Prostate cancer vaccines and uses thereof

Abstract

Disclosed herein is a method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject a therapeutic regimen comprising two or more vaccines comprising a great ape adenovirus serotype 20 (GAd20) virus, the vaccine comprising, in order, a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1, and one or more vaccines comprising a modified vaccinia Ankara (MVA) virus, the vaccine comprising, in order, a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3, thereby treating or preventing prostate cancer.

Description

[Technical field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Patent Application No. 63 / 290,164, filed December 16, 2021, the disclosure of which is incorporated by reference in its entirety herein.

[0002] (Sequence Listing) The contents of the electronic sequence listing (JBI6595WOPCT1_Sequence Listing.xml, size: 111,774 bytes, and creation date: November 23, 2022) are incorporated herein by reference in their entirety. [Background technology]

[0003] Prostate cancer is the most common non-cutaneous malignancy in men and the second leading cause of cancer deaths in men in the Western world. Prostate cancer results from the uncontrolled growth of abnormal cells in the prostate. Once a prostate cancer tumor develops, androgens (male hormones from the testes), such as testosterone, promote the growth of the prostate cancer. In its early stages, localized prostate cancer is often treatable by localized therapies, including, for example, surgical removal of the prostate and radiation therapy. However, if localized therapies do not cure the prostate cancer, as is the case in up to one-third of men, the disease progresses to incurable metastatic disease.

[0004] For many years, the established standard of care for men with malignant castration-resistant prostate cancer (mCRPC) has been docetaxel chemotherapy. More recently, abiraterone acetate (ZYTIGA®) in combination with prednisone has been approved to treat metastatic castration-resistant prostate cancer. Androgen receptor (AR)-targeted agents, such as enzalutamide (XTANDI®), have also entered the market to treat metastatic castration-resistant prostate cancer. Platinum-based chemotherapy has been tested in numerous clinical studies in molecularly unselected prostate cancer patients with limited results and significant toxicity. However, there remains a small population of patients who do not respond early or are refractory (or resistant) to these treatments. There are no approved treatment options available for such patients. Summary of the Invention [Means for solving the problem]

[0005] Disclosed herein is a method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject a therapeutic regimen comprising two or more vaccines comprising great ape adenovirus serotype 20 (GAd20) virus, the vaccine comprising, in order, a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1, and one or more vaccines comprising modified vaccinia Ankara (MVA) virus, the vaccine comprising, in order, a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3, thereby treating or preventing prostate cancer.

[0006] Also provided is a method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject 1×10 11 10 viral particles (VP) of GAd20 virus, wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8A vaccine comprising infectious units (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 11 Vaccine containing GAd20 virus in VP; 1 × 10 at approximately week 24 8 a vaccine containing MVA virus at IFU; and 1 x 10 8 Methods are also provided that include administering a vaccine that includes an IFU of an MVA virus.

[0007] 1. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject 1×10 11 10 viral particles (VP) of GAd20 virus, wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8 A vaccine comprising infectious units (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 11 Vaccine containing GAd20 virus in VP; 1 × 10 at approximately week 24 8 a vaccine containing MVA virus at IFU; and 1 x 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0008] Also provided is a method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject 1×10 11 10 viral particles (VP) of GAd20 virus, wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8 A vaccine comprising infectious units (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 8a vaccine containing MVA virus at IFU; and 1 x 10 8 Methods are also provided that include administering a vaccine that includes an IFU of an MVA virus.

[0009] Provided herein is a method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject 1×10 11 a vaccine comprising 1×10 viral particles (VP) of the GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8 A vaccine comprising infectious units (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 8 a vaccine containing MVA virus at IFU; and 1 x 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0010] A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject at week 0 and at about week 3 1 mg / kg to 3 mg / kg of an anti-CTLA4 antibody and 1×10 11 an anti-CTLA4 antibody and vaccine comprising 1 x 10 viral particles (VP) of GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8 an anti-CTLA4 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 11 A vaccine containing VP GAd20 virus; at approximately week 24, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8a vaccine containing the MVA virus at IFU; and 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 x 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0011] A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject at week 0 and at about week 3 1 mg / kg to 3 mg / kg of an anti-PD1 antibody and 1×10 11 an anti-PD1 antibody and a vaccine comprising 1×10 viral particles (VP) of GAd20 virus, wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8 an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 11 A vaccine containing VP GAd20 virus; at approximately week 24, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 a vaccine containing the MVA virus at IFU; and 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 The method includes administering a vaccine comprising an IFU of the MVA virus.

[0012] Also provided is a method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject at week 0 and about week 3, 1 mg / kg to 3 mg / kg of an anti-CTLA4 antibody and 1×10 11 an anti-CTLA4 antibody and vaccine comprising 1 x 10 viral particles (VP) of GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8an anti-CTLA4 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 11 A vaccine containing VP GAd20 virus; at approximately week 24, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 a vaccine containing the MVA virus at IFU; and 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 x 10 8 Methods are also provided that include administering a vaccine that includes an IFU of an MVA virus.

[0013] Provided herein is a method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject at week 0 and at about week 3 1 mg / kg to 3 mg / kg of an anti-PD1 antibody and 1×10 11 an anti-PD1 antibody and a vaccine comprising 1×10 viral particles (VP) of GAd20 virus, wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8 an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 11 A vaccine containing VP GAd20 virus; at approximately week 24, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 a vaccine containing the MVA virus at IFU; and 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0014] Provided herein is a method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject at week 0 and at about week 3 1 mg / kg to 3 mg / kg of an anti-CTLA4 antibody and 1×10 11 an anti-CTLA4 antibody and vaccine comprising 1 x 10 viral particles (VP) of GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8 an anti-CTLA4 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 8 a vaccine containing the MVA virus at IFU; and 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 x 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0015] Provided herein is a method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject at week 0 and at about week 3 1 mg / kg to 3 mg / kg of an anti-PD1 antibody and 1×10 11 an anti-PD1 antibody and a vaccine comprising 1×10 viral particles (VP) of GAd20 virus, wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8 an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 8 a vaccine containing the MVA virus at IFU; and 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8Methods are provided that include administering a vaccine that includes an IFU of the MVA virus.

[0016] A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject at week 0 and at about week 3 1 mg / kg to 3 mg / kg of an anti-CTLA4 antibody and 1×10 11 an anti-CTLA4 antibody and vaccine comprising 1 x 10 viral particles (VP) of GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8 an anti-CTLA4 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 8 a vaccine containing the MVA virus at IFU; and 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 x 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0017] Provided herein is a method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject at week 0 and at about week 3 1 mg / kg to 3 mg / kg of an anti-PD1 antibody and 1×10 11 an anti-PD1 antibody and a vaccine comprising 1×10 viral particles (VP) of GAd20 virus, wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; 8 an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 8a vaccine containing the MVA virus at IFU; and 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 Methods are provided that include administering a vaccine that includes an IFU of the MVA virus. [Brief description of the drawings]

[0018] The summary, as well as the detailed description which follows, will be better understood when read in conjunction with the appended drawings. For the purpose of illustrating the disclosed method, there are shown in the drawings exemplary embodiments of the method. However, the method is not limited to the particular embodiments disclosed. [Figure 1A] FIG. 1 illustrates an exemplary dosing schedule. [Figure 1B] FIG. 1 illustrates an exemplary dosing schedule. [Diagram 2] FIG. 1 shows the number of cynomolgus macaques (responders) exhibiting prostate cancer neoantigen-specific IFNγ+ T cells (SFU / 106 cells) at weeks 2 and 6 after receiving a single dose ("single prime") or two doses ("dual prime") of the disclosed GAd20 vaccine. [Diagram 3] FIG. 1 shows antigen-specific T cell responses in cynomolgus macaques after a dual GAd20 vaccine prime followed by an MVA vaccine boost at the indicated time points. [Figure 4] FIG. 1 shows the number of cynomolgus monkeys (responders) exhibiting prostate cancer neoantigen-specific IFNγ+ T cells (SFU / 106 cells) 4 weeks after receiving a single dose of GAd20 ("single prime") alone or in combination with anti-CTLA4 antibody (αCTLA4). [Diagram 5] FIG. 1 shows prostate cancer neoantigen-specific IFNγ+ T cell (SFU / 106 cells) responses in cynomolgus monkeys up to 22 weeks after receiving the indicated dosing schedules. [Figure 6]FIG. 1 shows polyfunctional (IFNγ / TNFα double positive) CD8+ T cell responses in cynomolgus monkeys after administration of GAd20 / GAd20 / MVA (weeks 0, 4, 12) alone (circles) or in combination with anti-CTLA4 antibody (squares). [Figure 7] FIG. 1 shows effector memory neo-antigen-specific CD8+ T cell responses in cynomolgus monkeys after administration of GAd20 / GAd20 / MVA (weeks 0, 4, 12) alone (circles) or in combination with anti-CTLA4 antibody at week 21 (squares). [Figure 8] FIG. 1 shows the breadth of CD8+ T cell responses in cynomolgus monkeys following administration of GAd20 / GAd20 / MVA (weeks 0, 4, 12) alone or in combination with an anti-CTLA4 antibody. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] The methods of the present disclosure may be more readily understood by reference to the following detailed description taken in conjunction with the accompanying drawings, which form a part of this disclosure: It is to be understood that the methods of the present disclosure are not limited to the specific methods described and / or illustrated herein, and further, the terminology used herein is for the purpose of describing particular embodiments by way of example only, and is not intended to limit the methods claimed.

[0020] Unless otherwise stated, any description of possible mechanisms or modes of operation or reasons for improvement is intended to be illustrative only, and the methods of the present disclosure are not limited by the merits or demerits of the proposed mechanisms or modes of operation or reasons for improvement.

[0021] When a range of numerical values ​​is recited or established herein, the range includes its endpoints, and all individual integers and fractions within the range, and also includes each of the narrower ranges formed by all the various possible combinations of these endpoints and internal integers and fractions, forming subgroups of the larger group of values ​​within the recited range, as if each of the narrower ranges were explicitly recited. When a range of numerical values ​​is recited herein as being greater than the recited value, the range is nevertheless finite, and its upper limit is defined by a value that is operable within the context of the method described herein. When a range of numerical values ​​is recited herein as being less than the recited value, the range is nevertheless defined by a non-zero value at its lower limit. It is not intended that the scope of the method be limited to the specific values ​​recited in defining the range. All ranges are inclusive and combinable.

[0022] When values ​​are expressed as approximations, by use of the antecedent "about," it is to be understood that the particular value forms another embodiment. Reference to a particular numerical value is intended to include at least the particular value unless the context dictates otherwise.

[0023] It should be understood that several features of the methods of the present disclosure that are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the methods of the present disclosure that are, for brevity, described in a single embodiment, may also be provided separately or in any subcombination.

[0024] As used herein, the singular forms "a," "an," and "the" are intended to include plurals.

[0025] Various terms relating to the embodiments of the present specification are used throughout the specification and claims. Unless otherwise indicated, such terms are to be given their ordinary meaning in the art. Other specifically defined terms are to be interpreted in a manner consistent with the definitions provided herein.

[0026] The term "about" is used to encompass a variation of no more than ±10%, a variation of no more than ±5%, a variation of no more than ±1%, a variation of no more than ±0.5%, or a variation of no more than ±0.1% from the stated value.

[0027] Similarly, the term "comprising" is intended to include examples encompassed by the terms "consisting essentially of" and "consisting of." Similarly, the term "consisting essentially of" is intended to include examples encompassed by the term "consisting of."

[0028] By "administered together" it is meant that two or more therapeutic agents (such as a virus and an antibody) can be administered to a subject together in a mixture, simultaneously as single agents, or sequentially as single agents in any order.

[0029] "Treating", "treatment" and similar terms refer to both therapeutic treatment and prophylactic or preventative measures, including reducing the severity and / or frequency of symptoms of prostate cancer, eliminating symptoms of prostate cancer and / or the underlying causes of symptoms, reducing the frequency or likelihood of symptoms of prostate cancer and / or the underlying causes of symptoms, ameliorating or repairing damage caused directly or indirectly by prostate cancer. Treatment also includes prolonging survival compared to the expected survival of a subject not receiving treatment. Subjects to be treated include those with prostate cancer, as well as those prone to having prostate cancer or those in whom prostate cancer is to be prevented.

[0030] Disclosed herein is a method of treating or preventing prostate cancer in a subject, where the prostate cancer comprises one or more prostate cancer neo-antigens encoding the amino acid sequence of SEQ ID NO: 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, or 93. The disclosed vaccines (e.g., those comprising a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 1 or 3) can be used to treat or prevent prostate cancer expressing one or more of these neo-antigens. The method of treating or preventing prostate cancer includes administering to a subject: Two or more vaccines comprising a great ape adenovirus serotype 20 (GAd20) virus, the nucleotide sequence of which in order encodes the amino acid sequence of SEQ ID NO:1; and one or more vaccines comprising a Modified Vaccinia Ankara (MVA) virus comprising, in order, a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3, This can include treating or preventing prostate cancer.

[0031] GAd20 is an adenovirus that infects gorillas and can be isolated from gorilla fecal samples. GAd20 can be engineered to have at least one of the gene products essential for viral replication, such as one or more of the adenoviral regions E1, E2, and E4, functionally deleted or completely removed, thereby rendering the adenovirus unable to replicate. The deletion of the El region can include deletion of EIA, EIB 55K, or EIB 21K, or any combination thereof. Replication-deficient adenoviruses are propagated by providing in trans the proteins encoded in the deleted region by the producer cell using a helper plasmid, or by engineering the producer cell to express the required proteins. Adenoviral vectors can also have a deletion in the E3 region that is not required for replication, and therefore such a deletion does not need to be complemented. The GAd20 of the present disclosure can include a functional deletion or complete removal of at least a portion of the E1 and E3 regions. GAd20 may further comprise a functional deletion or complete removal of the E4 and / or E2 regions. Suitable production cells that can be used are human retinal cells immortalized by El, such as 911 or PER.C6 cells (see, e.g., U.S. Pat. No. 5,994,128), El-transformed amniotic cells (see, e.g., European Patent No. 1230354), E1-transformed A549 cells (see, e.g., WO 1998 / 39411, U.S. Pat. No. 5,891,690). The nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1 may be inserted into a site or region (insertion region) in the viral genome that does not affect the viral viability of the resulting recombinant virus. The nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1 may be inserted in a parallel (transcribed 5'→3') or antiparallel (transcribed 3'→5' relative to the vector backbone) orientation to the deleted E1 region. In addition, suitable transcriptional regulatory elements capable of directing expression of the nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1 in a mammalian host cell in which the virus is prepared for use may be operably linked to the nucleotide sequence."Operably linked" sequences include both expression control sequences adjacent to the nucleic acid sequences they regulate, and regulatory sequences at a distance to act in trans or control the nucleic acid sequence that is regulated.

[0032] Recombinant GAd20 particles may be prepared and grown according to any conventional technique in the art (e.g., WO 1996 / 17070) using a complementing cell line or helper virus that provides in trans the defective viral genes necessary for viral replication. Cell lines 293 (Graham et al., 1977, J. Gen. Virol. 36:59-72), PER.C6 (see, e.g., U.S. Pat. No. 5,994,128), E1 A549, and 911 are commonly used to complement E1 deletions. Other cell lines have been genetically engineered to complement defective vectors (Yeh, et al., 1996, J. Virol. 70:559-565; Kroughak and Graham, 1995, Human Gene Ther. 6:1575-1586; Wang, et al., 1995, Gene Ther. 2:775-783; Lusky, et al., 1998, J. Virol. 72:2022-203; European Patent No. 919627; and International Publication No. WO 1997 / 04119). GAd20 particles may be recovered from the culture supernatant as well as from the cells after lysis and, optionally, further purified according to standard techniques (e.g., chromatography, ultracentrifugation as described in WO 1996 / 27677, WO 1998 / 00524, WO 1998 / 26048, and WO 2000 / 50573). Methods for constructing and propagating adenoviral vectors, such as GAd20, are also described in, for example, U.S. Patent Nos. 5,559,099, 5,837,511, 5,846,782, 5,851,806, 5,994,106, 5,994,128, 5,965,541, 5,981,225, 6,040,174, 6,020,191, and 6,113,913.

[0033] MVA originates from the cutaneous vaccinia strain Ankara (Chorioallantois vaccinia Ankara, CVA virus), which has been maintained for many years at the Vaccination Institute (Ankara, Turkey) and used as the basis for human vaccination. However, the high incidence of severe post-vaccination complications associated with vaccinia virus (VACV) led to several attempts to generate a more attenuated and safer smallpox vaccine.

[0034] MVA has been generated by serial passage of CVA virus in chicken embryo fibroblasts 516 times (see Meyer et al., J. Gen. Virol., 72:1031-1038 (1991) and U.S. Patent No. 10,035,832). As a result of these long-term passages, the resulting MVA virus has lost approximately 31 kilobases of its genomic sequence and has therefore been described as highly host-restricted to avian cells (Meyer, H. et al.,; Meisinger-Henschel et al., J. Gen. Virol. 88, 3249-3259, 2007). Comparison of the MVA genome with its parent CVA revealed six major deletions (deletions I, II, III, IV, V, and VI) totaling 31,000 base pairs of genomic DNA (Meyer et al., J. Gen. Virol. 72:1031-8 (1991)). The resulting MVA has been shown to be significantly attenuated in various animal models (Mayr, A. & Danner, K. Vaccination against pox diseases under immunosuppressive conditions, Dev. Biol. Stand. 41:225-34, 1978). Using many passages to attenuate MVA, there are many different strains or isolates depending on the number of passages in CEF cells, such as MVA 476 MG / 14 / 78, MVA-571, MVA-572, MVA-574, MVA-575, and MVA-BN. MVA 476 MG / 14 / 78 is described, for example, in WO 2019 / 115816 (A1). The MVA-572 strain was deposited at the European Collection of Animal Cell Cultures ("ECACC"), Health Protection Agency, Microbiology Services, Porton Down, Salisbury SP4 0JG, United Kingdom ("UK"), on January 27, 1994 under deposit number ECACC 94012707. The MVA-575 strain was deposited at the ECACC on December 7, 2000 under deposit number ECACC 00120707.The MVA-Bavarian Nordic ("MVA-BN") strain was deposited at the ECACC under accession number V00080038 on August 30, 2000. The genomic sequences of MVA-BN and MVA-572 are available in GenBank (accession numbers DQ983238 and DQ983237, respectively). The genomic sequences of other MVA strains can be obtained using standard sequencing methods.

[0035] MVA can be derived from any MVA strain or further derivatives of MVA strains. A further exemplary MVA strain is deposit VR-1508 deposited with the American Type Culture collection (ATCC) (Manassas, Va. 20108, USA). A "derivative" of MVA refers to a virus that exhibits essentially the same characteristics as the parent MVA, but exhibits differences in one or more portions of its genome. In some embodiments, the MVA vector is derived from MVA 476 MG / 14 / 78. In some embodiments, the MVA vector is derived from MVA-571. In some embodiments, the MVA vector is derived from MVA-572. In some embodiments, the MVA vector is derived from MVA-574. In some embodiments, the MVA vector is derived from MVA-575. In some embodiments, the MVA vector is derived from MVA-BN.

[0036] The nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3 may be inserted into a site or region (insertion region) in the MVA virus genome that does not affect the viral viability of the resulting recombinant virus. Such regions can be easily identified by testing segments of viral DNA for regions that allow recombinant formation without seriously affecting the viral viability of the recombinant virus. The thymidine kinase (TK) gene is an insertion region that can be used and is present in many viruses, for example in all examined poxvirus genomes. In addition, MVA contains six naturally occurring deletion sites, each of which can be used as an insertion site (e.g., deletions I, II, III, IV, V, and VI; see, e.g., U.S. Pat. Nos. 5,185,146 and 6,440,442). One or more intergenic regions (IGRs) of MVA may be used as insertion sites, such as IGRs IGR07 / 08, IGR 44 / 45, IGR 64 / 65, IGR 88 / 89, IGR 136 / 137, and IGR 148 / 149 (see, e.g., U.S. Patent Application Publication No. 2018 / 0064803). Further suitable insertion sites are described in WO 2005 / 048957.

[0037] MVA viruses can be prepared as previously described (see Piccini, et al., 1987, Methods of Enzymology 153:545-563; U.S. Pat. Nos. 4,769,330, 4,772,848, 4,603,112, 5,100,587, and 5,179,993). In an exemplary method, the DNA sequence to be inserted into the viral genome can be placed into an E. coli plasmid construct into which DNA homologous to a portion of the MVA DNA has been inserted. Separately, the DNA sequence to be inserted can be ligated to a promoter. The promoter gene linkage can be positioned within the plasmid construct such that it is flanked on both ends by DNA homologous to DNA sequences that flank the MVA DNA region containing the non-essential gene locus. The resulting plasmid construct can be amplified and isolated by growth in E. coli bacteria. The isolated plasmid containing the DNA gene sequence to be inserted may be transfected into a cell culture, for example, chicken embryo fibroblast (CEF), and the culture is simultaneously infected with MVA. Recombination between the homologous MVA DNA in the plasmid and the viral genome, respectively, can generate MVA modified by the presence of the foreign DNA sequence. MVA particles may be recovered from the culture supernatant or from the cultured cells after a lysis step (e.g., chemical lysis, freeze / thaw, osmotic shock, sonication, etc.). Successive purification rounds of plaques can be used to remove contaminating wild-type virus. The viral particles can then be purified using techniques known in the art (e.g., chromatographic methods or ultracentrifugation in cesium chloride or sucrose gradients).

[0038] The method may further include administering one or more vaccines comprising the GAd20 virus, one or more vaccines comprising the MVA virus, or one or more vaccines comprising the GAd20 virus and one or more vaccines comprising the MVA virus. In some embodiments, the method may further include administering the treatment regimen two or more times. After the initial treatment regimen, for example, the method may include administering two vaccines comprising the GAd20 virus and one vaccine comprising the MVA virus. In some embodiments, the method may further include administering one vaccine comprising the GAd20 virus and one vaccine comprising the MVA virus. In some embodiments, the method may further include administering one or more vaccines comprising the MVA virus. The method may further include, for example, administering three vaccines comprising the MVA virus. The method may further include, for example, administering two vaccines comprising the MVA virus.

[0039] The appropriate amount of GAd20 virus is approximately 1 × 10 9 Virus particles (VP) ~ approx. 1 × 10 13 The VP may include the GAd20 virus.

[0040] A suitable amount of MVA virus is about 1×10 6 Infectious unit (IFU) ~ approx. 1 x 10 10 It may contain an IFU of MVA virus.

[0041] The method can further include administering an anti-CTLA4 antibody. The anti-CTLA4 antibody can be administered with a vaccine comprising GAd20 virus, with a vaccine comprising MVA virus, or both. Suitable amounts of anti-CTLA4 antibody include about 0.5 mg / kg to about 5 mg / kg. Any antagonistic anti-CTLA4 antibody can be used in the disclosed method. Anti-CTLA4 antibodies suitable for use in the disclosed method include, but are not limited to, human anti-CTLA4 antibodies, murine anti-CTLA4 antibodies, mammalian anti-CTLA4 antibodies, humanized anti-CTLA4 antibodies, monoclonal anti-CTLA4 antibodies, polyclonal anti-CTLA4 antibodies, and chimeric anti-CTLA4 antibodies. Non-limiting examples of anti-CTLA4 antibodies include ipilimumab and tremelimumab.

[0042] The method can further include administering an anti-PD-1 antibody. The anti-PD-1 antibody can be administered with the vaccine comprising the GAd20 virus, with the vaccine comprising the MVA virus, or both. Suitable amounts of the anti-PD-1 antibody include from about 0.5 mg / kg to about 5 mg / kg. Any antagonistic anti-PD-1 antibody can be used in the disclosed methods. Anti-PD-1 antibodies suitable for use in the disclosed methods include, but are not limited to, human anti-PD-1 antibodies, murine anti-PD-1 antibodies, mammalian anti-PD-1 antibodies, humanized anti-PD-1 antibodies, monoclonal anti-PD-1 antibodies, polyclonal anti-PD-1 antibodies, and chimeric anti-PD-1 antibodies. Non-limiting examples of anti-PD-1 antibodies include setremab, pembrolizumab, nivolumab, sintilimab, cemiplimab, toripalimab, canrelizumab, tislelizumab, dostralimab, spartalizumab, progolimab, balstilimab, budidjalimab, sasunlimab, avelumab, atezolizumab, durvalumab, embafolimab, and iodapolimab.

[0043] The method includes administering a vaccine comprising a GAd20 virus at week 0 and about week 3, and administering a vaccine comprising an MVA virus at about week 9; administering a vaccine comprising the GAd20 virus at about week 15 and about week 18, and administering a vaccine comprising the MVA virus at about week 24; administering a vaccine containing the GAd20 virus at about week 15 and a vaccine containing the MVA virus at about week 24; administering a vaccine comprising the MVA virus at about week 15, at about week 18, and at about week 24; or This can include administering a vaccine comprising the MVA virus at about 15 weeks and at about 24 weeks.

[0044] In some embodiments, the method further comprises administering a vaccine comprising the MVA virus at about week 36, about week 48, and about week 60. The method can further comprise administering one or more additional vaccines comprising the MVA virus.

[0045] The method is to administer 1 × 10 11 VP containing GAd20 virus, and at approximately week 9, 1 × 10 8 administering a vaccine containing an IFU MVA virus; 1×10 at approximately 15 and 18 weeks 11 VP GAd20 virus vaccine and 1 × 10 8 administering a vaccine containing an IFU MVA virus; At about week 15, 1 x 10 11 VP GAd20 virus vaccine and 1 × 10 8 administering a vaccine containing an IFU MVA virus; 1 × 10 at approximately 15 weeks, 18 weeks, and 24 weeks 8 administering a vaccine containing an MVA virus in IFU; or 1×10 at approximately 15 and 24 weeks 8 This may include administering a vaccine comprising an IFU of the MVA virus.

[0046] In some embodiments, the method includes administering 1×10 8 The method can further include administering a vaccine comprising an IFU of MVA virus. 8 It may further comprise administering one or more additional vaccines comprising an IFU of the MVA virus.

[0047] The disclosed methods can include exemplary treatment schedules provided in Table 1 below.

[0048] [Table 1] * Cycle 1 is referred to herein as the "treatment regimen."

[0049] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: 1×10 at week 0 and about week 3 11 a vaccine comprising one viral particle (VP) of the GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 x 10 8 a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 1×10 at approximately 15 and 18 weeks 11 Vaccines containing VP GAd20 virus; At approximately week 24, 1 x 10 8 A vaccine containing an MVA virus in IFU; and 1 x 10 at approximately 36 weeks, 48 ​​weeks, and 60 weeks 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0050] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: 1×10 at week 0 and about week 311 a vaccine comprising one viral particle (VP) of the GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 x 10 8 a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about week 15, 1 x 10 11 Vaccines containing VP GAd20 virus; At approximately week 24, 1 x 10 8 A vaccine containing the MVA virus in IFU; and 1 x 10 at approximately 36 weeks, 48 ​​weeks, and 60 weeks 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0051] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: 1×10 at week 0 and about week 3 11 a vaccine comprising one viral particle (VP) of the GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 x 10 8 a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 1 × 10 at approximately 15 weeks, 18 weeks, and 24 weeks 8 A vaccine containing the MVA virus in IFU; and 1 x 10 at approximately 36 weeks, 48 ​​weeks, and 60 weeks 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0052] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: 1×10 at week 0 and about week 3 11a vaccine comprising one viral particle (VP) of the GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 x 10 8 a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 1×10 at approximately 15 and 24 weeks 8 A vaccine containing the MVA virus in IFU; and 1 x 10 at approximately 36 weeks, 48 ​​weeks, and 60 weeks 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0053] Any of the above methods can further include administering an anti-CTLA4 antibody. The anti-CTLA4 antibody can be administered with a vaccine comprising the GAd20 virus, with a vaccine comprising the MVA virus, or both. Suitable amounts of anti-CTLA4 antibody include from about 1 mg / kg to about 3 mg / kg.

[0054] Any of the above methods can further include administering an anti-PD-1 antibody. The anti-PD-1 antibody can be administered with the vaccine comprising the GAd20 virus, with the vaccine comprising the MVA virus, or both. Suitable amounts of the anti-PD-1 antibody include from about 1 mg / kg to about 3 mg / kg.

[0055] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 an anti-CTLA4 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8anti-CTLA4 antibodies and vaccines comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15 and 18, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 Vaccines containing VP GAd20 virus; At approximately 24 weeks, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 A vaccine containing the MVA virus in IFU; and At about weeks 36, 48, and 60, patients received 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0056] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 an anti-PD1 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At approximately 15 and 18 weeks, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 Vaccines containing VP GAd20 virus; At approximately 24 weeks, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 A vaccine containing the MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0057] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 an anti-CTLA4 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 anti-CTLA4 antibodies and a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At approximately 15 weeks, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 Vaccines containing VP GAd20 virus; At approximately 24 weeks, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 A vaccine containing an MVA virus in IFU; and At about weeks 36, 48, and 60, patients received 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0058] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 an anti-PD1 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At approximately 15 weeks, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 Vaccines containing VP GAd20 virus; At approximately 24 weeks, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 A vaccine containing the MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0059] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 an anti-CTLA4 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 anti-CTLA4 antibodies and vaccines comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15, 18, and 24, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 A vaccine containing the MVA virus in IFU; and At about weeks 36, 48, and 60, patients received 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0060] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 an anti-PD1 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15, 18, and 24, subjects received 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 A vaccine containing the MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0061] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 an anti-CTLA4 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 anti-CTLA4 antibodies and vaccines comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At approximately 15 and 24 weeks, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8A vaccine containing the MVA virus in IFU; and At about weeks 36, 48, and 60, patients received 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0062] Provided herein are methods of treating or preventing prostate cancer in a subject, the methods comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 an anti-PD1 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At approximately 15 weeks and approximately 24 weeks, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 A vaccine containing the MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 Methods are disclosed that include administering a vaccine that includes an IFU of the MVA virus.

[0063] Each of the one or more GAd20 viruses can comprise the nucleotide sequence of SEQ ID NO:2. In some embodiments, the nucleotide sequence further comprises an N-terminal TCE, a C-terminal His tag, or both. The TCE can comprise the amino acid sequence of SEQ ID NO:5. The TCE can be encoded by the nucleotide sequence of SEQ ID NO:6. The His tag can comprise the amino acid sequence of SEQ ID NO:7. The His tag can be encoded by the nucleotide sequence of SEQ ID NO:8. The GAd20 virus can comprise a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:9. In some embodiments, the GAd20 virus can comprise the nucleotide sequence of SEQ ID NO:10.

[0064] Each of the one or more MVA viruses can comprise the nucleotide sequence of SEQ ID NO:4. In some embodiments, the nucleotide sequence further comprises an N-terminal TCE, a C-terminal His tag, or both. The TCE can comprise the amino acid sequence of SEQ ID NO:5. The TCE can be encoded by the nucleotide sequence of SEQ ID NO:6. The His tag can comprise the amino acid sequence of SEQ ID NO:7. The His tag can be encoded by the nucleotide sequence of SEQ ID NO:8. The MVA virus can comprise a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:11. In some embodiments, the MVA virus can comprise the nucleotide sequence of SEQ ID NO:12.

[0065] The methods can include screening the subject for the presence of one or more prostate cancer neoantigens encoding the amino acid sequence of SEQ ID NO: 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, or 93 prior to administering the one or more vaccines. In some embodiments, the method includes screening for the presence of one or more neoantigens comprising the nucleotide sequence of SEQ ID NO: 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, or 94 prior to administering the vaccine. For example, the disclosed methods of treating or preventing prostate cancer in a subject include: screening for the presence of one or more prostate cancer neoantigens encoding the amino acid sequence of SEQ ID NO: 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, or 93; If one or more neoantigens are detected, the subject is Two or more vaccines comprising a great ape adenovirus serotype 20 (GAd20) virus, the nucleotide sequence of which in order encodes the amino acid sequence of SEQ ID NO:1; and one or more vaccines comprising a Modified Vaccinia Ankara (MVA) virus comprising, in order, a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; Thereby, prostate cancer is treated or prevented.

[0066] Screening can include assaying for the presence of a neo-antigen protein or for the presence of a nucleic acid sequence encoding a neo-antigen protein. Exemplary screening techniques include, for example, genotyping, PCR, and protein analysis. EXAMPLES

[0067] The following examples are provided to further illustrate some of the embodiments disclosed herein and are intended to be illustrative and not limiting of the embodiments of the present disclosure.

[0068] Immunogenicity study of Gad20 / MVA in combination with αCTLA4 and / or αPD1 antibodies in cynomolgus monkeys The primary objective of this study was to determine whether GAd20 / GAd20 / MVA treatment could generate prostate neoantigen-specific T cell responses in non-human primates (NHPs), which could be further augmented using anti-CTLA4 antibody (αCTLA4) and anti-PD-1 antibody (αPD-1). A secondary objective was to assess whether two consecutive primings with GAd20 at weeks 0 and 4 increased the number of responding animals by week 6.

[0069] The dosing sequence used in this study is shown in Figure 1B. Briefly, animals were administered 5 x 10 10 VP at GAd20 and at 12 weeks, 1 × 10 8MVA of IFU was administered (groups 1 to 4). Group 2 received 3 mg / kg of anti-CTLA4 antibody (Ipi) intravenously at weeks 0, 4, and 12. Group 3 received 3 mg / kg of anti-CTLA4 antibody (Ipi) subcutaneously at weeks 0, 4, and 12, and 10 mg / kg of anti-PD-1 antibody intravenously at weeks 4, 8, 12, and 16. Group 4 received 3 mg / kg of anti-CTLA4 antibody (Ipi) subcutaneously at weeks 0 and 4, 1 mg / kg of anti-CTLA4 antibody (Ipi) subcutaneously at week 12, and 10 mg / kg of anti-PD-1 antibody intravenously at weeks 4, 8, 12, and 16.

[0070] NHP-derived PBMCs were tested using an overnight peptide recall IFNγ ELISpot assay. As shown in Figure 2, the GAd20 vaccine is immunogenic in NHPs (4 of 8 responders in group 1 at week 2). GAd20 double priming increased the number of responding animals (8 of 8 responders in group 1 at week 6) and increased the frequency of antigen-specific T cell responses. These results confirmed that GAd20 is immunogenic and that double priming enhances vaccine-specific T cell responses at week 6.

[0071] As shown in Figure 3, the MVA vaccine increased antigen-specific T cell responses by 2.4-fold at week 18 compared to week 12. The average of all time points after week 12 was significantly elevated compared to responses at week 12 (P=0.005). These results confirmed that MVA is immunogenic and significantly enhances antigen-specific T cell responses primed with 2xGAd20.

[0072] As shown in Figure 4, co-administration of αCTLA4 significantly increased the number of responding animals and the magnitude of the antigen-specific T cell response 2 weeks after the initial GAd20 immunization. With GAd20 alone, the mean was 215 SFU / 1e6 PBMC at week 4, with only 6 of 8 animals responding (Group 1). Co-administration of αCTLA4 increased the mean to 682 SFU / 1e6 PBMC, with all animals responding (Group 2).

[0073] αCTLA4 was shown to increase the magnitude of prostate neoantigen-specific T cell responses over the course of the study (Figure 5). As shown in Figure 5, MVA enhanced immune responses by week 16; αCTLA4 increased the magnitude of immune responses over the entire course of the study; αPD-1 (groups 3 and 4) did not significantly decrease antigen-specific immune responses. These results confirmed that (1) coadministration of αCTLA4 improved the magnitude of T cell responses over all time points compared to vaccination alone, and (2) the combination of anti-PD-1 and αCTLA4 with GAd20 / MVA heterologous multi-dose vaccination also improved the magnitude of antigen-specific T cell responses over all time points compared to vaccination alone.

[0074] NHP PBMCs were tested in an overnight peptide recall intracellular cytokine staining (ICS) assay. As shown in Figure 6, NHPs receiving vaccine + αCTLA4 generated more neo-antigen-specific polyfunctional (IFNγ / TNFα double positive) CD8+ T cells than NHPs receiving vaccine alone. These results indicate that co-administration of αCTLA4 improves the quality of antigen-specific T cells as evidenced by an increased frequency of polyfunctional CD8+ T cells.

[0075] As shown in FIG. 7, effector memory neo-antigen-specific CD8+ T cells are increased by co-administration of αCTLA4 compared to vaccine alone at week 21.

[0076] NHP PBMCs were tested in an ICS assay utilizing four subpools of peptides, each consisting of approximately 1 / 4 of the total neoantigens in the vaccine. As shown in Figure 8, vaccine alone generated primarily subpool 3 responses, whereas animals administered both αCTLA4 and vaccine generated high frequencies of T cell responses specific for all four subpools. Thus, coadministration of αCTLA4 expands the breadth of unique vaccine antigen-specific T cells.

[0077] conclusion · GAd20 could effectively prime prostate neoantigen immune responses in NHPs, and the use of "double priming" with GAd20 increased the number of responding animals. · MVA effectively boosted animals that had received a 2 × GAd20 immunization. Addition of αCTLA4 to the vaccine resulted in increased magnitude and sustained immune responses, both in terms of total immune responses as measured by ELISpot and in the frequency of antigen-specific polyfunctional CD8 T cell responses. · αCTLA4 can increase the frequency of both neoantigen-specific effector CD8+ T cells and increase the breadth of neoantigen-specific immune responses. · Addition of αPD-1 does not significantly reduce neoantigen-specific immune responses.

[0078] Those skilled in the art will appreciate that numerous changes and modifications may be made to the preferred embodiments of the present invention, and that such changes and modifications may be made without departing from the spirit of the present invention. It is therefore intended in the appended claims to cover all such equivalent variations that fall within the true spirit and scope of the present invention.

[0079] The disclosures of each patent, patent application, and publication cited or described in this specification are hereby incorporated by reference in their entirety.

[0080] [Table 2-1]

[0081] [Table 2-2]

[0082] [Table 2-3]

[0083] [Table 2-4]

[0084] [Table 2-5]

[0085] [Table 2-6]

[0086] [Table 2-7]

[0087] [Table 2-8]

[0088] [Table 2-9]

[0089] [Table 2-10]

[0090] [Table 2-11]

[0091] [Table 2-12]

[0092] [Table 2-13]

[0093] Embodiment The following list of embodiments is intended to complement, not replace or supplant, the foregoing description.

[0094] Embodiment 1. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: Two or more vaccines comprising a great ape adenovirus serotype 20 (GAd20) virus, the nucleotide sequence of which in order encodes the amino acid sequence of SEQ ID NO:1; and one or more vaccines comprising a Modified Vaccinia Ankara (MVA) virus comprising, in order, a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3, thereby treating or preventing prostate cancer.

[0095] Embodiment 2. The method of embodiment 1, further comprising administering the treatment regimen two or more times.

[0096] Embodiment 3. one or more vaccines comprising the GAd20 virus; one or more vaccines comprising the MVA virus; or further comprising administering one or more vaccines comprising a GAd20 virus and one or more vaccines comprising an MVA virus; 3. The method according to embodiment 1 or 2.

[0097] Embodiment 4. The method of embodiment 3, comprising administering two vaccines comprising the GAd20 virus and one vaccine comprising the MVA virus.

[0098] Embodiment 5. The method of embodiment 3, comprising administering one vaccine comprising a GAd20 virus and one vaccine comprising an MVA virus.

[0099] Embodiment 6 The method of embodiment 3, comprising administering three vaccines comprising the MVA virus.

[0100] Embodiment 7. The method of embodiment 3, comprising administering two vaccines comprising the MVA virus.

[0101] Embodiment 8. The method of any one of the preceding embodiments, further comprising administering one or more vaccines comprising an MVA virus.

[0102] Embodiment 9. Each of the vaccines comprising the GAd20 virus comprises about 1 x 10 9 Virus particles (VP) ~ approx. 1 × 10 13 The method of any one of the preceding embodiments, comprising the VP GAd20 virus.

[0103] Embodiment 10. Each of the vaccines comprising the MVA virus comprises about 1x10 6 Infectious unit (IFU) ~ approx. 1x10 10 The method of any one of the preceding embodiments, comprising an MVA virus in an IFU.

[0104] Embodiment 11 The method of any one of the preceding embodiments, further comprising administering an anti-CTLA4 antibody.

[0105] Embodiment 12. The anti-CTLA4 antibody is Vaccines containing the GAd20 virus; A vaccine containing the MVA virus; or 12. The method of embodiment 11, comprising administering together

[0106] Embodiment 13. The method of embodiment 11 or 12, comprising administering 0.5 mg / kg to 5 mg / kg of an anti-CTLA4 antibody.

[0107] Embodiment 14. The method of any one of embodiments 1 to 10, further comprising administering an anti-PD-1 antibody.

[0108] Embodiment 15. The anti-PD-1 antibody is Vaccines containing the GAd20 virus; A vaccine containing the MVA virus; or 15. The method of embodiment 14, comprising administering together

[0109] Embodiment 16. The method of embodiment 14 or 15 comprising administering 0.5 mg / kg to 5 mg / kg of an anti-PD-1 antibody.

[0110] Embodiment 17. The method of any one of the preceding embodiments, wherein each of the one or more GAd20 viruses comprises the nucleotide sequence of SEQ ID NO:2.

[0111] Embodiment 18. The method of embodiment 17, wherein the nucleotide sequence further comprises an N-terminal TCE, a C-terminal His tag, or both.

[0112] Embodiment 19. The method of any one of the preceding embodiments, wherein each of the one or more MVA viruses comprises the nucleotide sequence of SEQ ID NO:4.

[0113] Embodiment 20. The method of embodiment 19, wherein the nucleotide sequence further comprises an N-terminal TCE, a C-terminal His tag, or both.

[0114] Embodiment 21. The method of any one of the preceding embodiments, comprising administering a vaccine comprising a GAd20 virus at weeks 0 and about 3, and administering a vaccine comprising an MVA virus at about week 9.

[0115] Embodiment 22. The method of embodiment 21, comprising administering a vaccine comprising a GAd20 virus at about week 15 and about week 18, and administering a vaccine comprising an MVA virus at about week 24.

[0116] Embodiment 23. The method of embodiment 21, comprising administering a vaccine comprising a GAd20 virus at about week 15 and administering a vaccine comprising an MVA virus at about week 24.

[0117] Embodiment 24. The method of embodiment 21, comprising administering a vaccine comprising an MVA virus at about week 15, about week 18, and about week 24.

[0118] Embodiment 25. The method of embodiment 21, comprising administering a vaccine comprising an MVA virus at about week 15 and about week 24.

[0119] Embodiment 26 The method of any one of the preceding embodiments, comprising administering a vaccine comprising an MVA virus at about week 36, about week 48, and about week 60.

[0120] 27. 1×10 at week 0 and about week 3 11 VP GAd20 virus vaccine and 1 × 10 8 Administering a vaccine comprising an MVA virus in an IFU.

[0121] Embodiment 28. 1 x 10 at about week 15 and about week 18 11 VP GAd20 virus vaccine and 1 × 10 8 28. The method of embodiment 27, comprising administering a vaccine comprising an IFU of the MVA virus.

[0122] Embodiment 29. 1 × 10 at approximately week 15 11 VP GAd20 virus vaccine and 1 × 10 8 28. The method of embodiment 27, comprising administering a vaccine comprising an IFU of the MVA virus.

[0123] Embodiment 30. 1×10 at about week 15, about week 18, and about week 24 8 28. The method of embodiment 27, comprising administering a vaccine comprising an MVA virus in an IFU.

[0124] Embodiment 31. 1 x 10 at about week 15 and about week 24 828. The method of embodiment 27, comprising administering a vaccine comprising an MVA virus in an IFU.

[0125] Embodiment 32. 1×10 at about week 36, about week 48, and about week 60 8 The method of any one of the preceding embodiments, comprising administering a vaccine comprising an MVA virus in an IFU.

[0126] Embodiment 33.1 x 10 8 33. The method of embodiment 32, comprising administering one or more additional vaccines comprising an IFU of an MVA virus.

[0127] Embodiment 34. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: 1×10 at week 0 and about week 3 11 a vaccine comprising 10 viral particles (VP) of a GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 x 10 8 a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 1×10 at approximately 15 and 18 weeks 11 Vaccines containing VP GAd20 virus; At approximately week 24, 1 x 10 8 A vaccine containing an MVA virus in IFU; and 1 x 10 at approximately 36 weeks, 48 ​​weeks, and 60 weeks 8 Administering a vaccine comprising an IFU of an MVA virus.

[0128] Embodiment 35. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: 1×10 at week 0 and about week 3 11 a vaccine comprising 10 viral particles (VP) of a GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 x 10 8 a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about week 15, 1 x 10 11 Vaccines containing VP GAd20 virus; At approximately week 24, 1 x 10 8 A vaccine containing an MVA virus in IFU; and 1 x 10 at approximately 36 weeks, 48 ​​weeks, and 60 weeks 8 Administering a vaccine comprising an IFU of an MVA virus.

[0129] Embodiment 36. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: 1×10 at week 0 and about week 3 11 a vaccine comprising 10 viral particles (VP) of a GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 x 10 8 a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 1 × 10 at approximately 15 weeks, 18 weeks, and 24 weeks 8 A vaccine containing an MVA virus in IFU; and 1 x 10 at approximately 36 weeks, 48 ​​weeks, and 60 weeks 8 Administering a vaccine comprising an IFU of an MVA virus.

[0130] Embodiment 37. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: 1×10 at week 0 and about week 3 11 a vaccine comprising 10 viral particles (VP) of a GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 x 10 8a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 1×10 at approximately 15 and 24 weeks 8 A vaccine containing the MVA virus in IFU; and 1 x 10 at approximately 36 weeks, 48 ​​weeks, and 60 weeks 8 Administering a vaccine comprising an IFU of an MVA virus.

[0131] Embodiment 38. The method of any one of embodiments 34 to 37, further comprising administering an anti-CTLA4 antibody.

[0132] Embodiment 39. An anti-CTLA4 antibody is administered Vaccines containing the GAd20 virus; A vaccine containing the MVA virus; or 39. The method of embodiment 38, comprising administering together

[0133] Embodiment 40. The method of embodiment 38 or 39, comprising administering 1 mg / kg to 3 mg / kg of an anti-CTLA4 antibody.

[0134] Embodiment 41 The method of any one of embodiments 34 to 37, further comprising administering an anti-PD-1 antibody.

[0135] Embodiment 42. An anti-PD-1 antibody is administered Vaccines containing the GAd20 virus; A vaccine containing the MVA virus; or 42. The method of embodiment 41, comprising administering together

[0136] Embodiment 43. The method of embodiment 41 or 42 comprising administering 1 mg / kg to 3 mg / kg of the anti-PD-1 antibody.

[0137] Embodiment 44 The method of any one of embodiments 34 to 43, wherein the GAd20 virus comprises the nucleotide sequence of SEQ ID NO:2.

[0138] Embodiment 45. The method of embodiment 44, wherein the nucleotide sequence further comprises an N-terminal TCE, a C-terminal His tag, or both.

[0139] Embodiment 46 The method of any one of embodiments 34 to 43, wherein the MVA virus comprises the nucleotide sequence of SEQ ID NO:4.

[0140] Embodiment 47. The method of embodiment 46, wherein the nucleotide sequence further comprises an N-terminal TCE, a C-terminal His tag, or both.

[0141] Embodiment 48. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 an anti-CTLA4 antibody and a vaccine comprising one viral particle (VP) of GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 anti-CTLA4 antibodies and a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15 and 18, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 Vaccines containing VP GAd20 virus; At approximately 24 weeks, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 A vaccine containing an MVA virus in IFU; and At about weeks 36, 48, and 60, patients received 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 Administering a vaccine comprising an IFU of an MVA virus.

[0142] Embodiment 49. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 an anti-PD1 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At approximately 15 and 18 weeks, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 Vaccines containing VP GAd20 virus; At approximately 24 weeks, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 A vaccine containing an MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 Administering a vaccine comprising an IFU of an MVA virus.

[0143] Embodiment 50. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 an anti-CTLA4 antibody and a vaccine comprising one viral particle (VP) of GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 anti-CTLA4 antibodies and a vaccine comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At approximately 15 weeks, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 Vaccines containing VP GAd20 virus; At approximately 24 weeks, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 A vaccine containing an MVA virus in IFU; and At about weeks 36, 48, and 60, patients received 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 Administering a vaccine comprising an IFU of an MVA virus.

[0144] Embodiment 51. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 an anti-PD1 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At approximately 15 weeks, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 Vaccines containing VP GAd20 virus; At approximately 24 weeks, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 A vaccine containing an MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 Administering a vaccine comprising an IFU of an MVA virus.

[0145] Embodiment 52. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 an anti-CTLA4 antibody and a vaccine comprising one viral particle (VP) of GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 anti-CTLA4 antibodies and vaccines comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15, 18, and 24, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 A vaccine containing the MVA virus in IFU; and At about weeks 36, 48, and 60, patients received 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 Administering a vaccine comprising an IFU of an MVA virus.

[0146] Embodiment 53. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 an anti-PD1 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15, 18, and 24, subjects received 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 A vaccine containing the MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 Administering a vaccine comprising an IFU of an MVA virus.

[0147] Embodiment 54. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 11 an anti-CTLA4 antibody and a vaccine comprising one viral particle (VP) of GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 anti-CTLA4 antibodies and vaccines comprising an infectious unit (IFU) of MVA virus, the MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At approximately 15 and 24 weeks, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 A vaccine containing the MVA virus in IFU; and At about weeks 36, 48, and 60, patients received 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1 × 10 8 Administering a vaccine comprising an IFU of an MVA virus.

[0148] Embodiment 55. A method of treating or preventing prostate cancer in a subject, the method comprising administering to the subject: At week 0 and approximately week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 11 an anti-PD1 antibody and a vaccine comprising a GAd20 virus of 10 viral particles (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At approximately week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of MVA virus, wherein the MVA virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At approximately 15 weeks and approximately 24 weeks, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 A vaccine containing the MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1 × 10 8 Administering a vaccine comprising an IFU of an MVA virus.

Claims

1. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: two or more vaccines comprising a great ape adenovirus serotype 20 (GAd20) virus, the nucleotide sequence of which in order encodes the amino acid sequence of SEQ ID NO:1; one or more vaccines comprising a modified vaccinia Ankara (MVA) virus comprising, in order, a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; A treatment regimen including thereby treating or preventing said prostate cancer. method.

2. 10. The method of claim 1, further comprising administering the treatment regimen two or more times.

3. one or more vaccines comprising the GAd20 virus; one or more vaccines comprising said MVA virus; or further comprising administering one or more vaccines comprising the GAd20 virus and one or more vaccines comprising the MVA virus. The method according to claim 1 or 2.

4. 4. The method of claim 3, comprising administering two vaccines comprising the GAd20 virus and one vaccine comprising the MVA virus.

5. 4. The method of claim 3, comprising administering one vaccine comprising the GAd20 virus and one vaccine comprising the MVA virus.

6. 4. The method of claim 3, comprising administering three vaccines comprising the MVA virus.

7. 4. The method of claim 3, comprising administering two vaccines comprising the MVA virus.

8. 13. The method of any one of the preceding claims, further comprising administering one or more vaccines comprising the MVA virus.

9. Each of the vaccines comprising the GAd20 virus is about 1×10 ９ Virus particles (VP) ~ about 1 x 10 １３ 2. The method according to any one of the preceding claims, comprising said GAd20 virus of VP.

10. Each of the vaccines containing the MVA virus is about 1 x 10 ６ Infectious units (IFU) ~ approximately 1 x 10 １０ 10. The method according to any one of the preceding claims, comprising the MVA virus in IFU.

11. 2. The method of any one of the preceding claims, further comprising administering an anti-CTLA4 antibody.

12. The anti-CTLA4 antibody, the vaccine comprising the GAd20 virus; the vaccine comprising the MVA virus; or both and administering The method of claim 11.

13. 13. The method of claim 11 or 12, comprising administering 0.5 mg / kg to 5 mg / kg of the anti-CTLA4 antibody.

14. The method of any one of claims 1 to 10, further comprising administering an anti-PD-1 antibody.

15. The anti-PD-1 antibody, the vaccine comprising the GAd20 virus; the vaccine comprising the MVA virus; or both and administering The method of claim 14.

16. 16. The method of claim 14 or 15, comprising administering 0.5 mg / kg to 5 mg / kg of the anti-PD-1 antibody.

17. 2. The method of any one of the preceding claims, wherein each of the one or more GAd20 viruses comprises the nucleotide sequence of SEQ ID NO:

2.

18. 18. The method of claim 17, wherein the nucleotide sequence further comprises an N-terminal TCE, a C-terminal His tag, or both.

19. 10. The method of any one of the preceding claims, wherein each of the one or more MVA viruses comprises the nucleotide sequence of SEQ ID NO:

4.

20. 20. The method of claim 19, wherein the nucleotide sequence further comprises an N-terminal TCE, a C-terminal His tag, or both.

21. 2. The method of any one of the preceding claims, comprising administering a vaccine comprising the GAd20 virus at week 0 and about week 3, and administering a vaccine comprising the MVA virus at about week 9.

22. 22. The method of claim 21, comprising administering a vaccine comprising the GAd20 virus at about week 15 and about week 18, and administering a vaccine comprising the MVA virus at about week 24.

23. 22. The method of claim 21, comprising administering a vaccine comprising the GAd20 virus at about week 15 and administering a vaccine comprising the MVA virus at about week 24.

24. 22. The method of claim 21, comprising administering a vaccine comprising the MVA virus at about week 15, about week 18, and about week 24.

25. 22. The method of claim 21, comprising administering a vaccine comprising the MVA virus at about week 15 and about week 24.

26. 10. The method of any one of the preceding claims, comprising administering a vaccine comprising the MVA virus at about week 36, about week 48, and about week 60.

27. 1 x 10 at week 0 and about week 3 １１ VP and administering a vaccine containing the GAd20 virus to the 1×10 ８ 2. The method of any one of the preceding claims, comprising administering a vaccine comprising IFU of said MVA virus.

28. At about the 15th and 18th weeks, １１ and administering a vaccine containing the GAd20 virus to 1×10 ８ and administering a vaccine comprising the MVA virus in IFU.

29. At about the 15th week, 1x10 １１ and administering a vaccine containing the GAd20 virus to 1×10 ８ and administering a vaccine comprising the MVA virus in IFU.

30. 1 x 10 at about week 15, about week 18, and about week 24 ８ 28. The method of claim 27, comprising administering a vaccine comprising IFU of the MVA virus.

31. At about the 15th and 24th weeks, ８ 28. The method of claim 27, comprising administering a vaccine comprising IFU of the MVA virus.

32. 1 x 10 at about week 36, about week 48, and about week 60 ８ 13. The method of any one of the preceding claims, comprising administering a vaccine comprising IFU of the MVA virus.

33. 1x10 ８ 33. The method of claim 32, comprising administering one or more additional vaccines comprising IFU of the MVA virus.

34. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: 1 x 10 at week 0 and about week 3 １１ a vaccine comprising a GAd20 virus of viral particles (VP), said GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about the 9th week, 1 x 10 ８ A vaccine comprising an infectious unit (IFU) of MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about the 15th and 18th weeks, １１ a vaccine comprising said GAd20 virus of VP; At about the 24th week, 1 x 10 ８ A vaccine comprising said MVA virus in IFU; and 1 x 10 at about week 36, about week 48, and about week 60 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

35. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: 1 x 10 at week 0 and about week 3 １１ a vaccine comprising a GAd20 virus of viral particles (VP), said GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about the 9th week, 1 x 10 ８ A vaccine comprising an infectious unit (IFU) of MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about the 15th week, 1x10 １１ a vaccine comprising said GAd20 virus of VP; At about the 24th week, 1 x 10 ８ A vaccine comprising said MVA virus in IFU; and 1 x 10 at about week 36, about week 48, and about week 60 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

36. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: 1 x 10 at week 0 and about week 3 １１ a vaccine comprising a GAd20 virus of viral particles (VP), said GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about the 9th week, 1 x 10 ８ A vaccine comprising an infectious unit (IFU) of MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; 1 x 10 at about week 15, about week 18, and about week 24 ８ A vaccine comprising said MVA virus in IFU; and 1 x 10 at about week 36, about week 48, and about week 60 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

37. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: 1 x 10 at week 0 and about week 3 １１ a vaccine comprising a GAd20 virus of viral particles (VP), said GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about the 9th week, 1 x 10 ８ A vaccine comprising an infectious unit (IFU) of MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about the 15th and 24th weeks, ８ A vaccine comprising said MVA virus in IFU; and 1 x 10 at about week 36, about week 48, and about week 60 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

38. The method of any one of claims 34 to 37, further comprising administering an anti-CTLA4 antibody.

39. The anti-CTLA4 antibody, the vaccine comprising the GAd20 virus; the vaccine comprising the MVA virus; or both 40. The method of claim 38, comprising administering

40. 40. The method of claim 38 or 39, comprising administering 1 mg / kg to 3 mg / kg of the anti-CTLA4 antibody.

41. The method of any one of claims 34 to 37, further comprising administering an anti-PD-1 antibody.

42. The anti-PD-1 antibody, the vaccine comprising the GAd20 virus; the vaccine comprising the MVA virus; or both 42. The method of claim 41 , comprising administering

43. 43. The method of claim 41 or 42, comprising administering 1 mg / kg to 3 mg / kg of the anti-PD-1 antibody.

44. The method of any one of claims 34 to 43, wherein the GAd20 virus comprises the nucleotide sequence of SEQ ID NO:

2.

45. 45. The method of claim 44, wherein the nucleotide sequence further comprises an N-terminal TCE, a C-terminal His tag, or both.

46. The method according to any one of claims 34 to 43, wherein the MVA virus comprises the nucleotide sequence of SEQ ID NO:

4.

47. 47. The method of claim 46, wherein the nucleotide sequence further comprises an N-terminal TCE, a C-terminal His tag, or both.

48. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: At week 0 and about week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 １１ an anti-CTLA4 antibody and a vaccine comprising a single viral particle (VP) of a GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ anti-CTLA4 antibodies and vaccines comprising an infectious unit (IFU) of MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15 and 18, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 １１ a vaccine comprising said GAd20 virus of VP; At about week 24, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ A vaccine comprising said MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

49. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: At week 0 and about week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 １１ an anti-PD1 antibody and a vaccine comprising a GAd20 virus of one viral particle (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 ８ an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of an MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15 and 18, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 １１ a vaccine comprising said GAd20 virus of VP; At about week 24, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 ８ A vaccine comprising said MVA virus in IFU; and 1 mg / kg to 3 mg / kg of anti-PD1 antibody at about week 36, about week 48, and about week 60, and 1×10 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

50. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: At week 0 and about week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 １１ an anti-CTLA4 antibody and a vaccine comprising a single viral particle (VP) of a GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ anti-CTLA4 antibodies and vaccines comprising an infectious unit (IFU) of MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about week 15, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 １１ a vaccine comprising said GAd20 virus of VP; At about week 24, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ A vaccine comprising said MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

51. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: At week 0 and about week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 １１ an anti-PD1 antibody and a vaccine comprising a GAd20 virus of one viral particle (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 ８ an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of an MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about week 15, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 １１ a vaccine comprising said GAd20 virus of VP; At about week 24, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 ８ A vaccine comprising said MVA virus in IFU; and 1 mg / kg to 3 mg / kg of anti-PD1 antibody at about week 36, about week 48, and about week 60, and 1×10 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

52. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: At week 0 and about week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 １１ an anti-CTLA4 antibody and a vaccine comprising a single viral particle (VP) of a GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ anti-CTLA4 antibodies and vaccines comprising an infectious unit (IFU) of MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15, 18, and 24, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ A vaccine comprising said MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

53. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: At week 0 and about week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 １１ an anti-PD1 antibody and a vaccine comprising a GAd20 virus of one viral particle (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 ８ an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of an MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15, 18, and 24, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 ８ A vaccine comprising said MVA virus in IFU; and 1 mg / kg to 3 mg / kg of anti-PD1 antibody at about week 36, about week 48, and about week 60, and 1×10 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

54. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: At week 0 and about week 3, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 １１ an anti-CTLA4 antibody and a vaccine comprising a single viral particle (VP) of a GAd20 virus, the GAd20 virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ anti-CTLA4 antibodies and vaccines comprising an infectious unit (IFU) of MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15 and 24, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ A vaccine comprising said MVA virus in IFU; and At about weeks 36, 48, and 60, 1 mg / kg to 3 mg / kg of anti-CTLA4 antibody and 1×10 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

55. 1. A method of treating or preventing prostate cancer in a subject, comprising: The method includes administering to the subject: At week 0 and about week 3, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 １１ an anti-PD1 antibody and a vaccine comprising a GAd20 virus of one viral particle (VP), wherein the GAd20 virus comprises a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:1; At about week 9, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 ８ an anti-PD1 antibody and a vaccine comprising an infectious unit (IFU) of an MVA virus, said MVA virus comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:3; At about weeks 15 and 24, 1 mg / kg to 3 mg / kg of anti-PD1 antibody and 1×10 ８ A vaccine comprising said MVA virus in IFU; and 1 mg / kg to 3 mg / kg of anti-PD1 antibody at about week 36, about week 48, and about week 60, and 1×10 ８ Vaccine containing the MVA virus in IFU The method of claim 1, further comprising administering

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