4-specific rabbit monoclonal IgG antibody (Abcam; catalog no. ab183685) or a 1:500 dilution of primary CD8-specific rabbit monoclonal IgG antibody (Abcam; catalog no. ab217344) followed by HRP-conjugated goat anti-rabbit IgG secondary antibody (Abcam; catalog no. ab214880). Visualization was performed using a DAB substrate kit (BD Pharmingen, San Diego, CA; catalog no. 550880) followed by counterstaining with hematoxylin, and sections were mounted in mounting medium (Ricard-Allan Scientific, Kalamazoo, MI; catalog no. 4112) and examined by light microscopy.

[0069] 2.5. Real-time quantitative RT-PCR (qRT-PCR). Tissues were excised and cryopreserved in RNAlater stabilizing solution (Invitrogen; Cat. No. AM7021). RNA was extracted from each tissue by homogenization in TRIZOL Reagent (Invitrogen: Cat. No. 15596026). qRT-PCR was performed using SYBR Green PCR mix (Applied Biosystems, Carlsbad, CA; Cat. No. 4309155) with gene-specific primer pairs (Invitrogen) including: Mouse AMHR2-CD primer: forward-CTGAGCCGCTGTTCCGATTTGA (SEQ ID NO: 1); reverse-ATGTTGGGGCGCTTCCTCTCCT (SEQ ID NO: 2); Mouse AMHR2-ED primer: forward-GCGGGGAAGCACAAAGACACT (SEQ ID NO: 3); reverse-CCGGCCATGGGTAAGATTCC (SEQ ID NO: 4); Mouse β-actin primer: forward-GGT CAT CAC TAT TGG CAA CG (SEQ ID NO: 5); reverse-ACG GAT GTC AAC GTC ACA CT (SEQ ID NO: 6). Relative gene expression was determined by normalization of each gene of interest to β-actin expression in each sample.

[0070] 2.6. Enzyme-linked immunosorbent assay (ELISA). ELISA was performed as previously reported

[29] . Briefly, serum samples were collected from vaccinated mice 8 weeks after vaccination, and each serum sample was diluted 1:10,000 and then added to triplicate wells on 96-well plates precoated with AMHR2-ED or control antigens, ovalbumin (Sigma-Aldrich; Cat. No. A7641-250MG), and recombinant mouse β-casein, a protein produced in our laboratory in Escherichia coli (E. coli) in a manner similar to the production of AMHR2-ED

[17] . After completing the incubation at each step, the absorbance at 405 nm was determined according to the manufacturer's instructions, as previously described

[17] . Isotype-specific serum antibody responses to AMHR2-ED at a 1:10,000 dilution were determined using the Mouse Typer Isotyping Panel (Bio-Rad, Hercules, Calif., catalog no. 1722051) according to the manufacturer's instructions.

[0071] 2.7. Biostatistical analysis. Differences between mRNA expression levels were compared using Student's t-test and two-way analysis of variance. Differences between tumor growth curves were compared using two-way analysis of variance. Differences in mouse survival were identified by log-rank and chi-square using correlated samples. All experiments were independently repeated at least three times. 3.Results 3.1. Elevated AMHR2 expression in mouse ovarian tumor cell lines. We determined the relative gene expression of both AMHR2-ED and AMHR2-CD in ID8 and MOVCAR murine EOC cells. We found that while MOVCAR and ID8 cells express high levels of both AMHR2 domains, the expression levels of each domain were consistently approximately two-fold higher in MOVCAR cells compared to ID8 cells (Figure 1).

[0072] 3.2. AMHR2-ED Vaccination Formulated with ADDAVAX™ Adjuvant Induces Robust IgG Responses in Female C57BL / 6 Mice. To determine whether a robust IgG response to AMHR2-ED could be induced when ADDAVAX™ was used as an adjuvant, we immunized 6-7 week old C57BL / 6 female mice with 100 μg of recombinant mouse AMHR2-ED using both CFA and ADDAVAX™ as adjuvants. Eight weeks after immunization, serum IgG responses at 1:10,000 dilution were evaluated in response to AMHR2-ED as well as in response to the irrelevant antigens ovalbumin and recombinant mouse β-casein.

[0073] The absorbance obtained using ADDAVAX™ as an adjuvant was found to be similar to that obtained using CFA as an adjuvant (Figure 2a). The IgG isotype profile generated using ADDAVAX™ as an adjuvant was also found to be similar to that obtained using CFA as an adjuvant (Figure 2b).

[0074] 3.3.AMHR2-ED vaccination formulated with ADDAVAX™ effectively inhibits the growth of murine EOC. We next investigated whether AMHR2-ED vaccination using ADDAVAX™ as an adjuvant was comparable to the inhibition of murine EOC tumor growth induced by AMHR2-ED vaccine incorporating CFA as an adjuvant. We found that prophylactic vaccination against AMHR2-ED using ADDAVAX™ adjuvant 15 days prior to inoculation of TgMISIIR-Tag(low) female mice with MOVCAR cells significantly inhibited the growth of MOVCAR-derived tumors when compared to vaccination with CFA alone or vaccination with AMHR2-ED using CFA as an adjuvant (Figure 3a; P<0.0001, respectively). Furthermore, prophylactic vaccination against AMHR2-ED using ADDAVAX™ as an adjuvant resulted in a statistically significant increase in overall survival when compared to vaccination with CFA alone (Fig. 3b, P<0.009) or when compared to vaccination against AMHR2-ED using CFA as an adjuvant (Fig. 3b; P<0.006). It was also found that therapeutic vaccination against AMHR2-ED at the time MOVCAR tumors were palpable was similarly effective using either CFA or ADDAVAX™ as an adjuvant when compared to vaccination with CFA alone (Fig. 3c, P<0.0001 for each). However, perhaps the best way to understand the efficacy of ADDAVAX™ is most clearly evident when female C57BL / 6 mice were vaccinated with either ADDAVAX™ alone or AMHR2-ED adjuvanted with ADDAVAX™ after inoculated ID8 EOC cells had become palpable tumors (Figure 3d, P<0.0001).

[0075] 3.4. AMHR2-ED vaccination with ADDAVAX™ as an adjuvant results in T cell infiltration of murine EOC. We next investigated immune cell infiltration of ID8 tumors 175 days after tumor inoculation and 168 days after AMHR2-ED vaccination with ADDAVAX™ as an adjuvant. We observed substantial infiltration of CD3+, CD4+, and CD8+ T cells (Figure 4). The observation of infiltration of CD8+ T cells into ID8 tumors was rather surprising, since infiltration of CD8+ T cells in C57BL / 6 female mice vaccinated against AMHR2-ED when CFA was used as an adjuvant has never been observed before

[17] .

[0076] This example shows that our formulation of the AMHR2-ED vaccine with ADDAVAX™ as an adjuvant induces robust IgG responses and inhibits the growth of murine EOC in a similar and generally superior manner to the use of CFA as an adjuvant. Indeed, in some comparisons, ADDAVAX™ appears surprisingly superior to CFA as an adjuvant. Most notably, it induces much improved overall survival in the transplantable MOVCAR EOC mouse model and there was a T cell infiltration of CD8+ T cells in ID8-derived EOC tumor beds that did not occur in a previous study using CFA as an adjuvant

[17] . Thus, our data support the substitution of ADDAVAX™ for CFA as an adjuvant for the prevention of EOC in women at high genetic risk of developing EOC. Studies have shown that 39%-44% of women with deleterious BRCA1 variants and 11%-17% of women with deleterious BRCA2 variants will develop ovarian cancer by age 70-80 years.[30-32] The availability of such a high-risk population of women with high incidence of EOC and who are most in need of an EOC preventive vaccine makes it important to treat women with the adjuvant and AMHR2-ED peptide herein. References 1. Siegel,et al.,Jemal,A.Cancer Statistics,2017.CA.Cancer J.Clin.2017,67,7-30. 2. Howlader,N.;Noone,AM;Krapcho,M;Miller,D;Brest,A;Yu,M;Ruhl,J;Tatalovich,Z;Mariotto,A;Lewis,DR;et al.(eds).SEER Cancer Statistics Review,1975-2018,National Cancer Institute.Bethesda,MD.https: / / seer.cancer.gov / csr / 197 5_2018 / 2020 on 11 / 11 / 2020 and SEER on 4 / 4 / 2021.

[0077] 3. Pignata,et al.,Treatment of recurrent epithelial ovarian cancer.Cancer 2019;125,Suppl 24,4609-4615. 4. Pfisterer,J.;Ledermann,JManagement of platinum-sensitive recurrent ovarian cancer.Semin.Oncol.2006,33,(2 Suppl 6),S12-16. 5. Fung-Kee-Fung, et al., Optimal chemotherapy treatment for women with recurrent ovarian cancer.Curr.Oncol.2007,14,195-208. 6. Montemorano,L.;Lightfoot,MD;Bixel,K.Role of Olaparib as maintenance treatment for ovarian cancer:The evidence to date.Onco.Targets Ther.2019,12,11497-11506. 7. Kubalanza,K.;Konecny,G.E.Mechanisms of PARP inhibitor resistance in ovarian cancer.Curr.Opin.Obstet.Gynecol.2020,32,36-41. 8. Zhang,et al.Intratumoral T cells,recurrence,and survival in epithelial ovarian cancer.N.Engl.J.Med.2003,348,203-213. 9. Masiakos,et al.Human ovarian cancer,cell lines,and primary as-cites cells express the human Mullerian inhibiting substance(MIS) type II receptor,bind,and are re-sponsive to MIS.Clin.Cancer.Res.1999,5,3488-3499. 10. Bakkum-Gamez,et al.,Mullerian inhibiting substance type II receptor(MISIIR):a novel,tissue-specific target expressed by gynecologic cancers.Gynecol.Oncol.2008,108,141-148. 11. Song,et al.,The expression of Mullerian inhibiting substance / anti-Mullerian hormone type II receptor protein and mRNA in benign,borderline and malignant ovarian neoplasia.Int.J.Oncol.2009,34,1583-1591. 12. Mazumder,et al.,Immunotherapy of ovarian cancer with a monoclonal antibody specific for the extracellular domain of anti-Mullerian hormone receptor II.Oncotarget 2020,11,1894-1910. 13. Kristensen,et al.,Expression of TGF-beta superfamily growth factors,their receptors,the associated SMADs and antagonists in five isolated size-matched populations of pre-antral follicles from normal human ovaries.Mol.Hum.Reprod.2014,20,293-308. 14. Josso,et al.,Anti-Mullerian hormone and its receptors. Mol.Cell. Endocrinol.2001,179,25-32. 15. Gruijters,et al.,Anti-Mullerian hormone and its role in ovarian function. Mol.Cell. Endocrinol.2003,211,85-90. 16. Uhlen,et al.Towards a knowledge-based Human Protein Atlas.Nat.Biotechnol.2010,28,1248-1250. 17. Mazumder,et al.,Primary immunoprevention of epithelial ovarian carcinoma by vaccination against the extracellular domain of anti-Mullerian hormone receptor II.Cancer Prev.Res.(Phila).2017,10,612-624. 18. Tuohy,V.K.;Johnson,J.M.;Mazumder,S.Primary immunoprevention of adult onset cancers by vaccinating against retired tissue-specific self proteins.Semin.Immunol.2020,47,1-7. 19. Chapel,et al.,Report of nine cases of accidental injury to man with Freund’s complete adjuvant.Clin.Exp.Immunol.1976,24,538-541. 20. Stills,H.F.,Jr. Adjuvants and antibody production:dispelling the myths associated with Freund’s complete and other adjuvants.ILAR J.2005,46,280-293. 21. Apostolico,et al.,Adjuvants:Classification,Modus Operandi,and Licensing. J.Immunol. Res.2016,2016,1459394. 22. Ott,et al.,MF59.De-sign and evaluation of a safe and potent adjuvant for human vaccines.Pharm.Biotechnol.1995,6,277-296. 23. Black,S.Safety and effectiveness of MF-59 adjuvanted influenza vaccines in children and adults.Vac-cine 2015,33,Suppl 2:B3-5. 24. Bernstein,et al.Safety and efficacy of a cytomegalovirus glycoprotein B(gB) vaccine in adolescent girls:A randomized clinical trial.Vaccine 2016,34,313-319. 25. Essink,et al.,Immunogenicity and safety of MF59-adjuvanted quadrivalent influenza vaccine versus standard and al-ternate B strain MF59-adjuvanted trivalent influenza vaccines in older adults.Vaccine 2020,38,242-250. 26. Gray,et al.Vaccine efficacy of ALVAC-HIV and bivalent subtype C gp120-MF59 in adults.N.Engl.J.Med.2021,384,1089-1100. 27. Connolly,et al.,Female mice chimeric for expression of the simian virus 40 TAg under control of the MISIIR promoter develop epithelial ovarian cancer.Cancer Res.2003,63,1389-1397. 28. Quinn,et al.,Development of a syngeneic mouse model of epithelial ovarian cancer.J.Ovarian Res.2010,3,24. 29. Sakalar,et al.,Regulation of murine ovarian epithelial carcinoma by vaccination against the cytoplasmic domain of anti-Mullerian hormone receptor II.J.Immunol.Res.2015,2015,630287. 30. Antoniou,et al.Average risks of breast and ovarian cancer associated with BRCA1 or BRCA2 mutations detected in case series unselected for family history:a combined analysis of 22 studies.Am.J.Hum.Genet.2003,72,1117-1130. 31. Chen,S.;Parmigiani,G.Meta-analysis of BRCA1 and BRCA2 penetrance.J.Clin.Oncol.2007,25,1329-1333. 32. Kuchenbaecker,et al.Risks of breast,ovarian,and contralateral breast cancer for BRCA1 and BRCA2 mutation carriers.JAMA.2017,317,2402-2416. 33. Strasser-Weippl,et al.,Suitable trial designs and cohorts for preventive breast cancer agents.Nat.Rev.Clin.Oncol.2013,10,677-687.doi:10.1038 / nrclinonc.2013.174 34. Temkin,et al.,Ovarian cancer prevention in high-risk women.Clin.Obstet.Gynecol.2017,60,738-757. 35. National Comprehensive Cancer Network,NCCN clinical practice guidelines in oncology.Ovarian Cancer Including Fallopian Tube Cancer and Primary Peritoneal Cancer,Version 1.2020-March 11,2020;https: / / www.nccn.org / professionals / physician_gls / pdf / ovarian.pdf 36. Manoukian, et al., Risk-reducing surgery in BRCA1 / BRCA2 mutation carriers:Are there factors associated with the choice?Psychooncology 2019,28,1871-1878.Psychooncology 2019,28,1871-1878. Incorporation by Reference All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference in their entirety, as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. [Brief description of the drawings]

[0078] [Figure 1] Figure 1 shows AMHR2 gene expression in mouse EOC. qRT-PCR analysis of ID8 and MOVCAR EOC cell lines indicates that both cell lines express the AMHR2-ED and AMHR2-CD domains, but expression of each AMHR2 domain in MOVCAR cells was consistently approximately twice the expression level measured in ID8 cells. Error bars indicate ±SD. [Diagram 2]Figure 1 shows that formulation of AMHR2-ED vaccine with ADDAVAX™ induces high IgG responses. ELISA analysis shows that the use of ADDAVAX™ as an adjuvant when immunizing C57BL / 6 female mice resulted in (A) antigen-specific IgG responses to the AMHR2-ED immunogen, and (B) antigen-specific IgG isotype responses similar to those obtained using CFA as the adjuvant. Serum was analyzed 8 weeks after immunization using a dilution of 1:10,000. Error bars indicate ±SD. [Diagram 3] Figure 1 shows that AMHR2-ED vaccination with ADDAVAX™ as an adjuvant is highly effective in inhibiting the growth of mouse EOC. Prophylactic AMHR2-ED vaccination with ADDAVAX™ as an adjuvant resulted in significant inhibition of (A) MOVCAR growth and significant increase in (B) overall survival in TgMISIIR-Tag(low) female mice compared to vaccination with CFA alone or vaccination against AMHR2-ED in CFA. AMHR2-ED vaccination in ADDAVAX™ was also effective in inhibiting the growth of (C) MOVCAR and (D) ID8 tumors when vaccination was administered when tumors were palpable. Error bars = ± SD, asterisks indicate significance. [Figure 4] Figure 1 shows that AMHR2-ED vaccination with ADDAVAX™ adjuvant results in T cell infiltration of murine EOC. AMHR2-ED vaccination in ADDAVAX™ resulted in infiltration of CD3+ T cells (arrow, bottom row, left), CD4+ T cells (arrow, bottom row, center), and CD8+ T cells (arrow, bottom row, right) compared to corresponding EOC tissues from mice vaccinated with ADDAVAX™ alone (arrow, top row). Tissues are representative of several examined from several different mice. Tissue magnification = 20x. [Diagram 5]Exemplary sequences are shown: Panel A shows an exemplary human AMHR2-ED amino acid sequence (SEQ ID NO:7). Panel B shows an exemplary N-terminally truncated human AMHR2-ED amino acid sequence (SEQ ID NO:8). Panel C shows an exemplary C-terminally truncated human AMHR2-ED amino acid sequence (SEQ ID NO:9). Panel D shows an exemplary N- and C-terminally truncated human AMHR2-ED amino acid sequence (SEQ ID NO:10).

Claims

1. 1. A composition comprising: a) a polypeptide comprising at least a portion of the anti-Müllerian hormone receptor type II extracellular domain (AMHR-ED); b) an adjuvant, i) squalene oil; ii) a non-ionic surfactant; and iii) an emulsifier; and iv) a buffer solution; The adjuvant comprising The composition comprising:

2. The composition of claim 1 , wherein the emulsifier comprises sorbitan trioleate and / or the non-ionic surfactant comprises polysorbate 80.

3. The composition of claim 1 or claim 2, wherein the polypeptide is derived from human AMHR-ED.

4. The composition of any one of claims 1 to 3, wherein the buffer comprises a citrate buffer or a phosphate buffered saline.

5. The composition according to any one of claims 1 to 4, wherein the pH of the composition is from 5.5 to 6.

9.

6. The composition according to any one of claims 1 to 5, wherein the composition is in the form of an emulsion.

7. 7. The composition of claim 6, wherein the emulsion is a nanoemulsion of a first component and a second component, the first component comprising the emulsifier and squalene oil, and the second component comprising the non-ionic surfactant and the buffer.

8. The composition of claim 7, wherein the emulsifier is present in the first component at about 0.4 to 0.6 weight / volume.

9. 9. The composition of claim 7 or claim 8, wherein the squalene oil is present in the first component at about 4-6% weight / volume.

10. The composition of any one of claims 7 to 9, wherein the non-ionic surfactant is present in the second component in an amount of about 0.4 to 0.6 weight / volume.

11. The composition of any one of claims 7 to 10, wherein the buffer is present in the second component at about 4-6% weight / volume.

12. The composition according to any one of claims 7 to 11, wherein the nanoemulsion is composed of particles having an average size of about 150 to 170 nm.

13. The composition of any one of claims 1 to 12, wherein the adjuvant further comprises at least one of the following: saponin, cholesterol, phospholipid, phosphatidylcholine, and a second buffer.

14. The composition of any one of claims 1 to 13, wherein the adjuvant further comprises α-tocopherol.

15. The composition of any one of claims 1 to 14, wherein the adjuvant further comprises monophosphoryl lipid A.

16. The composition of any one of claims 1 to 15, wherein the adjuvant further comprises an immunomodulatory agent.

17. 17. The composition of claim 16, wherein the immunomodulatory agent comprises synthetic trehalose dicorynomycolate.

18. The composition of any one of claims 1 to 17, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 7, 8, 9, or 10.

19. The composition of any one of claims 1 to 17, wherein the polypeptide comprises the entire human AMHR-ED or nearly the entire human AMHR2-ED.

20. The composition of any one of claims 1 to 17, wherein the polypeptide comprises at least 20 consecutive amino acids from SEQ ID NO:

7.

21. The composition of any one of claims 1 to 17, wherein the polypeptide comprises at least 40 consecutive amino acids from SEQ ID NO:

7.

22. The composition of any one of claims 1 to 17, wherein the polypeptide comprises at least 60 consecutive amino acids from SEQ ID NO:

7.

23. The composition of any one of claims 1 to 17, wherein the polypeptide comprises at least 100 contiguous amino acids from SEQ ID NO:

7.

24. The composition of any one of claims 1 to 17, wherein the polypeptide comprises at least 115 consecutive amino acids from SEQ ID NO:

7.

25. The composition of any one of claims 1 to 17, wherein the polypeptide comprises at least 122 consecutive amino acids from SEQ ID NO:

7.

26. The composition of any one of claims 1 to 17, wherein the polypeptide has an amino acid sequence comprising 100 contiguous amino acids that are at least 80% identical to the amino acid sequence of the extracellular domain of AMHR2.

27. The composition of any one of claims 1 to 26, wherein administration of the composition to a subject activates CD4+ T cells in the subject.

28. The composition of any one of claims 1 to 27, wherein administration of the composition to a subject induces the production of AMHR2 ED-specific IgG in the subject.

29. 29. The composition of claim 27 or 28, wherein the subject is a human female.

30. A method for treating ovarian or endometrial cancer tumors in a female subject, comprising administering to said female subject a composition according to any one of claims 1 to 29.

31. 31. The method of claim 30, wherein the subject has an ovarian cancer tumor, and the ovarian cancer tumor is a primary ovarian cancer tumor or a metastatic tumor.

32. 32. The method of claim 30 or 31, wherein the female subject is a human.

33. The method of any one of claims 30 to 32, wherein the subject has an ovarian cancer tumor, and the ovarian cancer tumor is an epithelial ovarian cancer (EOC) tumor.

34. The method of any one of claims 30 to 33, wherein administration of the composition induces an immune response in the subject against the ovarian cancer and / or the endometrial cancer tumor.

35. The method of any one of claims 30 to 34, wherein the ovarian cancer and / or the endometrial cancer tumor expresses AMHR2-ED.

36. The method of any one of claims 30 to 35, further comprising determining whether the ovarian cancer and / or the endometrial cancer tumor expresses AMHR2-ED.

37. 37. The method of any one of claims 30-36, further comprising determining whether said administration of said composition induces an immune response in said subject against said ovarian cancer or said endometrial cancer tumor.

38. 38. The method of any one of claims 30 to 37, further comprising repeating said administering of said composition to said subject.

39. The method of any one of claims 30 to 38, wherein administration of the composition induces expression of anti-AMHR2-ED IgG antibodies by the subject.

40. 40. The method of any one of claims 30 to 39, further comprising administering to the subject an additional anti-cancer agent.

41. 41. The method of claim 40, wherein the additional anticancer agent is paclitaxel, cisplatin, topotecan, gemcitabine, bleomycin, etoposide, carboplatin, docetaxel, doxorubicin, topotecan, cyclophosphamide, trabectedin, olaparib, tamoxifen, letrozole, bevacizumab, an anti-CTLA4 antibody, an anti-PD-1 antibody, or an anti-PD-L1 antibody.

42. The method of any one of claims 30 to 41, wherein the subject has undergone surgery to remove at least a portion of the ovarian cancer and / or endometrial cancer tumor.

43. A method for preventing ovarian and / or endometrial cancer, comprising administering to a female subject a composition according to any one of claims 1 to 29.

44. 44. The method of claim 43, wherein the subject has no detectable ovarian and / or endometrial cancer.

45. 45. The method of claim 43 or 44, wherein the subject is at increased risk of developing ovarian and / or endometrial cancer.

46. 1. A system or kit comprising: a) a polypeptide comprising at least a portion of the anti-Müllerian hormone receptor type II extracellular domain (AMHR-ED); b) an adjuvant, i) squalene oil; ii) a non-ionic surfactant; and iii) an emulsifier; and iv) a buffer solution; The adjuvant comprising The system or the kit comprising:

47. The system or kit of claim 46, wherein the polypeptide is derived from human AMHR-ED.

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