Methods for treating multiple cancers

Abstract

Embodiments include a method of treating (preventing or reducing the occurrence of) multiple cancers by administering to the solitary cancerous lesion a therapeutically effective amount of a composition comprising a pharmaceutically active ingredient capable of inducing necrosis of the solitary cancerous tumor, wherein the administration reduces the occurrence of multiple cancers, the grade of multiple cancers, and the progression (worsening) of multiple cancers in an organ or an entire organism.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Patent Application No. 16 / 528,390, filed July 31, 2019, the subject matter of which is incorporated herein by reference in its entirety.

[0002] 1. Field of the embodiment

[0003] Embodiments include methods for treating multiple cancers in a mammal afflicted with multiple cancers, more particularly, methods for preventing and / or reducing the development and progression of multiple cancers in an affected organ or entire organism by directly administering to a single (solitary) cancer tumor a composition comprising a pharmaceutically active ingredient capable of inducing tumor necrosis and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutically active ingredient is fexapotide triflutate ("FT"), and the multiple cancer is prostate cancer. The method includes, but is not limited to, administering the composition intramuscularly, orally, intravenously, intraperitoneally, intraprostatically, intracerebrally (intraparenchymal), intraventricularly, intralesionally, intraocularly, intraarterially, intrathecally, intratumorally, intranasally, topically, transdermally, subcutaneously, or intradermally to a patient in need thereof, wherein targeted administration of the composition to the solitary cancer tumor reduces or prevents the occurrence of multiple cancers, the grade of multiple cancers, and the progression (worsening) of multiple cancers. [Background technology]

[0003] 2. Description of Related Technology Many medical treatments and procedures essentially involve the removal or destruction of harmful or unwanted tissue. Examples of such treatments include the surgical removal of cancerous or precancerous growths, the destruction of metastatic tumors with chemotherapy, and the reduction of glandular (e.g., prostate) hyperplasia. Other examples include the removal of unwanted facial hair, the removal of warts, and the removal of unwanted fatty tissue.

[0004] There is a need for effective compositions that destroy harmful or unwanted cells and tissues, thus facilitating their removal or inhibiting their further growth, while having primarily local effects with minimal or no systemic toxicity. There is also a need to reduce the need for invasive surgical intervention, radiation and / or chemotherapy, and other invasive procedures.

[0005] Agents known to have the ability to destroy harmful or unwanted cells and tissues, thus facilitating their removal or inhibiting their further growth, are disclosed in U.S. Patent Application No. 14 / 808,713, filed July 24, 2015 ("Method for reducing the need for surgery for patients with benign prostatic hyperplasia"), U.S. Patent Application No. 14 / 606,683, filed January 27, 2015 ("Method for treating diseases requiring the destruction or removal of cells"), U.S. Patent Application No. 14 / 738,551, filed June 12, 2015 ("Combined compositions for treating diseases requiring the removal or destruction of unwanted cell growths"), U.S. Patent Application Publication No. 2007 / 0237780 (abandoned), U.S. Patent Application Publication No. No. 2003 / 0054990 (now U.S. Pat. No. 7,172,893), U.S. Patent Application Publication No. 2003 / 0096350 (now U.S. Pat. No. 6,924,266), U.S. Patent Application Publication No. 2003 / 0096756 (now U.S. Pat. No. 7,192,929), U.S. Patent Application Publication No. 2003 / 0109437 (now U.S. Pat. No. 7,241,738), U.S. Patent Application Publication No. 2003 / 0166569 (now U.S. Pat. No. 7,317,077), U.S. Patent Application Publication No. 2005 / 0032704 (now U.S. Pat. No. 7,408,021), and U.S. Patent Application Publication No. 2015 / 0148303 (now U.S. Pat. No. 9,243,035), the disclosures of each of which are incorporated herein by reference in their entireties.

[0006] One of the drugs disclosed in these documents is fexapotide triflutate (FT). FT has been shown to reduce prostate cells, improve or reduce LUTS, and treat BPH in men with benign prostatic hyperplasia. FT has also been disclosed as being useful for reducing the incidence of prostate cancer by treating PPH in mammals with BPH, wherein a composition containing FT is administered to the mammal in the transition zone (central) prostate. See, for example, U.S. Patent No. 10,183,058, the disclosure of which is incorporated herein by reference in its entirety. FT is also the subject of a clinical trial to evaluate the effectiveness of FT in the treatment of low-grade, low-risk, localized (T1c) prostate cancer tumors, wherein the clinical trial is designed to evaluate the effectiveness of FT in eliminating localized tumors within 45 days after treatment.

[0007] Prostate cancer is known as a disease with a very high prevalence compared to its clinical incidence in the population. Prostate cancer has a high asymptomatic incidence rate and a long asymptomatic period. Prostate cancer has an average preclinical interval of 7 to 14 years, during which cancer is present but not detected by typical clinical or laboratory tests (see Etzioni et 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)).

[0008] Prostate cancer is often considered a multifocal disorder in which the prostate gland contains multiple adenocarcinoma foci of varying heterogeneity. This makes it difficult to effectively treat the cancer and often leads to radical prostatectomy, which causes many life-altering problems for men, including erectile dysfunction and urinary incontinence. However, some prostate cancers are considered low-grade, low-risk, localized (T1c) prostate cancer, which is reported in approximately 20 to 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, pp. 11816-11829 (2013).

[0009] Due to the severity of radical prostatectomy, recent studies have reported on focal therapy, in which part of the prostate is preserved, but the effectiveness of focal therapy for prostate cancer and prevention of cancer progression remains unclear. 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). Identifying, targeting, and locally destroying specific tumors has not yet been achieved (Mazzucchelli et al., 2010). To date, focal therapy has involved ablation of large portions of the prostate (e.g., entire lobes or hemi-ablation). Therefore, it was not previously known or expected that treating low-grade, low-risk isolated prostate cancer by targeting only the solitary tumor would be effective in reducing cancer incidence, cancer grade, and cancer progression (worsening) throughout the prostate. As a result, clinical trials were not designed to evaluate FT for treating only solitary prostate cancer, and it was not expected to be effective in treating the entire prostate.

[0010] A common technique used in identifying and monitoring prostate cancer is assessing PSA levels in conjunction with biopsy evaluation. A typical prostate biopsy usually involves taking multiple samples through the prostate and evaluating the tissue using a Gleason score. The Gleason score measures how abnormal cancer cells appear under a microscope and is a good indicator of how quickly the cells may grow and spread. The Gleason score is calculated by adding together the two grades of cancer that make up the largest area of ​​the biopsied tissue sample and is often expressed as two numbers, e.g., 3 + 3, along with a total score, e.g., 6. The Gleason score for the primary pattern is listed first, and the secondary score is listed second. A biopsy with a Gleason grade of 3 + 4 (total 7) is considered less severe than a biopsy with a Gleason grade of 4 + 3 (total 7), because the primary pattern is greater than 3. Therefore, when assessing tumor severity, clinicians review not only the total Gleason score but also the primary and secondary pattern values. The following table provides a classification of prostate cancer groups and their associated risks. [Table 1]

[0011] For low-grade, low-risk localized (T1c) prostate cancer with a Gleason score of ≦6, active surveillance (AS) is the usual course of treatment. This is because, although some may very well mature into high-risk cancer (e.g., Gleason score ≧8) requiring radical prostatectomy, many do not. As mentioned above, up to approximately 35% of surgically removed prostates contain only low-grade, low-risk localized (T1c) prostate cancer. Therefore, these patients whose prostates have been surgically removed but who do not yet have high-risk cancerous tumors unnecessarily suffer the harmful effects of radical prostatectomy. Therefore, the typical standard of care is for patients with a Gleason score of 7 or higher, especially those with a primary pattern of 4, to undergo corrective treatment, either by removing most of the prostate, chemotherapy or radiation, or radical prostatectomy. Therefore, it is desirable to find a safe and effective method of treating low and / or low and intermediate risk prostate cancer patients with localized treatment of a single lesion, where the treatment is effective in improving, reducing and / or preventing the progression of cancer throughout the prostate.

[0012] Like prostate cancer, other cancers are known to be multifocal in nature, preceded by and associated with solitary tumors or small cancerous tumors that are not significant enough to justify treatment. Such multifocal cancers include, but are not limited to, one or more of the following: squamous cell head and neck cancer (HNSCC), cutaneous squamous cell carcinoma (cutaneous SCC), breast cancer, adenocarcinoma and SCC of the lung, esophageal cancer, gastric cancer, colon cancer, bladder cancer, cervical cancer, melanoma, brain cancer, pancreatic cancer, ovarian cancer, bone marrow cancer, and leukemia.

[0013] Throughout this description, including the foregoing description of the related art, all publicly available documents (including all U.S. patents and published patent applications) mentioned herein are specifically incorporated herein by reference in their entirety. The foregoing description of the related art is not an admission in any way that any of the above documents, including pending U.S. patent applications, are prior art to the present disclosure. Furthermore, any statements herein about disadvantages associated with the described products, methods, and / or apparatus are not intended to limit the embodiments in any way. Indeed, aspects of the embodiments may include certain features of the described products, methods, and / or apparatus without suffering from the described disadvantages. Summary of the Invention

[0014] There remains a need in the art for new, less toxic, less frequent, and essentially non-invasive treatments for preventing or reducing the progression or incidence of multiple cancers. There also remains a need in the art for such treatments that reduce the incidence of multiple cancers. Embodiments meet these needs.

[0015] This disclosure is premised, in part, on the discovery that pharmaceutically active ingredients capable of inducing necrosis of a solitary cancer tumor can be administered to the tumor, while still having the unexpected effect of reducing the incidence of multiple cancers, reducing the grade of multiple cancers, and reducing the progression (worsening) of multiple cancers in the affected organ or entire organism. Suitable pharmaceutically active ingredients capable of inducing necrosis of such tumors include, for example, fexapotide triflutate (FT), the 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 ( These include leuprolide acetate), Erida (apalutamide), Farmagon (degarelix), flutamide, goserelin acetate, Jevtana (cabazitaxel), mitoxantrone hydrochloride, Nilandrone (nilutamide), Provenzi (sipuleucel-T), sipuleucel-T, Taxotere (docetaxel), Xofigo (radium-223 dichloride), Xtandi (enzalutamide), Zoladex (goserelin acetate), and mixtures and combinations thereof. Such administration has unexpectedly been found to be effective in reducing the incidence of multiple cancers, reducing the grade of multiple cancers, and reducing the progression (worsening) of multiple cancers in affected organs or throughout the organism after several years of continuous follow-up, and has not been limited to local effectiveness. Thus, embodiments can greatly improve the quality of life for many men with multiple cancers who would otherwise undergo more aggressive treatments such as ablation of large parts of the organ, chemotherapy, radiation, or complete removal of the gland (e.g., radical prostatectomy, colectomy, lung removal or transplant, etc.).

[0016] Some embodiments are directed to a method of reducing the incidence of multiple cancers, reducing the grade of multiple cancers, and reducing the progression (worsening) of multiple cancers in a mammal with low-grade or low-risk multiple cancers (i.e., for prostate cancer, having a Gleason score of ≦6) 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 cancer tumors. The method comprises administering a therapeutically effective amount of the composition to a single cancer focus (solitary tumor) and reducing the percentage of mammals exhibiting one or more new lesions in an organ or whole organism.

[0017] In certain embodiments, the method comprises administering a therapeutically effective amount of FT to a single cancerous lesion (sole tumor) in the prostate of a mammal, and reducing the percentage of mammals exhibiting one or more new lesions with an increase in Gleason grade throughout the prostate (new solitary with Gleason upgrading) by about 40% to about 100% compared to active surveillance when measured at least 18 months after treatment, or by about 30% to about 90% compared to active surveillance when measured at least 36 months after treatment, or by about 5% to about 85% compared to active surveillance when measured at least 48 months after treatment. The method also includes administering a therapeutically effective amount of the composition to a single cancerous lesion (solitary tumor) in the prostate of a mammal, and reducing the percentage of mammals exhibiting one or more new lesions in the entire prostate with an increase in the primary Gleason grade pattern (new solitary lesions with a primary Gleason pattern ≧4) by an amount of about 40% to about 100% compared to active surveillance when measured at least 18 months after treatment, or about 30% to about 100% compared to active surveillance when measured at least 36 months after treatment, or about 20% to about 100% compared to active surveillance when measured at least 48 months after treatment.

[0018] The compositions can be administered intramuscularly, orally, intravenously, intraperitoneally, intracerebrally (intraparenchymal), intraventricularly, intratumorally, intralesionally, intradermally, intrathecally, intranasally, intraocularly, intraarterially, topically, transdermally, via aerosol, by infusion, bolus injection, implantable device, sustained release system, and the like.

[0019] In another embodiment, the composition comprises a therapeutically effective amount of FT administered in an amount ranging from about 2.0 mg to about 20 mg.

[0020] In another embodiment, the method comprises administering a therapeutically effective amount of FT to a single cancerous lesion (solitary tumor) in the prostate of a mammal, and reducing the percentage of mammals that have had conventional cancer treatment (surgery, radiation, or chemotherapy) and exhibit one or more new lesions in the entire prostate with an increase in Gleason grade (new solitary with Gleason upgrade) by about 50% to about 100% compared to active surveillance when measured at least 18 months after treatment, or by about 40% to about 90% compared to active surveillance when measured at least 36 months after treatment, or by about 35% to about 85% compared to active surveillance when measured at least 48 months after treatment. The method also includes administering a therapeutically effective amount of FT to a single cancerous lesion (solitary tumor) in the prostate of a mammal, and reducing the percentage of mammals that have had conventional cancer treatment (surgery, radiation, or chemotherapy) and exhibit one or more new lesions in the entire prostate with an increase in the primary Gleason grade pattern (new solitary lesions with a primary Gleason pattern of ≧4) by about 70% to about 100% when measured at least 18 months after treatment, compared to active surveillance, or by about 60% to about 100% when measured at least 36 months after treatment, compared to active surveillance, or by about 50% to about 100% when measured at least 48 months after treatment, compared to active surveillance.

[0021] Both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the claimed embodiments. Other objects, advantages and features will become readily apparent to those skilled in the art from the following detailed description of each embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0022] Before reviewing the proteins, nucleotide sequences, peptides, compositions, active agents, etc., and methods of the present invention, it is to be understood that the present invention is not limited to the particular methodology, protocols, cell lines, vectors, and reagents. 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 each embodiment of the present invention, which is limited only by the appended claims.

[0023] The terms and phrases used herein are defined as follows, unless otherwise indicated. Throughout this specification, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, a reference to a "host cell" includes a plurality of host cells, and a reference to an "antibody" is a reference to one or more antibodies and equivalents thereof known to those skilled in the art.

[0024] The amino acids and amino acid residues described herein may be referred to according to the commonly accepted one-letter or three-letter codes shown in the table below. [Table 2]

[0025] The expression "suitable pharmaceutically active ingredient capable of inducing necrosis of such tumors" includes, for example, fexapotide triflutate (FT), the 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 Leu

[0013] 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). 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 which are incorporated herein by reference in their entireties. FT is expressed as follows: SEQ ID NO. 1 (SEQ ID NO: 1): IDQQVLSRIKLEIKRCL or Ile-Asp-Gln-Gln-Val-Leu-Ser-Arg-Ile-Lys-Leu-Glu-Ile-Lys-Arg-Cys-Leu

[0026] The term "fragment" refers to a protein or polypeptide consisting of a contiguous subsequence of the amino acid sequence of a protein or peptide, including naturally occurring fragments such as splice variants and fragments resulting from naturally occurring in vivo protease activity. Such fragments may be truncated at the amino terminus, carboxy terminus, and / or internally (such as by natural splicing). Such fragments may be prepared with or without an amino-terminal methionine. The term "fragment" includes identical or different fragments from the same protein or peptide, having common or non-common contiguous amino acid sequences linked directly or via a linker. Using the guidelines and procedures outlined herein, one of skill in the art will be able to select suitable fragments for use in embodiments without undue experimentation.

[0027] The term "variant" refers to a protein or polypeptide in which one or more amino acid substitutions, deletions, and / or insertions are present when compared to the amino acid sequence of the protein or peptide described herein, and includes naturally occurring allelic or alternatively spliced ​​variants of the proteins or peptides so described. The term "variant" includes the substitution of one or more amino acids in a peptide sequence with similar or homologous amino acid(s) or dissimilar amino acid(s). There are many criteria for which amino acids can be identified as similar or homologous. (Gunnar von Heijne, Sequence Analysis in Molecular Biology, pp. 123-39 (Academic Press, New York, NY, 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 shown in Table 2 below. [Table 3] Table 3 shows an alternative scheme for amino acid substitution. [Table 4]

[0028] Other variants can consist of less conservative amino acid substitutions, such as selecting residues with more significant differences in their impact on (a) the structure of the polypeptide backbone in the region of the substitution, e.g., as a sheet or helix conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) maintaining the bulk of the side chain. In general, substitutions expected to have a more significant effect on function are those in which (a) glycine and / or proline are substituted for another amino acid or deleted or inserted; (b) hydrophilic residues, such as seryl or threonyl, are substituted for (or by) hydrophobic residues, such as leucyl, isoleucyl, phenylalanyl, valyl, or alanyl; (c) cysteine ​​residues are substituted for (or by) any other residue; (d) residues with electropositive side chains, such as lysyl, arginyl, or histidyl, are substituted for (or by) negatively charged residues, such as glutamyl or aspartyl; or (e) residues with large side chains, such as phenylalanine, are substituted for (or by) one lacking such a side chain, such as glycine. Other mutants include those designed to either generate new glycosylation and / or phosphorylation sites or to delete 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 containing additional amino acid residues before or after the protein or peptide amino acid sequence on a linker peptide. For example, cysteine ​​residues can be added to both the amino and carboxy termini of a peptide to allow for cyclization of the peptide by disulfide bond formation. The term "variant" also includes polypeptides having the amino acid sequence of a peptide with at least one and up to 25 or more additional amino acids adjacent to either the 3' or 5' end of the peptide.

[0029] The term "derivative" refers to a chemically modified protein or polypeptide that has been chemically modified not only by natural processes, such as processing and other post-translational modifications, but also by chemical modification techniques, such as the addition of one or more polyethylene glycol molecules, sugars, phosphates, and / or other such molecules when such molecules are not naturally attached to the wild-type protein or peptide. Derivatives include salts. Such chemical modifications are well described in basic textbooks and in more detailed monographs, as well as in a voluminous research literature, and are well known to those skilled in the art. It will be recognized that the same type of modification may be present in the same or varying degrees at several sites in a given protein or polypeptide. Also, a given protein or polypeptide may contain many types of modifications. Modifications can occur anywhere in the protein or polypeptide, including the peptide backbone, amino acid side chains, and the amino or carboxy 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 nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of 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, hydroxylation and ADP-ribosylation of glutamic acid residues, selenoylation, sulfation, transfer RNA-mediated addition of amino acids to proteins, e.g., arginylation, and ubiquitination.See, e.g., Proteins--Structure And Molecular Properties, 2nd ed., TECreighton, W.H. Freeman and Company, New York (1993); and Wold, F., "Posttranslational Protein Modifications: Perspectives and Prospects," in Posttranslational Covalent Modification Of Proteins, pp. 1-12, 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. NY Acad. Sci. 663:48-62 (1992). The term "derivative" includes chemical modifications that result in proteins or polypeptides that are branched, or branched and cyclic, or cyclic without branching. Cyclic, branched and branched circular proteins or polypeptides may result from posttranslation natural processes and may also be made by entirely synthetic methods.

[0030] The term "homologue" refers to a protein that is at least 60% identical in amino acid sequence to a peptide, as determined by commonly used standard methods for comparing the similarity in amino acid positions of two polypeptides. The degree of similarity or identity between two proteins can be measured using, but is not limited to, 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.), 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.

[0031] Preferred computer program methods useful in 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 program is 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 (University of Wisconsin Genetics Computer Group, Madison, Wisconsin), the two proteins or polypeptides for which the percent sequence identity is to be determined are aligned so that their respective amino acids are optimally matched (the "matched span" as determined by the algorithm).

[0032] The algorithm is used in conjunction with a gap opening penalty (calculated as three times the average diagonal; "average diagonal" is the average of the diagonals of the comparison matrix used; "diagonal" is the score or number assigned to each perfect amino acid match by a particular comparison matrix) and a gap extension penalty (usually 1 / 10 of the gap opening penalty), as well as a comparison matrix such as PAM250 or BLOSUM62. Standard comparison matrices (for the PAM250 comparison matrix, see Dayhoff et al.: Atlas of Protein Sequence and Structure, vol. 5, supp. 3; for the BLOSUM62 comparison matrix, see Henikoff et al., Proc. Natl. Acad. Sci USA, 89: 10915-10919) can also be used with the algorithm. The algorithm then calculates percent identity. Homologues will typically have one or more amino acid substitutions, deletions, and / or insertions when compared with the corresponding protein or peptide, as the case may be.

[0033] The term "fusion protein" refers to a protein in which one or more peptides are recombinantly fused or chemically conjugated (including covalently and non-covalently) to a protein, such as (but not limited to) an antibody or an antibody fragment, such as a Fab fragment or a single-chain Fv. The term "fusion protein" also refers to multimers (i.e., dimers, trimers, tetramers, and higher multimers) of peptides. Such multimers include homomeric multimers comprising one peptide; heteromeric multimers comprising two or more peptides; and heteromeric multimers comprising at least one peptide and at least one other protein. Such multimers may be the result of hydrophobic, hydrophilic, ionic, and / or covalent association, binding, or linkage; may be formed by cross-linking using linker molecules; or may be linked indirectly, for example, by liposome formation.

[0034] 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 peptidic in nature, i.e., no longer contains any peptide bonds (i.e., amide bonds between amino acids). The term peptidomimetic is used in a broader sense herein to include molecules that are no longer completely peptidic, such as pseudopeptides, semi-peptides, and peptoids. Examples of peptidomimetics in this broader sense (where portions of a peptide are replaced with structures lacking peptide bonds) are described below. Whether completely non-peptide or partially non-peptide, peptidomimetics according to each embodiment provide a spatial arrangement of reactive chemical moieties that closely mimics the three-dimensional arrangement of the active groups of the peptide on which they are based. As a result of this similarity in the geometry of the active site, peptidomimetics have effects on biological systems that resemble the biological activity of peptides.

[0035] The peptidomimetics of the present invention are preferably substantially similar to the peptides described herein in both three-dimensional shape and biological activity. Examples of methods for structurally modifying peptides to generate peptidomimetics known in the art include inversion of the main-chain chiral center to introduce D-amino acid residue structures, particularly at the N-terminus, which result in enhanced stability against proteolytic degradation without adversely affecting activity. One example is described in the article "Tritiated D-ala1-Peptide T Binding," Smith CS et al., Drug Development Res., 15, 371-379 (1988). A second method is the modification of cyclic structures for 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). An example of this is provided by conformationally restricted 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 approach is to replace peptide bonds within the peptide with pseudopeptide bonds that confer resistance to proteolysis.

[0036] Several pseudopeptide bonds have been described that generally do not affect peptide structure and biological activity. One example of this approach is to replace them with retro-inverso pseudopeptide bonds ("Biologically active retroinverso analogues of thymopentin," Sisto A. et al., Rivier JE and Marshall GR (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). Following this modification, the amino acid sequence of the peptide may be identical to that of the above-mentioned peptide, except that one or more peptide bonds are replaced with retro-inverso pseudopeptide bonds. Preferably, the most N-terminal peptide bond is substituted, since such a substitution will confer resistance to proteolysis by exopeptidases acting on the N-terminus. Further modifications can be made by replacing the amino acid's chemical group with another chemical group of similar structure. Another suitable pseudopeptide bond known to increase stability against enzymatic cleavage with little or no 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).

[0037] Thus, the amino acid sequences of these peptides can be identical to those of peptides except that one or more peptide bonds have been replaced by isostere pseudopeptide bonds. Preferably, the most N-terminal peptide bond is replaced, as this would confer resistance to proteolysis by exopeptidases acting on the N-terminus. The synthesis of peptides with one or more reduced isostere pseudopeptide bonds is known in the art (Couder et al. (1993), supra). Other examples include the replacement of a peptide bond by the introduction of a ketomethylene or methylsulfide bond.

[0038] Peptoid derivatives of the peptides described herein represent another type of peptidomimetic, retaining structural determinants important for biological activity but eliminating peptide bonds, thereby conferring resistance to proteolysis (Simon et al., 1992, Proc. Natl. Acad. Sci. USA, 89:9367-9371, incorporated herein by reference in its entirety). Peptoids are oligomers of N-substituted glycines. Several N-alkyl groups have been described, each corresponding to the side chain of a natural amino acid (Simon et al. (1992), cited above). Some or all of the amino acids of a peptide can be replaced with N-substituted glycines corresponding to the replaced amino acid.

[0039] The term "peptidomimetic" or "mimetic" also includes reverse-D peptides and enantiomers, as defined below.

[0040] The term "reverse D peptide" refers to a biologically active protein or peptide consisting of D-amino acids arranged in reverse order as compared to the L-amino acid sequence of the peptide. Thus, the carboxy terminal residue of the L-amino acid peptide becomes the amino terminal residue of the D-amino acid peptide, etc. For example, the peptide ETESH has the structure H d S d E d T d E d where Ed , H d , S d , and T d are the D-amino acids corresponding to the L-amino acids E, H, S, and T, respectively.

[0041] 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.

[0042] As used herein, the term "composition" broadly refers to any composition containing the recited peptide or amino acid sequence and, optionally, additional active agents. The composition can include a dry formulation, an aqueous solution, or a sterile composition. Compositions containing peptides can be used as hybridization probes. The probes can be stored in lyophilized form and can be associated with stabilizers such as carbohydrates. For hybridization, the probes can be placed in aqueous solutions containing salts, e.g., NaCl; detergents, e.g., sodium dodecyl sulfate (SDS); and other components, e.g., Denhardt's solution, powdered milk, salmon sperm DNA, etc.

[0043] The term "low-grade prostate cancer" refers to prostate tissue with a highest Gleason grade of ≦6 or 3+3 detected by biopsy, i.e., prostate cancer representing a single or multiple lesion biopsy. It will be understood that in many cases, biopsy procedures that take multiple samples from the prostate do not sample the entire gland, and therefore other lesions may exist that were not detected. The term "low-grade isolated prostate cancer" refers to a single cancerous lesion with a Gleason grade of ≦6 or 3+3. The term "progressing prostate cancer" typically refers to a higher Gleason grade in any single lesion (the highest grade in all biopsies taken together is the grade), but also refers to a greater amount of cancer in the biopsy (i.e., a higher percentage of cancer in the lesions of a given biopsy, for example, if it is more than 50%; or more lesions positive for cancer). For example, if a patient progresses from one positive core sample with 5% tumor at one time point to one with five cores each with 40% tumor (all the same Gleason grade) at a later time point, this would not be a "Gleason grade progression," but would be considered progression. On the other hand, a "Gleason grade progression" exists if a patient progresses from four cores with 40% tumor each, grade 6, and then has only one positive core with 5% tumor, but with Gleason grade 7; therefore, the progression would be considered a "Gleason grade progression."

[0044] When referring to a "biopsy," one skilled in the art will recognize that a typical biopsy consists of multiple "quadrant" samples, usually at least 10 or 12, sampling all areas of the gland (left and right; apex, middle, and bottom for each; and middle and lateral, and L and R transitions for each), thus equating to 14 zones. Thus, reference to "biopsying" or "biopsy" refers to 10-15 biopsies taken simultaneously, each reported separately.

[0045] Throughout this specification, the term "multiple cancers" refers to one or more cancers selected from prostate cancer, squamous cell head and neck cancer (HNSCC), cutaneous squamous cell carcinoma (cutaneous SCC), breast cancer, lung adenocarcinoma and SCC, esophageal cancer, stomach cancer, colon cancer, bladder cancer, cervical cancer, melanoma, brain cancer, pancreatic cancer, ovarian cancer, bone marrow cancer, and leukemia. Throughout this specification, the term "prevention" or variations thereof does not necessarily mean complete prevention in all cases, but instead refers to preventing the development or occurrence of multiple cancers compared to subjects treated with active surveillance, etc. For example, assuming that 60% of patients with solitary cancer or low-grade, low-risk cancer develop multiple cancers over a 36-month period without treatment (or treated with a control), only 20% develop multiple cancers over a 36-month period when treated according to the embodiments. Thus, the treatments described herein will be effective in preventing from developing multiple cancers in many patients who would otherwise have developed multiple cancers if not treated according to the embodiments.

[0046] In embodiments in which an additional active agent is used in conjunction with the composition, the term "active agent" is used to refer to any agent capable of removing unwanted cell and / or tissue growths. Suitable active agents may include, but are not limited to, (i) anti-cancer active agents (such as alkylating agents, topoisomerase I inhibitors, topoisomerase II inhibitors, RNA / DNA antimetabolites, and antimitotic agents), (ii) active agents for treating benign growths, for example, anti-acne and anti-wart active agents, (iii) anti-androgenic compounds (cyproterone acetate (1α,2β-methylene-6-chloro-17α-acetoxy-6-dehydroprogesterone), tamoxifen, aromatase inhibitors), (iv) alpha 1-adrenergic receptor blockers (tamsulosin, terazosin, doxazosin, prazosin, bunazosin, indoramin, alfuzosin, silodosin), (v) 5α-reductase inhibitors (finasteride, dutasteride), (vi) phosphodiesterase type 5 (PDE5) inhibitors (tadalafil) and combinations thereof.

[0047] Some embodiments relate to a method for reducing the incidence of prostate cancer, reducing the grade of prostate cancer, and reducing the progression (worsening) of prostate cancer by administering a composition comprising at least one pharmaceutically active ingredient capable of inducing necrosis of low-grade, low-risk localized prostate cancer tumors to low-grade, low-risk localized prostate cancer, and in the entire prostate where the solitary tumor is initially located and treated. Suitable pharmaceutically active ingredients capable of inducing necrosis of such tumors 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 ( The compositions include leuprolide acetate, Erida (apalutamide), Farmagon (degarelix), flutamide, goserelin acetate, Jevtana (cabazitaxel), mitoxantrone hydrochloride, Nilandrone (nilutamide), Provenzi (sipuleucel-T), sipuleucel-T, Taxotere (docetaxel), Xofigo (radium-223 dichloride), Xtandi (enzalutamide), Zoladex (goserelin acetate), and mixtures and combinations thereof. In certain embodiments, the composition is administered two or more times. Thus, embodiments provide a method for reducing the incidence, grade, and progression of prostate cancer in a non-invasive manner by administering the composition to a mammal that would typically not be treated. Generally, mammals with low-grade solitary or multiple prostate cancers with a Gleason grade of ≦6 are permitted to undergo active surveillance (AS) or no treatment. 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).

[0048] The inventor has unexpectedly discovered that administering such compositions to a single lesion of multiple cancers significantly reduces the occurrence of multiple cancers, the grade of cancer, and the progression of cancer in organs or the whole body.Therefore, the method of the embodiment provides a non-invasive method of reducing multiple cancers compared with organ removal or transplantation, or even lesion ablation, resection, chemotherapy or radiation.Even active surveillance requires multiple and repeated prostate biopsies and evaluations, which places a heavy burden on medical systems.Therefore, the method described herein is useful for delaying the occurrence, appearance and progression of multiple cancers in a non-invasive manner.

[0049] With respect to prostate cancer, in contrast to published literature, mammals treated with the compositions of the present invention showed a dramatic reduction in the incidence of multiple prostate cancers, a dramatic reduction in the increase in multiple Gleason scores, and a dramatic reduction in the progression of multiple prostate cancers throughout the prostate. Embodiment methods can reduce multiple prostate cancers with Gleason upgrade (patients with one or more new lesions throughout the prostate with an increased Gleason grade) by about 40% to about 100%, or about 50% to about 90%, or about 50% to about 80%, or any value therebetween, when measured at least 18 months after treatment, compared to active surveillance controls. Embodiment methods can reduce multiple prostate cancers with Gleason upgrade by about 30% to about 90%, or about 45% to about 90%, or about 47% to about 80%, or any value therebetween, when measured at least 36 months after treatment, compared to active surveillance controls. Methods of the embodiments can reduce whole prostate multifocal cancers with Gleason upgrade by about 5% to about 85%, or about 10% to about 70%, or about 12% to about 65%, or any amount therebetween, compared to active surveillance controls, when measured at least 48 months after treatment.

[0050] Methods of the embodiments can reduce multiple whole prostate cancers with a primary Gleason grade pattern of ≥ 4 (patients with one or more new lesions in the whole prostate with an increasing primary Gleason grade pattern) by about 40% to about 100%, or about 70% to about 100%, or about 80% to about 100%, or any value therebetween, when measured at least 18 months after treatment, compared to an active surveillance control. Methods of the embodiments can reduce multiple whole prostate cancers with a primary Gleason grade pattern of ≥ 4 by about 30% to about 100%, or about 70% to about 100%, or about 80% to about 100%, or any value therebetween, when measured at least 36 months after treatment, compared to an active surveillance control. The method of the embodiments can reduce whole prostate cancers with a primary Gleason grade pattern ≧4 by an amount of 20% to about 100%, or about 50% to about 100%, or about 60% to about 100%, or any value therebetween, compared to an active surveillance control, when measured at least 48 months after treatment.

[0051] The method of the embodiment can reduce multiple prostate cancers in the lobe (or hemi-prostate) where lesions with Gleason upgrade were originally treated (patients with one or more new lesions in the treated lobe or hemi-prostate with an increased Gleason grade) by 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, compared to an active surveillance control. The method of the embodiment can reduce multiple hemi-prostate cancers with Gleason upgrade by 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, compared to an active surveillance control. Methods of the embodiments can reduce hemiprostatic multifocal carcinoma with Gleason upgrade by about 15% to about 80%, or about 15% to about 75%, or about 16% to about 72%, or any value therebetween, compared to active surveillance controls, when measured at least 48 months after treatment.

[0052] The method of the embodiment can reduce multiple prostate cancers in the lobe (or hemi-prostate) where lesions with a primary Gleason pattern ≥ 4 were initially treated (patients with one or more new lesions in the treated lobe or hemi-prostate with an increase in the primary Gleason pattern grade) by about 50% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any value therebetween, when measured at least 18 months after treatment, compared to an active surveillance control. The method of the embodiment can reduce multiple hemi-prostate cancers with a primary Gleason pattern ≥ 4 by about 45% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any value therebetween, when measured at least 36 months after treatment, compared to an active surveillance control. Methods of embodiments can reduce hemiprostatic multifocal cancers with Gleason upgrading (patients with one or more new lesions throughout the prostate with an increased Gleason grade) by 45% to about 100%, or about 60% to about 100%, or about 70% to about 100%, or any amount therebetween, compared to active surveillance controls, when measured at least 48 months after treatment.

[0053] The method of the embodiment can reduce the percentage of mammals undergoing conventional cancer treatment (surgery, radiation, or chemotherapy) that exhibit one or more new lesions in the whole prostate with an increase in Gleason grade (cancer treatment with new lesions and Gleason upgrade) by about 50% to about 100%, or about 55% to about 90%, or about 60% to about 80%, or any value therebetween, when measured at least 18 months after treatment, compared to an active surveillance control. The method of the embodiment can reduce the percentage of mammals undergoing conventional cancer treatment (surgery, radiation, or chemotherapy) that exhibit one or more new lesions in the whole prostate with an increase in Gleason grade (cancer treatment with new lesions and Gleason upgrade) by about 40% to about 90%, or about 45% to about 90%, or about 50% to about 80%, or any value therebetween, when measured at least 36 months after treatment, compared to an active surveillance control. Methods of the embodiments can reduce cancer treatment with new foci and Gleason upgrades throughout the prostate by about 35% to about 85%, or about 40% to about 85%, or about 48% to about 84%, or any amount therebetween, compared to an active surveillance control, when measured at least 48 months after treatment.

[0054] The method of the embodiment can reduce the percentage of mammals who have had conventional cancer treatment (surgery, radiation, or chemotherapy) and who exhibit prostate-wide multiple cancers with a primary Gleason grade pattern of ≥ 4 (cancer treatment with new multiples and increasing primary Gleason grade patterns) by about 70% to about 100%, or about 70% to about 100%, or about 80% to about 100%, or any value therebetween, when measured at least 18 months after treatment, compared to an active surveillance control. The method of the embodiment can reduce cancer treatment with new multiples and increasing primary Gleason grade patterns by about 60% to about 100%, or about 70% to about 100%, or about 80% to about 100%, or any value therebetween, when measured at least 36 months after treatment, compared to an active surveillance control. Methods of the embodiments can reduce cancer treatment associated with new multifocality and an increase in primary Gleason grade pattern by 50% to about 100%, or about 70% to about 100%, or about 80% to about 100%, or any amount therebetween, compared to an active surveillance control, when measured at least 48 months after treatment.

[0055] The method of the embodiment can reduce the percentage of mammals with conventional cancer treatment (surgery, radiation, or chemotherapy) and multiple prostate cancers in the lobe (or hemi-prostate) where lesions with Gleason upgrades were initially treated (cancer treatment with new multiples with increased Gleason grade in the hemi-prostate) by about 40% to about 100%, or about 50% to about 90%, or about 60% to about 75%, or any value therebetween, when measured at least 18 months after treatment compared to an active surveillance control. The method of the embodiment can reduce the percentage of cancer treatment with new multiples with increased Gleason grade in the hemi-prostate by about 50% to about 90%, or about 55% to about 75%, or about 50% to about 80%, or any value therebetween, when measured at least 36 months after treatment compared to an active surveillance control. The method of the embodiments can reduce new multifocal cancer treatments with increased Gleason grade in the hemiprostate by about 15% to about 80%, or about 35% to about 75%, or about 40% to about 75%, or any amount therebetween, compared to an active surveillance control, when measured at least 48 months after treatment.

[0056] The method of the embodiment can reduce the percentage of mammals with multiple prostate cancers in the lobe (or hemi-prostate) that has undergone conventional cancer treatment (surgery, radiation, or chemotherapy) and in which lesions with a primary Gleason pattern of ≥ 4 were initially treated (cancer treatment associated with new multifoci and an increase in the primary Gleason grade in the hemi-prostate) by about 65% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any value therebetween, when measured at least 18 months after treatment, compared to an active surveillance control. The method of the embodiment can reduce the percentage of mammals with multiple prostate cancers in the lobe (or hemi-prostate) that have undergone conventional cancer treatment (surgery, radiation, or chemotherapy) and in which lesions with a primary Gleason pattern of ≥ 4 were initially treated (cancer treatment associated with new multifoci and an increase in the primary Gleason grade in the hemi-prostate) by about 65% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any value therebetween, when measured at least 36 months after treatment, compared to an active surveillance control. The method of the embodiments can reduce cancer treatment associated with new multifocality and an increase in primary pattern of Gleason grade in the hemiprostate by 60% to about 100%, or about 65% to about 100%, or about 75% to about 100%, or any value therebetween, compared to an active surveillance control, when measured at least 48 months after treatment.

[0057]

[0010] Embodiments include methods of treating a mammal with low-grade, solitary prostate cancer, comprising administering a composition comprising FT, either alone or in combination with an additional active agent, one or more times to a single low-grade, low-risk prostate cancer lesion present in the prostate of the mammal. Methods include, but are not limited to, administering the composition, either alone or in combination with a carrier, intramuscularly, orally, intravenously, intraperitoneally, intracerebrally (intraparenchymal), intraventricularly, intralesionally, intraocularly, intraarterially, intrathecally, intratumorally, intranasally, topically, transdermally, subcutaneously, or intradermally.

[0058] The present invention is effective for any mammal, including humans, mice, rabbits, dogs, sheep, and other domestic animals, mammals that are or can be treated by veterinarians, zookeepers, or wildlife conservation personnel. Preferred mammals include humans, sheep, and dogs. The terms "mammal" and "patient" are used interchangeably herein.

[0059] It will be apparent to those skilled in the art that other smaller fragments of FT can be selected so that these peptides have the same or similar biological activity. Those skilled in the art can also select other fragments of FT so that these peptides have the same or similar biological activity. Thus, the term "FT" as used in the embodiments encompasses these other fragments. Generally, the peptides of each embodiment have at least four amino acids, preferably at least five amino acids, and more preferably at least six amino acids.

[0060] Additionally, embodiments of the present invention include therapeutic methods comprising administering a composition comprising FT, which is comprised of two or more linked FT sequences, together with an additional active agent. As long as FT has the desired biological activity, the two or more FT sequences will also have the desired biological activity.

[0061] FT and its fragments, variants, derivatives, homologs, fusion proteins, and mimetics encompassed by embodiments of the present invention can be produced using methods known to those of skill in the art. For example, methods such as recombinant DNA technology, protein synthesis, and isolation of naturally occurring peptides, proteins, variants, derivatives, and homologs can be used. FT and its fragments, variants, derivatives, homologs, fusion proteins, and mimetics can be produced from other peptides, proteins, and their fragments, variants, derivatives, and homologs using methods known to those of skill in the art. Such methods include, but are not limited to, the use of proteases to cleave peptides or proteins to FT. For example, any of the 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, the disclosures of which are incorporated herein by reference in their entireties, can be used to prepare the FT peptides described herein.

[0062] When an additional active agent is used in addition to one or more pharmaceutically active ingredients, the additional active agent can be one or more active agents selected from the following: (i) anti-cancer active agents (such as alkylating agents, topoisomerase I inhibitors, topoisomerase II inhibitors, RNA / DNA antimetabolites, and mitotic inhibitors), (ii) active agents for treating benign growths, for example, anti-acne and anti-wart active agents (salicylic acid), (iii) anti-androgenic compounds (cyproterone acetate (1α,2β-methylene-6-chloro-17α-acetoxy-6-dehydroprogesterone), tamoxifen, aromatase inhibitors), (iv) α1-adrenergic receptor blockers (tamsulosin, terazosin, doxazosin, prazosin, bunazosin, indoramin, alfuzosin, silodosin), (v) 5α-reductase inhibitors (finasteride, dutasteride), (vi) phosphodiesterase type 5 (PDE5) inhibitors (tadalafil) and combinations thereof. Preferably, the additional agent is an anti-cancer agent, particularly an agent useful in the treatment of prostate cancer.

[0063] The therapeutic compositions described herein may contain a certain amount of a pharmaceutically active ingredient mixed 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, while in other embodiments, the composition containing the pharmaceutically active ingredient is administered as an injection, while the additional active agent is formulated in the form of an oral medication (gel, capsule, tablet, liquid, etc.). The carrier material can be water for injection, preferably water supplemented with other materials typically contained in solutions for administration to mammals. Typically, when the pharmaceutically active ingredient is FT, FT is administered in the form of a composition containing purified FT peptide (or chemically synthesized FT peptide) in combination with one or more physiologically acceptable carriers, excipients, or diluents. Neutral buffered saline or saline mixed with serum albumin are examples of suitable carriers. Preferably, the product is formulated as a lyophilizate using appropriate excipients (e.g., sucrose). Other standard carriers, diluents, and excipients may be included if desired. The compositions of the embodiments may also contain a buffer solution having a pH value in a suitable range known to those skilled in the art, such as a Tris buffer solution having a pH of about 7.0 to 8.5 or an acetate buffer solution having a pH of about 4.0 to 5.5, which may further contain sorbitol or a suitable substitute thereof.

[0064] 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 may be mixed 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 extenders such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; (c) binders such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (d) humectants such as glycerol; and (e) agar, calcium carbonate. (f) disintegrating agents such as potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (f) solution retarding agents such as paraffin; (g) absorption accelerators such as quaternary ammonium compounds; (h) wetting agents such as acetyl alcohol and glycerol monostearate; (i) adsorbents such as kaolin and bentonite; and (j) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0065] Oral liquid dosage forms include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs.In addition to the active compound, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers.Examples of emulsifiers include ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils such as cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols, fatty acid esters of sorbitan, or mixtures of these substances.

[0066] Besides such inert diluents, the composition can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0067] The actual dosage level of the active ingredient in the compositions of the present invention may be varied to obtain an effective amount of the pharmaceutically active ingredient and additional active agent to obtain the desired therapeutic response for the particular composition. Thus, the selected dosage level will depend on the desired therapeutic effect, the route of administration, the desired duration of treatment, and other factors.

[0068] For mammals, including humans, effective amounts can be administered based on body surface area. The correlation of doses (mg / M) for animals of various sizes and species, and for humans, is 2 Body surface area (based on body surface area) is reported by E. J. Freireich et al., Cancer Chemother. Rep., 50(4):219 (1966). Body surface area can be approximately determined from an individual's height and weight (see, e.g., Scientific Tables, Geigy Pharmaceuticals, Ardsley, NY, pp. 537-538 (1970)).

[0069] It will be understood that the specific dosage level for any particular patient will vary depending on a variety of factors, such as body weight, general health, sex, diet, time and route of administration, potency of the administered drug, absorption and excretion rates, combination with other drugs, and the severity of the particular disease being treated.

[0070] Methods of administering compositions comprising a pharmaceutically active ingredient according to embodiments include, but are not limited to, administering the composition intramuscularly, orally, intravenously, intraperitoneally, intracerebrally (intraparenchymal), intraventricularly, intratumorally, intralesionally, intradermally, intrathecally, intranasally, intraocularly, intraarterially, topically, rectally, transperitoneally, transdermally, via aerosol, by infusion, bolus injection, implantable device, sustained release system, and the like. 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.

[0071] The use of FT is a preferred embodiment. Based on tissue culture genetic array data, FT is a novel molecule that stimulates the caspase pathway (activation of caspases 7, 8, 10, caspase recruitment domains 6, 11, 14, and DIABLO), tumor necrosis factor pathway (activation of TNF1, TNFSF6, TNFSF8, TNFSF9, CD70 ligand, TNFRSF19L, TNFRSF25, TRAF2, TRAF3, TRAF4, and TRAF6 receptor), and BCL pathway (activation of BIK, HRK, BCL2L10, and BCL3) in prostate epithelial cells in vitro. FT selectively induces loss of cell membrane integrity, mitochondrial metabolic arrest, RNA depletion, DNA dissolution and aggregation, cell fragmentation, and cell loss. The apoptotic process is characterized by progressive ultrastructural changes, including membrane disruption and swelling, progressively deeper nuclear infiltration, and ultimately membrane blebbing, followed by cell death and fragmentation into apoptotic bodies. Histologically, typical apoptotic changes, including immunohistochemical positivity for apoptotic markers, are evident throughout the injection site for several weeks after treatment.

[0072] FT has been extensively tested in patients with BPH. This compound and a placebo control were administered rectally in over 1,700 procedures in nine human clinical trials. In these large-scale, long-term clinical trials in patients with BPH, FT was administered at a concentration of 0.25 mg / ml (2.5 mg of FT corresponds to approximately 15-20% of the prostate volume). See, for example, 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). Therefore, it is preferred that a composition containing FT contains at least 2.5 mg of FT and can be administered in a single dose of up to 25 mg of FT. In another embodiment, FT is administered in an amount ranging from about 2.5 mg to about 200 mg, or from about 2.5 mg to about 15 mg. In one embodiment, FT is administered in an amount of 15 mg.

[0073] The following examples are provided to illustrate various embodiments of the present invention. However, it should be understood that various embodiments are not limited to the specific conditions or details described in these examples. Throughout this specification, any and all references to publicly available documents, including U.S. patents, are specifically incorporated herein by reference. In particular, embodiments of the present invention 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, and 2016 / 0215031, each of which discloses that certain peptides specified therein are effective agents for inducing cell death in vivo in normal rodent tissues such as muscle tissue, subcutaneous connective tissue, and dermis. [Example]

[0074] In a series of clinical studies, a total of 146 men with low-grade prostate cancer (Gleason grade ≦6) were treated in the following manner: Patients were randomized to receive a single intrarectal intraprostatic injection of a composition containing 2.5 mg of FT (n=49) or 15 mg of FT (n=48), or to receive controlled active surveillance (n=49). After the first follow-up biopsy 45 days after randomization, 18 patients in the controlled active surveillance group were crossed over to receive a single dose of the composition: 10 patients receiving 2.5 mg of FT and 8 patients receiving 15 mg of FT. Patients were followed for 5 years, including biopsies at baseline, 45 days, 18, 36, and 48 months, and urological evaluations with PSA every 6 months. Patients whose Gleason grade increased or who elected surgical or radiotherapy intervention were excluded from the study but were still included in the data analysis. Baseline lesion quadrant and median tumor grade and the percentage of normal biopsies in volume were assessed, and progression was measured by clinical and pathological outcomes, including Gleason grade and primary Gleason grade pattern, for the whole sampled prostate and treated prostate lobes. The total incidence of interventions associated with an increase in Gleason grade and any conventional surgical interventions (e.g., surgery, radiation, and / or chemotherapy) was assessed.

[0075] Example 1 This example evaluates the percentage of patients who showed two or more new lesions throughout the prostate (i.e., progressed from solitary to multiple), where the new lesions were accompanied by an increase in the total score of Gleason grade ("new multiple with Gleason upgrade"). The data in the following table for the follow-up time represent the percentage of patients who showed progression or worsening. The results are shown in Table 3 below. [Table 5]

[0076] The results from Example 1 demonstrate that embodiments can reduce the percentage of patients who had multiple new cancers of the whole prostate with Gleason upgrade by about 40% to about 100%, or about 50% to about 90%, or about 50% to about 80%, or any amount therebetween, when measured at least 18 months after treatment, compared to an active surveillance control. Methods of embodiments can reduce the percentage of patients who had multiple new cancers of the whole prostate with Gleason upgrade by about 30% to about 90%, or about 45% to about 90%, or about 47% to about 80%, or any amount therebetween, when measured at least 36 months after treatment, compared to an active surveillance control. Methods of embodiments can reduce the percentage of patients who had new multiple cancers throughout the prostate with Gleason upgrade by about 5% to about 85%, or about 10% to about 70%, or about 12% to about 65%, or any amount therebetween, compared to active surveillance controls, when measured at least 48 months after treatment.

[0077] Example 2 This example evaluated the percentage of patients who developed two or more new lesions throughout the prostate (i.e., progressed from solitary to multiple), where the new lesions were accompanied by an increase in Gleason primary pattern of ≥ 4 ("new multiple with increase in Gleason primary pattern"). The results are shown in Table 4 below. [Table 6]

[0078] The results from Example 2 demonstrate that embodiments can reduce the percentage of patients who had multiple new cancers throughout the prostate with an increasing primary Gleason pattern by about 40% to about 100%, or about 70% to about 100%, or about 80% to about 100%, or any amount therebetween, when measured at least 18 months after treatment compared to an active surveillance control. Methods of embodiments can reduce the percentage of patients who had multiple new cancers throughout the prostate with an increasing primary Gleason pattern by about 30% to about 100%, or about 70% to about 100%, or about 80% to about 100%, or any amount therebetween, when measured at least 36 months after treatment compared to an active surveillance control. The method of the embodiments can reduce the percentage of patients who had whole prostate multiple cancers with a primary pattern of increasing Gleason grade by an amount of 20% to about 100%, or about 50% to about 100%, or about 60% to about 100%, or any value therebetween, compared to an active surveillance control, when measured at least 48 months after treatment.

[0079] Example 3 This example evaluates the percentage of patients who developed two or more new lesions (i.e., progressed from solitary to multiple) in a hemiprostate that was initially treated for a solitary tumor, and the new lesions were accompanied by an increase in the total Gleason grade score ("multiple cancers of the hemiprostate with Gleason upgrade"). The results are shown in Table 5 below. [Table 7]

[0080] The results from Example 3 demonstrate that embodiments can reduce the percentage of patients who had multiple prostate cancers in the lobe (or hemi-prostate) where lesions with Gleason upgrade were initially treated (patients with one or more new lesions in the treated lobe or hemi-prostate with an increased Gleason grade) by about 40% to about 100%, or about 50% to about 90%, or about 60% to about 80%, or any amount therebetween, when measured at least 18 months after treatment, compared to an active surveillance control. Embodiment methods can reduce the percentage of patients who had multiple hemi-prostate cancers with Gleason upgrades by about 50% to about 90%, or about 50% to about 85%, or about 50% to about 80%, or any amount therebetween, when measured at least 36 months after treatment, compared to an active surveillance control. Methods of the embodiments can reduce the percentage of patients who had hemiprostatic multifocal carcinoma with Gleason upgrade by about 15% to about 80%, or about 15% to about 75%, or about 16% to about 72%, or any amount therebetween, compared to active surveillance controls, when measured at least 48 months after treatment.

[0081] Example 4 This example evaluates the percentage of patients who developed two or more new lesions in a hemiprostate gland initially treated for a solitary tumor (i.e., progressed from solitary to multiple), where the new lesions were accompanied by an increase in Gleason grade of the primary pattern of ≥ 4 ("multiple carcinomas of the hemiprostate gland with an increase in Gleason grade of the primary pattern"). The results are shown in Table 6 below. [Table 8]

[0082] The results from Example 4 demonstrate that embodiments can reduce the percentage of patients who had multiple hemiprostatic carcinomas with an increasing primary Gleason pattern by about 50% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any amount therebetween, when measured at least 18 months after treatment compared to an active surveillance control. Methods of embodiments can reduce the percentage of patients who had multiple hemiprostatic carcinomas with an increasing primary Gleason pattern by about 45% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any amount therebetween, when measured at least 36 months after treatment compared to an active surveillance control. Methods of the embodiments can reduce the percentage of patients who had hemiprostatic multiple carcinomas with an increased Gleason primary pattern by between 45% and about 100%, or between about 60% and about 100%, or between about 70% and about 100%, or any amount therebetween, compared to active surveillance controls, when measured at least 48 months after treatment.

[0083] Example 5 This example evaluates the percentage of patients who have received conventional cancer treatment (e.g., surgery, radiation and / or chemotherapy) and have two or more new lesions throughout the prostate (i.e., progress from solitary to multiple), where the new lesions are accompanied by an increase in the total Gleason grade score ("cancer treatment with new multiple lesions with Gleason upgrade").The results are shown in Table 7 below. [Table 9]

[0084] The results from Example 5 demonstrate that embodiments can reduce the percentage of patients with cancer treatment who had new multifocalities with Gleason upgrade in the whole prostate by about 50% to about 100%, or about 55% to about 90%, or about 60% to about 80%, or any amount therebetween, when measured at least 18 months after treatment, compared to an active surveillance control. Methods of embodiments can reduce the percentage of patients with cancer treatment who had new multifocalities with Gleason upgrade in the whole prostate by about 40% to about 90%, or about 45% to about 90%, or about 50% to about 80%, or any amount therebetween, when measured at least 36 months after treatment, compared to an active surveillance control. Methods of the embodiments can reduce the percentage of patients who had cancer treatment with new multifocality with Gleason upgrade throughout the prostate by about 35% to about 85%, or about 40% to about 85%, or about 48% to about 84%, or any amount therebetween, compared to active surveillance controls, when measured at least 48 months after treatment.

[0085] Example 6 This example evaluates the percentage of patients who have received conventional cancer treatment (e.g., surgery, radiation, and / or chemotherapy) and have developed two or more new lesions throughout the prostate (i.e., progressed from solitary to multiple lesions), where the new lesions are accompanied by an increase in the primary Gleason grade pattern of ≥ 4 ("cancer treatment with new multiple lesions and primary Gleason grade pattern"). The results are shown in Table 8 below. [Table 10]

[0086] Results from Example 6 demonstrate that embodiments can reduce the percentage of patients who have cancer treatment with new whole prostate tumors and an increase in primary pattern of Gleason grade by about 70% to about 100%, or about 75% to about 100%, or about 80% to about 100%, or any amount therebetween, when measured at least 18 months after treatment, compared to an active surveillance control. Methods of embodiments can reduce the percentage of patients who have cancer treatment with new whole prostate tumors and an increase in primary pattern of Gleason grade by about 60% to about 100%, or about 70% to about 100%, or about 80% to about 100%, or any amount therebetween, when measured at least 36 months after treatment, compared to an active surveillance control. The method of the embodiments can reduce the percentage of patients who have had cancer treatment with new multifocalities and an increase in primary pattern of Gleason grade throughout the prostate by 50% to about 100%, or about 70% to about 100%, or about 80% to about 100%, or any amount therebetween, compared to an active surveillance control, when measured at least 48 months after treatment.

[0087] Example 7 This example evaluated the percentage of patients who had received conventional cancer treatment (e.g., surgery, radiation, and / or chemotherapy) and had multiple prostate cancers in the lobe (or hemi-prostate) in which lesions with Gleason grade upgrades were initially treated (those who had new multiple lesions with increased Gleason grade in the hemi-prostate). The results are shown in Table 9 below. [Table 11]

[0088] The results from Example 7 demonstrate that embodiments can reduce the percentage of patients who have had cancer treatment with new cancers with Gleason upgrade in the hemi-prostate by about 40% to about 100%, or about 50% to about 90%, or about 60% to about 75%, or any amount therebetween, when measured at least 18 months after treatment, compared to an active surveillance control. Embodiment methods can reduce the percentage of patients who have had cancer treatment with new cancers with an increased Gleason grade in the hemi-prostate by about 50% to about 90%, or about 55% to about 75%, or about 50% to about 80%, or any amount therebetween, when measured at least 36 months after treatment, compared to an active surveillance control. The method of the embodiments can reduce the percentage of patients who have had cancer treatment with new multifocality with increased Gleason grade in the prostate hemisphere by about 15% to about 80%, or about 35% to about 75%, or about 40% to about 75%, or any amount therebetween, compared to active surveillance controls, when measured at least 48 months after treatment.

[0089] Example 8 This example evaluated the percentage of patients with multiple prostate cancers in a lobe (or hemi-prostate) that had received conventional cancer treatment (surgery, radiation, or chemotherapy) and in which a lesion with a primary Gleason pattern of ≥ 4 was initially treated (cancer treatment associated with new multiple lesions in the hemi-prostate and an increase in the primary Gleason pattern). The results are shown in Table 10 below. [Table 12]

[0090] The results from Example 8 demonstrate that embodiments can reduce the percentage of patients who have had cancer treatment with new multiple cancers in the hemi-prostate with an increase in the primary pattern of Gleason grade by about 65% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any amount therebetween, when measured at least 18 months after treatment, compared to an active surveillance control. Methods of embodiments can reduce the percentage of patients who have had cancer treatment with new multiple cancers in the hemi-prostate with an increase in the primary pattern of Gleason grade by about 65% to about 100%, or about 70% to about 100%, or about 75% to about 100%, or any amount therebetween, when measured at least 36 months after treatment, compared to an active surveillance control. The method of the embodiments can reduce the percentage of patients who have had cancer treatment with new multifocality in the hemiprostate and an increase in the primary pattern of Gleason grade by 60% to about 100%, or about 65% to about 100%, or about 75% to about 100%, or any amount therebetween, compared to an active surveillance control, when measured at least 48 months after treatment.

[0091] The results from the foregoing examples demonstrate the unexpectedly superior efficacy of the pharmaceutically active ingredients, particularly FT, in reducing the incidence of multifocal cancer, reducing the grade of multifocal cancer, and reducing the progression (worsening) of multifocal cancer in whole or hemi-prostates initially harboring low-grade tumors and treated. It will be apparent to those skilled in the art that various modifications and variations can be made in the methods and compositions of the present invention without departing from the spirit or scope of the invention.

Claims

1. 1. A composition for reducing the incidence of multiple cancers, reducing the grade of multiple cancers, and reducing the progression (worsening) of multiple cancers in the entire prostate of a mammal with solitary prostate cancer, comprising: The composition comprises an intraprostatic intratumoral injectable solution comprising 15 mg of fexapotide triflutate (FT) and neutral buffered saline; The injectable solution is 15 mg of FT lyophilisate dissolved in neutral buffered saline; A composition for identifying a mammal having a solitary prostate cancer tumor, wherein when the composition is administered by direct injection into the solitary prostate cancer tumor, the incidence of multiple cancers throughout the prostate, the grade of multiple cancers, and the progression (worsening) of multiple cancers are reduced.

2. When the composition is administered, new multiple prostate cancers associated with an increase in Gleason grade throughout the prostate gland are a) a 40% to 100% reduction compared to active surveillance when measured at least 18 months after treatment; b) a 30% to 90% reduction compared to active surveillance when measured at least 36 months after treatment; c) a 5% to 85% reduction compared to active surveillance when measured at least 48 months after treatment; The composition of claim 1.

3. When the composition is administered, new multiple prostate cancers associated with a primary pattern of increasing Gleason grade throughout the prostate gland are a) a 40% to 100% reduction compared to active surveillance when measured at least 18 months after treatment; b) a 30% to 100% reduction compared to active surveillance when measured at least 36 months after treatment; c) a 20% to 100% reduction compared to active surveillance when measured at least 48 months after treatment; The composition of claim 1.

4. When the composition is administered, new multiple prostate cancers associated with conventional cancer treatments and an increase in Gleason grade throughout the prostate are a) a 50% to 100% reduction compared to active surveillance when measured at least 18 months after treatment; b) a 40% to 90% reduction compared to active surveillance when measured at least 36 months after treatment; c) a 35% to 85% reduction compared to active surveillance when measured at least 48 months after treatment; The composition of claim 1.

5. When the composition is administered, new multiple prostate cancers associated with conventional cancer treatments and an increase in the primary pattern of Gleason grade throughout the prostate gland are a) a 70% to 100% reduction compared to active surveillance when measured at least 18 months after treatment; b) a 60% to 100% reduction compared to active surveillance when measured at least 36 months after treatment; c) a 50% to 100% reduction compared to active surveillance when measured at least 48 months after treatment; The composition of claim 1.

6. When the composition is administered, new multiple prostate cancers associated with conventional cancer treatments and an increase in the primary pattern of Gleason grade in the prostate lobes where low-grade solitary prostate cancer lesions were located are a) a 65% to 100% reduction compared to active surveillance when measured at least 18 months after treatment; b) a 65% to 100% reduction compared to active surveillance when measured at least 36 months after treatment; c) A 60% to 100% reduction compared to active surveillance when measured at least 48 months after treatment; The composition of claim 1.

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