Lesion treatment of prostate cancer

Abstract

Embodiments include methods of treating prostate cancer by administering to a low-grade single-lesion prostate cancer tumor a composition comprising a therapeutically effective amount of a pharmaceutically active ingredient capable of inducing necrosis of the low-grade single-lesion prostate cancer tumor, wherein the administration reduces the incidence of cancer, cancer level, and cancer progression (exacerbation) throughout the semi-prostate in which the initial lesion is located and treated.

Description

[0001] This application is a continuation of International Application No. PCT / US2020 / 044192, International Filing Date, July 30, 2020, Chinese Application No. 202080055228.4, entitled “Lesion Treatment of Prostate Cancer,” the subject matter of which is incorporated by reference herein in its entirety.

[0002] Cross Reference to Related Applications

[0003] This application claims priority to U.S. Utility Application No. 16 / 528,326, filed July 31, 2019, the subject matter of which is incorporated by reference herein in its entirety. BACKGROUND

[0004] 1. Field of Implementation

[0005] Embodiments include methods of treating prostate cancer in a mammal having prostate cancer, and more particularly, methods of preventing and / or reducing the development and progression of multifocal prostate cancer by administering to a low-grade, low-risk, localized (Tlc) prostate cancer tumor a composition comprising a pharmaceutically active ingredient capable of inducing necrosis of the low-grade, low-risk, localized prostate cancer tumor and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutically active ingredient is fexapotide triflutate (“FT”). The methods include, but are not limited to, intramuscular, oral, intravenous, intraperitoneal, intraprostatic, intracerebral (intra-parenchymal), intraventricular, intralesional, intraocular, intraarterial, intrathecal, intratumoral, intranasal, topical, transdermal, subcutaneous, or intradermal administration of the composition to a patient in need thereof, wherein the composition is targeted for administration to a low-grade, low-risk, localized (Tlc) prostate cancer tumor, reducing the grade of prostate cancer, the progression (worsening) of prostate cancer, and the incidence of prostate cancer throughout the entire hemi-prostate where the initial single lesion tumor was located and treated.

[0006] 2. Description of Related Art

[0007] The nature of many medical and surgical procedures involves the removal or destruction of harmful or unwanted tissue. Examples of such treatment include surgical removal of cancerous or precancerous growths, destruction of metastatic tumors by chemotherapy, and reduction of glandular (e.g., prostate) hyperplasia. Other examples include removal of unwanted facial hair, removal of warts, and removal of unwanted adipose tissue.

[0008] There is a need for an effective composition that will destroy and thus facilitate the removal of, or inhibit the further growth of, harmful or unwanted cells and tissue, but which has primarily local effects and minimal or no systemic toxicity. There is also a need to reduce the need for invasive surgical intervention, even after treatment with an effective composition.

[0009] Some agents known to have the ability to destroy and thus facilitate the removal or inhibit the further growth of harmful or unwanted cells and tissues are disclosed in U.S. Patent Application No. 14 / 808,713, filed July 24, 2015, entitled METHODS OF REDUCING THE NEED FOR SURGERY IN PATIENTS SUFFERING FROM BENIGN PROSTATIC HYPERPLASIA; U.S. Patent Application No. 14 / 606,683, filed January 27, 2015, entitled METHOD OF TREATING DISORDERS REQUIRING OR REMOVAL OF CELLS, U.S. Patent Application No. 14 / 738,551, filed June 12, 2015, entitled COMBINATION COMPOSITIONS FOR TREATING DISORDERS REQUIRING REMOVAL OR DESTRUCTION OF UNWANTED CELLULAR PROLIFERATIONS, U.S. Patent Application Publication No. 2007 / 0237780 (now abandoned); 2003 / 0054990 (now U.S. Patent No. 7,172,893); 2003 / 0096350 (now U.S. Patent No. 6,924,266); 2003 / 0096756 (now U.S. Patent No. 7,192,929); 2003 / 0109437 (now U.S. Patent No. 7,241,738); 2003 / 0166569 (now U.S. Patent No. 7,317,077); 2005 / 0032704 (now U.S. Patent No. 7,408,021); and 2015 / 0148303 (now U.S. Patent No. 9,243,035), the disclosures of each of which are incorporated by reference herein in their entireties.

[0010] One of the agents disclosed in these documents is fexapotide triflutate, or FT. FT has been shown to reduce prostate cells, improve or reduce LUTS, and treat BPH in men suffering from prostatic enlargement. FT is also disclosed for use in reducing the incidence of prostate cancer by treating BPH in a mammal suffering from BPH, wherein a composition containing FT is administered to the transitional zone (central) prostate in the mammal. See, e.g., U.S. Patent No. 10,183,058, the disclosure of which is incorporated by reference herein in its entirety.

[0011] Prostate cancer is known to be a disease with a very high prevalence relative to its clinical incidence in the population. Prostate cancer has a high rate of asymptomatic onset and long duration of asymptomatic. The average interval for prostate cancer is 7 to 14 years during which the cancer is present but in a preclinical stage because it cannot be detected by typical clinical or laboratory tests (see Etzioni, Ret al., Am J Epidemiol. Vol. 148, pp. 775-85 (1998); and Gulati, R, et al., Cancer Epidemiol Biomarkers Prev; Vol. 20(5), pp. 740-50 (2011)). These preclinical asymptomatic prostate cancers are a reasonable target for treatment before they become clinically apparent or otherwise detected, which can benefit the patient before the cancer becomes clinically apparent or otherwise detected.

[0012] Prostate cancer is generally considered to be a multifocal disease, with the prostate comprising multiple adenocarcinoma lesions with different heterogeneity. This makes the cancer difficult to treat effectively, often resulting in a radical prostatectomy, which can cause a number of life-altering problems for men, including erectile dysfunction and urinary incontinence. However, some prostate cancers, or unifocal prostate cancers, have been reported in about 20% to about 35% of radical prostatectomy specimens. Mazzucchelli, et al.,“Pathology of Prostate Cancer and Focal Therapy (‘Male Lumpectomy’),” Anticancer Research, Vol. 29, pp. 5155-5162 (2009); Ibeawuchi, et al.,“Genome-Wide Investigation of Multifocal and Unifocal Prostate Cancer— Are They Genetically Different?” Int. J. Mol. Sci., Vol. 14, ppp. 11816-11829 (2013).

[0013] Due to the severity of radical prostatectomy, recent studies have reported focal therapy of the prostate sparing portions of the prostate, although the efficacy of focal therapy of prostate cancer and prevention of cancer progression remains uncertain. Quann, et al.,“Current prostate biopsy protocols cannot reliably identify patients for focal therapy;…”, Int. J. Clin. Exp. Pathol., Vol. 3(4), pp. 401-407 (2010). To date, there is a need to achieve identification, targeting, and focal destruction of specific tumors (Mazzucchelli at 5159), and to date, focal therapy involves ablation of a large portion of the prostate (e.g., hemi-ablation). Therefore, to date, it is not known or expected that treatment of low-grade, low-risk, single-focal prostate cancer tumors by targeting only single-focal tumors will effectively reduce the cancer incidence, cancer grade, and cancer progression (worsening) of the entire lobe of the prostate in which the single-focal tumor resides. Therefore, there are no clinical trials designed in which FT treatment of single-focal prostate cancer is evaluated, and it is not expected to be effective in treating the prostate lobe in which the single-focal tumor resides.

[0014] A common technique for identifying and monitoring prostate cancer is the evaluation of PSA levels in conjunction with biopsies. A typical prostate biopsy generally involves taking a large sample through the prostate and evaluating the tissue using the Gleason score. The Gleason score measures the abnormality of the cancer cells under a microscope and is a good indicator of how fast the cells can grow and spread. The Gleason score is calculated by adding the two cancer grades that make up the largest area of the biopsy tissue sample, typically represented as two numbers, e.g., 3+3, and a total score, e.g., 6. The following table provides a classification of prostate cancer groups and associated risks:

[0015]

[0016] For low-grade, low-risk, localized (Tlc) prostate cancer with a Gleason score of < 6, active surveillance (AS) is the conventional course of treatment. This is because, while some can mature into high-risk cancer requiring radical prostatectomy (e.g., Gleason score of > 8), many do not. As noted above, up to about 35% of surgically removed prostates have only low-grade, low-risk, localized (Tlc) prostate cancer. Thus, those patients who have their prostate surgically removed but still do not have high-risk cancerous tumors unnecessarily suffer the deleterious effects of radical prostatectomy. Thus, the typical standard of care is to take corrective action when the Gleason score is 7 or above, particularly when the initial pattern is 4: by removing most of the prostate, chemotherapy or radiation, or radical prostatectomy. Thus, it would be desirable to find a safe and effective way to treat prostate cancer patients in the low and / or low and intermediate risk groups by lesion treatment of individual lesions, in which case the treatment can effectively improve, reduce and / or block cancer progression in the entire lobe of the prostate in which the individual lesion is located.

[0017] Throughout this specification, including the preceding description of the relevant art, any and all patents, patent applications, and other documents, including any and all U.S. Patent Application Publications, referred to in this disclosure are incorporated by reference in their entirety. The preceding description of the relevant art is not intended as an admission that any of the documents in the relevant art, including any and all U.S. Patent Application Publications, are prior art. Moreover, no admission is made that the preceding description of the relevant art is complete or wholly accurate or wholly relevant. Furthermore, the description of any disadvantages in relation to the described products, methods, and / or apparatus is not intended to limit the embodiments. Indeed, aspects of the embodiments can include certain features of the described products, methods, and / or apparatus without suffering from any described disadvantages.

[0018] SUMMARY

[0019] There remains a need in the art for new, less toxic, less frequent, and less invasive treatments for preventing or reducing the progression or incidence of prostate cancer. There also remains a need in the art for such treatments that reduce the incidence of multi-lesional prostate cancer at least in the lobe (or hemisphere) in which the initially treated lesion is located. The embodiments meet these needs.

[0020] The present disclosure is premised, in part, on the discovery that a pharmaceutically active ingredient capable of inducing necrosis of low-grade, low-risk, localized prostate cancer tumors can be administered to a single low-grade, low-risk prostate cancer lesion or tumor (i.e., Gleason score < 6) but has an unexpected effect in reducing multi-lesion cancer incidence, reducing multi-lesion cancer grade, and reducing multi-lesion progression (worsening) in the prostate lobe in which the low-grade, low-risk, localized prostate cancer tumor is located. Suitable pharmaceutically active ingredients capable of inducing such tumor necrosis include, for example, fexapotide triflutate (FT) - a peptide described by the amino acid sequence Ile-Asp-Gln-Gln-Val-Leu-Ser-Arg-Ile-Lys-Leu-Glu-Ile-Lys-Arg-Cys-Leu, Zytiga (abiraterone acetate), Apalutamide, abazitaxel, Casodex (bicalutamide), Eligard and Lupron, (leuprolide acetate), Erleada (apalutamide), Firmagon (degarelix), flutamide, goserelin acetate, Jevtana (carboplatin), mitoxantrone hydrochloride, Nilandron (nilutamide), Provenge (sipuleucel-T), sipuleucel-T, Taxotere (docetaxel), Xofigo (radium Ra 223 dichloride), Xtandi (enzalutamide), Zoladex (goserelin acetate), and mixtures and combinations thereof. Over several years of continued follow-up, it was discovered that such administration effectively reduces cancer incidence, reduces cancer grade, and reduces cancer progression (worsening) in the prostate lobe (half of the prostate) in which the initial single-lesion tumor is located and treated. Thus, embodiments can greatly improve the quality of life for many men suffering from prostate cancer who would otherwise undergo more aggressive treatments such as ablation of most of the prostate, chemotherapy, radiation, or radical prostatectomy.

[0021] Some embodiments are directed to a method of reducing cancer incidence, reducing cancer grade, and reducing cancer progression (worsening) in a mammal having low-grade or low-risk (i.e., Gleason score < 6) prostate cancer by administering to the mammal a therapeutically effective amount of a composition comprising at least one pharmaceutically active ingredient capable of inducing necrosis of low-grade, low-risk, localized prostate cancer tumors. The method comprises administering to a single cancer focus (single-lesion tumor) in the prostate of the mammal a therapeutically effective amount of the composition, and reducing the increase in Gleason grade of the treated-side half of the prostate when compared to active surveillance by an amount of about 15% to about 100% when measured at least 18 months after treatment.

[0022] The compositions can be administered intramuscularly, orally, intravenously, intraperitoneally, intracerebrally (intra-parenchymally), intracerebroventricularly, intratumorally, intralesionally, intradermally, intrathecally, intranasally, intraocularly, intraarterially, topically, transdermally, via aerosol, infusion, bolus, implantation device, slow release system, etc.

[0023] In another embodiment, the composition comprises a therapeutically effective amount of FT, which is administered in an amount of about 2.0 mg to about 20 mg. The method also comprises administering to a single cancer focus in the prostate of the mammal (single lesion tumor) a therapeutically effective amount of FT and reducing the percentage of mammals exhibiting >1 new lesion in the prostate lobe treated for the original lesion and an increase in Gleason grade (new multi-lesion with Gleason initial pattern >4) when measured at least 18 months after treatment, by an amount of about 50% to about 100% when compared to active surveillance, or about 45% to about 100% when measured at least 36 months after treatment, or about 45% to about 100% when measured at least 48 months after treatment, when compared to active surveillance. In another embodiment, the method reduces the increase in Gleason grade in the treated lateral half of the prostate by an amount of about 45% to about 70% when measured at least 36 months after treatment, when compared to active surveillance. In another embodiment, the method comprises administering to a single cancer focus in the prostate of the mammal (single lesion tumor) a therapeutically effective amount of FT and reducing the percentage of mammals exhibiting >1 new lesion in the prostate lobe treated for the original lesion and an increase in Gleason grade (new multi-lesion with Gleason grade up-regulation) when measured at least 18 months after treatment, by an amount of about 40% to about 100% when compared to active surveillance, or about 50% to about 90% when measured at least 36 months after treatment, or about 15% to about 80% when measured at least 48 months after treatment, when compared to active surveillance.

[0024] In another embodiment, the method comprises administering a therapeutically effective amount of FT to a single cancer focus in the prostate of the mammal (single lesion tumor) and reducing the percentage of mammals having conventional cancer therapy (surgery, radiation therapy, or chemotherapy) and exhibiting >1 new lesion and an increase in Gleason grade in the treated lobe of the prostate of the original lesion (new multi-lesion with an increase in Gleason grade) by an amount of about 40% to about 100% when measured at least 18 months after treatment, or about 50% to about 90% when compared to active monitoring when measured at least 36 months after treatment, or about 15% to about 80% when compared to active monitoring when measured at least 48 months after treatment. The method also comprises administering a therapeutically effective amount of FT to a single cancer focus in the prostate of the mammal (single lesion tumor) and reducing the percentage of mammals having conventional cancer therapy (surgery, radiation therapy, or chemotherapy) and exhibiting >1 new lesion and an increase in Gleason initial pattern in the treated lobe of the prostate of the original lesion (new multi-lesion with an increase in Gleason initial pattern >4) by an amount of about 65% to about 100% when measured at least 18 months after treatment, or about 65% to about 100% when compared to active monitoring when measured at least 36 months after treatment, or about 60% to about 100% when compared to active monitoring when measured at least 48 months after treatment.

[0025] The foregoing general description and the following detailed description are exemplary and explanatory only and are intended to provide further explanation of the embodiments claimed. Other objects, advantages and features of the present embodiments will become apparent to those skilled in the art from the following detailed description.

[0026] Detailed description of preferred embodiments

[0027] Before the proteins, nucleotide sequences, peptides, compositions, active agents, etc. and methods of the present application are described, it is to be understood that the application is not limited to the particular methodologies, protocols, cell lines, vectors, and reagents described as these can vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present embodiments which will be limited only by the appended claims.

[0028] The terms and phrases used herein are defined as follows unless otherwise indicated. Throughout the specification, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a host cell" includes a plurality of such host cells, reference to "the antibody" is a reference to one or more antibodies and equivalents thereof known to those skilled in the art, and so forth.

[0029] The amino acids and amino acid residues described herein can be referred to according to the accepted one-letter or three-letter codes provided in the following table.

[0030] Table 1

[0031]

[0032]

[0033] The expression “pharmaceutically active ingredients capable of inducing such tumor necrosis” refers to, for example, fexapotide triflutate (FT) - a peptide described by the amino acid sequence Ile-Asp-Gln-Gln-Val-Leu-Ser-Arg-Ile-Lys-Leu-Glu-Ile-Lys-Arg-Cys-Leu, Zytiga (abiraterone acetate), Apalutamide, abazitaxel, Casodex (bicalutamide), Eligard and Lupron, (leuprolide acetate), Erleada (apalutamide), Firmagon (degarelix), flutamide, goserelin acetate, Jevtana (carboplatin), mitoxantrone hydrochloride, Nilandron (nilutamide), Provenge (sipuleucel-T), sipuleucel-T, Taxotere (docetaxel), Xofigo (radium Ra 223 dichloride), Xtandi (enzalutamide), Zoladex (goserelin acetate), and mixtures and combinations thereof. Fexapotide Triflutate (“FT”), as used herein, refers to a 17-mer peptide having the following amino acid sequence: Ile-Asp-Gln-Gln-Val-Leu-Ser-Arg-Ile-Lys-Leu-Glu-Ile-Lys-Arg-Cys-Leu (SEQ ID NO. 1). FT is disclosed in U.S. Patent Nos. 6,924,266; 7,241,738; 7,317,077; 7,408,021; 7,745,572; 8,067,378; 8,293,703; 8,569,446; and 8,716,247, and U.S. Patent Application Publication Nos. 2017 / 0360885; 2017 / 0020957; 2016 / 0361380; and 2016 / 0215031. The disclosures of these patents and published applications are incorporated herein by reference in their entirety.

[0034] FT is represented by the following:

[0035] SEQ ID NO. 1: IDQQVLSRIKLEIKRCL or Ile-Asp-Gln-Gln-Val-Leu-Ser-Arg-Ile- Lys-Leu-Glu-Ile-Lys-Arg-Cys-Leu.

[0036] The term "fragment" refers to a protein or polypeptide consisting of a contiguous subsequence of the amino acid sequence of a protein or peptide, and includes naturally occurring fragments, such as splice variants and fragments produced by naturally occurring in vivo protease activity. Such fragments can be truncated at the amino terminus, carboxy terminus, and / or internally (e.g., by natural splicing). Such fragments can be prepared with or without the amino terminal methionine. The term "fragment" includes fragments from the same protein or peptide, whether identical or different, having common or different contiguous amino acid sequences, joined together directly or through a linker. One of ordinary skill in the art will be able to select appropriate fragments for use in the embodiments without undue experimentation using the guidelines and procedures outlined herein.

[0037] The term "variant" refers to a protein or polypeptide that has one or more amino acid substitutions, deletions, and / or insertions compared to the amino acid sequence of a protein or peptide, and includes naturally occurring allelic variants or alternatively spliced variants of a protein or peptide. The term "variant" includes replacement of one or more amino acids in a peptide sequence with similar or homologous amino acids or different amino acids. There are a number of scales that can be used to classify amino acids as similar or homologous. (Gunnar von Heijne, Sequence Analysis in Molecular Biology, p. 123-39 (Academic Press, New York, N.Y. 1987.) Preferred variants include alanine substitutions at one or more amino acid positions. Other preferred substitutions include conservative substitutions that have little or no effect on the overall net charge, polarity, or hydrophobicity of the protein. Conservative substitutions are listed in Table 2 below.

[0038] Table 2

[0039] Conservative amino acid substitutions

[0040]

[0041] Table 3 lists another amino acid substitution scheme:

[0042] Table 3

[0043]

[0044] Other variants can consist of substitutions of amino acids that are less conserved, e.g., residues that differ more significantly in their function to maintain (a) the polypeptide's main chain structure, e.g., as a folded or helical conformation, (b) the molecule's charge or hydrophobicity at the target site, or (c) the bulk of the side chain. Substitutions that are generally expected to have a more significant effect on function are those in which: (a) a glycine and / or proline is substituted for or deleted or inserted; (b) a hydrophilic residue, e.g., serine or threonine, is substituted for or by a hydrophobic residue, e.g., leucyl, isoleucyl, phenylalanyl, valyl, or alanyl (or vice versa); (c) a cysteine residue is substituted for or by any other residue (or vice versa); (d) a residue whose side chain bears a positive charge, e.g., lysyl, arginyl, or histidyl, is substituted for or by a residue whose side chain bears a negative charge (e.g., glutamyl or aspartyl) (or vice versa); or (e) a residue whose side chain has a large bulk, e.g., phenylalanine, is substituted for or by a residue whose side chain does not have such a bulk, e.g., glycine (or vice versa). Other variants include variants designed to create new glycosylation and / or phosphorylation sites, or variants designed to eliminate existing glycosylation and / or phosphorylation sites. Variants include at least one amino acid substitution at a glycosylation site, a proteolytic cleavage site, and / or a cysteine residue. Variants also include proteins and peptides having additional amino acid residues before or after the linker peptide of the protein or peptide amino acid sequence. For example, a cysteine residue can be added at both the amino terminal and carboxy terminal end of a peptide to cyclize the peptide by forming a disulfide bond. The term "variant" also encompasses polypeptides having the amino acid sequence of a peptide, wherein at least one and up to 25 or more additional amino acids are present flanking the 3' or 5' end of the peptide.

[0045] The term "derivative" refers to a chemically modified protein or polypeptide that has been chemically modified by natural processes (e.g., processing and other post-translational modifications) as well as chemical modification techniques (e.g., by the addition of one or more polyethylene glycol molecules, sugars, phosphates and / or other such molecules, where the molecule or molecules are not naturally attached to the wild-type protein or peptide). Derivatives include salts. Such chemical modifications are well described in the underlying texts and more detailed treatises and in the extensive body of research literature, and they are well known to those skilled in the art. It will be appreciated that the same type of modification can occur in several sites on a given protein or polypeptide, either independently or at the same time. In addition, a given protein or polypeptide can contain a variety of types of modifications. Modifications can occur anywhere on the protein or polypeptide, including the peptide backbone, the amino acid side chains and the amino or carboxyl termini. Modifications include, for example, acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleic acid derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of a phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, glycosylation, lipid attachment, sulfation, gamma-carboxylation of glutamic acid residues, hydroxylation and ADP-ribosylation, selenoylation, sulfation, transfer-RNA-mediated addition of amino acids to proteins, such as arginylation, and ubiquitination.See, e.g., Proteins— Structure And Molecular Properties, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993) and Wold, F., "Posttranslational Protein Modifications: Perspectives and Prospects," pgs. 1-12 in Posttranslational Covalent Modification Of Proteins, B. C. Johnson, Ed., Academic Press, New York (1983); Seifter et al., Meth. Enzymol. 182:626-646 (1990) and Rattan et al., "Protein Synthesis: Posttranslational Modifications and Aging," Ann. N.Y. Acad. Sci. 663: 48-62 (1992). The term "derivative" includes chemical modifications of a protein or polypeptide that result in branching or cyclization of the protein or polypeptide, with or without branching. Cyclized, branched, and branched cyclic proteins or polypeptides can be produced by post-translational processes in nature or can be prepared by entirely synthetic methods.

[0046] The term "homolog" refers to a protein having an amino acid sequence that is at least 60% identical to that of the peptide as determined by standard methods commonly used to compare the similarity in amino acid positions of two polypeptides. The degree of similarity or identity between two proteins can be readily calculated by known methods, including, but not limited to, those described in Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; and Carillo H. and Lipman, D., SIAM, J. Applied Math., 48:1073 (1988). Preferred methods to determine identity are designed to give the largest match between the sequences tested. Methods to determine identity and similarity are codified in publicly available computer programs.

[0047] Preferred computer program methods for determining identity and similarity between two sequences include, but are not limited to, the GCG program package (Devereux, J., et al., Nucleic Acids Research, 12(1): 387 (1984)), BLASTP, BLASTN, and FASTA, Atschul, S. F. et al., J. Molec. Biol., 215: 403-410 (1990). The BLAST X programs are publicly available from NCBI and other sources (BLAST Manual, Altschul, S., et al., NCBI NLM NIH Bethesda, Md. 20894; Altschul, S., et al., J. Mol. Biol., 215: 403-410 (1990). For example, using a computer algorithm, such as GAP (Genetic Computer Group, University of Wisconsin, Madison, Wis.), two proteins or polypeptides for which percent sequence identity is to be determined are aligned for optimal matching of their respective amino acids ("matched span", determined by the algorithm).

[0048] Gap open penalty (calculated as 3 times the average diagonal; "average diagonal" is the average of the diagonal of the comparison matrix used; "diagonal" is the score or number assigned by a particular comparison matrix to each perfect amino acid match) and gap extension penalty (usually a {fraction (1 / 10)} of the gap open penalty), and a comparison matrix such as PAM250 or BLOSUM 62 are used with the algorithm. Standard comparison matrices (see Dayhoff et al., in: Atlas of Protein Sequence and Structure, vol. 5, supp. 3 for the PAM250 comparison matrix; see Henikoff et al., Proc. Natl. Acad. Sci USA, 89: 10915-10919 for the BLOSUM 62 comparison matrix) can also be used with the algorithm. Percent identity is then calculated by the algorithm. Homologues will typically have one or more amino acid substitutions, deletions and / or insertions compared to the compared protein or peptide, as appropriate.

[0049] The term "fusion protein" refers to a protein in which one or more peptides are recombinantly fused or chemically conjugated (including covalent and non-covalent) to a protein, such as (but not limited to) an antibody or antibody fragment such as a Fab fragment or a short chain Fv. The term "fusion protein" also refers to multimers of peptides (i.e., dimers, trimers, tetramers, and higher multimers). Such multimers include homomultimers comprising one peptide, heteromultimers comprising more than one peptide, and heteromultimers comprising at least one peptide and at least one other protein. Such multimers can be the result of hydrophobic, hydrophilic, ionic, and / or covalent associations, bonds, or linkages, can be formed by cross-linking using a linking molecule, or can be indirectly linked by, for example, a liposome.

[0050] The term "peptidomimetic" or "mimetic" refers to a biologically active compound that mimics the biological activity of a peptide or protein but is no longer chemically a peptide, i.e., they no longer contain any peptide bonds (i.e., amide bonds between amino acids). Here, the term peptidomimetic is used in a broader sense to include molecules that are not entirely peptides in nature, such as pseudopeptides, semipeptides, and peptoids. Examples of peptidomimetics in the broadest sense are described below (wherein a portion of the peptide is replaced by a structure that lacks a peptide bond). Whether entirely or partially non-peptidic, the peptidomimetics according to the embodiments provide a spatial arrangement of reactive chemical moieties that closely resembles the three-dimensional arrangement of active groups in the peptide on which the peptidomimetic is based. Due to this similar active site geometry, the peptidomimetic has an influence on biological systems similar to that of the biological activity of the peptide.

[0051] Peptidomimetics of the embodiments are preferably substantially similar to the peptides described herein in three-dimensional shape and biological activity. Examples of methods for making structural modifications to peptides known in the art to produce peptidomimetics include inverting backbone chiral centers to produce D-amino acid residue structures that can (particularly at the N-terminus) result in enhanced stability to proteolytic degradation without adversely affecting activity. An example is given in the paper "Tritriated D-ala1-Peptide T Binding", Smith C. S. et al., Drug Development Res., 15, pp. 371-379 (1988). A second method is to alter cyclic structures for increased stability, such as N to C interchain imides and lactams (Ede et al. in Smith and Rivier (Eds.) "Peptides: Chemistry and Biology", Escom, Leiden (1991), pp. 268-270). Examples of this are given in conformationally constrained thymopentin-like compounds, such as those disclosed in U.S. Patent No. 4,457,489 (1985), Goldstein, G. et al., the disclosure of which is incorporated herein by reference in its entirety. A third method is to replace peptide bonds in the peptide with pseudopeptide bonds that confer resistance to proteolysis.

[0052] A number of pseudopeptide bonds that do not generally affect the structure and biological activity of the peptide have been described. One example of such an approach is the substitution of retro-inverso pseudopeptide bonds (Biologically active retroinverso analogues of thymopentin", Sisto A. et al in Rivier, J. E. and Marshall, G. R. (eds) "Peptides, Chemistry, Structure and Biology", Escom, Leiden (1990), pp. 722-773) and Dalpozzo, et al. (1993), Int. J. Peptide Protein Res., 41 :561-566, incorporated herein by reference). According to this modification, the amino acid sequence of the peptide can be identical to that of the peptide described above, except that one or more peptide bonds are replaced by retro-inverso pseudopeptide bonds. Preferably, the most N-terminal peptide bond is substituted, as such a substitution will confer resistance to proteolysis by exopeptidases acting on the N-terminus. Further modifications can also be made by replacing the chemical groups of the amino acids with other chemical groups of similar structure. Another suitable pseudopeptide bond known to enhance stability to enzymatic cleavage without or with little loss of biological activity is the reduced isostere pseudopeptide bond (Couder, et al. (1993), Int. J. Peptide Protein Res., 41 :181-184, incorporated herein by reference in its entirety).

[0053] Accordingly, the amino acid sequence of these peptides can be identical to that of the peptide, except that one or more peptide bonds are replaced by isostere pseudopeptide bonds. Preferably, the most N-terminal peptide bond is substituted, as such a substitution will confer resistance to proteolysis by exopeptidases acting on the N-terminus. The synthesis of peptides having one or more reduced isostere pseudopeptide bonds is known in the art (Couder, et al. (1993), supra). Other examples include the introduction of a ketomethylene or methylthio bond to replace a peptide bond.

[0054] Peptidomimetics of peptides represent another class of peptide mimics that retain important structural determinants of biological activity but eliminate the peptide bond, thereby conferring resistance to proteolysis (Simon, et al., 1992, Proc. Natl. Acad. Sci. USA, 89:9367-9371, incorporated by reference in its entirety). Peptidomimetics are oligomers of N-substituted glycines. A number of N-alkyl groups have been described, each corresponding to the side chain of a natural amino acid (Simon, et al. (1992), supra). Some or all of the amino acids of a peptide can be replaced by N-substituted glycines corresponding to the substituted amino acid.

[0055] The term "peptide mimetic" or "mimetic" also includes retro-D peptides and enantiomers as defined below.

[0056] The term "retro-D peptide" refers to a biologically active protein or peptide composed of D-amino acids arranged in reverse order compared to the L-amino acid sequence of the peptide. Thus, the carboxyl terminal residue of an L-amino acid peptide becomes the amino terminal of a D-amino acid peptide, and so on. For example, the peptide ETESH becomes H d S d E d T d E d where E d , H d , S d , and T d are D-amino acids corresponding to L-amino acids E, H, S, and T, respectively.

[0057] The term "enantiomer" refers to a biologically active protein or peptide in which one or more L-amino acid residues in the amino acid sequence of the peptide are replaced by the corresponding D-amino acid residue.

[0058] As used herein, "composition" refers broadly to any composition comprising the peptide or amino acid sequence and optionally an additional active agent. The composition can comprise a dry formulation, an aqueous solution, or a sterile composition. A composition comprising a peptide can be used as a hybridization probe. The probe can be stored in lyophilized form and can be combined with stabilizers such as carbohydrates. In hybridization, the probe can be deployed in an aqueous solution containing salts, e.g., NaCl, detergents, e.g., sodium dodecyl sulfate (SDS), and other ingredients, e.g., Denhardt's solution, milk powder, salmon sperm DNA, etc.

[0059] The expression "low grade prostate cancer" means prostate cancer that presents as a prostate tissue biopsy, i.e., a single lesion or multiple lesions, with a highest Gleason grade of < 6 or 3+3. The expression "low grade single lesion prostate cancer" means a single cancerous lesion with a Gleason grade < 6 or 3+3 detected by biopsy. It will be appreciated that the biopsy procedure, which typically takes a large number of samples from the prostate, does not sample the entire gland, so there can be other lesions that are not detected. The expression "prostate cancer progression" generally means a higher Gleason grade for any single lesion (the highest grade of that grade among all biopsies), but also a greater number of cancers in a biopsy (i.e., a given biopsy lesion has a higher percentage of cancer if, for example, more than 50%, or more, of the lesions are cancer positive). For example, if a patient has 5% tumor from one positive core sample at a certain point in time, to a later point in time, with 5 cores, each with 40% tumor (all with the same Gleason grade), this would be considered progression, although it is not "Gleason grade progression." On the other hand, if a patient starts with 4 cores at grade 6, each with 40% tumor, and then later has only one positive core with 5% tumor, but a Gleason grade of 7, then progression would be considered "Gleason grade progression."

[0060] When reference is made to "biopsy," those skilled in the art will appreciate that a typical biopsy consists of multiple "quadrant" samples, typically at least 10 or 12, sampling all regions of the gland (left and right of each; top, middle and bottom; middle and sides of each, as well as transitions L and R), thus equaling 14 regions. Accordingly, reference to "biopsy" or "one biopsy" means 10-15 biopsies taken at the same time, each reported separately.

[0061] In embodiments where additional active agents are used with the composition, the expression "active agent" is used to mean any agent capable of removing unwanted cell proliferation and / or tissue growth. Suitable active agents include, but are not limited to: (i) anti-cancer active agents (e.g., alkylating agents, topoisomerase I inhibitors, topoisomerase II inhibitors, RNA / DNA anti-metabolites, and anti-mitotic agents); (ii) active agents for treating benign growths, such as anti-acne and anti-wart active agents; (iii) anti-androgenic compounds, (cyproterone acetate (1 alpha, 2ß-methylene-6-chloro-17 alpha -acetoxy-6- dehydropregnenolone) tamoxifen, aromatase inhibitors); (iv) alpha 1 -adrenergic receptor blockers (tamsulosin, terazosin, doxazosin, prazosin, bunazosin, indoramin, alfuzosin, silodosin); (v) 5 alpha-reductase inhibitors (finasteride, dutasteride); (vi) 5 type phosphodiesterase (PDE5) inhibitors (tadalafil) and combinations thereof.

[0062] Embodiments relate to methods of administering a composition to low-grade, low-risk, localized prostate cancer tumors, the composition comprising at least one pharmaceutically active ingredient capable of inducing necrosis of low-grade, low-risk, localized prostate cancer tumors and reducing prostate cancer morbidity, reducing prostate cancer grade, and reducing prostate cancer progression (worsening) in the entire hemi-prostate where the initial single lesion tumor was located and treated. Suitable pharmaceutically active ingredients capable of inducing such tumor necrosis include, for example, fexapotidetriflutate (FT) - a peptide described by the amino acid sequence Ile-Asp-Gln-Gln-Val-Leu-Ser-Arg-Ile-Lys-Leu-Glu-Ile-Lys-Arg-Cys-Leu, Zytiga (abiraterone acetate), Apalutamide, abazitaxel, Casodex (bicalutamide), Eligard and Lupron, (leuprolide acetate), Erleada (apalutamide), Firmagon (degarelix), flutamide, goserelin acetate, Jevtana (carboplatin), mitoxantrone hydrochloride, Nilandron (nilutamide), Provenge (sipuleucel-T), sipuleucel-T, Taxotere (docetaxel), Xofigo (radium Ra 223 dichloride), Xtandi (enzalutamide), Zoladex (goserelin acetate), and mixtures and combinations thereof. In embodiments, the composition is administered more than once. Thus, embodiments provide methods of reducing prostate cancer morbidity, grade, and progression in a minimally invasive manner by administering the composition to a mammal that would not typically be treated. It is generally accepted that a mammal having low-grade single or multi-lesion prostate cancer with Gleason grade < 6 is placed on active surveillance (AS), or not treated. See, e.g., Ahmed, et al., “Do Low-Grade and Low-Volume Prostate Cancers Bear the Hallmarks of Malignancy,” www.thelancet.com / oncology, Vol. 13, pp e509-e517 (2012).

[0063] The inventors have unexpectedly discovered that administration of such compositions to a single lesion of a low-grade, low-risk, localized prostate cancer tumor significantly reduces the incidence of prostate cancer, the grade of prostate cancer, and the progression of prostate cancer in the lobe of the prostate where the initially treated tumor was located. Thus, the methods of the embodiments provide a non-invasive method of reducing multi-lesion prostate cancer compared to radical prostatectomy or even lesion ablation, resection, chemotherapy, or radiation. Even active surveillance requires multiple repeated prostate biopsies and assessments, which places a significant burden on the healthcare system. Thus, the methods described herein are used to delay the incidence, occurrence, and progression of prostate cancer in a non-invasive manner.

[0064] Mammals treated with the compositions of the invention exhibit a significant reduction in the incidence of prostate cancer, a significant reduction in the increase in Gleason grade, and a significant reduction in the progression of prostate cancer compared to published literature. The methods of the embodiments can reduce the increase in Gleason grade in the treated side lobe of the prostate by an amount of about 15% to about 100% when measured at least 18 months after treatment, or about 20% to about 95%, or about 25% to about 88%, or about 30% to about 85%, or about 40% to about 80%, or about 45% to about 95%, or about 45% to about 80%, or any value therebetween when compared to active surveillance controls when measured at 18 months, or 36 months, or 48 months, or 60 months.

[0065] The methods of the embodiments can reduce multi-lesion prostate cancer with Gleason upregulation (patient has >= 1 new lesion in the treated lobe or half of the prostate with an increase in Gleason grade) in the lobe (or half of the prostate) of the initially treated lesion by an amount of about 40% to about 100%, or about 50% to about 90%, or about 60% to about 80%, or any value therebetween when measured at least 18 months after treatment when compared to active surveillance controls. The methods of the embodiments can reduce half of the prostate multi-lesion cancer with Gleason upregulation by an amount of about 50% to about 90%, or about 50% to about 85%, or about 50% to about 80%, or any value therebetween when measured at least 36 months after treatment when compared to active surveillance controls. The methods of the embodiments can reduce half of the prostate multi-lesion cancer with Gleason upregulation by an amount of about 15% to about 80%, or about 15% to about 75%, or about 16% to about 72%, or any value therebetween when measured at least 48 months after treatment when compared to active surveillance controls.

[0066] When measured at least 18 months after treatment, the methods of the embodiments can reduce multi-focal prostate cancer in the lobe (or half of the prostate) of the initial treatment lesion with a Gleason initial pattern > 4 (patient has >= 1 new lesion in the treated lobe or half of the prostate with an increased Gleason grade initial pattern) by an amount of from about 50% to about 100%, or from about 70% to about 100%, or from about 75% to about 100%, or any value therebetween, when compared to an active surveillance control. When measured at least 36 months after treatment, the methods of the embodiments can reduce half prostate multi-focal cancer with a Gleason initial pattern > 4 by an amount of from about 45% to about 100%, or from about 70% to about 100%, or from about 75% to about 100%, or any value therebetween, when compared to an active surveillance control. When measured at least 48 months after treatment, the methods of the embodiments can reduce half prostate multi-focal cancer with a Gleason upregulation (patient has >= 1 new lesion in the entire prostate with an increased Gleason grade) by an amount of from about 45% to about 100%, or from about 60% to about 100%, or from about 70% to about 100%, or any value therebetween, when compared to an active surveillance control.

[0067] When measured at least 18 months after treatment, the methods of the embodiments can reduce the percentage of mammals with conventional cancer treatment (surgery, radiation therapy, or chemotherapy), and new multi-focal cancer treatment in the lobe (or half of the prostate) of the initial treatment lesion with a Gleason upregulation (new multi-focal cancer treatment with an increased Gleason grade in the half of the prostate) by an amount of from about 40% to about 100%, or from about 50% to about 90%, or from about 60% to about 75%, or any value therebetween, when compared to an active surveillance control. When measured at least 36 months after treatment, the methods of the embodiments can reduce new multi-focal cancer treatment with an increased Gleason grade in the half of the prostate by an amount of from about 50% to about 90%, or from about 55% to about 75%, or from about 50% to about 80%, or any value therebetween, when compared to an active surveillance control. When measured at least 48 months after treatment, the methods of the embodiments can reduce new multi-focal cancer treatment with an increased Gleason grade in the half of the prostate by an amount of from about 15% to about 80%, or from about 35% to about 75%, or from about 40% to about 75%, or any value therebetween, when compared to an active surveillance control.

[0068] The methods of the embodiments can reduce the percentage of mammals with conventional cancer treatment (surgery, radiation therapy, or chemotherapy), and multi-focal prostate cancer with a Gleason initial pattern > 4 in the lobe (or half-prostate) of the initial treatment lesion, by an amount of about 65% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any value therebetween, or about 65% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any value therebetween, of cancer treatment for new multi-focal and increased Gleason grade initial pattern in the half-prostate when compared to active surveillance controls when measured at least 18 months after treatment. The methods of the embodiments can reduce cancer treatment for new multi-focal and increased Gleason grade initial pattern in the half-prostate by an amount of about 65% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any value therebetween, or about 65% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any value therebetween, when compared to active surveillance controls when measured at least 36 months after treatment. The methods of the embodiments can reduce cancer treatment for new multi-focal and increased Gleason grade initial pattern in the half-prostate by an amount of 60% to about 100%, or about 65% to about 100%, or about 75% to about 100%, or any value therebetween, or about 60% to about 100%, or about 65% to about 100%, or about 75% to about 100%, or any value therebetween, when compared to active surveillance controls when measured at least 48 months after treatment.

[0069] Any mammal can benefit from the uses of the present application, including humans, mice, rabbits, dogs, sheep, and other livestock, any mammal treated or treatable by a veterinarian, zookeeper, or wildlife conservation employee. Preferred mammals are humans, sheep, and dogs. Throughout the specification, mammal and patient are used interchangeably.

[0070] It will be apparent to those skilled in the art that other smaller FT fragments can be selected to have the same or similar biological activity. Other FT fragments can be selected by those skilled in the art to have the same or similar biological activity. Thus, the term "FT" as used in the embodiments includes these other fragments. Typically, the peptides of the embodiments have at least 4 amino acids, preferably at least 5 amino acids, more preferably at least 6 amino acids.

[0071] The embodiments also include methods of treatment, including administering a composition comprising FT, the composition comprising two or more FT sequences linked together, and an additional active agent. The two or more FT sequences will also have the desired biological activity to the extent that the FT has the desired biological activity.

[0072] The FTs and fragments, variants, derivatives, homologues, fusion proteins and mimetics thereof encompassed by the present embodiments can be prepared using methods known to those of skill in the art, such as recombinant DNA technology, protein synthesis, and isolation of naturally occurring peptides, proteins, variants, derivatives, and homologues thereof. The Ft and fragments, variants, derivatives, homologues, fusion proteins and mimetics thereof can be prepared from other peptides, proteins, and fragments, variants, derivatives, and homologues thereof using methods known to those of skill in the art. Such methods include, but are not limited to, cleaving a peptide or protein with a protease to yield FT. Any of the methods disclosed in, e.g., U.S. Patent Nos. 6,924,266; 7,241,738; 7,317,077; 7,408,021; 7,745,572; 8,067,378; 8,293,703; 8,569,446; and 8,716,247, and U.S. Patent Application Publication Nos. 2017 / 0360885; 2017 / 0020957; 2016 / 0361380; and 2016 / 0215031, can be used to prepare the FT peptides described herein. The disclosures of these patent documents are incorporated herein by reference in their entireties.

[0073] If used in addition to one or more pharmaceutically active ingredients, the additional active agent can be one or more active agents selected from (i) anti-cancer active agents (e.g., alkylating agents, topoisomerase I inhibitors, topoisomerase II inhibitors, RNA / DNA anti-metabolites, and anti-mitotic agents); (ii) active agents for treating benign growths, such as anti-acne and anti-wart active agents (salicylic acid); (iii) anti-androgenic compounds, (cyproterone acetate (1 alpha, 2ß-methylene-6-chloro-17 alpha -acetoxy-6-dehydroprogesterone)) tamoxifen, aromatase inhibitors); (iv) alpha 1 -adrenergic receptor blockers (tamsulosin, terazosin, doxazosin, prazosin, bunazosin, indoramin, alfuzosin, silodosin); (v) 5 alpha-reductase inhibitors (finasteride, dutasteride); (vi) 5 type phosphodiesterase (PDE5) inhibitors (tadalafil) and combinations thereof. Preferably, the additional active agent is an anti-cancer agent, in particular an active agent for treating prostate cancer.

[0074] The therapeutic compositions described herein can comprise an amount of a pharmaceutically active ingredient in admixture with a pharmaceutically acceptable carrier. In some alternative embodiments, the additional active agent can be administered in the same composition as the pharmaceutically active ingredient, and in other embodiments, the composition comprising the pharmaceutically active ingredient is administered as an injection, while the additional active agent is formulated as an oral medication (gels, capsules, tablets, liquids, etc.). The carrier material can be water for injection, preferably supplemented with other materials commonly found in solutions for administration to mammals. Typically, when the pharmaceutically active ingredient is FT, it will be administered in the form of a composition comprising purified FT peptide (or chemically synthesized FT peptide) together with one or more physiologically acceptable carriers, excipients, or diluents. Neutral buffered saline or saline mixed with serum albumin are exemplary suitable carriers. Preferably, the product is formulated as a lyophilizate using suitable excipients (e.g., sucrose). Other standard carriers, diluents and excipients can be included as desired. The compositions of the embodiments can also comprise buffers with appropriate pH ranges known to those of ordinary skill in the art, including Tris buffers at about pH 7.0-8.5 or acetate buffers at about pH 4.0-5.5, which can also include sorbitol or suitable substitutes.

[0075] Solid dosage forms for oral administration include, but are not limited to, capsules, tablets, pills, powders, and granules. In such solid dosage forms, the additional active agent and / or pharmaceutically active ingredient can be admixed with at least one of the following: (a) one or more inert excipients (or carriers), such as sodium citrate or dicalcium phosphate; (b) fillers or supplements, such as starches, lactose, sucrose, glucose, mannitol, and silicic acid; (c) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (d) humectants, such as glycerol; (e) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (f) solution retarders, such as paraffin; (g) absorption accelerators, such as quaternary ammonium compounds; (h) wetting agents, such as acetyl alcohol and glycerol monostearate; (i) absorbents, such as kaolin and bentonite clay; (j) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. For capsules, tablets, and pills, the dosage form can also include buffering agents.

[0076] Liquid dosage forms for oral administration include pharmaceutically-acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, the liquid dosage forms can include inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers. Exemplary

[0077] In addition to such inert diluents, the composition can include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0078] The actual dose level of active ingredients in the compositions of the embodiments can be varied to obtain an amount of the pharmaceutically-active ingredient and additional active agent that is effective to obtain the desired therapeutic response for a particular composition. Thus, the selected dose level will depend on the desired therapeutic effect, on the route of administration, on the duration of the treatment desired, and on other factors.

[0079] For a mammal, including a human, an effective amount can be administered according to body surface area. E. J. Freireich et al., Cancer Chemother. Rep., 50 (4):219 (1966) describes the interrelationship of dose and the size and species of subjects in mice, rats, rabbits, dogs, monkeys, and humans. 2 Body surface area can be approximated from the height and weight of the individual (see, e.g., Scientific Tables, Geigy Pharmaceuticals, Ardsley, N.Y. pp. 537-538 (1970)).

[0080] It will be understood, that the specific dose level for any particular patient will depend upon a variety of factors including the body weight, general health, sex, diet, time and route of administration, potency, rate of absorption and excretion of the administered drug, combination with other drugs, and the severity of the particular disease being treated.

[0081] Methods of administering a composition comprising a pharmaceutically active ingredient according to an embodiment include, but are not limited to, intramuscular, oral, intravenous, intraperitoneal, intracerebral (intraparenchymal), intracerebroventricular, intratumoral, intralesional, intradermal, intrathecal, intranasal, intraocular, intraarterial, topical, transrectal, transperitoneal, transdermal, or by aerosol, infusion, bolus, implantation device, slow release system, etc. For example, any of the administration methods disclosed in U.S. Patent Nos. 6,924,266; 7,241,738; 7,317,077; 7,408,021; 7,745,572; 8,067,378; 8,293,703; 8,569,446; and 8,716,247, and U.S. Patent Application Publication Nos. 2017 / 0360885; 2017 / 0020957; 2016 / 0361380; and 2016 / 0215031 can be used.

[0082] The use of FT is a preferred embodiment. FT is a novel molecular entity that, based on tissue culture gene array data, stimulates the caspase pathway (activating caspases 7, 8 and 10, caspase recruitment domains 6, 11 and 14, and DIABLO), the tumor necrosis factor pathway (activating TNF1, TNFSF6, TNFSF8, TNFSF9, CD70 ligand and TNFRSF19L, TNFRSF25, TRAF2, TRAF3, TRAF4, TRAF6 receptors), and the BCL pathway (activating BIK, HRK, BCL2L10 and BCL3) in prostate epithelial cells in vitro. FT selectively leads to loss of cell membrane integrity, mitochondrial metabolic arrest, RNA depletion, DNA fragmentation and aggregation, and cell fragmentation and loss. The apoptotic process leads to progressive changes in the typical ultrastructure of membrane rupture and swelling, with gradual deepening of nuclear invagination, culminating in membrane blebbing, cell death and fragmentation into apoptotic bodies. Histologically, typical apoptotic changes were found throughout the injected area for up to several weeks after treatment, with positive immunohistochemical staining for markers of apoptosis.

[0083] FT has been extensively tested in BPH patients. In 9 human clinical trials, the compound and placebo controls have been administered via the transrectal route in over 1700 procedures. In these large long-term clinical trials of men with BPH, FT was administered at a concentration of 0.25 mg / ml (2.5 mg FT - equivalent to about 15-20% of the gland volume administered). See, e.g., Shore, et al., “The potential for NX-1207 in benign prostatic hyperplasia: an update for clinicians,” Ther Adv. Chronic Dis., 2(6), pp. 377-383 (2011). Compositions comprising FT are therefore preferred to include at least 2.5 mg FT, and can be administered in a single administration in an amount of up to 25 mg FT. In another embodiment, FT is administered in an amount ranging from about 2.5 mg to about 20 mg or about 2.5 mg to about 15 mg. In embodiments, FT is administered in an amount of 15 mg.

[0084] The following examples are provided to illustrate the embodiments. It should be understood, however, that the embodiments are not limited to the particular conditions or details described in these examples. Throughout this specification, any and all references including patents to publicly available documents are specifically incorporated by reference. In particular, the embodiments expressly incorporate by reference the examples contained in U.S. Patent Nos. 6,924,266; 7,241,738; 7,317,077; 7,408,021; 7,745,572; 8,067,378; 8,293,703; 8,569,446; and 8,716,247, as well as U.S. Patent Application Publication Nos. 2017 / 0360885; 2017 / 0020957; 2016 / 0361380; 2016 / 0215031; and 2016 / 0215031, each of which discloses certain peptides specified therein as effective agents for causing cell death in vivo in normal rodent muscle tissue, subcutaneous connective tissue, dermis, and other tissues. EXAMPLE

[0085] In a series of clinical studies, a total of 146 men with low grade prostate cancer (Gleason grade < 6) were treated as follows. Patients were randomized and received a single intraprostatic injection via the transrectal route of a composition comprising 2.5 mg FT (n = 49) or a composition comprising 15 mg FT (n = 48), or were subjected to control active monitoring (n = 49). Eighteen patients in the control active monitoring group were cross-overed to a single administration of a composition comprising 2.5 mg FT 45 days after the first follow-up biopsy. Patients were followed for 5 years, including biopsies at baseline, 45 days, 18, 36, and 60 months, and urological evaluations every 6 months for PSA. Patients with Gleason grade increase or who elected surgical or radiation therapy intervention were withdrawn from the study and were still included in the data analysis. The percentage of normal biopsies in the baseline lesion quadrant and the median tumor grade and volume were assessed; progression was measured by clinical and pathological outcomes, including Gleason grade of the entire prostate sampled and the treated lobe of the prostate. Intervention related to Gleason grade increase and the overall incidence of any intervention were assessed.

[0086] Example 1

[0087] This example assessed the percentage of patients who exhibited more than one new lesion (i.e., from single to multiple lesions) in the treated initial single lesion tumor half of the prostate, where the Gleason grade sum score of the new lesions increased ("Gleason upregulated half prostate multiple lesions"). The follow-up time data in the table below represent the percentage of patients who progressed or worsened. The results are shown in Table 3 below:

[0088] Table 3

[0089]

[0090] The results of Example 1 show that the implementation can reduce the percentage of patients with Gleason upstaged multi-focal prostate cancer in the lobe (or half of the prostate) of the initial treated lesion (patients with >= 1 new lesion in the treated lobe or half of the prostate with an increase in Gleason grade) by an amount of from about 40% to about 100%, or from about 50% to about 90%, or from about 60% to about 80%, or any value therebetween, when measured at least 18 months after treatment, when compared to an active surveillance control. The method of the implementation can reduce the percentage of patients with Gleason upstaged half-prostate multi-focal cancer by an amount of from about 50% to about 90%, or from about 50% to about 85%, or from about 50% to about 80%, or any value therebetween, when measured at least 36 months after treatment, when compared to an active surveillance control. The method of the implementation can reduce the percentage of patients with Gleason upstaged half-prostate multi-focal cancer by an amount of from about 15% to about 80%, or from about 15% to about 75%, or from about 16% to about 72%, or any value therebetween, when measured at least 48 months after treatment, when compared to an active surveillance control.

[0091] Example 2

[0092] This example evaluates the percentage of patients exhibiting more than one new lesion (i.e., changing from single to multi-focal) in the half of the prostate treated for the initial single lesion tumor, where the new lesion has an increase in Gleason grade of >= 4 from the initial pattern (“half-prostate multi-focal cancer with an increase in Gleason initial pattern”). The results are shown in Table 4 below:

[0093] Table 4

[0094]

[0095] The results of Example 2 show that the implementation can reduce the percentage of patients with half-prostate multi-focal cancer with an increase in Gleason initial pattern by an amount of from about 50% to about 100%, or from about 70% to about 100%, or from about 75% to about 100%, or any value therebetween, when measured at least 18 months after treatment, when compared to an active surveillance control. The method of the implementation can reduce the percentage of patients with half-prostate multi-focal cancer with an increase in Gleason initial pattern by an amount of from about 45% to about 100%, or from about 70% to about 100%, or from about 75% to about 100%, or any value therebetween, when measured at least 36 months after treatment, when compared to an active surveillance control. The method of the implementation can reduce the percentage of patients with half-prostate multi-focal cancer with an increase in Gleason initial pattern by an amount of from 45% to about 100%, or from about 60% to about 100%, or from about 70% to about 100%, or any value therebetween, when measured at least 48 months after treatment, when compared to an active surveillance control.

[0096] Example 3

[0097] This example evaluates the percentage of patients with multi-focal prostate cancer who receive conventional cancer treatment (e.g., surgery, radiation, and / or chemotherapy), and who have Gleason upgrading in the lobe (or half of the prostate) of the initial treated lesion (for cancer treatment of new multi-focal cancer with Gleason grade upgrading in half of the prostate). The results are shown in Table 5 below:

[0098] Table 5

[0099]

[0100] The results of Example 3 show that the implementation can reduce the percentage of patients with cancer treatment of new multi-focal cancer with Gleason upgrading in half of the prostate by an amount of about 40% to about 100%, or about 50% to about 90%, or about 60% to about 75%, or any value therebetween, when compared to an active monitoring control, when measured at least 18 months after treatment. The method of the implementation can reduce the percentage of patients with cancer treatment of new multi-focal cancer with Gleason grade increase in half of the prostate by an amount of about 50% to about 90%, or about 55% to about 75%, or about 50% to about 80%, or any value therebetween, when compared to an active monitoring control, when measured at least 36 months after treatment. The method of the implementation can reduce the percentage of patients with cancer treatment of new multi-focal cancer with Gleason grade increase in half of the prostate by an amount of about 15% to about 80%, or about 35% to about 75%, or about 40% to about 75%, or any value therebetween, when compared to an active monitoring control, when measured at least 48 months after treatment.

[0101] Example 4

[0102] This example evaluates the percentage of patients with multi-focal prostate cancer who receive conventional cancer treatment (e.g., surgery, radiation, and / or chemotherapy), and who have Gleason initial pattern > 4 in the lobe (or half of the prostate) of the initial treated lesion (for cancer treatment of new multi-focal and Gleason grade initial pattern increase in half of the prostate). The results are shown in Table 6 below:

[0103] Table 6

[0104]

[0105] The results of Example 4 show that the embodiments can reduce the percentage of cancer treated patients with new multifocal cancer with an initial pattern of Gleason grade increase in the hemi-prostate by an amount of from about 65% to about 100%, or from about 70% to about 100%, or from about 75% to about 100%, or any value therebetween, when compared to an active surveillance control, when measured at least 18 months after treatment. The methods of the embodiments can reduce the percentage of cancer treated patients with new multifocal and an initial pattern of Gleason grade increase in the hemi-prostate by an amount of from about 65% to about 100%, or from about 70% to about 100%, or from about 75% to about 100%, or any value therebetween, when compared to an active surveillance control, when measured at least 36 months after treatment. The methods of the embodiments can reduce the percentage of cancer treated patients with new multifocal and an initial pattern of Gleason grade increase in the hemi-prostate by an amount of from 60% to about 100%, or from about 65% to about 100%, or from about 75% to about 100%, or any value therebetween, when compared to an active surveillance control, when measured at least 48 months after treatment.

[0106] The results of the foregoing examples demonstrate the unexpected superior effect of the pharmaceutically active ingredient, in particular Ft, in reducing cancer incidence, reducing cancer grade, and reducing cancer progression (worsening) in the entire hemi-prostate where the initial low grade tumor was located and treated. It will be apparent to those skilled in the art that various modifications and variations can be made to the methods and compositions of the embodiments without departing from the spirit or scope of the embodiments.

Claims

1. Use of Fexapotide Triflutate (FT) in the preparation of an intraprostatic injection dosage form for reducing cancer incidence, reducing cancer grade, and reducing progression or worsening in a non-invasive manner throughout the hemi-prostate where an initial low-grade, unifocal prostate cancer tumor is located in a mammal with unifocal prostate cancer, wherein the intraprostatic injection dosage form comprises 15 mg of FT and neutral buffered saline, which, when injected directly into a low-grade, unifocal prostate cancer tumor in a mammal with unifocal prostate cancer, is sufficient to reduce cancer incidence, reduce cancer grade, and reduce cancer progression or worsening throughout the hemi-prostate where the initial low-grade, unifocal prostate cancer tumor is located.

2. The use according to claim 1, wherein the increase in hemi-prostate Gleason grade is reduced by an amount of 15% to 100% when compared to active surveillance when measured at least 18 months after treatment.

3. The use according to claim 2, wherein the increase in hemi-prostate Gleason grade is reduced by an amount of 45% to 70% when compared to active surveillance when measured at least 18 months after treatment.

4. Use of Fexapotide Triflutate (FT) in the preparation of an intraprostatic injection dosage form for reducing cancer incidence, reducing cancer grade, and reducing cancer progression (worsening) for at least 48 months in a non-invasive manner throughout the entire hemi-prostate where an initial low-grade unifocal prostate cancer tumor is located in a mammal with unifocal prostate cancer, wherein the intraprostatic injection dosage form comprises 15 mg of FT and a pharmaceutically acceptable carrier and, when directly injected into a low-grade unifocal prostate cancer tumor in a mammal with unifocal prostate cancer, reduces cancer incidence, reduces cancer grade, and reduces cancer progression or worsening throughout the entire hemi-prostate where the initial low-grade unifocal prostate cancer tumor is located.

5. The method according to claim 4, wherein the pharmaceutically acceptable carrier is neutral buffered saline.

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