Methods for treating prostate cancer

Abstract

The present disclosure is directed to methods of treating a prostate cancer in a subject using a virus, such as an adenovirus (e.g., aglatimagene besadenovec).

Description

Attorney Docket No.: CAND-020WOMETHODS FOR TREATING PROSTATE CANCERCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Provisional Application Number 63 / 730,427, filed December 10, 2024 and U.S. Provisional Application Number 63 / 813,872, filed May 29, 2025, the contents of each of which are incorporated by reference herein in their entirety.FIELD OF THE INVENTION

[0002] The present disclosure is directed to methods of treating prostate cancer in a subject using a virus such as an adenovirus (e.g., aglatimagene besadenovec).BACKGROUND

[0003] In the United States there are approximately 200,000 new annual diagnoses of prostate cancer. Current therapies provide an excellent 5-year survival prognosis, yet prostate cancer is still the second leading cause of cancer death in men in the U.S. Recurrence within 10 years occurs in 30-50% of patients, most occurring within 5 years of treatment. In the U.S., this translates into an annual incidence of prostate cancer recurrence of 60,000-100,000 and a prevalence of around 2.1 million in 2005. Prostate cancer is normally treated by surgical or pharmacologic castration (androgen deprivation therapy (ADT)), which has a significant negative impact on quality of life. As a consequence of better diagnostic tools and surveillance, the population of men affected by prostate cancer is shifting to younger, healthier men who may be most greatly impacted by recurrence and its treatment effects. The costs to society from treatment costs, lost productivity, and compromised quality of life due to recurrence are in the billions of dollars.

[0004] Despite the advances made to date for the treatment of prostate cancer, there remains a need for new and improved therapies for successfully treating prostate cancer.SUMMARY

[0005] The disclosure relates, in part, to the discovery that treatment of a subject with newly-diagnosed prostate cancer with a virus, such as aglatimagene besadenovec, and radiation therapy results in improved disease-free survival of the subject as compared to a subject with newly-diagnosed prostate cancer receiving only the radiation therapy (e.g., anAttorney Docket No.: CAND-020WO equivalent or same level of the radiation therapy received by the subject). Subjects with newly-diagnosed prostate cancer can include subjects that have been diagnosed with intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate -risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate -risk prostate cancer).

[0006] The disclosure relates, in part, to the discovery that treatment of a subject with newly-diagnosed prostate cancer with a virus, such as aglatimagene besadenovec, and radiation therapy results in an improved prostate -specific outcome of the subject as compared to a subject with newly-diagnosed prostate cancer receiving only the radiation therapy (e.g., an equivalent or same level of the radiation therapy received by the subject). Subjects with newly-diagnosed prostate cancer can include subjects that have been diagnosed with intermediate- to high-risk prostate cancer.

[0007] The disclosure also relates, in part, to the discovery that treatment of a subject with intermediate- to high-risk prostate cancer with a virus, such as aglatimagene besadenovec, and radiation therapy results in reduced prostate-specific antigen (PSA) level of the subject. The PSA level can be reduced to a nadir of < 0.2 ng / mL, for a prolonged period of time (e.g., months to years e.g., two months, three months, four months, five months, six months, eight months, ten months, one year, two years, three years, or more).

[0008] The disclosure also relates, in part, to the discovery that delivery of a virus, such as aglatimagene besadenovec, to a subject with prostate cancer who is contraindicated for androgen depletion therapy (ADT) or elects not to undergo ADT, and who is receiving radiation therapy may provide significant benefits over administration of the radiation therapy alone, for example, in terms of disease free survival.

[0009] The disclosure also relates, in part, to the discovery that delivery of a virus, such as aglatimagene besadenovec, to a subject with prostate cancer who is receiving radiation therapy may reduce at least one side effect associated with prostate cancer. The side effect(s) associated with prostate cancer can include pollakiuria, micturition urgency, and urinary tract pain.

[0010] Accordingly, in one aspect, the disclosure provides a method of increasing disease-free survival in a subject with prostate cancer in need thereof. The method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy, so that the subject’sAttorney Docket No.: CAND-020WO disease-free survival is increased as compared to a subject with prostate cancer receiving only the radiation therapy (e.g., an equivalent or same level of the radiation therapy received by the subject). Depending upon the circumstances, the method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec and a therapeutically effective amount of a radiation therapy.

[0011] In certain embodiments, the subject is newly-diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0012] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec. In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0013] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days.

[0014] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0015] Depending upon the circumstances, the method increases the subject’s disease- free survival to at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec. Moreover, the method increases the subject’s probability of disease-free survival to at least about 0.55, 0.6, 0.65, 0.75, 0.8, or 0.85 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.Attorney Docket No.: CAND-020WO

[0016] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT.

[0017] In certain embodiments, the subject achieves a PSA nadir of < 0.2 ng / mL (e.g., undetectable).

[0018] In another aspect, the disclosure provides a method of reducing a PSA level to a nadir of < 0.2 ng / mL in a subject with intermediate- to high-risk prostate cancer in need thereof. The method includes administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy.Depending upon the circumstances, the method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec and radiation therapy.

[0019] In certain embodiments, the subject is newly-diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0020] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0021] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0022] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days.

[0023] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0024] Depending upon the circumstances, the method increases the subject’s disease- free survival to at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years,Attorney Docket No.: CAND-020WO11 years, or 12 years following the initial administration of the aglatimagene besadenovec. For example, the method increases the subject’s probability of disease-free survival to at least about 0.55, 0.6, 0.65, 0.75, 0.8, or 0.85 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0025] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT.

[0026] In another aspect, the disclosure provides a method of treating a subject with prostate cancer who does not receive ADT. The method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy. Depending upon the circumstances, the method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec and a therapeutically effective amount of a radiation therapy.

[0027] In certain embodiments, the subject is newly -diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0028] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0029] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0030] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days.

[0031] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 GyAttorney Docket No.: CAND-020WO fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0032] Depending upon the circumstances, the method increases the subject’s disease- free survival to at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec. For example, the method increases the subject’s probability of disease-free survival to at least about 0.55, 0.6, 0.65, 0.75, 0.8, or 0.85 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0033] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT.

[0034] In certain embodiments, the subject achieves a PSA nadir of < 0.2 ng / mL (e.g., undetectable).

[0035] In another aspect, the disclosure provides a method of reducing a PSA level to a nadir of < 0.2 ng / mL in a subject with intermediate- to high-risk prostate cancer in need thereof. The method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy. Depending upon the circumstances, the method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec and radiation therapy.

[0036] In certain embodiments, the subject is newly-diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0037] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0038] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0039] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, theAttorney Docket No.: CAND-020WO prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days. In certain embodiments, the prodrug is valacyclovir.

[0040] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0041] In certain embodiments, the method increases the subject’s disease-free survival to at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following administration of the aglatimagene besadenovec. For example, the method increases the subject’s probability of disease-free survival to at least about 0.55, 0.6, 0.65, 0.75, 0.8, or 0.85 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0042] In certain embodiments, the subject achieves a PSA nadir of < 0.2 ng / mL (e.g., undetectable).

[0043] In another aspect, the disclosure provides a method of reducing at least one symptom associated with prostate cancer. The method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy, thereby reducing the at least one symptom associated with prostate cancer. Depending upon the circumstances, the method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec and an effective amount of radiation therapy.

[0044] Depending upon the circumstances, the method results in a reduction in tumor volume, thereby reducing the at least one symptom associated with prostate cancer. For example, at least one symptom associated with prostate cancer is selected from pollakiuria, micturition urgency, and urinary tract pain.

[0045] In certain embodiments, the subject is newly -diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).Attorney Docket No.: CAND-020WO

[0046] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0047] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0048] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days.

[0049] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0050] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT.

[0051] In another aspect, the disclosure provides a method of treating a subject with prostate cancer in need thereof. The method includes administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy. The method results in an increased likelihood that the subject achieves a pathological complete response (pCR) as compared to a subject with prostate cancer receiving only the radiation therapy (e.g., an equivalent or same level of the radiation therapy received by the subject).

[0052] In certain embodiments, the subject is newly-diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0053] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection isAttorney Docket No.: CAND-020WO administered 2-8 weeks after the first injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0054] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0055] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days.

[0056] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0057] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT.

[0058] In certain embodiments, the subject achieves a PSA nadir of < 0.2 ng / mL (e.g., undetectable).

[0059] In another aspect, the disclosure provides a method of increasing time to metastasis, biochemical failure, treatment failure, or new treatment in a subject with intermediate-risk prostate cancer. The method comprises administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy, so that the subject’s time to metastasis, biochemical failure, treatment failure, or new treatment is increased as compared to a subject with intermediaterisk prostate cancer receiving only the radiation therapy (e.g., an equivalent or same level of the radiation therapy received by the subject).

[0060] In certain embodiments, the subject is newly -diagnosed with intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediaterisk prostate cancer). In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.Attorney Docket No.: CAND-020WO

[0061] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0062] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days.

[0063] In certain embodiments, radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0064] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT.

[0065] In another aspect, the disclosure provides a method of treating a subject with prostate cancer in need thereof. The method includes administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy. The method results in an increased likelihood that the subject achieves a pCR as compared to a subject with prostate cancer receiving only radiation therapy.

[0066] In certain embodiments, the subject is newly-diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0067] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0068] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0069] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, theAttorney Docket No.: CAND-020WO prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days.

[0070] In certain embodiments, radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0071] In certain embodiments, the method increases the subject’s time to metastasis to at least 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0072] In certain embodiments, the method increases the subject’s time to biochemical failure to at least 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0073] In certain embodiments, the method increases the subject’s time to treatment failure to at least 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0074] In certain embodiments, the method increases the subject’s time to new treatment to at least 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0075] In certain embodiments, the method increases the subject’s probability of disease- free survival to at least about 0.75 for at least 2 years, 3, years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0076] In certain embodiments, the method increases the subject’s probability of prostatespecific disease-free survival to at least about 0.75 for at least 2 years, least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0077] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT.

[0078] In certain embodiments, the subject achieves a PSA nadir of < 0.2 ng / mL (e.g., undetectable).Attorney Docket No.: CAND-020WO

[0079] These and other aspects and features of the invention are described in the following detailed description and claims.BRIEF DESCRIPTION OF THE DRAWINGS

[0080] The invention can be more completely understood with reference to the following drawings.

[0081] FIG. 1 is a schematic representation of a randomized, double-blind, placebo- controlled, multicenter phase 3 clinical trial of CAN-2409 (aglatimagene besadenovec) + valacyclovir (prodrug) in combination with radiation therapy in patients with newly- diagnosed intermediate- to high-risk prostate cancer. N refers to sample size. Abbreviations: AE, adverse events; ITT, intended-to-treat population.

[0082] FIG. 2 is a graph showing the probability of disease-free survival in the CAN- 2409 + valacyclovir (prodrug) vs. placebo + valacyclovir (prodrug) arms. Abbreviation: CI, confidence interval.

[0083] FIG. 3 is a graph showing the hazard ratio and 95% confidence interval (CI) of the probability of disease-free survival in the phase 3 clinical trial described in FIG. 1, subdivided by sub-group with or without androgen deprivation therapy (ADT) and in subpopulations with intermediate- to high-risk prostate cancer. N refers to sample size. Abbreviations: ITT, intended-to-treat population; NCCN, National Comprehensive Cancer Network.

[0084] FIG. 4 is a graph showing the probability of prostate-specific disease-free survival in the CAN-2409 + valacyclovir (prodrug) vs. placebo + valacyclovir (prodrug) arms. Abbreviation: CI, confidence interval.

[0085] FIG. 5 is a graph showing the probability of disease-free survival in the CAN- 2409 + valacyclovir (prodrug) without ADT Arm vs. placebo + valacyclovir (prodrug) Arm + ADT Arm.

[0086] FIG. 6 is a graph showing the probability of disease-free survival in the CAN- 2409 + valacyclovir (prodrug) vs. placebo + valacyclovir (prodrug) amis in subjects with intermediate-risk. Related data is provided in tables accompanying the graph. Abbreviation: CI, confidence interval.

[0087] FIG. 7 is a graph showing the probability of prostate-specific disease-free survival in the CAN-2409 + valacyclovir (prodrug) vs. placebo + valacyclovir (prodrug) anus inAttorney Docket No.: CAND-020WO subjects with intermediate-risk. Related data is provided in tables accompanying the graph. Abbreviation: CI, confidence interval.

[0088] FIG. 8 is a graph showing the probability of time to new therapy in the CAN- 2409 + valacyclovir (prodrug) vs. placebo + valacyclovir (prodrug) arms in subjects with intermediate-risk. Related data is provided in tables accompanying the graph. Abbreviation: CI, confidence interval.

[0089] FIG. 9 is a graph showing the probability of time to metastasis in the CAN-2409 + valacyclovir (prodrug) vs. placebo + valacyclovir (prodrug) arms in subjects with intermediate-risk. Related data is provided in tables accompanying the graph. Abbreviation: CI, confidence interval.

[0090] FIG. 10 is a graph showing the probability of time to biochemical failure in the CAN-2409 + valacyclovir (prodrug) vs. placebo + valacyclovir (prodrug) arms in subjects with intermediate-risk. Related data is provided in tables accompanying the graph. Abbreviation: CI, confidence interval.

[0091] FIG. 11 is a graph showing the probability of time to all-treatment failure in the CAN-2409 + valacyclovir (prodrug) vs. placebo + valacyclovir (prodrug) arms in subjects with intermediate-risk. Related data is provided in a table accompanying the graph. Abbreviation: CI, confidence interval.DETAILED DESCRIPTIONI. Definitions

[0092] Unless defined otherwise, technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs. For example, nomenclatures utilized in connection with, and techniques of, e.g., molecular and immunology, etc. described herein are those well-known and commonly used in the art.

[0093] As used herein, the singular forms “a,” “an,” and “the” include plural referents unless context clearly dictates otherwise.

[0094] The use of any and all examples, or exemplary language herein, for example, “such as” or “including,” is intended merely to illustrate better the present disclosure and does not pose a limitation on the scope of any invention unless claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of any invention disclosed herein.Attorney Docket No.: CAND-020WO

[0095] It should be understood that the expression “at least one of’ includes individually each of the recited objects after the expression and the various combinations of two or more of the recited objects unless otherwise understood from the context and use.

[0096] The use of the term “include,” “includes,” “including,” “have,” “has,” “having,” “contain,” “contains,” or “containing,” including grammatical equivalents thereof, should be understood generally as open-ended and non-limiting, for example, not excluding additional unrecited elements or steps, unless otherwise specifically stated or understood from the context.

[0097] As used herein, the expression “and / or” in connection with two or more recited objects includes individually each of the recited objects and the various combinations of two or more of the recited objects, unless otherwise understood from the context and use.

[0098] Where the use of the term “about” is before a quantitative value, the present invention also includes the specific quantitative value itself, unless specifically stated otherwise. As used herein, the term “about” refers to a ± 10% variation from the nominal value unless otherwise indicated or inferred.

[0099] The term “determine,” “determined,” or “determining” includes any means of detecting, including direct and indirect detection. Depending upon the circumstances, the foregoing terms also include receiving results from a test or assessment.

[0100] The term “gene” encompasses both the regions coding a gene product as well as regulatory regions for that gene, such as a promoter or enhancer, unless otherwise indicated.

[0101] The term “virus,” as used herein, can refer to a virus that can be used therapeutically, such an adenovirus (e.g., an Ad5 adenovirus). Any instance of the term “virus” herein can include aglatimagene besadenovec.

[0102] The terms “patient” and “subject” are used interchangeably herein and may include any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, non-human primates, and humans. The compositions of the disclosure can be administered to a mammal, such as a human, but can also be administered to other mammals such as an animal in need of veterinary treatment, e.g., domestic animals (e.g., dogs, cats, and the like), farm animals (e.g., cows, sheep, pigs, horses, and the like) and laboratory animals (e.g., rats, mice, guinea pigs, non-human primates, and the like).

[0103] As used herein, the terms “administering” and “administration” refer to any method of providing an agent to the subject. Such methods are known to those skilled in the art, and include, but are not limited to, intraprostatic administration, intratumoral administration, oralAttorney Docket No.: CAND-020WO administration, transdermal administration, administration by inhalation, nasal administration, topical administration, intravaginal administration, ophthalmic administration, intra-aural administration, intracerebral administration, administration to spinal cord, administration to intracerebral fluid, rectal administration, parenteral administration, intravenous administration, intra-arterial administration, intramuscular administration, intrathecal administration, systemic administration, and subcutaneous administration. Administration can be continuous or intermittent. In some instances a virus described herein can be administered therapeutically or prophylactically, such as administered for treatment of a disease or condition (e.g., a prostate cancer) in a subject or for improvement of one or more functions in a subject.

[0104] The term “therapeutically effective amount” as used herein refers to the amount of an active agent (e.g., a virus according to the present disclosure) sufficient to effect beneficial or desired results in a subject. A therapeutically effective amount can be an amount of an active agent to treat a tumor (e.g., a prostate tumor) in a subject in need thereof. A therapeutically effective amount can be an amount of an active agent sufficient to effect treatment of a tumor, e.g., amounts that are effective to reduce a targeted tumor size or load, or otherwise hinder the growth rate of tumor cells. More particularly, such terms refer to amounts of the virus that is effective, at the necessary dosages and periods of treatment, to achieve a desired result. For example, in the context of treating a cancer, an effective amount of the compositions described herein is an amount that induces remission, reduces tumor burden, and / or prevents or slows tumor spread or growth of the cancer. Effective amounts may vary according to factors such as the subject’s disease state, age, gender, and weight, as well as the pharmaceutical formulation, the route of administration, and the like, but can nevertheless be routinely determined by one skilled in the art. A therapeutically effective amount is not intended to be limited to a particular formulation or administration route.

[0105] As used herein, “treat,” “treating,” and “treatment” refer to the treatment of a disease, disorder, or symptom or manifestation of such (e.g., prostate cancer) in a subject, e.g., in a human. This includes: (a) preventing a disease or disorder, (b) inhibiting the disease, disorder, etc., i.e., slowing or arresting its progress or development; and (b) relieving the disease, disorder, etc., i.e., causing regression of the disease state.

[0106] Throughout the description, where compositions are described as having, including, or comprising specific components, or where processes and methods are described as having, including, or comprising specific steps, it is contemplated that, additionally, there areAttorney Docket No.: CAND-020WO compositions of the present disclosure that consist essentially of, or consist of, the recited components, and that there are processes and methods according to the present disclosure that consist essentially of, or consist of, the recited processing steps. Similarly, throughout the description, where compositions are described as consisting essentially of specific components, or where processes and methods are described as consisting essentially of specific steps, it is contemplated that, additionally, there are compositions of the present disclosure that consist of the recited components, and that there are processes and methods according to the present disclosure that consist of the recited processing steps.

[0107] Throughout the disclosure, where an element or component is said to be included in and / or selected from a list of recited elements or components, it should be understood that the element or component can be any one of the recited elements or components, or the element or component can be selected from a group consisting of two or more of the recited elements or components.

[0108] Further, it should be understood that elements and / or features of a composition or a method described herein can be combined in a variety of ways without departing from the spirit and scope of the present disclosure, whether explicit or implicit herein. For example, where reference is made to a particular compound, that compound can be used in various embodiments of compositions of the present disclosure and / or in methods of the present disclosure, unless otherwise understood from the context. In other words, within this application, embodiments have been described and depicted in a way that enables a clear and concise application to be written and drawn, but it is intended and will be appreciated that embodiments may be variously combined or separated without parting from the present teachings and any invention provided herein. For example, it will be appreciated that all features described and depicted herein can be applicable to all aspects of any invention described.

[0109] It should be understood that the order of steps or order for performing certain actions is immaterial so long as disclosed invention(s) remain operable. Moreover, two or more steps or actions may be conducted simultaneously.

[0110] As used herein, all numerical values or numerical ranges include whole integers within or encompassing such ranges and fractions of the values or the integers within or encompassing ranges unless the context clearly indicates otherwise. Thus, for example, reference to a range of 90%-100%, includes 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.Attorney Docket No.: CAND-020WOIL Aglatimagene Besadenovec

[0111] Aglatimagene besadenovec (also referred to as CAN-2409) is an adenoviral replication-defective engineered gene construct encoding the thymidine kinase gene derived from herpes simplex virus (HSV). See, e.g., Chen et al. (1994) PROC. NATL. ACAD. Set. USA 91 :3054-2057; Trask et al. (2000) MOLECULAR THERAPY 1(2): 195-203. Aglatimagene besadenovec can be injected directly into tumor or target tissue. It is contemplated that localized injection (e.g., intraprostatic) can minimize the chance of development of anti-drug antibodies and systemic toxicities associated with systemic administration.

[0112] The adenoviral construct serves as a vector to transport the HSV-thymidine kinase gene into tumor cells at the site of injection. These tumor cells can then express HSV- thymidine kinase, which converts orally administered anti-herpes prodrugs, such as valacyclovir, ganciclovir, acyclovir, penciclovir, valganciclovir, and famciclovir, into a toxic nucleotide analogue, which blocks DNA synthesis in dividing cells. Cells exposed to the toxic nucleotide analogue in the tumor microenvironment have been observed to undergo immunogenic cell death. At the same time, the adenoviral serotype 5 capsid protein elicits a strong pro-inflammatory signal in the tumor microenvironment. This induces a CD8-positive T cell-mediated response against the injected tumor and uninjected distant metastases for broad and systemic anti-tumor activity.III. Pharmaceutical Compositions, Medicaments, and Routes of Administration

[0113] For therapeutic use, a virus described herein is preferably combined with a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers include buffers, carriers, and excipients suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable carriers include any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water, emulsions (e.g., such as an oil / water or water / oil emulsions), and various types of wetting agents.

[0114] The compositions also can include stabilizers and preservatives. For examples of carriers, stabilizers and adjuvants, see, e.g., Adejare (2020) REMINGTON: THE SCIENCE AND PRACTICE OF PHARMACY. 23rd ed. Pharmaceutically acceptable earners include buffers, solvents, dispersion media, coatings, isotonic and absorption delaying agents, and the like,Attorney Docket No.: CAND-020WO that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is known in the art.

[0115] In some embodiments, a pharmaceutical composition may contain formulation materials for modifying, maintaining or preserving, for example, the pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, rate of dissolution or release, adsorption, or penetration of the composition. In such embodiments, suitable formulation materials include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine, or lysine); antimicrobials; antioxidants (such as ascorbic acid, sodium sulfite, or sodium hydrogen-sulfite); buffers (such as borate, bicarbonate, Tris-HCl, citrates, phosphates, or other organic acids); bulking agents (such as mannitol or glycine); chelating agents (such as ethylenediamine tetraacetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin, or hydroxypropyl-beta-cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as glucose, mannose, or dextrins); proteins (such as serum albumin, gelatin, or immunoglobulins); coloring, flavoring and diluting agents; emulsifying agents; hydrophilic polymers (such as polyvinylpyrrolidone); low molecular weight polypeptides; salt-forming counterions (such as sodium); preservatives (such as benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid, or hydrogen peroxide); solvents (such as glycerin, propylene glycol, or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as pluronics, PEG, sorbitan esters, polysorbates such as polysorbate 20, polysorbate, triton, tromethamine, lecithin, cholesterol, or tyloxapal); stability enhancing agents (such as sucrose or sorbitol); tonicity enhancing agents (such as alkali metal halides, preferably sodium or potassium chloride, mannitol, or sorbitol); delivery vehicles; diluents; excipients; and / or pharmaceutical adjuvants see, Gennaro (1990) REMINGTON’S PHARMACEUTICAL SCIENCES, 18thed. (Mack Publishing Company).

[0116] In some embodiments, a pharmaceutical composition may contain nanoparticles, e.g., polymeric nanoparticles, liposomes, or micelles {see, Anselmo el al. (2016) BIOENG. TRANSL. MED. 1: 10-29).

[0117] In some embodiments, a pharmaceutical composition may contain a sustained- or controlled-delivery formulation. Techniques for formulating sustained- or controlled- delivery means, such as liposome carriers, bio-erodible microparticles or porous beads and depot injections, are also known to those skilled in the art. Sustained-release preparationsAttorney Docket No.: CAND-020WO may include, e.g., porous polymeric microparticles or semipermeable polymer matrices in the form of shaped articles, e.g., films, or microcapsules. Sustained release matrices may include polyesters, hydrogels, polylactides, copolymers of L-glutamic acid and gamma ethyl-L- glutamate, poly (2-hydroxyethyl-inethacrylate), ethylene vinyl acetate, or poly-D(-)-3- hydroxybutyric acid. Sustained release compositions may also include liposomes that can be prepared by any of several methods known in the art.

[0118] Pharmaceutical compositions containing a virus disclosed herein can be presented in a dosage unit form and can be prepared by any suitable method. A pharmaceutical composition should be formulated to be compatible with its intended route of administration. Examples of routes of administration are intraprostatic, intratumoral, intravenous (IV), intradermal, inhalation, transdermal, topical, transmucosal, intrathecal, rectal administration, oral, buccal, intranasal, intradermal, intrathecal, intracisternal, and intralesional. One of skill in the art would readily appreciate that the various routes of administration described herein would allow for the viruses or compositions to be delivered on, in, or near the tumor or targeted cancer cells. One of skill in the art would also readily appreciate that various routes of administration described herein will allow for the vectors and compositions described herein to be delivered to a region in the vicinity of the tumor or individual cells to be treated. “In the vicinity” can include any tissue or bodily fluid in the subject that is in sufficiently close proximity to the tumor or individual cancer cells such that at least a portion of the vectors or compositions administered to the subject reach their intended targets and exert their therapeutic effects. A virus can be administered to a site of surgical resection in a subject by, for example, injection directly into the surgical bed after resection of a primary tumor, either before or after closing of the surgical site. Alternatively, as is discussed above, the viruses can be injected directly into the prostate, tumors, or lymph nodes.

[0119] Useful formulations can be prepared by methods known in the pharmaceutical art. For example, see Adejare (2020) REMINGTON: THE SCIENCE AND PRACTICE OF PHARMACY. 23rd ed. Formulation components suitable for parenteral administration include a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol, or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as EDTA; buffers such as acetates, citrates, or phosphates; and agents for the adjustment of tonicity such as sodium chloride or dextrose.Attorney Docket No.: CAND-020WO

[0120] Pharmaceutical formulations preferably are sterile. Sterilization can be accomplished by any suitable method, e.g., filtration through sterile filtration membranes. Where the composition is lyophilized, filter sterilization can be conducted prior to or following lyophilization and reconstitution.

[0121] It is to be understood that the pharmaceutical compositions described herein can be included in a container, pack, or dispenser together with instructions for administration.IV. Methods of Treating Prostate Cancer with a Virus Such as Aglatimagene Besadenovec

[0122] The majority of newly-diagnosed prostate cancers are considered localized disease. Treatment options for local disease include radiation therapy (RT) and radical prostatectomy (RP). Radiation therapy may be delivered as external beam radiation therapy (EBRT), brachytherapy (radioactive seed implantation), or a combination of both. For patients with higher risk prostate cancer, the addition of androgen deprivation therapy (ADT) to primary radiation has been demonstrated to improve disease-free survival and overall survival. However, even with ADT, 20-50% of these patients suffer recurrence within 5 years and this rate is expected to rise with longer term follow up, as recurrences are known to continue 5-15 years after treatment.

[0123] Patients with localized prostate cancer are divided into prognostic groups based on pretreatment factors, which predict future risk of suffering morbidity or mortality as a result of recurrence. The three main factors are expression of pretreatment prostate-specific antigen (PSA), as well as the combined Gleason score (GS) and clinical stage (T stage). Various commonly used systems divide patients into low, intermediate, and high-risk categories using these three factors, with slight variations in the definitions used. An example from the National Comprehensive Cancer Network (NCCN) guidelines (Scherr et al. 2003) (www.nccn.org) is as follows:Low risk: PSA < 10 ng / mL, Gleason score (GS) < 6, and Tl-T2a*;Intermediate-risk: at least 1 of the following: PSA 10-20 ng / mL, GS = 7, or T2b-2c*; Favorable intermediate-risk: having less than 50% positive biopsy cores and a single intermediate-risk feature;Unfavorable intermediate-risk: having greater than 50% positive biopsy cores and two or more intermediate-risk features;High-risk: At least 1 of the following: PSA > 20 ng / mL, GS > 7, or T3a; andAttorney Docket No.: CAND-020WOLocally advanced / very high: T3b-T4.*2002 American Joint Committee on Cancer (AJCC) TNM classification: T2a: half of 1 lobe, T2b: 1 lobe, T2c: both lobes of prostate. TNM classification relates to cancer staging based on local tumor extend (T stage), dissemination to the regional lymph nodes (N stage), and metastatic spread to distant sites (M stage) (TNM staging).

[0124] Efforts to improve the outcome of primary therapy in prostate cancer have mostly focused on manipulating the current standard therapies, by increasing dose or approach, while minimizing the side effects. For radiation therapy, advances have been made through better delivery methods that allow increasing the radiation dose without a significant increase in side effects and by the use of adjuvant ADT. Multiple groups have evaluated radiation dose escalation leading to the employment of higher doses in many treatment centers. For example, a study of 1,325 patients at 9 institutions, with a median follow up of 5.7 years, revealed an improvement in freedom from failure (FFF), from 68.4% at < 72 Gray (the unit of absorbed radiation dose; “Gy”) to 75.6% for > 72 Gy. Separated by risk groups, for the patients treated with > 72Gy, the FFF rate was 79% for low risk, 72% for intermediate-risk, and 46% for high-risk.

[0125] Regardless of the patient risk stratification, the potential benefits of higher radiation doses are tempered by the risk of increased toxicity; and even at higher radiation doses, the recurrence rate is still substantial. Newer radiation therapy techniques, such as intensity modulated radiation therapy (IMRT), have made it possible to deliver higher doses while limiting the risk of increased toxicity. An alternative radiation therapy approach using proton beams has the potential to reduce exposure of structures outside the tumor target even further. However, for prostate cancer, this approach has not yet proven effective in improving the outcome.

[0126] In addition to dose increases, the addition of ADT to radiation therapy has also been evaluated, though several studies have raised concern about toxicity of ADT outweighing the benefit especially in men with lower risk of death from prostate cancer and in men with moderate-to-severe co-morbidities, of which most common was cardiovascular disease.

[0127] Based on these studies, currently many high-risk patients are treated with ADT in combination with radiation therapy, although the appropriate duration of ADT is still debated. The benefit of adjuvant ADT for intermediate-risk disease is still unclear. Nonetheless, most clinicians are treating these patients with short term ADT for 4-6 months starting 2 monthsAttorney Docket No.: CAND-020WO before and then concomitant with EBRT (NCCN guidelines for Prostate Cancer 2009 at www.nccn.org). As described herein, ADT include but are not limited to goserelin, leuprolide, triptorelin, degarelix, bicalutamide, cyproterone, flutamide, nilutamide, abiraterone, and enzalutamide.

[0128] Despite the use of optimized combined modality therapy for prostate cancer, the rate of recurrence and the incidence of toxicity is still substantial, highlighting the need for continued exploration of additional therapies.

[0129] Accordingly, in one aspect, the disclosure provides a method of increasing disease-free survival in a subject with prostate cancer in need thereof. The method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy, so that the subject’s disease-free survival is increased as compared to a subject with prostate cancer receiving only the radiation therapy (e.g., an equivalent or same level of the radiation therapy received by the subject). Depending upon the circumstances, the method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec and a therapeutically effective amount of a radiation therapy.

[0130] In certain embodiments, the subject is newly-diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer). In some embodiments, a patient who is newly diagnosed with prostate cancer has not received treatment for prostate cancer.

[0131] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the second injection is administered 2-7 weeks after the first injection. In certain embodiments, the third injection is administered 2-3 weeks after the second injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0132] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0133] In certain embodiments, a prodrug is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug is administeredAttorney Docket No.: CAND-020WOI day after administration of the aglatimagene besadenovec. For example, the prodrug is administered daily for 14 days. In certain embodiments, the prodrug is valacyclovir.

[0134] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0135] Depending upon the circumstances, the method increases the subject’s disease- free survival to at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years,I I years, or 12 years (e.g., 3-12 years, 5-10, or 7-8 years) following the initial administration of the aglatimagene besadenovec. For example, the method increases the subject’s probability of disease-free survival to at least about 0.55, 0.6, 0.65, 0.75, 0.8, or 0.85 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec. In certain embodiments, the method increases the subject’s probability of prostate-specific disease-free survival to at least about 0.75 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0136] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT (e.g., because of the side effect profile of ADT, because ADT is contraindicated, due to a lack of tolerance, or by personal choice).

[0137] In certain embodiments, the subject achieves a (PSA nadir of < 0.2 ng / mL (e.g., undetectable).

[0138] In another aspect, the disclosure provides a method of reducing a PSA level to a nadir of < 0.2 ng / mL in a subject with intermediate- to high-risk prostate cancer in need thereof. The method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy. Depending upon the circumstances, the method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec and a therapeutically effective amount of a radiation therapy.

[0139] In certain embodiments, the subject is newly-diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certainAttorney Docket No.: CAND-020WO embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0140] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the second injection is administered 2-7 weeks after the first injection. In certain embodiments, the third injection is administered 2-3 weeks after the second injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0141] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0142] In certain embodiments, a prodrug is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug is administeredI day after administration of the aglatimagene besadenovec. For example, the prodrug is administered daily for 14 days. In certain embodiments, the prodrug is valacyclovir.

[0143] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0144] Depending upon the circumstances, the method increases the subject’s disease- free survival to at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years,I I years, or 12 years (e.g., 3-12 years, 5-10, or 7-8 years) following the initial administration of the aglatimagene besadenovec. For example, the method increases the subject’s probability of disease-free survival to at least about 0.55, 0.6, 0.65, 0.75, 0.8, or 0.85 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 1 1 years, or 12 years following the initial administration of the aglatimagene besadenovec. In certain embodiments, the method increases the subject's probability of prostate-specific disease-free survival to at least about 0.75 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0145] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments,Attorney Docket No.: CAND-020WO the subject does not receive ADT (e.g., because of the side effect profile of ADT, because ADT is contraindicated, due to a lack of tolerance, or by personal choice).

[0146] In another aspect, the disclosure provides a method of treating a subject with prostate cancer who does not receive ADT. The method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy. Depending upon the circumstances, the method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec and radiation therapy.

[0147] In certain embodiments, the subject is newly -diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0148] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the second injection is administered 2-7 weeks after the first injection. In certain embodiments, the third injection is administered 2-3 weeks after the second injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0149] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0150] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days. In certain embodiments, the prodrug is valacyclovir.

[0151] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0152] Depending upon the circumstances, the method increases the subject’s disease- free survival to at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years,Attorney Docket No.: CAND-020WO11 years, or 12 years (e.g., 3-12 years, 5-10, or 7-8 years) following the initial administration of the aglatimagene besadenovec. For example, the method increases the subject’s probability of disease-free survival to at least about 0.55, 0.6, 0.65, 0.75, 0.8, or 0.85 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 1 1 years, or 12 years following the initial administration of the aglatimagene besadenovec. In certain embodiments, the method increases the subject's probability of prostate-specific disease-free survival to at least about 0.75 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0153] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT (e.g., because of the side effect profile of ADT, because ADT is contraindicated, due to a lack of tolerance, or by personal choice).

[0154] In certain embodiments, the subject achieves a PSA nadir of < 0.2 ng / mL (e.g., undetectable).

[0155] In another aspect, the disclosure provides a method of reducing a PSA level to a nadir of < 0.2 ng / mL in a subject with intermediate- to high-risk prostate cancer in need thereof. The method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy. Depending upon the circumstances, the method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec and radiation therapy.

[0156] In certain embodiments, the subject is newly-diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0157] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the second injection is administered 2-7 weeks after the first injection. In certain embodiments, the third injection is administered 2-3 weeks after the second injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.Attorney Docket No.: CAND-020WO

[0158] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0159] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days.

[0160] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0161] In certain embodiments, the method increases the subject’s disease-free survival to at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years (e.g., 3-12 years, 5-10, or 7-8 years) following administration of the aglatimagene besadenovec. For example, the method increases the subject’s probability of disease-free survival to at least about 0.55, 0.6, 0.65, 0.75, 0.8, or 0.85 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec. In certain embodiments, the method increases the subject's probability of prostate-specific disease-free survival to at least about 0.75 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

[0162] In certain embodiments, the subject achieves a PSA nadir of < 0.2 ng / mL (e.g., undetectable).

[0163] In another aspect, the disclosure provides a method of reducing at least one symptom associated with prostate cancer. The method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy, thereby reducing the at least one symptom associated with prostate cancer. Depending upon the circumstances, the method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec and radiation therapy.

[0164] Depending upon the circumstances, the method results in a reduction in tumor volume, thereby reducing the at least one symptom associated with prostate cancer. ForAttorney Docket No.: CAND-020WO example, at least one symptom associated with prostate cancer is selected from pollakiuria, micturition urgency, and urinary tract pain.

[0165] In certain embodiments, the subject is newly-diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0166] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the second injection is administered 2-7 weeks after the first injection. In certain embodiments, the third injection is administered 2-3 weeks after the second injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0167] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0168] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days.

[0169] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0170] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT (e.g., because of the side effect profile of ADT, because ADT is contraindicated, due to a lack of tolerance, or by personal choice).

[0171] In another aspect, the disclosure provides a method of treating a subject with prostate cancer in need thereof. The method includes administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy. The method results in an increased likelihood that the subject achieves a pathological complete response (pCR) as compared to a subject with prostateAttorney Docket No.: CAND-020WO cancer receiving only the radiation therapy (e.g., an equivalent or same level of the radiation therapy received by the subject).

[0172] In certain embodiments, the subject is newly-diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certain embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0173] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0174] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0175] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days. In certain embodiments, the prodrug is valacyclovir.

[0176] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0177] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT (e.g., because of the side effect profile of ADT, because ADT is contraindicated, due to a lack of tolerance, or by personal choice).

[0178] In certain embodiments, the subject achieves a PSA nadir of < 0.2 ng / mL (e.g., undetectable).

[0179] In another aspect, the disclosure provides a method of increasing time to metastasis, biochemical failure, treatment failure, or new treatment in a subject with intermediate-risk prostate cancer. The method can include administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject alsoAttorney Docket No.: CAND-020WO receives radiation therapy, so that the subject’s time to metastasis, biochemical failure, treatment failure, or new treatment is increased as compared to a subject with prostate cancer receiving only the radiation therapy (e.g., an equivalent or same level of the radiation therapy received by the subject).

[0180] In certain embodiments, the subject is newly-diagnosed with intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediaterisk prostate cancer). In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0181] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0182] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug (e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days.

[0183] In certain embodiments, radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0184] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT.

[0185] In another aspect, the disclosure provides a method of treating a subject with prostate cancer in need thereof. The method includes administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy. The method results in an increased likelihood that the subject achieves a pCR as compared to a subject with prostate cancer receiving only radiation therapy.

[0186] In certain embodiments, the subject is newly -diagnosed with the prostate cancer. In certain embodiments, the subject has intermediate- to high-risk prostate cancer. In certainAttorney Docket No.: CAND-020WO embodiments, the subject has intermediate-risk prostate cancer. In certain embodiments, the subject has favorable- or unfavorable-risk prostate cancer (e.g., favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer).

[0187] In certain embodiments, the aglatimagene besadenovec is administered as a first, second, and third injection. Depending upon the circumstances, the second injection is administered 2-8 weeks after the first injection. In certain embodiments, the subject receives no more than three injections of aglatimagene besadenovec.

[0188] In certain embodiments, the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

[0189] In certain embodiments, a prodrug (e.g., valacyclovir) is administered following administration of aglatimagene besadenovec. Depending upon the circumstances, the prodrug e.g., valacyclovir) is administered 1 day after administration of the aglatimagene besadenovec. For example, the prodrug (e.g., valacyclovir) is administered daily for 14 days.

[0190] In certain embodiments, the radiation therapy begins 0-3 days after the second injection. Depending upon the circumstances, the radiation therapy begins on the same day as the second injection. For example, the subject receives 78 Gy of radiation in 2 Gy fractions. In certain embodiments, the subject receives hypofractionated radiation. For example, the hypofractionated radiation includes 60 Gy in 3 Gy fractions.

[0191] In certain embodiments, the method increases the subject's time to metastasis to at least 10 years, 11 years, 12 years, 13 years, 14 years, or 15 years following the initial administration of the aglatimagene besadenovec.

[0192] In certain embodiments, the method increases the subject’s time to biochemical failure to at least 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years (e.g., 4-12 years, 5-10, or 7-8 years) following the initial administration of the aglatimagene besadenovec.

[0193] In certain embodiments, the method increases the subject’s time to treatment failure to at least 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years (e.g., 2-12 years, 5-10, or 7-8 years) following the initial administration of the aglatimagene besadenovec.

[0194] In certain embodiments, the method increases the subject’s time to new treatment to at least 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years (e.g., 2-12 years, 5-10, or 7-8 years) following the initial administration of the aglatimagene besadenovec.Attorney Docket No.: CAND-020WO

[0195] In certain embodiments, the method increases the subject’s probability of disease- free survival to at least about 0.75 for at least 2 years, 3, years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 (e.g., 2-12 years, 5-10, or 7-8 years) years following the initial administration of the aglatimagene besadenovec.

[0196] In certain embodiments, the method increases the subject’s probability of prostatespecific disease-free survival to at least about 0.75 for at least 2 years, least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 (e.g., 2-12 years, 5-10, or 7- 8 years) years following the initial administration of the aglatimagene besadenovec.

[0197] Depending upon the circumstances, the subject also receives ADT. In certain embodiments, the subject receives no more than 6 months of ADT. In certain embodiments, the subject does not receive ADT.

[0198] In certain embodiments, the subject achieves a PSA nadir of < 0.2 ng / mL (e.g., undetectable).

[0199] The viruses described herein can be administered intraprostatically (e.g., to one or more lobes of the prostate or to all four lobes of the prostate), intratumorally, intravenously, intradermally, transdermally, transmucosally, intrathecally, rectally, orally, or buccally. One of skill in the art would readily appreciate that the various routes of administration described herein would allow for the virus to be delivered on, in, or near the tumor or targeted cancer cells. One of skill in the art would also readily appreciate that various routes of administration described herein will allow for the virus described herein to be delivered to a region in the vicinity of the tumor or individual cells to be treated. “In the vicinity” can include any tissue or bodily fluid in the subject that is in sufficiently close proximity to the tumor or individual cancer cells such that at least a portion of the virus administered to the subject reach their intended targets and exert their therapeutic effects. A virus can be administered to a site of surgical resection in a subject by, for example, injection directly into the surgical bed after resection of a primary tumor, either before or after closing of the surgical site. Alternatively, as discussed in the Examples herein, the viruses can be injected directly into the prostate or into preselected quadrants of the prostate.

[0200] The amount of virus administered will depend on variables such as the type and extent of disease or indication to be treated, the overall health of the subject, the in vivo potency of the active component, the pharmaceutical formulation, and the route of administration. The initial dosage can be increased beyond the upper level in order to rapidly achieve the desired blood-level or tissue-level. Alternatively, the initial dosage can beAttorney Docket No.: CAND-020WO smaller than the optimum, and the daily dosage may be progressively increased during the course of treatment. Human dosage can be optimized, e.g., in a conventional Phase I dose escalation study. Dosing frequency can vary, depending on factors such as route of administration, dosage amount, and the disease being treated. In certain embodiments, the virus (e.g., aglatimagene besadenovec) is administered in an amount between about 106PFU and about IO10PFU per subject. In certain embodiments, the virus is administered in an amount between about 5 x 109and about 5 x 1013viral particles (vp) per subject. In certain embodiments, the vims is administered in an amount between about 5 x IO10and about 5 x 1012vp per subject. In certain embodiments, the vims is administered in an amount of about 5 x 1011vp per subject. In certain embodiments, three doses of vims are administered, wherein administration of each dose is separated by at least 2-6 weeks. In certain embodiments, administration of a vims (e.g., aglatimagene besadenovec) is followed by administration of an effective amount of a substrate for HSV-TK (e.g., valacyclovir). In certain embodiments, the substrate for HSV-TK is administered at a dose of about two grams, three times a day for 14 days after each administration of a vims (e.g., aglatimagene besadenovec).EXAMPLES

[0201] The disclosure is further illustrated by the following Examples. The Examples are provided for illustrative purposes only, and are not to be constmed as limiting the scope or content of the disclosure in any way.Example 1: A Randomized Controlled Trial of CAN-2409 As Adjuvant to Up-Front Radiation Therapy For Localized Prostate Cancer

[0202] The overall objective for this study was to determine if adjuvant therapy with CAN-2409 improved the clinical outcome of radiation therapy for newly-diagnosed prostate cancer.Materials and MethodsPatient SelectionInclusion Criteria

[0203] Inclusion criteria included:- Histologically confirmed adenocarcinoma of the prostate.Attorney Docket No.: CAND-020WO- Patients meeting any of the NCCN intermediate-risk criteria or subset of NCCN high- risk group that had a single high-risk feature defined as shown in TABLE 1.TABLE 1. NCCN Risk Criteria*Stage was based on the 2010 AJCC TNM clinical staging.Patients were planning and medically able to undergo standard prostate-only external beam radiation therapy and were able to tolerate multiple transrectal ultrasound guided injections.18 years of age or older.Performance status was ECOG 0-2.The following laboratory criteria were met: o SGOT (AST) < 3x upper limit of normal; o Serum creatinine < 2 mg / dL; o Calculated creatinine clearance > 30 mL / min. Cr clearance formula =[(140 - age in yr) x (weight in kg)] -? [72 x serum Cr in mg / dL]; o WBC > 3000 / mm3; and o Platelets > 100,000 / mm3.- Patients agreed to have prostate biopsy two years after end of radiation.Attorney Docket No.: CAND-020WO- Patients gave study-specific informed consent prior to enrollment.Exclusion Criteria

[0204] Exclusion criteria included:- Had active liver disease, including known cirrhosis or active hepatitis.- Patient was on systemic corticosteroids (> 10 mg prednisone per day) or other immunosuppressive drug.Known HIV-positive patients.- Regional lymph node involvement or distant metastases.- Patient planned to receive whole pelvic irradiation.- Other current malignancy (except squamous or basal cell skin cancers).Other serious co-morbid illness or compromised organ function that, in the opinion of the Investigator, would have interfered with treatment or follow-up. For example, patients with diseases that precluded radiation therapy to the prostate, such as severe prostatitis and inflammatory bowel disease.Prior treatment for prostate cancer except TURP or ADT, if ADT given for a maximum of 6 months. If prior TURP, patients were deemed able to receive multiple intraprostatic injections by the Investigator.Patients who had or planned to have orchiectomy as the form of hormonal ablation. Known sensitivity or allergic reactions to acyclovir or valacyclovir.Pretreatment Evaluation

[0205] The following evaluations were performed within the specified time prior to study enrollment:- Within 12 months, prostate biopsy with histologic evaluation and assignment of Gleason score; o When feasible, slides from the diagnostic biopsy were provided to Sponsor for central archiving.- Within 6 months and prior to ADT start, prostate-specific antigen (PSA; the specific assay used was recorded on the CRF).The following evaluations were performed < 4 weeks prior to first CAN-2409 (aglatimagene besadenovec) or placebo injection:Attorney Docket No.: CAND-020WO o Clinical assessment included history and physical exam, DRE, and ECOG performance status assessment. o Quality of Life assessment by questionnaire. o CBC, differential, platelets. o Creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ASP), and bilirubin. o Optional blood collection for immunology research studies.All patients also underwent all standard of care and clinically indicated evaluations and the data was submitted to Sponsor. This included a bone scan if PSA > 20 ng / mL or Gleason > 7 or stage T3a or if patient was symptomatic. Pelvic CT or MRI were performed prior to enrollment for high-risk patients.Registration Procedures

[0206] This was a double-blind placebo-controlled trial. Clinical site personnel determined eligibility of prospective subjects and completed an Eligibility Checklist (EC) form (CRF EC) according to the Patient Registration SOP (SOP PR). The completed form EC was sent to Sponsor where a case number was assigned, randomization was performed, and blinded treatment assignments were made. This information was communicated back to the clinical site via the Patient Registration form (CRF PR).Treatment PlanOverview

[0207] Patients were randomized to test or control at a 2:1 ratio; two identical test Anns and one Control Arm:- Test Arms: CAN-2409 + radiation therapy ± ADT.Control Arm: Placebo + valacyclovir + radiation therapy ± ADT (FIG. 1).

[0208] Patients on the Test Arms received three courses of CAN-2409 each consisting of CAN-2409 injection + oral valacyclovir. Patients in the Control Arm received corresponding courses of placebo injection + oral valacyclovir. CAN-2409 or placebo injection was delivered to the prostate via transrectal ultrasound (TRUS)-guided injection as follows:1. 1st injection was at least 2 weeks prior to initiation of radiation (to allow 14- day valacyclovir course completion) and not more than 8 weeks before starting radiation.Attorney Docket No.: CAND-020WO2. 2nd injection was on the day of or up to 3 days before initiation of radiation therapy and at least 2 weeks after injection #1 (to allow 14-day valacyclovir course completion). Injection may have been on the last day of valacyclovir for the previous course.3. 3rd injection was 2-3 weeks after injection #2 (to allow 14-day valacyclovir course completion). Injection may have been on the last day of valacyclovir for the previous course.

[0209] The prodrag, valacyclovir, was administered at a fixed dose for 14 days after each CAN-2409 or placebo injection starting the day after the injection.CAN -2409 Administration

[0210] Delivery of CAN-2409 or placebo injection included:- TRUS was used to visualize the prostate and guide delivery to four sites, one apical and one basal in each lobe.- For the Test Arms, the dose of CAN-2409 was 5 x 1011vector particles (vp) total for each course. The dose was in 2 mL total volume and approximately 0.5 mL was injected into each of the four sites.For the Control Anu, the placebo solution appeared the same as the vector solution and consisted of 2 mL total volume and approximately 0.5 mL was injected into each of the four sites.- For prostate injections, the same prophylactic antibiotic and other preparations as appropriate for prostate biopsy were used.Prophylactic medications

[0211] Prophylactics included:Prophylactic treatment with acetaminophen alternating with ibuprofen given for the first 24-48 hours following injection to prevent pain, fever, and flu-like symptoms possibly associated with injections.- In addition, standard prophylactic antibiotic treatment, such as ciprofloxacin, was recommended with the biopsy and injection procedures.Attorney Docket No.: CAND-020WOValacyclovir Administration- On the day of each CAN-2409 or placebo injection, the subject was given a bottle containing the appropriate number of caplets and a diary to record each dose taken.- The subject was instructed to begin taking the caplets on the day following the CAN- 2409 or placebo injection and continued for 14 days.- The dose of valacyclovir was 2 grams orally three times per day for 14 days if the calculated creatinine clearance was greater than 50 mL / min. The dose was adjusted for renal impairment based on the calculated creatinine clearance as provided inTABLE 2TABLE 2. Valacyclovir Dosage for Calculated Creatinine Clearance

[0212] Creatinine clearance was calculated at the time of trial enrollment and recalculated if creatinine became abnormal, using the following formula:Cr clearance (mL / min) = [(140 - age in yr) x (weight in kg)] - [72 x serum Cr in mg / dL]Treatment Escape RulesIf fever occurred due to the local inflammatory and immune response to CAN-2409, then the patient was treated with anti-pyrectics, as necessary. If local inflammation was extensive or infection developed, valacyclovir was stopped and antibiotics and / or anti-inflammatory therapy was given, as appropriate.- If a patient experienced side effects related to valacyclovir, valacyclovir was stopped at the discretion of the Investigator. The Sponsor was notified.- If a patient experienced toxicity or side effects with the first or second course of treatment, subsequent courses were withheld at the discretion of the Investigator. The Sponsor was notified.Attorney Docket No.: CAND-020WORadiation TherapyTechnical Factors

[0213] Megavoltage equipment was required with effective photon energies of at least 6 MV.Localization, Simulation, and Immobilization

[0214] A urethrogram was recommended, but not required, to establish the most inferior portion of the prostate. A treatment planning CT scan was required to define tumor, clinical, and planning target volumes as well as the critical normal structures. The treatment planning CT was acquired with the patient set up in the same position as for daily treatments. The CT scan of the pelvis started at or above the iliac crest down to the perineum. All tissues that were irradiated were included in CT scan. CT scan thickness was not more than 0.5 cm through the region that contained the target volumes (i.e., from the bottom of the sacroiliac joints down to the penile urethra). The regions above and below the target volume region were scanned with slice thickness not more than 1.0 cm.

[0215] Patients who did not have pre-enrollment CT / MR1 and were found to have locally advanced disease on the radiation therapy treatment planning CT / MRI, continued on study as planned, and were evaluated as per intent to treat. The GTV, CTV, and PTV and relevant normal tissues were outlined on all CT slices in which the structures existed. Daily target localization (fiducial markers, transabdominal ultrasound or kV- or MV-CT) were required. Prostate gland immobilization with rectal balloon were allowed for those sites routinely employing such techniques. Image Guided Radiation Therapy was required if using a hypofractionation regimen.Treatment Planning / Target Volumes (The Specific Doses Listed In This Section Were For a Total of 78 Gy to The PTV In 2 Gy Fractions But Were Modified As Per Standard of Care To Accommodate The Range of Prescription Doses Allowed, Including Moderate Hypofractionation With 60 Gy to The PTV in 3 Gy Fractions)

[0216] The Gross Tumor Volume (GTV) was defined by the physician as all known disease as defined by the planning CT, urethrogram, and clinical information. The GTV for the purposes of this protocol was the prostate only. If a urethrogram was used, the GTV encompassed a volume inferiorly 10 mm superior to the tip of the dye and no less than the entire prostate. Prostate dimensions were defined as visualized on CT scan. The ClinicalAttorney Docket No.: CAND-020WOTarget Volume-1 (CTV-1) was the GTV (the prostate) plus the seminal vesicles. The Planning Target Volume-1 (PTV-1) was defined as the PTV for the initial 46 Gy. The PTV-1 provided a margin around the CTV-1 to compensate for the variability of treatment set up and internal organ motion. A minimum of 5 mm around the CTV-1 was required to define the PTV-1. Up to 10 mm was used in all directions. The Clinical Target Volume-2 (CTV-2) was the GTV (the prostate) only. The Planning Target Volume-2 (PTV-2) was defined as the second or ‘cone-down’ PTV and was treated after 46 Gy for an additional 32 Gy. The PTV-2 provided a margin around the CTV-2 to compensate for the variability of treatment set up and internal organ motion. A minimum of 5 mm around the CTV-2 was required to define the PTV-2. The posterior margin was 5-7 mm. Treatment was given only to PTV-1 and PTV-2 using IMRT to exclude as much of the bladder and rectum as possible. Field arrangements were determined by inverse-based planning to produce the optimal plan in accordance with volume definitions.Critical Normal Structures

[0217] Dose-volume histograms (DVHs) were generated for all critical normal structures.

[0218] Portions of the bladder and rectum, by necessity, received the full dose to the PTV; however, careful planning was performed to ensure that the volume of the bladder and rectum receiving the full dose was kept to a minimum.

[0219] Based upon a review of patient dosimetry, the following normal tissue guidelines were followed:Rectum (fractionation up to 2 Gy): o No more than 15% volume of the rectum received more than 75 Gy. o No more than 25% volume of the rectum received more than 70 Gy. o No more than 35% volume of the rectum received more than 65 Gy. o No more than 50% volume of the rectum received more than 60 Gy.- Rectum (moderate hypofractionation, > 2.4 Gy): o No more than 3% volume of the rectum received more than 60 Gy. o No more than 15% volume of the rectum received more than 57 Gy. o No more than 30% volume of the rectum received more than 53 Gy. o No more than 50% volume of the rectum received more than 49 Gy. o No more than 60% volume of the rectum received more than 41 Gy.- Bladder (fractionation up to 2 Gy):Attorney Docket No.: CAND-020WO o No more than 15% volume of the bladder received more than 80 Gy. o No more than 25% volume of the bladder received more than 75 Gy. o No more than 35% volume of the bladder received more than 70 Gy. o No more than 50% volume of the bladder received more than 65 Gy.- Bladder (moderate hypofractionation, > 2.4 Gy): o No more than 5% volume of the rectum received more than 60 Gy. o No more than 25% volume of the rectum received more than 49 Gy. o No more than 50% volume of the rectum received more than 41 Gy.

[0220] Care was taken not to shield the penile bulb at the expense of adequate coverage of the PTV in this study.

[0221] The normal tissue volume contoured included bladder, rectum, bilateral femora (to the level of ischial tuberosity), penile bulb, and skin. The normal tissues were contoured and considered as solid organs. The bladder was contoured from its base to the dome, and the rectum from the anus (at the level of the ischial tuberosities) for a length of 15 cm or to the rectosigmoid flexure.Prescription Dose

[0222] At least 95% of the PTV received the prescribed dose. The maximum dose to the PTV did not exceed the prescription dose by more than 7%.

[0223] Prescription dose to the PTV was 78 Gy in 2 Gy dose fractions, 60 Gy in 3 Gy dose fractions, or similar local standard (e.g., 79.2 Gy in 1.8 Gy dose fractions, 81 Gy in 1.8 Gy fractions, or 70 Gy in 2.5 Gy fractions). All fields treated once daily, 5 fractions per week.Dose Heterogeneity

[0224] Maximum dose to the PTV volume did not exceed the prescription dose by more than 7%.

[0225] The maximum point dose to critical normal structures outside the PTV did not exceed the prescription dose.

[0226] The treating physician carefully considered the tolerance dose / volume to each critical normal structure and unspecified tissue.Attorney Docket No.: CAND-020WOAndrogen Deprivation Therapy (ADT)

[0227] ADT was optional but administration was decided before enrollment to allow for stratification at the time of randomization.

[0228] Patients who chose to receive ADT received standard of care short-term ADT not exceeding 6 months. The duration of anti-androgen use was recorded on the CRF.Evaluation During and After Treatment

[0229] All patients underwent all standard of care and clinically indicated evaluations in addition to any study specific evaluations. Data was collected for these non-specified evaluations.

[0230] Clinical assessment with history and physical, including toxicity assessment, was performed prior to each CAN-2409 or placebo injection at follow up visits 1-2 weeks after each injection (which, in some instances, corresponded to the pre-injection visit at some time points) and at 3, 6, 12, 18, and 24 months after completion of radiation therapy. After this, clinical assessment of late toxicity and disease status was conducted every 6 months through year 5 after which long-term follow up of general health status was continued yearly. Longterm survival and prostate cancer-specific survival follow-up continued for at least 10 years after completion of radiation. If disease progression / recurrence occurred or new serious health problems developed, this was reported immediately to the Sponsor.

[0231] CBC, differential, platelets, creatinine, AST, ALT, ASP, and bilirubin were monitored 1-2 weeks after each CAN-2409 or placebo injection and repeated at subsequent visits if abnormal until they returned to baseline.

[0232] PSA and DRE were monitored as per standard of care at follow up visits after completion of radiation. For PSA measurements, the specific assay used was recorded on the CRF. As per current NCCN practice guidelines, the standard of care for evaluation of a positive DRE or rising PSA, defined as 2 ng / mL > nadir, required confirmation by prostate biopsy, bone-scan, or abdomen / pelvic CT / MRI. Similar confirmation was required for patients that were symptomatic. Patients with a positive bone scan did not need further evaluation but patients with local recurrence were restaged with a bone scan as well as abdomen / pelvic CT / MRI.

[0233] Quality of life (QOL) was assessed at 3, 6, 12, 18, and 24 months after completion of radiation therapyAttorney Docket No.: CAND-020WO

[0234] Prostate biopsy was performed 22-26 months after completion of radiation therapy.

[0235] Optional blood for immunology research studies (40-60 cc) was collected before treatment initiation and 1-2 weeks after each CAN-2409 or placebo injection, and at 3 and 12 months after completion of radiation.

[0236] Long-term follow-up of general health status and any side effects was continued for life. This was done by telephone contact or letter with the patient or their medical provider. Medical records were also reviewed to assess for any potential side effects and for general health status.Toxicity

[0237] Toxicity reporting was recorded.

[0238] Acute toxicity was defined as side effects occurring until the completion of radiation and was graded using the NCI Common Toxicity Criteria CTCAE ver4.0.

[0239] The Late Toxicity Monitoring period began after the Acute Toxicity Monitoring period and continued for 5 years from the end of radiation and after that, long-term follow up continued. Data was collected on new medical conditions according to categories including allergy / rheumatology, cardiovascular, endocrine, hematology / oncology, and neurologic. Genito-urinary and gastrointestinal toxicities, which are expected after prostate radiation therapy, were graded using CTCAEver4.0.Pathology

[0240] Central pathologic review: inclusion and randomization were done using the local pathology report. Central pathology review was performed retrospectively from archived samples.

[0241] Archival tissue collection: the Sponsor coordinated and implement collection and archival of pretreatment diagnostic material and post-treatment 2-year biopsy material. This consisted of the slides and / or paraffin blocks from the two biopsy encounters.

[0242] Post-treatment biopsies: all patients who had not already had declared failure had post-treatment prostate biopsy done at 24 months (± 2 months) after completion of radiation. The preferred protocol for the 2-year biopsy was to sample at least 6 areas of the prostate (base, mid, and apex on both sides). If the organ was deemed too small to physically allow for 6 cores, fewer cores were taken as long as the rationale was provided, and size of prostateAttorney Docket No.: CAND-020WO was specified. All biopsies were fixed in 10% neutral buffered formalin as soon as possible after the biopsy procedure. Regardless of the sampling technique employed, the location of the cancer in the original biopsy was included. At least one representative core or H&E slide of each core was submitted for central archive. Biopsy results were classified as negative (benign, high-grade PIN, or suspicious for adenocarcinoma) or positive (adenocarcinoma present, with or without treatment effect).

[0243] Central pathology review was performed for post-treatment biopsies and retrospective review of baseline diagnostic pathologies. Sensitivity analyses were performed to determine if imbalances of baseline characteristic deviations biased the results.Quality of Life (QOL)

[0244] Patient-reported Health Related Quality of Life outcomes were collected using the Expanded Prostate Cancer Index Composite (EPIC-26) questionnaire. Data were collected at baseline and 3, 6, 12, 18, and 24 months after completion of radiation therapy.CAN-2409

[0245] The study agent was CAN-2409, a replication-defective adenoviral vector based on the dl309 serotype 5 vims. The vector had a deletion in the El region of the adenoviral genome substituted by a herpes simplex vims thymidine kinase (HSV-tk) expression cassette. The dl309 backbone also had a deletion in the E3 region substituted by non-coding DNA. The vector for this study was prepared for clinical use under current Good Manufacturing Practice (cGMP) conditions for biologies.Study Design

[0246] This was a multicenter, randomized, placebo-controlled, double-blind study in newly-diagnosed prostate cancer. The aim of the study was to assess the efficacy of CAN- 2409 + valacyclovir in combination with the standard of care (Test Arm) compared to placebo + valacyclovir + standard of care (Control Ami), where the standard of care (SOC) was external beam radiation therapy with or without short-term ADT. Eligible patients (per inclusion / exclusion criteria) were randomly assigned onto the Test Arm or the Control Arm with a 2:1 randomization ratio. A stratification procedure was used in the randomization on the variables of NCCN risk category and short-term ADT.Attorney Docket No.: CAND-020WOPrimary Endpoints and Sample Size Calculations

[0247] The primary endpoint for the study was disease-free survival (DFS), which was evaluated on the intent-to-treat (ITT) population. Enrollment was at a rate of 10 subjects a month for the first 6 months and 30 patients a month during the next 22 months, 745 patients (about 474 subjects on the Test Arm and about 237 on the Control Arm) provided 98 events within 58 months of the first patient enrollment to the study. A total sample size of 745 subjects yielded approximately 90% power to detect a 15% relative improvement at an a level of 0.05 to be statistically significant. The sample size estimation took into account randomizing in favor of the Test Arm by a 2:1 ratio and up to 20% dropout rate. This calculation further assumed that, over 58 months of follow-up, there would be a DFS rate of 75% in the Control Arm and 86.5% in the Test Arm, or equivalently a hazard ratio of 0.5, along with a 4% dropout rate per year on each arm due to loss to follow-up. Patients were stratified by the variable of ADT (present or absent) and NCCN risk category (intermediate- or high-risk with only one high-risk feature) prior to study arm allocation.

[0248] An interim analysis after the first 49 events, giving an information fraction of 0.5, took place. The Lan-DeMets Family was chosen to formalize the error spending function for stopping the trial early to reject the null hypothesis at an a level of 0.003. There was 25% estimated probability of rejecting the null at such early stage. However, it was decided not to perform an interim analysis prior to the final analysis since the number of events necessary to support the interim analysis was reached only after all patients had already received treatment. The sample size calculations were done using East 5.2 (Cytel, Cambridge, MA).Disease-Free Survival ( DFS )

[0249] The disease-free survival (DFS) was defined as time from randomization to Endpoint Adjudication Committee (EAC) adjudicated local failure, regional failure, distant metastases, or death from any cause. The time to DFS was measured from the date of randomization to the earliest event or to the date of the most recent follow-up if no event occurred. The earliest event date was defined as the date when the event was confirmed. For example, for a positive DRE detected recurrence, the date of the event was the date of the confirmatory biopsy if medically possible, otherwise, it was the date recurrence was considered definitive by the Investigator. For a symptomatic patient, the date of confirmatory image or biopsy was considered the date of the event, even if the patient had missed followup visits. Analyses of DFS were based on the intent to treat (ITT) population. If a patientAttorney Docket No.: CAND-020WO was lost to follow-up, the data was censored at the time of last follow-up when patient was event-free. If patient refused to continue with the protocol, every attempt was made to continue follow-up and analyze as per ITT. If they refused follow-up, they were censored at the last follow-up.

[0250] Patients who did not have pre-enrollment CT / MRI and were found to have locally advanced disease on the radiation therapy treatment planning CT / MRI, continued on the study as planned and were evaluated as per ITT.

[0251] Local failure was defined as:1. Increased tumor size by 50% or more as measured by palpation (product of length and width).2. Reappearance of palpable tumor after clearing.3. Biopsy revealing adenocarcinoma of the prostate at least 2 years after randomization. Prostate biopsy were performed at 2 years in all subjects except those who already experienced a DFS event.4. Urinary obstruction secondary to tumor confirmed by biopsy.

[0252] Evaluation of a positive DRE or urinary obstruction as the basis for declaration of local failure required confirmation by prostate biopsy or clear medical justification of why this was not medically possible.

[0253] Regional failure was defined as clinical with radiographic evidence of tumor in the pelvis.

[0254] Distant metastases were defined as clinical with radiographic evidence of disease beyond the pelvis.

[0255] The first site of suspected metastatic disease to the pelvis or distantly were biopsied, if medically indicated, for confirmation in addition to radiographic and PSA documentation.

[0256] The trial had ADT as optional, based on the clinical judgment of the treating physicians. ADT was used as a stratification criterion and for subset analysis.

[0257] The DFS result for the Test Arm was conservatively projected at 86.25%. The DFS for the control group was estimated at 75% based on data from multiple recent studies.

[0258] A blinded EAC was used to provide an objective medical review of the deaths and treatment failures that occurred in the trial. Additional details, including estimates, sensitivity and supporting analyses, were provided in the SAP.Attorney Docket No.: CAND-020WOKey Secondary EndpointProstate specific DFS

[0259] Prostate cancer-specific DFS was defined as time from randomization until the earliest of biopsy proven recurrent prostate cancer, EAC-adjudicated treatment failures, or EAC-adjudicated prostate cancer associated death. The EAC adjudicated all treatment failures and deaths. Deaths were adjudicated to be deaths due to prostate cancer, deaths not associated with prostate cancer, and deaths due to unknow causes.Secondary EndpointsPSA Nadir

[0260] A secondary endpoint was PSA nadir. This endpoint was followed but was not a primary endpoint for the study and thus was not used for sample size determination. The proportion of patients with PSA nadir < 0.2 ng / mL in the CAN-2409 Arm were compared to the Control Arm using Chi-square test. Logistic regression model was used to estimate the odds of PSA nadir < 0.2 ng / mL between the two arms to adjust for other covariates, as needed.Freedom From Biochemical Failure, Prostate Cancer-Specific Survival and Overall Survival

[0261] Biochemical failure was defined using the PSA nadir plus 2 ng / mL definition.

[0262] Freedom from biochemical failure was defined as the date from randomization to the date of the earliest confirmed biochemical failure. Prostate cancer-specific survival was defined as the date from randomization to date of death due to prostate cancer. Overall, survival was defined as the date from randomization to date of death due to all causes. The data was censored at the time of last follow-up when the patient was event-free. If the patient refused to continue with the protocol, every attempt was made to continue follow-up and analyze as per ITT. If they refused follow-up, they were censored at the last follow-up. These endpoints were followed but were not the primary endpoints for the study and thus were not used for sample size determination. The data analyses were performed on all randomized patients. The probability distribution of all these endpoints was estimated using Kaplan-Meier method. The comparison between treatment groups used a Cox multivariate regression model adjusted for stratification factors.Attorney Docket No.: CAND-020WOQuality of Life Assessment

[0263] Patient-reported Health Related Quality of Life outcomes were collected using the Expanded Prostate Cancer Index Composite (EPIC-26). Data were collected at baseline and 3, 6, 12, 18, and 24 months after completion of radiation. This was not a primary endpoint for the study and was not used for sample size determination.

[0264] Quality of life was evaluated including all patients entered in the study. All components and single items were scored on categorical scales and were summarized as a global health / quality of life scale at each time point. All data were analyzed using descriptive statistics for the subscales and single items for each study group at each of the assessment points. The global health status / QoL scale was used as an overall measure for QoL. Quality of Life between the two treatment groups was evaluated in a longitudinal fashion using the longitudinal mixed model (PROC Mixed in SAS). The model allowed the change of QoL in the two groups over time to be investigated.Exploratory and Subset Analyses

[0265] As described herein, radiation therapy in combination with CAN-2409 without ADT may be comparable to radiation therapy in combination with ADT in patients with defined intermediate-risk (data not shown). There was also evidence that the overall impact of CAN-2409 varied depending on the patients’ specific risk categorization. The studies herein further explored such associations using regression models and Chi-square statistics. Exploratory subset analysis by geographical region of treatment (country) in addition to the stratification factors were evaluated using stratified log rank test. Blood and tissue were collected from patients agreeing to have their samples archived for future immune studies.Results

[0266] The safety profile of CAN-2409 was generally consistent with previous studies, with no new safety signals identified. The treatment-related adverse events (AEs) were > 5% in either arm and are reported in TABLE 3. Chills, fever, and flu-like symptoms were commonly mild-to-moderate and self-limited. The incidence of treatment discontinuation due to AEs was lower on the CAN-2409 Arm, specifically 5.4% on the CAN-2409 + SoC Arm, while 6.0% on the placebo + SoC Arm. The incidence of severe AEs (SAEs) were also lower on the CAN-2409 Arm, specifically 5.8% on the CAN-2409 + SoC Arm, while 7.3% on the placebo + SoC Arm. Also, the incidence of treatment-related SAEs were lower on theAttorney Docket No.: CAND-020WOCAN-2409 Ann, specifically 1.7% on the CAN-2409 + SoC Arm, while 2.2% on the placebo + SoC Arm.

[0267] Meanwhile, the demographics / baseline characteristics of the enrolled, randomized patients are provided in TABLE 4.TABLE 3.TABLE 4.Attorney Docket No.: CAND-020WO

[0268] It was observed that there was a statistically significant improvement in DFS for CAN-2409 plus radiation therapy (n = 496) vs. radiation therapy alone (n=249) (p = 0.0155;Attorney Docket No.: CAND-020WOHazard Ratio (HR) 0.7) in ITT analysis (FIG. 2). A 14.5% relative improvement in DFS at 54 months for the CAN-2409 Treatment Arm compared to the placebo Control Arm was observed. This finding was also observed in biopsies, as shown in TABLE 5, where the percentage of positive biopsies was smaller in the CAN-2409 Arm. DFS improvement was observed both in patients receiving short-course ADT and in patients not receiving ADT (FIG. 3 and FIG. 5). CAN-2409 treatment improved DFS in both favorable and unfavorable intermediate-risk prostate cancer (TABLE 6). CAN-2409 showed a highly significant effect (p = 0.0046) on prostate cancer-specific outcomes (FIG. 4). There was a significant increase in the proportion of patients achieving a PSA nadir (<0.2 ng / mL) in the Treatment Arm compared to the placebo Control Arm (respectively 67.1% vs. 58.6%, p = 0.0164). CAN- 2409 induced 80.4% pathological complete responses (pCRs) in the 2-year post-treatment biopsies compared to 63.6% observed in the Control Arm (p=0.0015) (TABLE 7). The freedom from PSA failure was numerically lower in the CAN-2409 Treatment Arm (HR 0.84). Overall survival was similar by Treatment Arm, with only 2 deaths due to prostate cancer over 10+ years, while 50 patients died due to other causes, unrelated to treatment.TABLE 5.TABLE 6.Attorney Docket No.: CAND-020WO*use of ADT is not recommended in favorable disease by NCCN guidelinesTABLE 7.Difference between arms chi-square test p=0.0015Example 2: A Randomized Controlled Trial of CAN-2409 As Adjuvant to Up-Front Radiation Therapy For Intermediate-Risk Localized Prostate Cancer

[0269] The overall objective for this study was to determine if adjuvant therapy with CAN-2409 improved the clinical outcome of radiation therapy for newly-diagnosed prostate cancer in subjects with intermediate-risk.Materials and Methods

[0270] See Example 1.ResultsIn subjects with intermediate-risk, it was observed that there was a statistically significant improvement in DFS for CAN-2409 plus radiation therapy (n=635) vs. radiation therapy alone (p = 0.0091; HR 0.7) in ITT analysis (FIG. 6). CAN-2409 showed a highly significant effect (p = 0.0020) on prostate cancer-specific outcomes (FIG. 7). Additionally, it was observed that there was a statistically significant improvement in time to new therapy (FIG. 8), time to metastasis (FIG. 9), time to biochemical failure (FIG. 10), and time to alltreatment failure (FIG. 11) for CAN-2409 plus radiation therapy (n = 411, 421, 415, and 412, respectively) vs. radiation therapy alone (n=200, 208, 204, and 200, respectively) (p = 0.0202, 0.0079, 0.0248, and 0.0107, respectively; HR 0.4, 0.1, 0.3, and 0.4, respectively).Attorney Docket No.: CAND-020WOExample 3: Phase 2b Clinical Trial of CAN-2409 in Patients with Prostate Cancer: Active Surveillance Population

[0271] 190 subjects were fully enrolled and chose active surveillance. In a 2: 1 randomization ratio, CAN-2409 + prodrug (2 injections) or Active Surveillance + Placebo + Valacyclovir (2 injections) were administered. Primary endpoints included PFS, while secondary endpoints included progression to radical treatment, pathological response / PSA kinetics, and quality of life.

[0272] A numerical improvement in the percentage of patients with negative biopsies at 1 year and time-to-radical treatment for the CAN-2409 Treatment Arm compared to placebo, was observed, although it had not yet reached statistical significance.INCORPORATION BY REFERENCE

[0273] The entire disclosure of each of the patent documents and scientific articles cited herein is incorporated by reference for all purposes.EQUIVALENTS

[0274] The disclosure can be embodied in other specific forms without departing from the essential characteristics thereof. The foregoing embodiments therefore are to be considered illustrative rather than limiting on the disclosure described herein. The scope of the disclosure is indicated by the appended claims rather than by the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.

Claims

1. Attorney Docket No.: CAND-020WOCLAIMS1. A method of increasing disease-free survival in a subject with prostate cancer in need thereof, the method comprising administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy, so that the subject’s disease-free survival is increased as compared to a subject with prostate cancer receiving only the radiation therapy.

2. The method of claim 1, wherein the subject is newly-diagnosed with the prostate cancer.

3. The method of claim 1 or 2, wherein the subject has intermediate- to high-risk prostate cancer, optionally wherein the subject has intermediate-risk prostate cancer, and optionally wherein the subject has favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer.

4. The method of any one of claims 1-3, wherein the aglatimagene besadenovec is administered as a first, second, and third injection.

5. The method of claim 4, wherein the second injection is administered 2-8 weeks after the first injection.

6. The method of claim 5, wherein the second injection is administered 2-7 weeks after the first injection.

7. The method of any one of claims 4-6, wherein the third injection is administered 2-3 weeks after the second injection.

8. The method of any one of claims 4-7, wherein the subject receives no more than three injections of aglatimagene besadenovec.

9. The method of any one of claims 1-8, wherein the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

10. The method of any one of claims 1-9, wherein a prodrug is administered following administration of aglatimagene besadenovec.

11. The method of claim 10, wherein the prodrug is administered 1 day after administration of the aglatimagene besadenovec.

12. The method of claim 10 or 11, wherein the prodrug is administered daily for 14 days.

13. The method of any one of claims 10-12, wherein the prodrug is valacyclovir.Attorney Docket No.: CAND-020WO14. The method of any one of claims 4-13, wherein the radiation therapy begins 0-3 days after the second injection.

15. The method of claim 14, wherein the radiation therapy begins on the same day as the second injection.

16. The method of any one of claims 1-15, wherein the subject receives 78 Gy of radiation in 2 Gy fractions.

17. The method of any one of claims 1-15, wherein the subject receives hypofractionated radiation.

18. The method of claim 17, wherein the hypofractionated radiation comprises 60 Gy in 3 Gy fractions.

19. The method of any one of claims 1-18, wherein the method increases the subject’s disease-free survival to at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

20. The method of any one of claims 1-18, wherein the method increases the subject’s probability of disease-free survival to at least about 0.60 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

21. The method of any one of claims 1-18, wherein the method increases the subject’s probability of prostate-specific disease-free survival to at least about 0.75 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

22. The method of any one of claims 1-21, wherein the subject also receives androgen deprivation treatment (ADT), optionally wherein the subject receives no more than 6 months of ADT.

23. The method of any one of claims 1-21, wherein the subject does not receive ADT.

24. The method of any one of claims 1-23, wherein the subject achieves a prostate-specific antigen (PSA) nadir of < 0.2 ng / mL.

25. A method of reducing a prostate-specific antigen (PSA) level to a nadir of < 0.2 ng / mL in a subject with intermediate- to high-risk prostate cancer in need thereof, the methodAttorney Docket No.: CAND-020WO comprising administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy.

26. The method of claim 25, wherein the subject is newly -diagnosed with the prostate cancer.

27. The method of claim 25 or 26, wherein the subject has intermediate-risk prostate cancer, and optionally wherein the subject has favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer.

28. The method of any one of claims 25-27, wherein the aglatimagene besadenovec is administered as a first, second, and third injection.

29. The method of claim 28, wherein the second injection is administered 2-8 weeks after the first injection.

30. The method of claim 29, wherein the second injection is administered 2-7 weeks after the first injection.

31. The method of any one of claims 28-30, wherein the third injection is administered 2-3 weeks after the second injection.

32. The method of any one of claims 28-31, wherein the subject receives no more than three injections of aglatimagene besadenovec.

33. The method of any one of claims 25-32, wherein the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

34. The method of any one of claims 25-33, wherein a prodrug is administered following administration of aglatimagene besadenovec.

35. The method of claim 34, wherein the prodrug is administered 1 day after administration of the aglatimagene besadenovec.

36. The method of claim 34 or 35, wherein the prodrug is administered for 14 days.

37. The method of any one of claims 34-36, wherein the prodrug is valacyclovir.

38. The method of any one of claims 28-37, wherein the radiation therapy begins 0-3 days after the second injection.

39. The method of claim 38, wherein the radiation therapy begins on the same day as the second injection.Attorney Docket No.: CAND-020WO40. The method of any one of claims 25-39, wherein the subject receives 78 Gy of radiation in 2 Gy fractions.

41. The method of any one of claims 25-39, wherein the subject receives hypofractionated radiation.

42. The method of claim 41, wherein the hypofractionated radiation comprises 60 Gy in 3 Gy fractions.

43. The method of any one of claims 25-42, wherein the subject also receives ADT, optionally wherein the subject receives no more than 6 months of ADT.

44. The method of any one of claims 25-43, wherein the subject does not receive ADT.

45. A method of treating a subject with prostate cancer who does not receive androgen depletion therapy (ADT), the method comprising administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy.

46. The method of claim 45, wherein the subject is newly-diagnosed with the prostate cancer.

47. The method of claim 45 or 46, wherein the subject has intermediate- to high-risk prostate cancer, optionally wherein the subject has intermediate-risk prostate cancer, and optionally wherein the subject has favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer.

48. The method of any one of claims 45-47, wherein the aglatimagene besadenovec is administered as a first, second, and third injection.

49. The method of claim 48, wherein the second injection is administered 2-8 weeks after the first injection.

50. The method of claim 49, wherein the second injection is administered 2-7 weeks after the first injection.

51. The method of any one of claims 48-50, wherein the third injection is administered 2-3 weeks after the second injection.

52. The method of claim 51, wherein the subject receives no more than three injections of aglatimagene besadenovec.Attorney Docket No.: CAND-020WO53. The method of any one of claims 45-52, wherein the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

54. The method of any one of claims 45-53, wherein a prodrug is administered following administration of aglatimagene besadenovec.

55. The method of claim 54, wherein the prodrug is administered 1 day after administration of the aglatimagene besadenovec.

56. The method of claim 54 or 55, wherein the prodrug is administered daily for 14 days.

57. The method of any one of claims 54-56, wherein the prodrug is valacyclovir.

58. The method of any one of claims 48-57, wherein the radiation therapy begins 0-3 days after the second injection.

59. The method of claim 58, wherein the radiation therapy begins on the same day as the second injection.

60. The method of any one of claims 45-59, wherein the subject receives 78 Gy of radiation in 2 Gy fractions.

61. The method of any one of claims 45-59, wherein the subject receives hypofractionated radiation.

62. The method of claim 61, wherein the hypofractionated radiation comprises 60 Gy in 3 Gy fractions.

63. The method of any one of claims 45-62, wherein the method increases the subject’s disease-free survival to at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following administration of the aglatimagene besadenovec.

64. The method of any one of claims 45-62, wherein the method increases the subject’s probability of disease-free survival to at least about 0.60 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

65. The method of any one of claims 45-62, wherein the method increases the subject’s probability of prostate-specific disease-free survival to at least about 0.75 for at least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.Attorney Docket No.: CAND-020WO66. The method of any one of claims 45-65, wherein the subject achieves a PSA nadir of < 0.2 ng / mL.

67. A method of reducing at least one symptom associated with prostate cancer, the method comprising administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy, thereby reducing the at least one symptom associated with prostate cancer.

68. The method of claim 67, wherein the method results in a reduction in tumor volume, thereby reducing the at least one symptom associated with prostate cancer.

69. The method of claim 67 or 68, wherein the at least one symptom associated with prostate cancer is selected from pollakiuria, micturition urgency, and urinary tract pain.

70. The method of any one of claims 67-69, wherein the subject is newly-diagnosed with the prostate cancer.

71. The method of any one of claims 67-70, wherein the subject has intermediate- to high- risk prostate cancer, optionally wherein the subject has intermediate-risk prostate cancer, and optionally wherein the subject has favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer.

72. The method of any one of claims 67-71, wherein the aglatimagene besadenovec is administered as a first, second, and third injection.

73. The method of claim 72, wherein the second injection is administered 2-8 weeks after the first injection.

74. The method of claim 73, wherein the second injection is administered 2-7 weeks after the first injection.

75. The method of any one of claims 72-74, wherein the third injection is administered 2-3 weeks after the second injection.

76. The method of any one of claims 72-75, wherein the subject receives no more than three injections of aglatimagene besadenovec.

77. The method of any one of claims 67-76, wherein the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

78. The method of any one of claims 67-77, wherein a prodrug is administered following administration of aglatimagene besadenovec.Attorney Docket No.: CAND-020WO79. The method of claim 78, wherein the prodrug is administered 1 day after administration of the aglatimagene besadenovec.

80. The method of claim 78 or 79, wherein the prodrug is administered daily for 14 days.

81. The method of any one of claims 78-80, wherein the prodrug is valacyclovir.

82. The method of any one of claims 72-80, wherein the radiation therapy begins 0-3 days after the second injection.

83. The method of any one of claims 67-82, wherein the subject receives 78 Gy of radiation in 2 Gy fractions.

84. The method of any one of claims 67-83, wherein the subject receives hypofractionated radiation.

85. The method of claim 84, wherein the hypofractionated radiation comprises 60 Gy in 3 Gy fractions.

86. The method of any one of claims 67-85, wherein the subject also receives ADT, optionally wherein the subject receives no more than 6 months of ADT.

87. The method of any one of claims 67-85, wherein the subject does not receive ADT.

88. A method of treating a subject with prostate cancer in need thereof, the method comprising administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy, wherein the method results in an increased likelihood that the subject achieves a pathological complete response (pCR) as compared to a subject with prostate cancer receiving only the radiation therapy.

89. The method of claim 88, wherein the subject is newly-diagnosed with the prostate cancer.

90. The method of claim 88 or 89, wherein the subject has intermediate- to high-risk prostate cancer, optionally wherein the subject has intermediate-risk prostate cancer, and optionally wherein the subject has favorable intermediate-risk prostate cancer or unfavorable intermediate -risk prostate cancer.

91. The method of any one of claims 88-90, wherein the aglatimagene besadenovec is administered as a first, second, and third injection.

92. The method of claim 91, wherein the second injection is administered 2-8 weeks after the first injection.Attorney Docket No.: CAND-020WO93. The method of claim 92, wherein the second injection is administered 2-7 weeks after the first injection.

94. The method of any one of claims 91-93, wherein the third injection is administered 2-3 weeks after the second injection.

95. The method of any one of claims 91-94, wherein the subject receives no more than three injections of aglatimagene besadenovec.

96. The method of any one of claims 88-95, wherein the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

97. The method of any one of claims 88-96, wherein a prodrug is administered following administration of aglatimagene besadenovec.

98. The method of claim 97, wherein the prodrug is administered 1 day after administration of the aglatimagene besadenovec.

99. The method of claim 97 or 98, wherein the prodrug is administered daily for 14 days.

100. The method of any one of claims 97-99, wherein the prodrug is valacyclovir.

101. The method of any one of claims 91-100, wherein the radiation therapy begins 0-3 days after the second injection.

102. The method of claim 101, wherein the radiation therapy begins on the same day as the second injection.

103. The method of any one of claims 88-102, wherein the subject receives 78 Gy of radiation in 2 Gy fractions.

104. The method of any one of claims 88-103, wherein the subject receives hypofractionated radiation.

105. The method of claim 104, wherein the hypofractionated radiation comprises 60 Gy in 3 Gy fractions.

106. The method of any one of claims 88-105, wherein the subject also receives ADT, optionally wherein the subject receives no more than 6 months of ADT.

107. The method of any one of claims 88-106, wherein the subject does not receive ADT.

108. The method of any one of claims 88-107, wherein the subject achieves a PSA nadir of < 0.2 ng / mL.Attorney Docket No.: CAND-020WO109. A method of increasing time to metastasis, biochemical failure, treatment failure, or new treatment in a subject with intermediate-risk prostate cancer, the method comprising: administering to the subject a therapeutically effective amount of aglatimagene besadenovec, wherein the subject also receives radiation therapy, so that the subject’s time to metastasis, biochemical failure, treatment failure, or new treatment is increased as compared to a subject with intermediate-risk prostate cancer receiving only the radiation therapy.

110. The method of claim 109, wherein the subject is newly-diagnosed with the intermediate-risk prostate cancer.

111. The method of claim 109 or 110, wherein the subject has favorable intermediate-risk prostate cancer or unfavorable intermediate-risk prostate cancer.

112. The method of any one of claims 109-111, wherein the aglatimagene besadenovec is administered as a first, second, and third injection.

113. The method of claim 112, wherein the second injection is administered 2-8 weeks after the first injection.

114. The method of claim 113, wherein the second injection is administered 2-7 weeks after the first injection.1 15. The method of any one of claims 112-114, wherein the third injection is administered 2- 3 weeks after the second injection.1 16. The method of any one of claims 112-115, wherein the subject receives no more than three injections of aglatimagene besadenovec.1 17. The method of any one of claims 109-116, wherein the aglatimagene besadenovec is administered intraprostatically to each of the four quadrants of the prostate.

118. The method of any one of claims 109-117, wherein a prodrug is administered following administration of aglatimagene besadenovec.

119. The method of claim 118, wherein the prodrug is administered 1 day after administration of the aglatimagene besadenovec.

120. The method of claim 118 or 119, wherein the prodrug is administered daily for 14 days.

121. The method of any one of claims 118-120, wherein the prodrug is valacyclovir.

122. The method of any one of claims 112-121, wherein the radiation therapy begins 0-3 days after the second injection.Attorney Docket No.: CAND-020WO123. The method of claim 122, wherein the radiation therapy begins on the same day as the second injection.

124. The method of any one of claims 109-123, wherein the subject receives 78 Gy of radiation in 2 Gy fractions.

125. The method of any one of claims 109-124, wherein the subject receives hypofractionated radiation.

126. The method of claim 125, wherein the hypofractionated radiation comprises 60 Gy in 3 Gy fractions.

127. The method of any one of claims 109-126, wherein the method increases the subject’s time to metastasis to at least 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

128. The method of any one of claims 109-126, wherein the method increases the subject’s time to biochemical failure to at least 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

129. The method of any one of claims 109-126, wherein the method increases the subject’s time to treatment failure to at least 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

130. The method of any one of claims 109-126, wherein the method increases the subject’s time to new treatment to at least 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

131. The method of any one of claims 109-130, wherein the method increases the subject’s probability of disease-free survival to at least about 0.75 for at least 2 years, 3, years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

132. The method of any one of claims 109-130, wherein the method increases the subject’s probability of prostate-specific disease-free survival to at least about 0.75 for at least 2 years, least 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years following the initial administration of the aglatimagene besadenovec.

130. The methodAttorney Docket No.: CAND-020WO of any one of claims 109-132, wherein the subject also receives ADT, optionally wherein the subject receives no more than 6 months of ADT.

131. The method of any one of claims 109-133, wherein the subject does not receive ADT.

132. The method of any one of claims 109-134, wherein the subject achieves a prostatespecific antigen (PSA) nadir of < 0.2 ng / mL.

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