Recent Patents on Live Bacteria and Their Products As Potential Anti- Cancer Agents

Recent Patents on Anti-Cancer Drug Discovery, 2012, 7, 31-55 31 Recent Patents on Live Bacteria and their Products as Potential Anti- cancer Agents

1, 1 2 Arsenio M. Fialho *, Nuno Bernardes and Ananda M. Chakrabarty

1Institute for Biotechnology and Bioengineering (IBB), Center for Biological and Chemical Engineering, Institute Supe- rior Tecnico, Lisbon, 1049-001, Portugal; 2Department of Microbiology & Immunology, University of Illinois College of Medicine, 835 South Wolcott Avenue, Chicago, IL 60612, USA

Received: November 10, 2010; Accepted: January 6, 2011; Revised: March 26, 2011

Abstract: This review intends to provide a comprehensive coverage of the various patents, published or issued, since 2007 on live or attenuated bacteria as potential anticancer agents, as well as microbial products including toxins, enzymes, antibiotics, various proteins and peptides as well as other small molecular weight products. Below is a list of such published/issued patents and a summary of the main contents of many such patents. Keywords: Anticancer drugs, bacteria, bacterial products, cancer, patents.

INTRODUCTION strains expressing genes encoding foreign enzymes that have the ability to convert non-toxic prodrugs to cytotoxic drugs. Each year, many patents covering the use of live bacteria and their products as anti-cancer agents are filed or pub- The remaining patents (376) cover a wide range of bacte- lished. A review on the patents covering the current and po- rial products as anti-cancer agents, including among others, tential use of bacterial proteins and peptides derived from proteins, enzymes, immunotoxins and secondary metabolites them in cancer therapy was published in 2007 [1]. Now, we (Part B, Tables 2 to 6). Overall, these patents represent 87 intend to update our previous analysis as well as to extend it percent of total covered patents. In recent years, such over- to live bacteria and their products. These searches were per- representation gained a special relevance resulting in the formed through a worldwide patent database maintained by disclosure of many bacterial products with cancer killing the European Patent Office, based on the search terms ability. (“name of the bacteria” or “name of the bacterial product” and “cancer”) appearing in the abstract and title. The search A. LIVE OR ATTENUATED BACTERIAL STRAINS over the period of January 2007-August 2011, resulted in a AS ANTICANCER AGENTS list of 433 patents. Each of these patents was examined, The use of live bacteria and their products has long been categorized and some of them cited in the text and Tables 1 to 6. investigated and recognized as potential cancer treatments. In the early 1890’s, William B. Coley, a surgeon in the Me- Among the patents selected, 57 cover the use of bacteria morial Hospital in New York, reported for the first time, (either wild-type or recombinant strains) as anti-cancer spontaneous tumour regression in patients suffering from agents. A special emphasis is given to attenuated strains of streptococcal infections. He developed a mixture consisting intracellular pathogens such as Salmonella, Mycobacterium, of extracts of killed bacteria and used it as a vaccine to treat Listeria and Clostridium acting as immunotherapy agents, advanced cancers. After the treatment, approximately 10% of pro-drug modification agents and delivering anti-tumor patients achieved a complete remission of tumour [2-4]. agents. Overall, such patents describe the use of facultative Such observation led to the emergence of immunotherapy as or obligate anaerobic attenuated bacteria that have the poten- a therapeutic approach to treat cancer. Many scientists since tial to specifically target the hypoxic core region of solid then know that certain live, attenuated and engineered mi- tumors, thereby causing cytotoxicity to the cancer cells (Part croorganisms such as Clostridium, Bifidobacterium, Salmo- A, Table 1). In general the disclosures of these patents are nella, Mycobacterium and Listeria have the ability to act as biological materials, experimental strategies and pharmaceu- anticancer agents Fig. (1) [5]. They act as effectors in cancer tical compositions involving the use of attenuated bacterial immunotherapy or are capable of selectively growing in hypoxic areas of tumors. Because of their selectivity for tumor micro-environment, these bacteria are also promising vectors for delivering therapeutic genes for anticancer therapies [5]. *Address correspondence to these authors at the Department of Microbiol- In this review, we presented an update of the various patents, ogy & Immunology, University of Illinois College of Medicine, 835 South published or issued, since 2007 on live bacteria as anticancer Wolcott Avenue, Chicago, IL 60612, USA; Tel: +1 312 9964586; Fax: +1 312 9966415; E-mails: [email protected]; [email protected] agents (Part A, Table 1).

Mycobacterium (IA)

LIVE OR ATTENUATED Clostridium BACTERIAL STRAINS AS ANTICANCER AGENTS (DATA)

Listeria (IA)

Bifidobacterium (DATA)

Fig. (1). - (A) - Live bacteria used to treat cancer. Wild-type or attenuated bacterial strains were used as immunotherapy agents (IA), pro-drug modification agents (PDMA) and delivering anti-tumor agents (DATA). (B) – Number of recent issued patents (2007-2011) covering the use of live bacteria as anticancer agents. A worldwide patent search was performed (http://worldwide.espacenet.com/; August 2011, maintained by the European Patent Office) based on the search terms (“name of the bacteria” and “cancer”) appearing in the abstract and title. Each of these patents was examined, categorized and some of them cited in Table 1.

Table 1. Selected List of Recent Worldwide Issued and Published Patents on Live or Attenuated Bacterial Strains as Anticancer Agents.

LIVE OR ATTENUATED BACTERIAL STRAINS AS ANTICANCER AGENTS Recent Issued and Published Patents (2007-2011)

Bacteria

Microorganism Strain CRC2631 of Salmonella typhimurium and its Use as a Cancer Therapeutic Publication number: US2010135973; Publication date: 2010-06-03; Inventors: Eisenstark Abraham, Kazmierczak Roberta DNA vaccines against tumor growth and methods of use thereof Publication number: US2010136058; Publication date: 2010-06-03; Inventors: Luo Yunping, Xiang Rong Salmonella Method of cancer treatment (5 out 17) Publication number: US2010098665; Publication date: 2010-04-22; Inventors: Leonard Arnold, Saltzman Daniel A New strain of Salmonella enterica S. typhimurium, its use and a method to obtain a therapeutic vaccine vector Publication number: WO2010095966; Publication date: 2010-08-26; Inventors: Michael Bereta, Chorobik Paulina Method of cancer treatment Publication number: CA2701052; Publication date: 2010-10-17; Inventors: Saltzman Daniel A, Leonard Arnold,Mueller Mark J

Vaccine adjuvant properties of liposomes formed at elevated temperatures from the polar chloroform extractable lipids from Mycobacterium bovis Publication number: US 2008248094; Publication date: 2008-10-09; Inventors:Sprott G Dennis, Krishnan Lakshmi, Sad Subash Process for obtaining a product containing polar lipids and mycobacterial hydrophilic substances from Mycobacterium tuberculosis used as a vaccine or immunostimulant

Publication number: MX2008001575; Publication date: 2009-07-3; Inventor:Garcia Ursino Pacheco Mycobacte- Combination therapy of hybrid cells with BCG injection for treating Cancer Patients rium Publication number: US2009087450; Publication date: 2009-04-02; Inventors:Wagner Thomas E, Wei Yanzhang (5 out of 18) Recombinant Mycobacterium strain expressing a mycobacterial FAP protein under the control of a promoter active under

hypoxia and its application for cancer therapy Publication number: HK 1112753; Publication date: 2010-10-22; Inventors: Marchal Gilles, Abolhassani Mohammad Pharmaceutical composition containing Mycobacterium tuberculosis-EIS recombinant protein as an active ingredient Publication number: KR20100119161; Publication date: 2010-11-09; Inventors:Jo Eun Kyeong, Kim Kwang Kyu, Shin Dong Min, Lee Hye Mi Bacterial Strains and Products as Anticancer Agents Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 33

(Table 1) Contd….

LIVE OR ATTENUATED BACTERIAL STRAINS AS ANTICANCER AGENTS Recent Issued and Published Patents (2007-2011)

Bacteria

Therapy with clostridium perfringens enterotoxin to treat ovarian and uterine cancer Publication number: US2009291047; Publication date: 2009-11-26; Inventor: Santin Alessandro D Clostridium botulinum C3 exotransferase compositions and methods for treating tumour spreading Publication number: US2009285833; Publication date: 2009-11-19; Inventors: Mckerracher Lisa, Lasko Dana

Growth inhibitor of colon cancer cells Clostridium Publication number: JP2009269836; Publication date: 2009-11-19; Inventor: Nakanishi Takasuke (5 out of 10) Compositions and methods comprising genetically enhanced obligate and facultative anaerobic bacteria for oncopathic

therapy Publication number: WO2009111177; Publication date: 2009-09-11; Inventors: Woo Savio L C, Li Zhiyu Use of Clostridium butyricum in preparing medicine composition for preventing and curing intestinal cancer Publication number: CN101306020; Publication date: 2008-11-19; Inventors: Yunlong Cui, Hongfu Li

Specific immunotherapy of cancer using live recombinant bacterial vaccine vector Publication number: JP 2007254485; Publication date: 2007-10-04 ; Inventors: Paterson Yvonne Dual delivery system for heterologous antigens Publication number: WO2009143167; Publication date: 2009-11-26; Inventors: Maciag Paulo, Wallecha Anu; Shahabi Vafa Listeria Compositions and methods for treatment of cervical cancer (5 out of 10) Publication number: US2009081250; Publication date: 2009-03-26; Inventors: Paterson Yvonne, Rothman John Compositions, methods and uses for stimulating immune responses Publication number: WO2011060093; Publication date: 2011-05-19; Inventors: Lenz Laurel L, Schmidt Rebecca L Live Listeria-based vaccines for central nervous system therapy Publication number: WO2011100754; Publication date: 2011-08-18; Inventors: Shahabi Vafa, Seavey Matthew, Paterson Yvonne

Compositions and methods comprising genetically enhanced obligate and facultative anaerobic bacteria for oncopathic Bifidobacte- therapy rium Publication number: WO2009111177; Publication date: 2009-09-11; Inventors: Woo Savio L C, Li Zhiyu (2 out of 2) Fermented fruit and anti-cancer pharmaceutical composition containing the same Publication number: KR20070024447; Publication date: 2007-03-02; Inventors: Kim Dong Hyun, Kim Sang Wook

constructs are introduced in an attenuated Salmonella typhi- Patents on Live Bacterial Strains as Anti-Cancer Agents murium strain (aroA- dam-). This patent also describes meth- i). Salmonella Strains as Anti-Cancer Agents ods of inhibiting tumor growth, how to vaccinate patients against cancer, and how to deliver the genetic material to a Microorganism Strain CRC2631 of Salmonella typhi- mammalian cell in vivo [7]. murium and its Use as a Cancer Therapeutic Method of Cancer Treatment This invention describes the construction of Salmonella strains with a disruption of at least one out of three genes This patent describes a method and a composition for the (aroA, rfaH, and thyA). It also provides a method to treat treatment of cancer, comprising the administration of a live cancer including the step of administering these Salmonella attenuated Salmonella typhimurium strain harbouring a strains [6]. plasmid carrying a gene that encodes a truncated form of the human interleukin-2 [8]. DNA Vaccines Against Tumor Growth and Methods of Use Thereof New Strain of Salmonella enterica S. typhimurium, its Use and a Method to Obtain a Therapeutic Vaccine Vector This patent discloses a DNA vaccine that elicits an immune response against cancer cells. It comprises, in a phar- This patent discloses the use of a bacterial strain of Sal- maceutically delivery system, a mixture containing two monella enterica serovar Typhimurium VNP20009 deposited genes, respectively encoding a Fra-1 protein and an interleu- in the Polish Collection of Microorganisms (access no. kin (IL-18). In a particular embodiment, these two genetic B/00024) efficient to be used as a vaccine against cancer 34 Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 Fialho et al. disease. Indeed, this patent describes the methodology used iii). Clostridium Strains as Anti-Cancer Agents to obtain this vaccine either from the wild-type strain or ge- Therapy with Clostridium perfringens Enterotoxin to Treat netically (attenuated) strains [9]. Ovarian and Uterine Cancer Method of Cancer Treatment This invention describes the existence of overexpressed The patent discloses the use of an attenuated Salmonella receptors on the surface of ovarian and uterine cancer cells typhimurium strain harbouring a plasmid that encodes a trun- that bind to Clostridium perfringens enterotoxin (CPE), cated form of the human interleukin-2. This strain is used namely claudin-3 and/or claudin-4 proteins. Moreover, this with success as a therapeutic agent against cancer diseases patent discloses the success of an in vivo treatment of a [10]. mouse model of ovarian cancer with intraperitoneal injection ii). Mycobacterium Strains as Anti-Cancer Agents of CPE. This invention also provides a method of treating ovarian cancer and uterine cancer through administration of Vaccine Adjuvant Properties of Liposomes Formed at Ele- CPE. Protection against CPE cell toxicity involves the ad- vated Temperatures from the Polar Chloroform Extract- ministration of a specific agent that binds to claudin-3 and/or able Lipids from Mycobacterium bovis claudin-4, thereby inhibiting CPE interaction [16]. This patent discloses methods and compositions to treat a Clostridium botulinum C3 Exotransferase Compositions cancer disease. The invention describes a liposome formula- and Methods for Treating Tumour Spreading tion prepared with polar lipids extracted from Mycobacte- This patent discloses pharmaceutical compositions com- rium bovis BCG. This pharmaceutical formulation is used to posed by a chimeric protein that conjugates a Clostridium modulate (increase) the immune system in a mammal, botulinum C3 exotransferase unit with a tumor-membrane thereby conferring cancer regression [11]. polypeptide. Spreading and migration of a metastatic cancer Process for Obtaining a Product Containing Polar Lipids cell can be inhibited by this pharmaceutical composition and Mycobacterial Hydrophilic Substances from Mycobac- [17]. terium tuberculosis Used as a Vaccine or Immunostimu- Growth Inhibitor of Colon Cancer Cells lant This patent provides a Clostridium butyricum cultivation This invention describes a method aiming to obtain a supernatant as an active ingredient to treat colon cancer [18]. microbial complex mixture derived from the bacterium My- cobacterium tuberculosis. It is composed by medium- and Compositions and Methods Comprising Genetically En- high-polarity lipids, proteins, peptides, polysaccharides and hanced Obligate and Facultative Anaerobic Bacteria for other hydrophilic molecules. This mixture could be obtained Oncopathic Therapy from any species of Mycobacterium and used as a vaccine This patent discloses genetically modified anaerobic bac- either in applied research, or in human or veterinary medi- teria useful to treat anoxic tumors. These bacteria contain cine [12]. one or more of the following gene mutations: superoxide Combination Therapy of Hybrid Cells with BCG Injection dismutase (sod), glutathione peroxidase (gpo), rubrerythrin for Treating Cancer Patients (rbr), alcohol dehydrogenase (ydaD) and phospholipase c (plc). Within other embodiments, genetically anaerobic bac- This invention discloses a method to treat patients with a teria expressing an inflammation suppressive gene such as a particular type of cancer (neuroblastoma), with a hybrid cell Staphylococcus aureus Panton-Valentine Leukocidin (PVL) preparation in combination with Mycobacterium bovis bacil- gene and/or aYersinia enterocolitica virulence factor (LcrV) lus Calmette-Guerin (BCG) [13]. gene, are also disclosed [19]. Recombinant Mycobacterium Strain Expressing a Myco- Use of Clostridium butyricum in Preparing Medicine Com- bacterial Fap Protein Under the Control of a Promoter position for Preventing and Curing Intestinal Cancer Active Under Hypoxia and its Application for Cancer Therapy This invention discloses a pharmaceutical composition used to prevent and treat intestinal cancers and chronic intes- This patent describes a recombinant vector expressing a mycobacterial gene encoding a FAP protein under the tran- tinal disease. It comprises a Clostridium butyricum mixture presented in the form of tablets, capsules, powders, liquid scriptional control of a hypoxia-sensitive promoter. Moreo- preparations or suppositories [20]. ver, this invention discloses the use of this genetic construction aiming to treat epithelial tumors [14]. iv). Listeria strains as anti-cancer agents Pharmaceutical Composition Containing Mycobacterium Specific Immunotherapy of Cancer Using Live Recombi- tuberculosis-EIS Recombinant Protein as an Active Ingre- nant Bacterial Vaccine Vector dient This invention discloses a vaccine containing a live re- This invention discloses an anti-cancer pharmaceutical combinant Listeria monocytogenes strain expressing a tu- composition containing a Mycobacterium tuberculosis strain mor-specific antigen. This vaccine proved to be effective expressing the EIS protein as the active ingredient [15]. against cancer [21].

Bacterial Strains and Products as Anticancer Agents Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 35

Dual Delivery System for Heterologous Antigens duction [5]. In this review, we presented an update of the various patents, published or issued, since 2007 on bacterial This patent describes the use of engineered Listeria products as anticancer agents (Part B, Tables 2 to 6). strains expressing a tumor-specific antigenic polypeptide and/or an angiogenic polypeptide. Genes are cloned in frame with a sequence encoding a PEST-containing polypeptide. 1. PATENTS ON BACTERIAL TOXINS AS ANTI- This patent also discloses methods of preparing same, and CANCER AGENTS methods of inducing an immune response, and treating, in- 1.1. Enterotoxin hibiting, or suppressing tumors [22]. Therapy with Clostridium Perfringens Enterotoxin to Treat Compositions and Methods for Treatment of Cervical Can- Ovarian and Uterine Cancer cer The summary of this invention is described in reference This invention provides methods of treating and inducing [16]. immune response against cervical cancer. It comprises the step of administering a genetically modified Listeria strain Claudins as Markers for Early Detection, Diagnosis, Prog- harbouring a foreign gene encoding a chimeric protein which nosis and as Targets of Therapy for Breast and Metastatic possesses a listeriolysin O fragment together with the E7 Brain or Bone Cancer and/or E6 proteins from Human papillomavirus (HPV). This invention discloses methods of diagnosis, prognosis, Moreover, this invention discloses methods for inducing an and treatment of breast and bone cancers, and of metastatic anti-E7 humoral response (anti-E7 antibody) [23]. brain cancer. The diagnostic and prognostic methods are Compositions, Methods and Uses for Stimulating Immune based on immunohistochemical detection of claudin recep- Responses tors, namely 1 and 7 (frequently underexpressed) and 3 and 4 (frequently overexpressed). The methods of treatment in- This patent describes the use for cancer treatment of a volve the use of Clostridium perfringens enterotoxin (or a novel pharmaceutical composition containing the p60 protein variant thereof) to lyse metastatic cancer cells [27]. from Listeria monocytogenes [24]. Antineoplastics Activities of Ellipticine and its Derivatives Live Listeria-Based Vaccines for Central Nervous System Therapy In this patent, gene expression assays together with hier- archical cluster data analysis are used to screen and distin- This patent discloses live Listeria-based recombinant guish between normal ovarian epithelial cells and papillary vaccines for cancer treatment [25]. carcinomas. The genes (over/underexpressed) used in this v). Bifidobacterium Strains as Anti-Cancer Agents screening are: i) - overexpressed in ovarian serous papillary carcinomas: larninin, tumor-associated calcium signal trans- Compositions and Methods Comprising Genetically En- ducer 1 and 2, claudin 3, claudin 4, ladinin I, SI00A2, SER- hanced Obligate and Facultative Anaerobic Bacteria for PIN2 (PAl-2), CD24, lipocalin 2, osteopontin, kallikrein 6, Oncopathic Therapy kallikrein JO, matriptase and stratifin; ii) - underexpressed in The summary of this invention is described in reference ovarian carcinoma: transforming growth factor beta receptor [19]. HI, platelet-derived growth factor receptor alpha, SE- MACAP3, ras homolog gene family, member I (ARHI) and Fermented Fruit and Anti-Cancer Pharmaceutical Compo- thrombospondin 2. The patent also provides methods of sition Containing the Same treating ovarian serous papillary carcinoma using the TROP- In this patent a treatment for multiple diseases including I /Ep-CAM monoclonal chimeric/humanized antibody. Fi- cancer is disclosed. As effective component, this pharmaceu- nally, in this invention, cancer treatment methods using the tical composition contains fermented fruit by lactic acid bac- enterotoxin CPE from Clostridium perfringens are also dis- teria or enterobacteria [26]. closed [28]. Mucosal Vaccine B. MICROBIAL PRODUCTS AS ANTICANCER This invention discloses a mucosal vaccine in which the AGENTS active compound is the C-terminal fragment of Bacillus Over the last decade, as stated above, several microor- welchii enterotoxin. Its administration in the surface of vari- ganisms have been used aiming the development of novel ous mucous membranes elicits an effective immunity (local therapies for treating cancer. Despite the encouraging results and systemic) thereby acting as an anti-cancer pharmaceuti- in some studies, a major disadvantage of their use has been cal composition [29]. the undesired infections caused by the bacteria themselves. Minigene In order to overcome this drawback, novel approaches for anticancer therapies are in course and include the use of puri- This invention describes the construction and use of vari- fied products from microbial origin such as proteins, en- ous chimeric genes susceptible to be used as vaccines againts zymes, immunotoxins, antibiotics and other secondary me- cancers and other diseases. As an example, one genetic con- tabolites Fig. (2). These approaches seek to overcome the struction contains i) a human tissue plasminogen signal pep- limitation of using live organisms and have been proved to tide; (ii) one T-cell epitope; and (iii) an E. coli heat labile be effectives in anticancer therapy causing tumor regression enterotoxin B subunit. Other embodiment of this invention through growth inhibition, cell cycle arrest or apoptosis in- 36 Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 Fialho et al.

A Enzymes: Antibiotics:

Toxins: Arginine deiminase Actinomycin

Cytosine deaminase Plicamycin Enterotoxin L-Methioninase Doxorubicin Diphtheria toxin Glutaminase Bleomycin Shiga toxin L-asparaginase Mitomycin Botulinum neurotoxin Rhizoxin Exotoxin A Staurosporine Listeriolysin S Borrelidin

BACTERIAL PRODUCTS AS ANTICANCER AGENTS Other proteins and Peptides:

Azurin and Laz p28 Plantaricin Other products:

Microcin Epothilones Pep27 Farnesyl transferase inhibitors Spiruchostatin Salinosporamide Romidepsin Belactosin Syringolin A Rapamycin

Fig. (2). - (A) - Purified bacterial products used to treat cancer. (B) – Number of recent issued patents (2007-2011) covering the use of bacterial products as anticancer agents. A worldwide patent search was performed (http://ep.espacenet.com; August 2011, maintained by the Euro- pean Patent Office) based on the search terms (“name of the bacterial product” and “cancer”) appearing in the abstract and title. Each of these patents was examined, categorized and some of them cited in Tables 2 to 6. describes methods of treating cancer by administering such variant that has activity of hindering an HB-EGF from bind- agents [30]. ing to an EGF receptor [33]. Activation of Recombinant Diphtheria Toxin Fusion Pro- 1.2. Diphtheria Toxin teins by Specific Proteases Highly Expressed on the Sur- Nucleic Acid Constructs, Pharmaceutical Compositions face of Tumor Cells and Methods of Using same for Treating Cancer This patent provides compositions and methods for inhib- This patent discloses the construction of a recombinant iting cancer cell growth. The invention provides nucleic acid immunotoxin based on the TNF alpha fused with the Diph- sequences encoding the diphtheria toxin (corresponding to theria toxin. This invention also describes the methods and the residues 1-388). Methods of treating cancer by adminis- uses of the immunotoxin as a novel anticancer agent [31]. tering such agent are described in this invention [34]. Diphtheria Toxin Variant Methods and Compositions Based on Diphtheria Toxin- Interleukin-3 Conjugates One embodiment of this invention describes the use of a recombinant diphtheria toxin (fused or not with another pep- This invention provides methods for inhibiting interleu- tide) in the treatment of cancer [32]. kin-3 receptor-expressing cells by using a diphtheria toxin- human interleukin-3 conjugate (DT-IL3). The DT-IL3 chi- Anticancer Agent mera comprises amino acids 1-388 of diphtheria toxin fused This patent provides an effective anticancer agent for with human interleukin-3. This recombinant protein was treating malignant tumors, particularly ovarian cancer. It used to inhibit the growth of cancer cells and/or cancer stem comprises as an active ingredient, a diphtheria toxin (DT) cells, such as myeloid leukemia cancer stem cells [35]. Bacterial Strains and Products as Anticancer Agents Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 37

1.3. Shiga toxin iniferatoxin, capsaicin, or tinyatoxin, and toxins, such as botulinum toxin. These formulations are used with success to Endoprostatic Srping (Stent) Impregnated with enhance the lifetime of a particular drug, e.g., antibiotics and Rapamycin/Sirolimus and Shiga Toxin Acting Against anticancer agents. This patent also discloses methods to pre- Cancer of Prostate and Benign Hyperplasias of the pare liposomes coated with antibodies that are useful for Urologic Tract targeting specific receptors for drug, peptide, polypeptide, or This patent discloses a novel pharmaceutical component nucleic acid delivery [42]. composed by the marketed Cypher (Cordis Company) and Methods and Compositions for the Treatment of Cancer Shiga-stx/stxA toxins. This mixture is used to impregnate an endoprostatic stent coated with rapamycin. The purpose of This patent discloses an anticancer pharmaceutical com- this invention is to impede the growth of hyperplasic benign position containing a botulinum toxin (BT) [43]. or malignant tissues [36]. Methods for Treating Cancers Treatment of Metastatic Colon Cancer with -Subunit of This patent provides methods for treating diverse cancers Shiga Toxin by local administration of a botulinum toxin [44]. This invention provides compositions and methods for treating invasive colon cancer through the use of the - 1.5. Exotoxin A subunit of shiga toxin as an immunotoxin [37]. Pseudomonas Exotoxin A Mutants and Uses Thereof Chimeric Polypeptide Comprising Fragment of Shiga Toxin and Peptide of Therapeutic Interest This invention discloses chimeric immunotoxin proteins that can be used as anticancer agents. They are constructed This patent describes the construction and use of a chi- based on antibodies or antibody fragments that have im- meric immunotoxin protein composed with the fragment of proved ability to bind the CD22 antigen (overexpressed in Shiga toxin as an active and cytotoxic compound. The cell cancer cells). In parallel, this invention describes the con- targeting moiety of this protein is one of various polypep- struction of a mutated Pseudomonas exotoxin A form that tides suitable to recognize a target, such as cancer cells [38]. has been shown to be highly cytotoxic. Finally, this patent, Shiga Toxin -Subunit as a Vector for Tumor Diagnosis based on the chimeric immunotoxins constructed, discloses and Drug Delivery to GB3 Expressing Tumors compositions and methods for inhibiting the growth and proliferation of cancers [45]. This invention describes the construction and use of a novel chimeric protein able to recognize Gb3 receptors, Chimeric Toxins for Targeted Therapy overexpressed in some tumors. The inventors disclose its use This invention describes chimeric toxins comprising of in cancer therapy and diagnosis. This chimeric protein con- cell targeting/killing moieties. The cell targeting and killing tains in its N-terminal the subunit of Shiga toxin linked by moieties correspond respectively to the hormone gonadotro- specific aminoacids to a particular molecule, selected in a pin and the Pseudomonas Exotoxin A. This patent also pro- group comprising: agents for in vivo diagnosis, cytotoxic vides a method for the production of these chimeric toxins agents, prodrugs, or enzymes for the conversion of a prodrug and their use to treat malignant carcinoma cells and benign to a drug [39]. hyperplasia including uterine leiomyoma cells, extrauterine Shiga Toxin -Subunit/Chemotherapeutics Conjugates endometrial island cells, benign hyperplasia of prostate and breast and pituitary tumor adenoma cells [46]. This invention discloses the use of an immunotoxin con- Methods for Treating Cancer Using an Immune-Toxin structed with the Shiga toxin -subunit moiety linked to a particular anti-cancer agent that requires intracellular uptake Comprising an Exotoxin a Moiety having a Furin Cleavage Site Replaced with a Cancer Associated Protease Site to exert anti-cancer activity. This patent also provides meth- Cleaved by MMP-2 or MMP-9 ods for treating cancer through the use of this chimeric protein [40]. Invention published in this patent provides a recombinant toxin (immunotoxin) having an ETA moiety that has the 1.4. Botulinum Neurotoxin furin site replaced with a cancer-associated protease site. Modifications in immunotoxins were made to provide Treating Neoplasms with Neurotoxin ligands that bind to cancer cells and attached to the ETA This invention discloses a method of treating a cancer moiety in which the furin site was replaced by a cancer- using the botylinum toxin (BTX). BTX inhibits the growth associated protease site. Methods for inhibiting or destroying of cancer cells and is also valuable in treating other diseases, mammalian cancer cells using these immunotoxins are also such as viral-induced growths, autoimmune disease, and HIV disclosed in this patent along with the compositions for treat- [41]. ing human cancer [47]. Application of Lipid Vehicles and use for Drug Delivery Tweak-Pseudomonas Exotoxin a Fusion Protein for Can- cer Therapy This invention discloses compositions and methods based on lipid pharmaceutical compositions to treat cancer and In this patent, TWEAK is fused with a modified (mutant) other diseases. In particular, this invention is related to Pseudomonas exotoxin (PE38). TWEAK is a member of liposome-based delivery of various compounds, such as res- soluble TNF superfamily ligand monomers, which spontane- ously associate into a homotrimeric structure that binds with 38 Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 Fialho et al. high affinity to a cell surface receptor(s). The resulting fu- ine deiminase achieves 1.6U/mL of fermentation broth. En- sion protein with the TWEAK domain binds with high affin- zyme produced by the strain and methods described in this ity to a fibroblast growth factor-inducible 14 (Fn14) cell sur- patent show ability to strongly inhibit a liver cancer cell line face receptor, acting as cytotoxins targeting various diseased [54]. cells, including cancer cells and cells undergoing cellular insult response [48]. 2.2. Cytosine Deiminase CD19-Specific Immunotoxin and Treatment Method Formulations of 5-Fluorocytosine and Uses Thereof This patent describes an immunotoxin construct com- This patent discloses pharmaceutical composition composed by: i) an anti-CD19 antibody lacking an Fc fragment, posed by the prodrug 5-fluorocytosine (5-FC) together with a and ii) a modified exotoxin A protein lacking domain I re- live bacteria overexpressing a gene encoding the cytosine sponsible for cell recognition and binding. These two do- deiminase enzyme. This enzyme is capable of converting the mains are held together by a peptide linker resistant to prote- prodrug 5-fluorocytosine to 5-fluorouracil to achieve a can- olytic degradation. Moreover, this immunotoxin can has a cer-treating effect [55]. modified C-terminal (KDEL sequence) aiming to promote its transport to the endoplasmic reticulum of cells. A method for Stem Cell Targeting of Cancer, Methods and Compositions inhibiting or destroying mammalian cancer cells using this Therefor recombinant immunotoxin is also disclosed [49]. Stem cells labeled with monocrystalline iron oxide nanoparticles are disclosed in this patent as a novel and 1.6. Listeriolysin S promising method for detecting and treating ovarian cancer. Magnetic resonance imaging (MRI) is the technology used to Specific Immunotherapy of Cancer Using Live Recombi- detect such cells. Treatment methods include administration nant Bacterial Vaccine Vector of stem cells overexpressing a gene encoding the cytosine In this patent, a vaccine is provided for the treatment of deiminase enzyme. This enzyme is capable of converting the cancer by means of a recombinant strain of Listeria monocy- prodrug 5-fluorocytosine to 5-fluorouracil to achieve a can- togenes expressing Listeriolysin (LLO) fused to a protein of cer-treating effect [56]. a fragment of a tumor-specific antigen, suppressing tumor Viral Vector Driven Mutant Bacterial Cytosine Deaminase formation in a host [50]. Gene and Uses Thereof Listeria Monocytogenes Cytotoxin Listeriolysin S This invention discloses a mutation in the gene encoding A Listeria monocytogenes cytotoxin, named Listeriolysin cytosine deaminase. The mutated form of cytosine deami- S, and a gene derived from Listeria pathogenicity island 3 nase interacts efficiently with its substrate, 5-fluorocytosine (LIPI-3) are disclosed in this patent as a therapeutic agent for (5-FC). In this invention, the mutant and wild-type genes are cancer treatment [51]. cloned into an appropriate vector. Moreover, the efficacy of the mutated cytosine deaminase form to convert the prodrug 2. PATENTS ON BACTERIAL ENZYMES AS ANTI- 5-FC into the toxic drug 5-fluorouracil (5-FU) was evalu- CANCER AGENTS ated. It is demonstrated that infection with the viral vector expressing the mutant gene resulted in increased 5-FC- 2.1. Arginine deiminase mediated in vitro cell killing. Furthermore, in vivo studies Mutated form of Arginine Deiminase proved the efficacy of this treatment. Finally, this invention discloses the success of a combined treatment using this en- This patent discloses pharmaceutical compositions based zyme/prodrug therapy together with radiotherapy [57]. on recombinant Arginine deiminase (ADI) that are used in the treatment of cancer and other diseases. ADI is genetically 2.3. L-Methioninase modified for a more efficient manufacturing and processing [52]. Conjugate for the Specific Targeting of Anticancer Agents to Tumor Cells or Tumor Vasculature and Production Use of Bacteria Endowed with Arginine Deiminase to In- Thereof duce Apoptosis and/or Reduce an Inflammatory Reaction and Pharmaceutical or Dietetic Compositions Containing This patent discloses a pharmaceutical composition con- Such Bacteria taining a specific ligand that interacts to an external overexpressed receptor or binding site on a tumor. In addition, this A bacteria endowed with arginine deiminase is used as a formulation is composed by an anti-cancer agent, such as the novel formulation to induce apoptosis and/or reduce an in- enzyme L-methioninase. The conjugate is used as an anti- flammatory reaction. Moreover, pharmaceutical or dietetic cancer agent [58]. compositions containing such bacteria are also dislosed [53]. A Strain Capable of Producing Arginine Deiminase and 2.4. Glutaminase the use Thereof Glutadon In this patent, an arginine deiminase producing bacterial strain is disclosed. This patent also describes the method of This invention is related to a pharmaceutical combination producing arginine deiminase by fermentation of Pseudo- product for a cancer therapy containing two active ingredi- monas plecoglossicida CGMCC No.2039. Activity of argin- ents: glutaminase and 6-diazo-5-oxo-L-norleucine (DON) Bacterial Strains and Products as Anticancer Agents Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 39

Table 2. Selected List of Recent Worldwide Issued and Published Patents on Bacterial Toxins as Anticancer Agents.

MICROBIAL PRODUCTS AS ANTICANCER AGENTS Toxins - Recent Issued and published patents (2007-2011)

Therapy with Clostridium perfringens enterotoxin to treat ovarian and uterine cancer Publication number: US2009291047; Publication date: 2009-11-26; Inventor: Santin Alessandro D Claudins as Markers for Early Detection, Diagnosis, Prognosis and as Targets of Therapy for Breast and Metastatic Brain or Bone Cancer Publication number: US2009155278; Publication date: 2009-06-18; Inventors: Sukumar Saraswati V, Kominsky Scott L Enterotoxin Antineoplastics activities of ellipticine and its derivatives (5 out of 5) Publication number: WO2007102869; Publication date: 2007-09-13; Inventor: Santin Alessandro D Mucosal vaccine Publication number: WO2010089940; Publication date: 2010-08-12; Inventors: Yagi Kiyohito, Kondoh Masuo, Isoda katsuhiro, Horigu- chi Yasuhiko Minigene Publication number: US2011182926; Publication date: 2011-07-28; Inventors: La Monica Nicola, Scarselli Elisa, Ciliberto Gennaro, Aurisicchio Luigi, Fridman Arthur, Bagchi Ansuman

Nucleic acid constructs, pharmaceutical compositions and methods of using same for treating cancer Publication number: US2009221681; Publication date: 2009-09-03; Inventor: Hochberg Avraham Diphtheria toxin variant Publication number: US2009156502; Publication date: 2009-06-18; Inventors: Harrison Robert J, Vanderspek Johanna C Diphtheria Anticancer agent toxin Publication number: JP2008285491; Publication date: 2008-11-27; Inventors: Mekata Eisuke, Miyamoto Shingo (5 out of 7) Activation of Recombinant Diphtheria Toxin Fusion Proteins by Specific Proteases Highly Expressed on the Surface of Tumor Cells Publication number: US2008166375; Publication date: 2008-07-10; Inventors: Leppla Stephen H, Avallone Jennifer, Bugge Thomas, liu Shi-Hui, Osorio Manuel Methods and compositions based on diphtheria toxin-interleukin-3 conjugates Publication number: CA2698595; Publication date: 2008-03-13; Inventor: Frankel Arthur E

Endoprostatic srping (Stent) impregnated with Rapamycin/Sirolimus and Shiga toxin acting against cancer of prostate and benign hyperplasias of the urologic tract Publication number: GR1006552; Publication date: 2009-10-02; Inventor: Dimofilos Nikolaos Christou Treatment of metastatic colon cancer with b-subunit of shiga toxin Publication number: US2007071772; Publication date: 2007-03-29; Inventors: Kovbasnjuk Olga N; Donowitz Mark

Chimeric polypeptide comprising fragment B of Shiga toxin and peptide of therapeutic interest Shiga toxin Publication number: JP2010180216; Publication date: 2010-08-19; Inventors: Goud Bruno, Johannes Ludger (5 out of 6) Shiga Toxin B-subunit as a vector for tumor diagnosis and drug delivery to GB3 expressing tumors

Publication number: US2010329992; Publication date: 2010-12-30; Inventors: Johannes Ludger, Grierson David, Robine Sylvie, Flor- ent jean-Claude, Maillard Philipe, Roger Jacky Shiga toxin B-subunit/chemotherapeutics conjugates Publication number: US 2011152252; Publication date: 2011-06-23; Inventors: Johannes Ludger, El Alaoui Abdessamed, Decaudin Didier, Robine Sylvie, Schmidt Frederic, Florent jean-Claude

Treating neoplasms with neurotoxin Publication number: US2010209456; Publication date: 2010-08-19; Inventor: Shaari Christopher Application of lipid vehicles and use for drug delivery Botulinum neurotoxin Publication number: US2007003610;Publication date: 2007-01-04; Inventors: Chancellor Michael B; Fraser Matthew O; Chuang Yao- Chi; De Groat William C; Huang Leaf; Yoshimura Naoki (4 out of 4) Methods and compositions for the treatment of cancer

Publication number: US2009232849; Publication date: 2009-09-17; Inventors: Gallez Bernard; Ansiaux Reginald Methods for treating cancers Publication number: CN101031317; Publication date: 2007-09-05; Inventors: Stephen Brin Mitchell F Donova 40 Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 Fialho et al.

(Table 2) Contd….

MICROBIAL PRODUCTS AS ANTICANCER AGENTS Toxins - Recent Issued and published patents (2007-2011)

Pseudomonas exotoxin A mutants and uses thereof Publication number: EP2204385; Publication date: 2010-07-07; Inventors: Pastan Ira H; Ho Mitchell; Bang Sook-Hee Chimeric toxins for targeted therapy Publication number: US2009181894; Publication date: 2009-07-16; Inventors: Yarkoni Shai; Nechushtan Amotz; Lorberboum-Galski Haya; Marianovsky Irina Exotoxin A Methods for treating cancer using an immune-toxin comprising an exotoxin a moiety having a furin cleavage site replaced with a (5 out of 10) cancer associated protease site cleaved by MMP-2 or MMP-9

Publication number: US2009214543; Publication date: 2009-08-27; Inventors: Zangemeister-Wittke Uwe; De Paolo Claudio Tweak-pseudomonas exotoxin a fusion protein for cancer therapy Publication number: US2008175896; Publication date: 2008-07-24; Inventors: Winkles Jeffrey; Brown Sharron CD19-specific immunotoxin and treatment method Publication number: WO2007085470; Publication date: 2007-08-02; Inventors: Fey Georg H; Peipp Matthias; Schwemmlein Michael

Specific immunotherapy of cancer using live recombinant bacterial vaccine vector Listeriolysin S Publication number: JP2007254485 (A); Publication date: 2007-10-04; Inventor: Paterson Yvonne (2 out of 2) Listeria monocytogenes cytotoxin Listeriolysin S

Publication number: IE20080050 (A1); Publication date: 2009-11-11; Inventors: Ross Paul; Hill Colin; Cotter Paul and the methods for their use as a combination product for Combinations of Antibodies Selective for a Tumor Necrosis the treatment of cancers [59]. Factor-Related Apoptosis-Inducing Ligand Receptor and Other Therapeutic Agents Genetically Engineered Glutaminase and its Use in Antivi- ral and Anticancer Therapy This patent discloses a monoclonal antibody that specifically binds the TRAIL receptor DR5. The inventors found A purified form of glutaminase, free of other proteins is that this antibody displays in vitro and in vivo activity on disclosed in this patent. The enzyme is suitable for a therapeutic use in cancer treatments [60]. human cancer cells expressing DR5. The formulation is intended to be administered with one or more therapeutic agents. The chemotherapeutic drugs are selected from the 2.5. Asparaginase following list: leflunomide, dactinomycin, tamoxifen, inter- Medicament for the Treatment of Cancer of the Pancreas feron alpha-2b, glutamic acid, plicamycin, mercaptopurine, 6-thioguanine, carmustine, BCNU, lomustine, CCNU, cyto- A suspension of red corpuscles encapsulating aspar- sine, araboside, estramustine, hydroxyurea, procarbazine, aginase as a pharmaceutical combination for the treatment of bulsulfan, medroxyprogesterone, estramustine phosphate so- pancreatic cancer is disclosed in this patent. It concerns the dium, ethinyl estradiol, estradiol, megestrol acetate, meth- composition and the effective amount of the suspension in- ylestosteraone, diethylstilbestrol diphosphate, chlorotrian- tended for the treatment of pancreatic cancer [61]. isene, testolactone, mephalen, chorambucil, mechloretha- Treatment with Anti-VEGF Antibody mine, thiotepa, bethamethasone sodium phosphate, dicarba- zine, asparaginase, mitotane, vincristine sulphate and vin- This invention discloses a method for treating cancer by blastine sulphate [63]. administering a mixture containing an anti-VEGF antibody in conjugation with a chemotherapeutic drug(s) (anti- Pegylated L-Asparaginase malignant tumor agent(s)). The following is a list of drug This patent discloses a pharmaceutical composition con- classes used in this pharmaceutical composition: an alkylat- taining the bacterial enzyme L-asparagine aminohydrolase as ing agent, an antimetabolite, a folic acid analogue, a pyri- the active ingredient. The purified enzyme is mixed with midine or a purine analogue, a anti-mitotic/anti-microtubule polyethylene glycol with a molecular weight less than or agent (vinca alkaloid), an epipodopyllotoxin, an antibiotic, equal to about 5000 Da. The conjugate has a high level of in L-asparaginase, a topoisomerase inhibitor, interferon, a vitro activity and an unexpected increase in half-life in vivo. platinum coordinated complex, an anthracenedione-substilu- In other embodiment, the inventors disclose the use of this ted urea, a methylhydrazine derivative, an adrenal cortex conjugate for the treatment of cancer [64]. depressor, an adrenocorticosteroid, a progestin, an estrogen, an antiestrogen, an androgen, an antiandrogen, and a go- nadotrophic hormone-releasing hormone analogue [62].

Bacterial Strains and Products as Anticancer Agents Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 41

Table 3. Selected List of Recent Worldwide Issued and Published Patents on Bacterial Enzymes as Anticancer Agents.

MICROBIAL PRODUCTS AS ANTICANCER AGENTS

Enzymes - Recent Issued and published patents (2007-2011)

Mutated form of arginine deiminase Publication number: PT1278868; Publication date: 2008-04-11; Inventors: Ensor Charles Mark; Holtsberg Frederick Wayne; Clark Mike A Arginine Use of bacteria endowed with arginine deiminase to induce apoptosis and/or reduce an inflammatory reaction and pharmaceu- deiminase tical or dietetic compositions containing such bacteria (3 out of 3) Publication number: SI1058725; Publication date: 2008-08-31; Inventor: De Simone Claudio

A strain capable of producing arginine deiminase and the use thereof Publication number: WO2008141523; Publication date: 2008-11-27; Inventors: Sun Zhihao; Zheng Pu; Ni Ye; liu Yongmei; Liu Yupeng

Formulations of 5-fluorocytosine and uses thereof Publication number: WO2010002937; Publication date: 2010-01-07; Inventors: Gruber Harry E; Jolly Douglas; Olmstead Kay Cytosine deaminase Stem cell targeting of cancer, methods and compositions therefor (3 out of 3) Publication number: WO2009052394; Publication date: 2009-04-23; Inventors: Cady Craig; Mcasey Mary Viral vector driven mutant bacterial cytosine deaminase gene and uses thereof

Publication number: US2007225245; Publication date: 2007-09-27; Inventors: Buchsbaum Donald J; Gillespie G Y; Markert J James M; Kaliberov Serget A

L-Methioninase Conjugate for the specific targeting of anticancer agents to tumor cells or tumor vasculature and production thereof (1 out of 1) Publication number: US2009304666; Publication date: 2009-12-10; Inventors: Harrison Roger G; Pento J Thomas; Lind Stuart

Glutadon Glutaminase Publication number: US2009169537;Publication date: 2009-07-02;Inventors: Bausch Michael; Wetzler Rainer; Mueller Christian (2 out of 2) Genetically engineered glutaminase and its use in antiviral and anticancer therapy Publication number: EP1854876; Publication date: 2007-11-14; Inventors: Roberts Joseph; Macallister Thomas W; Sethuramen Natarajan; Freeman Abbie G

Medicament for the treatment of cancer of the pancreas Publication number: WO2009080837; Publication date: 2009-07-02; Inventors: Dufour Emmanuelle-Cecile; Godfrin Yann Treatment with anti-VEGF antibody Publication number: JP2007326860; Publication date: 2007-12-20; Inventors: Fyfe Gwendolyn; Holmgren Eric; mass Robert D; L-asparaginase Novotny William (4 out of 4) Combinations of antibodies selective for a tumor necrosis factor-related apoptosis-inducing ligand receptor and other therapeutic agents Publication number: NZ533164; Publication date: 2008-09-26; Inventors: Zhou Tong; Ichikawa Kimihisa; Kimberly Robert P; Koopman William J; Oshumi Jun; Lobuglio Albert F; Buchsbaum Donald J Pegylated L-asparaginase Publication number: WO2011003886; Publication date: 2011-01-13; Inventor: Abribat Thierry

3. PATENTS ON OTHER BACTERIAL PROTEINS Compositions and Methods to Prevent Cancer with Cupre- AND PEPTIDES AS ANTICANCER AGENTS doxins 3.1. Azurin (Pseudomonas aeruginosa) This invention discloses methods and materials for inhibiting the growth of cancer cells through a preferential entry Compositions and Methods to Prevent and/or Treat Cancer of a cytotoxic compound into target cells (cancer cells). with Pa-CARD Preferential entry of the cytotoxic compound (p.e. bacterial This invention relates to methods and materials for kill- toxin) is accomplished by the use of protein transduction ing cancer cells with bacterial proteins. In particular this in- domains (PTD) derived from bacterial cupredoxins, includ- vention focused on five proteins, namely Azurin, Laz, Pa- ing the p18 and p28 truncations of azurin [66]. CARD, and fusion proteins Azu-H.8 and H.8-Azu, and their use in killing leukemia and/or ovarian cancer cells [65]. 42 Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 Fialho et al.

Compositions and Methods to Prevent Cancer with Cupre- Cytotoxic Factor for Modeling Cell Death doxins This patent provides a cytotoxic factor which stimulates This patent discloses compositions and methods compris- cell death by necrosis or apoptosis. This invention provides a ing chemopreventive agents that may be cupredoxin(s) or use of a truncated azurin for preparing a pharmaceutical structural related derivatives and at least one chemotherapeu- composition for increasing cell death of a cancer cell. The tic drug (Tamoxifen). These pharmaceutical compositions truncated azurin is at least one compound that is cytotoxic by may comprise the bacterial protein azurin from Pseudo- an ATP-dependent mechanism, and is the one obtained from monas aeruginosa, and/or its derivative peptide comprising pathogenic bacteria secreting at least one compound which is the residues (50-77) (named p28). The pharmaceutical com- cytotoxic by the ATP-nondependent mechanism [72]. positions of this invention may be used to prevent the development of premalignant lesions, thereby preventing cancer 3.2. p28 (Azurin-Derived Linear Peptide) [67]. Compositions and Methods to Prevent Cancer with Cupre- Compositions and Methods to Prevent Cancer with Cupre- doxins doxins This invention discloses methods and materials for kill- This invention discloses pharmaceutical compositions ing and/or inhibiting the growth of a cancer cell via preferen- comprising synthetic peptides derived from bacterial cupre- tial entry of a cytotoxic compound which is accomplished by doxins that may be used to prevent the development of pre- the use of protein transduction domains (p18 and p28) de- malignant lesions, thereby preventing cancer. Specifically, rived from a bacterial cupredoxin (azurin) [73]. these compositions may comprise the bacterial protein azurin from Pseudomonas aeruginosa, and/or its derivative peptide Compositions and Methods to Prevent Cancer with Cupre- comprising the residues (50-77) (named p28) [68]. doxins Compositions and Methods to Prevent Cancer by Stabiliz- This patent discloses compositions and methods comprising p53 Through Non MDM2-Mediated Pathways ing chemopreventive agents that may be cupredoxin(s) or structural related derivatives and at least one chemotherapeu- This patent discloses compositions and methods compris- tic drug (Tamoxifen). These pharmaceutical compositions ing chemopreventive agents that may be cupredoxin(s) or may comprise the bacterial protein azurin from Pseudo- structural related derivatives and at least one chemotherapeu- monas aeruginosa, and/or its derivative peptide comprising tic drug. Specifically, compositions may comprise azurin the residues (50-77) (named p28). The compositions de- from Pseudomonas aeruginosa, and/or the 50-77 residue scribed in this invention may be used to prevent the devel- region of azurin (p28). The pharmaceutical compositions of opment of premalignant lesions in mammalian cells, tissues this invention may be used to prevent the development of and animals, and thus prevent cancer [74]. premalignant lesions, thereby preventing cancer [69]. Compositions and Methods to Prevent Cancer with Cupre- Compositions and Methods to Treat Cancer with Cupre- doxins doxins and CpG rich DNA This invention relates to compositions comprising syn- This invention relates to compositions comprising CpG thetic peptides that are derivatives of bacterial cupredoxins. rich DNA sequences from Pseudomonas aeruginosa and, Specifically, these compositions may comprise the bacterial optionally a bacterial cupredoxin also derived from the same protein azurin from Pseudomonas aeruginosa, and/or its microrganism. Specifically, these pharmaceutical composi- derivative peptide comprising the residues (50-77) (named tions are useful for treating cancer. This invention is also p28). Possible uses of the compositions of the invention may related to methods to express proteins near cancer cells. be the prevention of the development of premalignant lesions These methods may be used to express therapeutic or diag- in mammalian cells, tissues and animals, and thus prevent nostic proteins in a patient suffering from cancer, and can cancer [75]. also be used for diagnosing cancer. The method described in this patent uses the gene enconding azurin from P. aerugi- Compositions and Methods to Prevent Cancer by Stabiliz- nosa as an expression system for azurin or heterologous pro- ing p53 Through Non MDM2-Mediated Pathways teins in P. aeruginosa or heterologous cells [70]. This invention relates to compositions and methods com- Compositions and Methods for Treating Conditions Re- prising chemopreventive agents that may be cupredoxin(s) or lated to Ephrin Signaling with Cupredoxins variants, derivatives and structural equivalents of cupredoxins, and at least one other chemopreventive agent. These This invention relates to compositions and methods based compositions may comprise azurin from Pseudomonas aeru- on bacterial cupredoxins or their derivatives that interfere ginosa, and/or the 50-77 residue region of azurin (p28) and with the ephrin signaling system in mammalian cells. Spe- may be used to prevent or inhibit the development of prema- cifically, this invention relates to compositions and methods lignant lesions in mammalian cells, tissues and animals, thus that use cupredoxins, such as azurin, rusticyanin and plasto- preventing cancer [76]. cyanin and structural equivalents thereof to treat cancer in mammals [71].

Bacterial Strains and Products as Anticancer Agents Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 43

3.3. Plantaricin (Two-Peptide Bacteriocin) (Lactobacillus alpha-1 antitrypsin deficiency. The patent establishes meth- plantarum) ods for assaying HDAC inhibition and pharmaceutical compositions for the derivative compounds [81]. Anti-Tumor Agent for Gastrointestinal Cancer Spiruchostatin Analogues and Their Therapeutic Use This patent discloses a short peptide derived from a pro- biotic (an enterobacterium which lives symbiotically in a The invention discloses pharmaceutical compositions human intestine). This peptide is safe, inexpensive and effec- composed by histone deacetylase (HDAC) inhibitors. Such tive against cancer cells. Moreover, this patent also discloses compounds are proposed to be used in the treatment of sev- an activator for an insulin-like growth factor-binding protein eral conditions such as cancer, cardiac dystrophy, chronic and an anti-tumor agent for gastrointestinal cancer including heart failure, skin inflammatory conditions, haemoglobi- colorectal cancer. Plantaricin-A, a polypeptide (26 amino nopathy, thalassemia, sickle cell disease, CNS disorders, acid residues) secreted from Lactobacillus plantarum, or autoimmune diseases, diabetes and other HDAC-mediated CSF (competence and sporulation factor), a 5 amino acid conditions. Methods for assaying HDAC inhibition are also residues secreted from Bacillus subtilis are the active com- provided [82]. pounds [77]. Spiruchostatin Derivatives and Their Therapeutic Use 3.4. Microcin (Klebsiella pneumoniae RYC492) As in previous patents, this invention provides Spi- ruchostatin derivatives and their possible therapeutic use. Microcin B17 Analogs and Methods for Their Preparation These compounds are used to treat several malignancies, and Use such as cancer, cardiac hypertrophy, chronic heart failure, an Synthetic analogues of Microcin B17, methods of mak- inflammatory condition, a cardiovascular disease, a haemo- ing and using them are presented in this patent. This inven- globinpathy, a thalassemia, a sickle cell disease, a CNS dis- tion also discloses the use of these synthetic compounds to order, an autoimmune disease, diabetes, osteoporosis, MDS, inhibit DNA gyrase activity in cells and also for possible benign prostatic hyperplasia, endometriosis, oral leu- therapies that target diseases affected by DNA gyrase activ- koplakia, a genetically related metabolic disorder, an infec- ity by reducing it, such as bacterial infections and cancer tion, Rubens-Taybi, fragile X syndrome or alpha-1 antitryp- [78]. sin. Methods for assaying the level of HDAC inhibition and pharmaceutical compositions comprising such compounds 3.5. Pep27 (Streptococcus pneumoniae) are also provided [83]. Peptide with Enlarged Hydrophobic Domain by Substitut- 3.7. Romidepsin (Cyclic Peptide) (Chromobacterium ing Hydrophilic Domain of pep-27 Peptide with Other violaceum No.968) Amino Acids, and Pharmaceutical Composition Containing the Same Method for Killing Tumor by Photosensitization Treatment Under Conditions where Histone is Highly Acetylated This patent provides a synthetic peptide which exhibits anticancer activity. The peptide or a derivative is modified in This patent discloses a therapeutic agent for cancer its hydrophobic domain by substituting is hydrophilic do- treatment based on a histone deacetylase inhibitor. Some of main with hydrophobic amino acids selected from trypto- the examples of the histone deacetylase inhibitor displayed phane, alanine, isoleucine, methionine, phenylalanine, in the patent are sodium butyrate, trichostatin A (TSA), proline, valine and leucine. The patent also provides a phar- suberoylanilide hydroxamic acid (Zolinza/Volinostat/ maceutical composition containing the original or the syn- SAHA), valproic acid, Belinostat/PXD101, ITF2357, CRA- thetic derivatives of this peptide [79]. 24781/PCI24781, Romidepsin/FK228/Depsipeptide, Phenyl- butyrate/VP-101/EL-532, Pivanex/AN-9, SB939, MS- 3.6. Spiruchostatin (Cyclic Peptide) (Pseudomonas sp.) 275/SNDX-275/entinostat, MGCD-0103 and JHJ-26481585. Another embodiment of this patent discloses a novel phar- Depsipeptides and Their Therapeutic Use maceutical composition composed by the histone deacetylase inhibitor together with a photosensitizing agent. This com- This patent provides analogues of Spiruchostatin, a po- pound is also intended to be used in combination with a tent histone deacetylase inhibitor [80]. PUVA therapy [84]. Spiruchostatin Derivatives and Their Therapeutic Use Romidepsin-Based Treatments for Cancer This patent provides spiruchostatin derivatives and its This invention provides an improved process for prepar- potential therapeutic use, and also pharmaceutically accept- ing romidepsin that involves its production, purification and able salts, useful in the inhibition of histone deacetylase storage. This patent also elucidate that the purification of (HDAC). Such compounds are proposed to be used in the following list of diseases: cancer, cardiac hypertrophy, romidepsin at pH lower than 6.0 prevents the reduction of the disulfide bonds formed between adjacent cysteines with chronic heart failure, an inflammatory condition, a cardio- the subsequent formation of oligomerized forms. Pharmaceu- vascular disease, a haemoglobinpathy, a thalassemia, a sickle tical compositions of monomeric romidepsin free of dimer- cell disease, a CNS disorder, an autoiminune disease, diabe- ized, oligomerized or polymerized adducts are also disclosed tes, osteoporosis, MDS, benign prostatic hyperplasia, endo- in this patent [85]. metriosis, oral leukoplakia, a genetically related metabolic disorder, an infection, Rubens-Taybi, fragile X syndrome or 44 Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 Fialho et al.

Combination Therapy Comprising Romidepsin and I.A. ethylene glycol/polylactic copolymer, EVAc, fatty acid and Bortezomib decanedioic copolymer. Microspheres are produced into sustained-release implant, and by injecting or positioning the This invention provides a therapy for treating cancer and sustained-release agent in tumor or tumor margin, drug re- other neoplastic diseases that comprises a mixture with ro- lease in local part of tumor can be performed for approxi- midepsin and a proteasome inhibitor. The cyclic peptide ro- mately 30 to 40 days. These dispositives can be used com- midepsin and a proteasome inhibitor (e.g., bortezomib) inter- bined with non-operative treatments such as chemical treat- act synergistically to selectively kill cancer cells. The effect ment and/or radiation treatment [90]. is particularly relevant in malignant hematological cells (e.g., leukemia, lymphoma, multiple myeloma), both in vitro and The Inhibitor on the Binding of DDR Proteins to Collagens in vivo. This combination proved to be useful in resistant Comprising an Actinomycin D as an Effective Compound neoplasms, such as treating bortezomib-resistant cancers and This invention discloses novel pharmaceutical com- steroid-resistant cancers. The patent includes the pharmaceu- pounds for the treatment of cancer, liver cirrhosis, rheuma- ticals compositions, preparations and kits that include the toid disease or arteriosclerosis caused by excessive activities above referred substantces [86]. of DDR proteins. The active compound of these formulations Treatment of Ras-Expressing Tumors is the antibiotic actinomycin D [91]. The use of at least one DAC inhibitor (e.g. romidepsin) Temperature Controlled Sustained-Release Injection Con- for the treatment of alpha-Ras expressing tumors and the taining Anti-Cancer Medicine compositions and methods to do so are disclosed in this pat- This invention discloses a pharmaceutical composition ent [87]. based on an anti-cancer drug that has been stabilized with an Combination Therapy for Cancer amphiphilic copolymer hydrogel. This injectable formulation is temperature sensitive and permits a controlled release of A novel anti-cancer pharmaceutical composition com- the cancer drug. This invention also discloses the method posed by romidepsin, a DAC inhibitor, and a tyrosine kinase used to inject the drug in the body thereby demonstrating the inhibitor (e.g., erlotinib) is disclosed in this patent. Together, accumulation of the drug in the tumor tissue and its effec- these pharmaceutical compounds interact to induce apoptosis tiveness for a plurality of weeks to a plurality of months. The which is particularly pronounced in wild type non-small cell injectable formulation contains one or more of the following lung cancers, particularly wild type EGFR and wild type antibiotics: vincristine, vinorelbine, navelbine, vindesine, KRAS non-small cell lung cancer cells in vitro and in vivo. vinleurosine, vinrosidine, cephalotaxine, bleomycin, dauno- This invention also provides the pharmaceutical composi- mycin, aclarubicin, epirubicin, idarubicin, pirarubicin, valru- tions, preparations, and kits of the compounds described bicin, mitomycin C, actinomycin D, losoxantrone, mitoxan- [88]. trone, mitozolomide, temozolomide [92]. 4. PATENTS ON ANTIBIOTICS FROM MICROBIAL Temperature-Sensitive Immunoliposomes, Preparation SOURCES AS ANTICANCER AGENTS Method and Uses Thereof 4.1 - Actinomycin (Streptomyces) This patent describes and discloses novel temperature- sensitive immunoliposomes aiming to improve their selectiv- Application of Actinomycin Compound Actinomycin V in ity for cancer cells. They are constructed based on antibodies Preparing Drugs for Anti Liver Cancer that have improved ability to recognize a specific epitope This invention discloses an anti-cancer pharmaceutical (antigen) overexpressed in cancer cells. Specifically, the composition based on the antibiotic actinomycin V. This temperature-sensitive immunoliposomes are vesicules com- antibiotic has molecular weight of 1268 Da and is extracted posed of a phospholipid membrane bilayer (dipalmitoyl- from the crude extract of the fermentation culture of Strep- phosphatidylcholine, dimyristoylphosphatidylcholine and tomyces zhapoensis H41-26. In particular, this antibiotic is distearoylphosphatidylcholine and dioleoylphosphatidyle- expected to have a wide application prospect when used as thanolamine) decorated with monoclonal antibody such as an liver cancer anti cancer agent [89]. epidermal growth factor receptor (EGFR). Liposome- encapsulated antibiotics are selected from the following list: Novel Anticancer Sustained-Release Injection gemcitabine, busulfan, chlorambucil, cyclophosphamide, This invention relates to novel sustained-release injection melphalan, cisplatin, ifosfamide, cytarabine, 5-fuorouracil, for cancer therapy comprising sustained-release micro- methotrexate, actinomycin-D, bleomycin, daunorubicin, spheres and dissolvent. Microspheres are composed of active adriamycin, mitomycin-C, vinblastine, vincristine, vindesine, ingredients for cancer therapy and sustained-release auxiliary procarbazine, tamoxifen, megestrol acetate, flutamide and materials. Active compounds for cancer therapy are made goserelin acetate [93]. based on a combination of the following drugs: fotemustine, bendamustine, tallimustine, carmustine, nimustine, lomus- 4.2. Plicamycin (Streptomyces plicatus) tine, semustine or ranimustine with cytotoxic drug of adria- A Cancer Sensitizer Comprising Chlorogenic Acid cin, epidoxorubicin, melphalan, 4H- cyclophosphamide per- oxide, actinomycin D, vinorelbine or tamoxifen. The sus- This patent discloses a chlorogenic acid derivative as a tained-release auxiliary materials comprise polylactic acid cancer sensitizer. The pharmaceutical composition comprises and the copolymer, monomethyl polyethylene glycol, poly- a chlorogenic acid derivative, a compatible carrier and a potent anti-cancer drug selected from the following list: Bacterial Strains and Products as Anticancer Agents Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 45

Table 4. Selected List of Recent Worldwide Issued and Published Patents on Bacterial Proteins and Peptides as Anticancer Agents.

MICROBIAL PRODUCTS AS ANTICANCER AGENTS

Other proteins and peptides - Recent Issued and Published Patents (2007-2011)

Compositions and methods to prevent and/or treat cancer with Pa-CARD Publication number: WO2010096754; Publication date: 2010-08-26; Inventors: Das Gupta Tapas; Chakrabarty Ananda; Fialho Arsenio Compositions and methods to prevent cancer with cupredoxins Publication number: WO2010080506; Publication date: 2010-07-15; Inventors: Taylor Brad; Metha Rajeshwari; Yamada Tohru; Beattie Craig; Das Gupta Tapas Compositions and Methods to Prevent Cancer with Cupredoxins Publication number: US2010087377; Publication date: 2010-04-08; Inventors: Das Gupta Tapas; Chakrabarty Ananda Compositions and methods to prevent cancer with cupredoxins Azurin and Laz Publication number: WO2010078042; Publication date: 2010-07-8; Inventors: Das Gupta Tapas; Metha Rajendra; Metha Ra- (8 out of 13) jeshwari; Beattie Craig; Compositions and methods to prevent cancer by stabilizing p53 through non MDM2-mediated pathways Publication number: US2010093057; Publication date: 2010-04-15; Inventors: Beattie Craig; Yamada Tohru; Das Gupta Tapas Compositions and Methods to treat cancer with cupredoxins and CpG rich DNA Publication number: MX2009005849; Publication date: 2009-08-12; Inventors: Chakrabarty Ananda; Das Gupta Tapas Compositions and Methods for treating conditions related to ephrin signaling with cupredoxins Publication number: US2008312413; Publication date: 2008-12-18; Inventors: Chakrabarty Ananda; Das Gupta Tapas; Yamada Tohru; Chaudhari Anita; Fialho Arsenio; Zhu Yonghua Cytotoxic factor for modeling cell death Publication number: JP2009091370; Publication date: 2009-04-30; Inventors: Das Gupta Tapas; Chakrabarty Ananda; Punj Vasu; Zaborina Olga

MICROBIAL PRODUCTS AS ANTICANCER AGENTS Other Proteins and Peptides - Recent Issued and Published Patents (2007-2011)

Compositions and methods to prevent cancer with cupredoxins Publication number: WO2010080506; Publication date: 2010-07-15; Inventors: Taylor Brad; Metha Rajeshwari; Yamada Tohru; Beattie Craig; Das Gupta Tapas p28 Compositions and methods to prevent cancer with cupredoxins (Azurin-derived lin- Publication number: WO2010078042; Publication date: 2010-07-8; Inventors: Das Gupta Tapas; Metha Rajendra; Metha Ra- ear peptide) jeshwari; Beattie Craig (4 out of 4) Compositions and Methods to Prevent Cancer with Cupredoxins

Publication number: US2010087377; Publication date: 2010-04-08; Inventors: Das Gupta Tapas; Chakrabarty Ananda Compositions and methods to prevent cancer by stabilizing p53 through non MDM2-mediated pathways Publication number: US2010093057; Publication date: 2010-04-15; Inventors: Beattie Craig; Yamada Tohru;; Das Gupta Tapas

Plantaricin Anti-tumor agent for gastrointestinal cancer (two-peptide bacte- Publication number: WO2009093447; Publication date: 2009-07-30; Inventors: Fujiya Mikihiro; Kohgo Yutaka; Okamoto Ko- riocin) taro (1 out of 1) Lactobacillus plantarum

Microcin Microcin B17 Analogs And Methods For Their Preparation And Use Klebsiella pneumo- Publication number: US2008234132, Publication date: 2008-09-25; Inventors: Coquin Laurence; Jourdan Fabrice; Pierrat Oliv- niae RYC492 ier; Lowe David John; Maxwell Anthony; Pickett Christopher John; Wall Melisa (1 out of 1) 46 Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 Fialho et al.

(Table 4) Contd….

MICROBIAL PRODUCTS AS ANTICANCER AGENTS Other Proteins and Peptides - Recent Issued and Published Patents (2007-2011)

Pep 27 Peptide with enlarged hydrophobic domain by substituting hydrophilic domain of pep-27 peptide with other amino acids, Streptococcus pneu- and pharmaceutical composition containing the same moniae Publication number: KR20030073833; Publication date: 2003-09-19; Inventors: Choi Cheol Hui; ham Gyeong Su; lee Dong (1 out of 1) Geon

Depsipeptides and Their Therapeutic Use Publication number: US2010056434; Publication date:2010-03-04; Inventors: Packhman Graham Keith; Ganesan Arasu; Cecil Alexander Richard Liam

Spiruchostatin Spiruchostatin derivatives and their therapeutic use Publication number: GB2460181; Publication date: 2009-11-25; Inventors: Shuttleworth Stephen Joseph; Silva Franck Alexan- (cyclic peptide) dre; Tomassi Cyrille Davy; Cecil Alexander Richard Liam; Hill Thomas James Pseudomonas sp. Spiruchostatin analogues and their therapeutic use (4 out of 4) Publication number: GB2460180; Publication date: 2009-11-25; Inventors: Shuttleworth Stephen Joseph; Silva Franck Alexan-

dre; Tomassi Cyrille Davy; Cecil Alexander Richard Liam; Hill Thomas James Spiruchostatin derivatives and their therapeutic use Publication number: GB2460178; Publication date: 2009-11-25; Inventors: Shuttleworth Stephen Joseph; Silva Franck Alexan- dre; Tomassi Cyrille Davy; Cecil Alexander Richard Liam; Ganesan Arasu; Hill Thomas James

Method for killing tumor by photosensitization treatment under conditions where histone is highly acetylated Publication number: WO2010064422; Publication date: 2010-06-10; Inventors: Toyooka Tasushi; Ibuki Yuko

Romidepsin-based treatments for cancer Romidepsin Publication number:US2010093610;Publication date:2010-04-15;Inventors: Vrolijk Nicholas H; Verdine Gregory L (cyclic peptide) Combination therapy comprising romidepsin and I.A. bortezomib Chromobacterium Publication number: MX2009007777; Publication date: 2009-12-16; Inventors: Keegan Mitchell; Grant Steven violaceum No.968 Treatment of Ras-Expressing Tumors (5 out of 7) Publication number: US2009305956; Publication date: 2009-12-10; Inventors: Mcculloch William; Keegan Mitchell

Combination therapy for cancer Publication number: WO2008127659; Publication date: 2008-10-23; Inventor: Frenkel Eugene P mechloethamine, chlorambucil, phenylalanine, mustard, cy- cin, busulfan, thiotepa, cisplatin, carboplatin, dactinomy- clophosphamide, ifosfamide, carmustine, lomustine, strepto- cin(actinomycin D), doxorubicin(adriamycin), daunorubicin, zotocin, busulfan, thiotepa, cisplatin, carboplatin, dactino- idarubicin, mitoxantrone, plicamycin, mitomycin, C bleomy- mycin, doxorubicin, daunorubicin, idarubicin, mitoxantrone, cin, vincristine, vinblastine, paclitaxel, docetaxel, etoposide, plicamycin, mitomycin, C Bleomycin, vincristine, vin- teniposide, topotecan and irinotecan; and iv) a method de- blastine, paclitaxel, docetaxel, etoposide, teniposide, topote- scribing the administration of the pharmaceutical composi- can and iridotecan [94]. tion [95]. Combinations of Antibodies Selective for a Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand Receptor and 4.3. Doxorubicin (Streptomyces peucetius) other Therapeutic Agents Immunocytokines for Cancer Treatment in Combination The summary of this invention is described in reference with Chemotherapeutic Agents [63]. This invention discloses a cancer treatment based on a A Cancer Sensitizer Comprising Glucosamine, Glucosa- pharmaceutical composition composed by doxorubicin or mine Derivatives or Salts Thereof paclitaxel, together with antibody-interleukin 2 (IL2) conjugates which target tenascin-C [96]. This patent discloses a cancer sensitizer (a glucosamine derivative) aiming to enhance the efficacy of a chemothera- Combination Therapy for Ovarian Cancer peutic agent. Moreover, this invention discloses: i) a phar- This invention provides a method to treat ovarian cancer maceutically compatible carrier; ii) an anti-cancer drug se- by the administration of a pharmaceutical composition com- lected from the following list: mechlorethamine, chloram- posed by picoplatin and pegylated liposomal doxorubicin bucil, phenylalanine, mustard, cyclophosphamide, ifos- [97]. famide, carmustine(BCNU), lomustine(CCNU), streptozoto- Bacterial Strains and Products as Anticancer Agents Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 47

Anti-Cancer Medicine Both for Diagnosing and Treating Temperature-Sensitive Immunoliposomes, Preparation Cancer Method and Uses Thereof This patent discloses an anti-cancer pharmaceutical com- The summary of this invention is described in reference position containing i) a end-functionalized polymer chain; ii) [93]. iron oxide as a contrast agent; iii) and a anti-cancer drug such as doxorubicin. This therapeutic composition has been 4.5. Mitomycin (Streptomyces caespitosus) found to increase the efficacy of the anti-cancer drug, per- forms diagnosis and reduces a cardiotoxic side effect of the Method for Treating Cervical Cancer doxorubicin. Moreover, the presence of the contrast agent in The summary of this invention is described in reference this formulation can be used to diagnose the cancer and [101]. monitor the progress of the disease (detected by magnetic resonance imaging - MRI) [98]. Use of Lymphotoxin for Sensitizing Tumor Cells to Che- motherapeutic Drugs A Synergistic Pharmaceutical Combination for the Treat- ment of Cancer This patent discloses the lymphokine cytokine (Lympho- toxin (LT)) used herein as a cancer sensitizer for epidemis This patent describes a pharmaceutical combination tumours. The pharmaceutical composition comprises the comprising a cytotoxic antineoplastic agent (paclitaxel, do- administration of LT together with a chemotherapeutic drug cetaxel, doxorubicin or gemcitabine) and one cyclin depend- selected from the following list: anthracene nucleus type ent kinase (CDK) inhibitor. This combination has synergistic medicine, 5-Fluorouracil (5-FU), mitomycin, and any com- effects when used as an anti-cancer formulation. This inven- bination thereof. The treatment has proved to be effective for tion also discloses a method for the treatment of cancer, us- lung cancer, breast cancer, laryngocarcinoma, colorectal ing this therapeutic agent [99]. cancer, carcinoma of uterine cervix, melanoma and gastric Methods of Treating Ovarian Cancer cancer [102]. This patent discloses methods of treating ovarian cancer Apparatus and Method for Delivery of Mitomycin Through (e.g., epithelial ovarian cancer) using a farnesyl-OH- an Eluting Biocompatible Implantable Medical Device transferase inhibitor (lonafarnib), a taxane (e.g., paclitaxel or This invention discloses a device and a method for a con- docetaxel) and a platinum coordinator complex (e.g., car- trolled release of the antibiotic mitomycin. The use of this boplatin, cisplatin or oxaliplatin). Moreover, this invention device permits a significant and constant concentration of the discloses a cancer treatment based on a formulation with drug in the tissues thereby inhibiting the proliferation of the lonafarnib in combination with a liposomal doxorubicin disease [103]. [100]. Mitomycin Double Sustained Release Film of Implanted Antineoplastic Agents and its Preparation Method 4.4. Bleomycin (Streptomyces verticillus) This invention discloses a sustained release film coated Method for Treating Cervical Cancer with the antineoplastic agent mitomycin. After implantation, This patent provides a method for treating cervical cancer the anti-cancer agent is released continuously on tumor le- or cells infected with human papilloma virus comprising sions thereby inhibiting the proliferation of the disease. Interleukin-20 (IL-20). This formulation could be adminis- Moreover, this invention discloses the method used to pre- tered alone or in conjunction with radiation or chemothera- pare the film (polymer) [104]. peutic agents or surgical excision of the lesion. The che- Magnetically Targetable Mitomycin C Compositions and motherapeutic drugs are selected from the following list: Methods of Their Use bleomycin, chlorambucil, epirubicin, 5-fluorouracil, ifosfamide, mitomycin, methotrexate, vincristine, cisplatin and This patent discloses a pharmaceutical composition com- vinblastine [101]. prising mitomycin C encapsulated into magnetic particles. The use of this composition permits the rapid localization of Temperature Controlled Sustained-Release Injection Con- mitomycin C within a patient due to their magnetic proper- taining Anti-Cancer Medicine ties [105]. The summary of this invention is described in reference [92]. 4.6. Rhizoxin (Burkholderia rhizoxinica) A Cancer Sensitizer Comprising Chlorogenic Acid Antimitotic Rhizoxin Deritivatives of Burkholderia rhizox- The summary of this invention is described in reference ina, Method for Producing Said Derivatives and Use [94]. Thereof A Cancer Sensitizer Comprising Glucosamine, Glucosa- This invention discloses the use of four secondary me- mine Derivatives or Salts Thereof tabolites of the Burkholderia rhizoxina endosymbiont. The inventors proved the action of these bacterial compounds as The summary of this invention is described in reference antimitotic agents. Moreover the invention discloses a [95]. method for the isolation and purification of these

48 Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 Fialho et al. compounds. These compounds exhibit potential antifungal, Cancer Treatment with Epothilones cytotoxic and antiproliferative properties [106]. This patent discloses anticancer pharmaceutical composi- Burkholderia rhizoxina Microorganisms, Novel Endosym- tions containing epothilones as the active substances [113]. bionts of Rhizopus sp. and Method for Producing Rhizoxin and/or Rhizoxin-Derivates Using Said Micro-Organisms Epothilone Derivative for the Treatment of Hepatoma and other Cancer Diseases The present invention describes the use of Burkholderia This patent discloses pharmaceutical compositions com- rhizoxina strains to produce rhizoxin and their derivatives prising derivatives of an epothilone. These formulations have [107]. been tested and found effective against many types of cancer [114]. 4.7. Staurosporine (Streptomyces staurosporeus) Staurosporine Derivatives for Treating Non-Small Cell 5.2. Farnesyl Transferase Inhibitors (Streptomyces Parvu- Lung Cancer lus) This invention discloses a pharmaceutical composition to Farnesyl Protein Transferase Inhibitors for Treating treat lung cancer. The formulation comprises a kinase inhibi- Breast Cancer tor (PKC412) together with an activator of permeablization This patent discloses the use of farnesyl transferase in- of the mitochondrial outer membrane, such as an activator of hibitors for preparing formulations for treating breast cancer BAK [108]. [115]. 4.8. Borrelidin (Streptomyces parvulus) Anticancer Agents Based on Regulation of Protein Preny- lation Novel Compounds This patent discloses the use of farnesyl transferase in- This patent discloses pharmaceutical compositions com- hibitors (FTI) as anti-cancers agents. FTI block farnesylation prising derivatives of borrelidin. These formulations have thereby preventing the functions of oncoproteins such as Ras been tested and found effective against cancer and other dis- and RhoB [116]. eases including diabetic retinopathy, macular degeneration, and corneal neovascularisation. Moreover the invention dis- Novel Farnesyl Protein Transferase Inhibitors as Antitu- closes methods for the production of these compositions and mor Agents their use [109]. This patent discloses novel tricyclic compounds as farnesyl transferase inhibitors (FTI). This invention also discloses 5. PATENTS ON OTHER PRODUCTS FROM MI- a method for the treatment of cancer, using these therapeutic CROBIAL SOURCES AS ANTICANCER AGENTS agents [117].

5.1. Epotholiones (Sorangium cellulosum) Farnesyl Protein Transferase Inhibitors Epothilone Derivative for the Treatment of Hepatoma and This patent discloses pharmaceutical compositions com- Other Cancer Diseases prising derivatives of an epothilone. Methods of treating cancer and inhibiting farnesyl protein transferase using these This patent discloses pharmaceutical compositions com- therapeutic agents are also disclosed [118]. prising derivatives of an epothilone. These formulations have Novel Farnesyl Protein Transferase Inhibitors and Their been tested and found to be effective against many types of Use to Treat Cancer cancer [110]. Combinations of Therapeutic Agents for Treating Cancer This invention provides compositions and methods for treating cancer through the use of farnesyl transferase inhibi- This patent provides a pharmaceutical composition com- tors [119]. posed by an anti-cancer drug selected from the following list: carboplatin, cisplatin, Epothilone B, everolimus, docetaxel 5.3. Salinosporamide (Salinispora trópica) and paclitaxel [111]. Use of Salinosporamide A and Analogs Thereof for the Anti-Cancer Sustained-Released Injection Containing Treatment of Cancer, Inflammation and Infectious Dis- Epothilone Derivate eases This invention discloses a pharmaceutical composition This patent discloses the use of salinosporamide A or a based on an epothilone derivative encapsulated into micro- derivative in the manufacture of a pharmaceutical composi- spheres. The epothilone derivative is selected from the fol- tion for treating liver cancer [120]. lowing list: epothilone B, epothilone D, iso epothilone D, BMS-247550, azaepothilone B, furan epothilone D or BMS- Salinosporamide Derivatives as Proteasome Inhibitors 310705. The administration of the proposed formulation acts This patent discloses compositions and methods compris- synergistically with other conventional treatments, such as ing anti-cancers agents that may be salinosporamide or struc- radiotherapy and chemotherapy [112]. tural related derivatives [121].

Bacterial Strains and Products as Anticancer Agents Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 49

Table 5. Selected List of Recent Worldwide Issued and Published Patents on Microbial Antibiotics as Anticancer Agents.

MICROBIAL PRODUCTS AS ANTICANCER AGENTS

Antibiotics - Recent Issued and Published Patents (2007-2011)

Application of actinomycin compound actinomycin V in preparing drugs for anti liver cancer Publication number: CN101601856; Publication date: 2009-12-16; Inventor: Mengfeng Li; Jie Yuan; Birun Lin; Xun Zhu; Jiahui Pan; Yongcheng Lin; Jueheng Wu; Li Jun; Zhenjian He; Minghui Gu Novel anticancer sustained-release injection Actinomycin Publication number: CN101427996; Publication date: 2009-05-13; Inventor: Qinglun Kong Streptomyces The inhibitor on the binding of DDR proteins to collagens comprising an actinomycin D as an effective compound (5 out of 7) Publication number: KR20090042398; Publication date: 2009-04-30; Inventors: Yang Beom Seok; Cho Seung Joo

Temperature controlled sustained-release injection containing anti-cancer medicine Publication number: CN101273965; Publication date: 2008-10-01; Inventors: Qingzhong Kong; Jie Zhang Temperature-sensitive immunoliposomes, preparation method and uses thereof Publication number: KR20080000007; Publication date: 2008-01-02; Inventor: Kim Jin Seok

Plicamycin A cancer sensitizer comprising chlorogenic acid Streptomyces Publication number: KR20090033699; Publication date: 2009-04-06; Inventors: Kin Soo Youl; Park Kang Seo; Jeong Kyung Chae plicatus A cancer sensitizer comprising glucosamine, glucosamine derivatives or salts thereof (3 out of 3) Publication number: KR20070120015; Publication date: 2007-12-21; Inventors: Kim Soo Youl; kim Dae Seok Combinations of antibodies selective for a tumor necrosis factor-related apoptosis-inducing ligand receptor and other therapeutic agents Publication number: NZ533164; Publication date: 2008-09-26; Inventors: Zhou Tong; Ichikawa Kimihisa; Kimberly Robert P; Koop- man William J; Oshumi Jun; Lobuglio Albert F; Buchsbaum Donald J

MICROBIAL PRODUCTS AS ANTICANCER AGENTS Antibiotics - Recent Issued and published patents (2007-2011)

Immunocytokines for cancer treatment in combination with chemotherapeutic agents Publication number: US2010215615; Publication date: 2010-08-26; Inventors: Trachsel Eveline; Kaspar Manuela; Neri Dario; Marlind Jessica Combination therapy for ovarian cancer Doxorubicin Publication number: US2010178328; Publication date: 2010-07-15; Inventors: Martell Ronald A; Karlin David A; Breitz hazel B; Streptomyces Weiden Paul L peucetius Anti-Cancer Medicine Both for Diagnosing and Treating Cancer (5 out of 135) Publication number: US2010158812; Publication date: 2010-06-24; Inventors: Shin Dong-Youn; Kim Jang-Seop A synergistic pharmaceutical combination for the treatment of cancer Publication number: MX2009012155; Publication date: 2010-01-15; Inventors: Joshi Kalpana; rathos Maggie; Sharma Somesh; Khanwalkar Harshal Methods of treating ovarian cancer Publication number: US2010112039; Publication date: 2010-05-06; Inventors: Estok Thomas M; Zaknoen Sara; Baum Charles M

Method for treating cervical cancer Publication number: JP2010013467; Publication date: 2010-01-21; Inventors: Chandrasekher Yasmin A; Mckernan Patricia A Temperature controlled sustained-release injection containing anti-cancer medicine Bleomycin Publication number: CN101273965; Publication date: 2008-10-01; Inventors: Qingzhong Kong; Jie Zhang Streptomyces A cancer sensitizer comprising chlorogenic acid verticillus Publication number: KR20090033699; Publication date: 2009-04-06; Inventors: Kim Soo Youl; Park Kang Seo; Jeong Kyung Chae (5 out of 6) A cancer sensitizer comprising glucosamine, glucosamine derivatives or salts thereof

Publication number: KR20070120015; Publication date: 2007-12-21; Inventors: Kim Soo Youl; kim Dae Seok Temperature-sensitive immunoliposomes, preparation method and uses thereof Publication number: KR20080000007; Publication date: 2008-01-02; Inventor: Kim Jin Seok 50 Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 Fialho et al.

(Table 5) Contd….

MICROBIAL PRODUCTS AS ANTICANCER AGENTS

Antibiotics - Recent Issued and published patents (2007-2011)

Method for treating cervical cancer Publication number: JP2010013467; Publication date: 2010-01-21; Inventors: Chandrasekher Yasmin A; Mckernan Patricia A Use of lymphotoxin for sensitizing tumor cells to chemotherapeutic drugs Publication number: WO2009062344; Publication date: 2009-05-22; Inventors: Liu Yanjun; Shen Yijun; Yang Tong; Wang Jing Mitomycin Apparatus and Method for Delivery of Mitomycin Through an Eluting Biocompatible Implantable Medical Device Streptomyces caespitosus Publication number: US2008161908; Publication date: 2008-07-03; Inventors: Granada F; Furnish Simon M (5 out of 32) Mitomycin double sustained release film of implanted antineoplastic agents and its preparation method Publication number: CN101204382; Publication date: 2008-06-25; Inventors: Qiqing Zhang; Qian Wang; Yange Wang; Zhenqing Hou; Shefang Ye Magnetically Targetable Mitomycin C Compositions and Methods of Their Use Publication number: US2007274907; Publication date: 2007-11-29; Inventors: Li Yuhua; Peterson Caryn; Rudge Scott R; Stevens Suzanne; Tapolky Gilles H

Antimitotic Rhizoxin Deritivatives of Burkholderia Rhizoxina, Method for Producing Said Derivatives and Use Thereof Rhizoxin Publication number: US2009233976; Publication date: 2009-09-17; Inventors: Scherlach Kirstin; Partida-Martinez Laila; Hertweck Burkholderia Christian rhizoxinica Burkholderia rhizoxina micro-organisms, novel endosymbionts of rhizopus sp. and method for producing rhizoxin and/or (2 out of 2) rhizoxin-derivates using said micro-organisms

Publication number: US2009209013; Publication date: 2009-08-20; Inventors: Hertweck Christian; Partida-Martinez Laila P

Staurosporine Staurosporine derivatives for treating non-small cell lung cancer Streptomyces Publication number: KR20080046161; Publication date: 2008-05-26; Inventor: Schuler Martin staurosporeus (1 out of 1)

MICROBIAL PRODUCTS AS ANTICANCER AGENTS Antibiotics - Recent Issued and published patents (2007-2011)

Borrelidin Novel compounds Streptomyces Publication number: US2009312316; Publication date: 2009-12-17; Inventors: Wilkinson Barrie; Moss Steven; Zhang Ming parvulus (1 out of 1)

Analogs of Salinosporamide A disease. Methods disclosed in this invention involve genetic manipulation, selection of reagents in the fermentation feed- This patent discloses a new generation of salinospora- stock, and selection of fermentation conditions [124]. mide A as potent proteasome inhibitors [122]. Methods of Sensitizing Cancer to Therapy-Induced Cyto- 5.4. Belactosin (Streptomyces sp.) toxicity Novel Belactosin Derivatives as Therapeutic Agents/ Bio- logical Probes and their Synthesis This patent discloses Salinosporamide A as a cancer sensitizer. A pharmaceutical composition based on Salinospo- This patent discloses methods for the synthesis of belac- ramide A acts as a therapeutic agent to kill or inhibit cancer tosin and their derivatives. These pharmaceutical composi- cells. Salinosporamide A induces the expression of Raf tions have anti-cancer, antiviral and antibacterial activity. kinase inihibitor (RKIP) and phosphatase and tensin ho- The synthetic steps comprise a tandem Mukaiyama aldol molog (PTEN) and inhibits the expression of YYl, a tran- lactonization reaction and data demonstrating the utility of scriptional regulator protein overexpressed in cancer cells some of the derivatives as proteasome inhibitors [125]. [123]. 5.5. Syringolin A (Pseudomonas syringae) Biosynthesis of Salinosporamide A and its Analogs Derivatives of Syringolin A This patent discloses methods of modulating biosynthesis of Salinosporamide A and its analogs, which are useful in This patent discloses methods for the synthesis of syrin- treating cancer, inflammatory conditions, and/or infectious golin A and their derivatives. The process involves a tandem Bacterial Strains and Products as Anticancer Agents Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 51

Mukaiyama aldol-lactonization recation. These pharmaceuti- Endoprostatic Spring (STENT) Impregnated with Rapamy- cal compositions are used to treat neurodegenerative diseases cin/Sirolimus and Shiga Toxin Acting Against Cancer of and proliferative diseases such as cancer [126]. Prostate and Benign Hyperplasias of the Urologic Tract Development of a Synthesis of Syringolin A and B and De- The summary of this invention is described in reference rivatives Thereof [36]. This invention is related to the synthesis of syringolin A Composition of mTOR Antagonist and Angiogenesis In- and B and derivatives. These pharmaceutical compositions hibitor for Treating Cancer are used for both prophylaxis and cancer treatment [127]. This invention discloses a pharmaceutical composition containing two active substances, one antagonist of mTOR 5.6. Rapamycin (Streptomyces hygroscopicus) activity (rapamycin) and one angiogenesis inhibitor (Bevaci- Nanoparticle Comprising Rapamycin and Albumin as zumab (Avastin)). This formulation may be used to treat or Anticancer Agent prevent cancer, such as hepatocellular carcinoma [129]. This invention provides methods for treating, stabilizing, Rapamycin Analogs as Anti-Cancer Agents preventing, and/or delaying cancer by administering rapamy- This invention is related to the synthesis of rapamycin cin nanoparticles. Methods of combination therapy compris- and derivatives/analogs. These compounds bind to FK-506 ing the use of an effective amount of rapamycin nanoparti- binding protein (FKBP), or inhibit the mTOR function of an cles and a second therapy are also provided in this applica- FKBP, or both. This patent also provides methods of using tion [128]. the rapamycin analogs and derivatives in treatment of cancer [130].

Table 6. Selected List of Recent Worldwide Issued and Published Patents on Other Products as Anticancer Agents.

MICROBIAL PRODUCTS AS ANTICANCER AGENTS Other products - Recent Issued and Published Patents (2007-2011)

Epothilone derivative for the treatment of hepatoma and other cancer diseases Publication number: HK1073601; Publication date: 2010-04-16; Inventor: Rothermel John David Combinations of therapeutic agents for treating cancer Publication number: NZ555370; Publication date: 2010-03-26; Inventors: Zaknoen Sara; Woo Margaret Ma; Versace Richard Wil- Epothilones liam; Pisano Claudio; Vesci Loredana Sorangium Anti-cancer sustained-released injection containing epothilone derivate cellulosum Publication number: CN101396340; Publication date: 2009-04-01; Inventors: Juan Sun; Yuyan Liu; Qingxin Kong (5 out of 45) Cancer treatment with epothilones Publication number: US2009069394; Publication date: 2009-03-12; Inventors: Dilea Clifford; Mcsheey Paul M J; Maira Sauveur- Michel; Tanaka Chiaki; Wartmann Markus Epothilone derivative for the treatment of hepatoma and other cancer diseases Publication number: US2008161369; Publication date: 2008-07-03; Inventor: Rothermel John David

Farnesyl protein transferase inhibitors for treating breast cancer Publication number: US2009018164; Publication date: 2009-01-15; Inventors: Palmer Peter Albert; Horak Ivan David Anticancer agents based on regulation of protein prenylation Farnesyl Publication number: US2009143467; Publication date: 2009-06-04; Inventors: Rose Seth D; Lefler Scott R; Ottersberg Steven R; Kim transferase Ann Y; Okolotowicz Karl J; Hartman Rosemarie F inhibitors Novel farnesyl protein transferase inhibitors as antitumor agents Streptomyces Publication number: MX2007007611; Publication date: 2009-06-04; Inventors: Wang James J-S; Cooper Alan; Zhu Hugh; Desai parvulus Jagdish (5 out of 13) Farnesyl protein transferase inhibitors

Publication number: US2007213340; Publication date: 2007-09-13; Inventors: Kelly Joseph M; Afonso Adriano Novel farnesyl protein transferase inhibitors and their use to treat cancer Publication number: MX2007007074; Publication date: 2007-09-04; Inventor: Rane Dinanath

52 Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 Fialho et al.

(Table 6) Contd….

MICROBIAL PRODUCTS AS ANTICANCER AGENTS Other products - Recent Issued and Published Patents (2007-2011)

Use of salinosporamide A and analogs thereof for the treatment of cancer, inflammation and infectious diseases Publication number: NZ544588; Publication date: 2010-06-25; Inventors: Palladino Michael; Neuteboom Saskia Theodora Cornelia; Macherla Venkata Rami reddy; Potts Barbara Christine Salinosporamide derivatives as proteasome inhibitors Salinosporamide Publication number: WO2009140287; Publication date: 2009-11-19; Inventors: Macherla Venkat Rami Reddy; Potts Barbara Chris- Salinispora tro- tine; Manan Rama Rao; Mcarthur Katherine A; Chao Tahsiang; Neuteboom Saskia Theodora Cornelia pica Analogs of salinosporamide A (5 out of 9) Publication number: US2009054665; Publication date: 2009-02-26; Inventors: Myers Andrew G; Sun Binyan; Jackson Stona R

Methods of sensitizing cancer to therapy-induced cytotoxicity Publication number: WO2007056335; Publication date: 2007-05-18; Inventors: Bonavida Benjamin; Palladino Michael Biosynthesis of salinosporamide A and its analogs Publication number: WO2007030662; Publication date: 2007-03-15; Inventors: Lam kin Sing; Palladino Michael

Belactosin Novel belactosin derivatives as therapeutic agents/biological probes and their synthesis Streptomyces sp Publication number: WO2008006113; Publication date: 2008-01-10; Inventors: Romo Daniel; Cho Sung Wook; Smith Jeffrey W; (1 out of 1) Richardson Robyn D

Syringolin A Derivatives of syringolin A Pseudomonas Publication number: EP2210881; Publication date: 2010-07-28; Inventors: Kaiser Markus; Clerc Jerome syringae Development of a synthesis of syringolin A and B and derivatives thereof (2 out of 2) Publication number: US2010022767; Publication date: 2010-01-28; Inventors: Kaiser Markus; Clerc Jerome

MICROBIAL PRODUCTS AS ANTICANCER AGENTS Other products - Recent Issued and Published Patents (2007-2011)

Nanoparticle comprising rapamycin and albumin as anticancer agent Publication number: US2010183728; Publication date: 2010-07-22; Inventors: Desai Neil P; Soon-Shiong Patrick; Trieu Vuong Endoprostatic spring (STENT) impregnated with rapamycin/sirolimus and shiga toxin acting against cancer of prostate and benign hyperplasias of the urologic tract Rapamycin Publication number: GR1006552; Publication date: 2009-10-02; Inventor: Dimofilos Nikolaos Christou Streptomyces Composition of mTOR antagonist and angiogenesis inhibitor for treating cancer hygroscopicus Publication number: CN101583357; Publication date: 2009-11-18; Inventors: Hung Huynh The; Chee Soo Khee (5 out of 41) Rapamycin analogs as anti-cancer agents

Publication number: WO2009131631; Publication date: 2009-10-29; Inventors: Sun Connie L; Li Xiaoyuan Improved antitumoral treatments Publication number: WO2009109649; Publication date: 2009-09-11; Inventors : Ramon Y Cajal Agueeras Santiago; Hernandez Losa Javier; Jimeno Donaque Jose Ma; Raymond Eric

Improved Antitumoral Treatments ucts, thereby acting as anticancer agents. The list is varied, covering various bacteria and their products such as proteins, This invention provides combinations of PM02734 (elis- peptides and small molecule compounds. The list also in- idepsin) with another anticancer drug selected from the fol- cludes the use of bacterial DNA, such as CpG-rich DNA, in lowing list: Cisplatin, Gemcitabine, Paclitaxel, Oxaliplatin, potential cancer therapy, see for example the cited patent 5-Fluorouracil, Trabectedin, Rapamycin, and Sunitinib. The MX2009005849 Table 4 published on August 12, 2009. The use of these combinations in the treatment of cancer is also issuance and publication of such patents represent the provided in this application [131]. breadth and intensity with which anticancer drug development, and protection of such developments through patent- CURRENT & FUTURE DEVELOPMENTS ing, are proceeding. Undoubtedly such activities will con- In this article, we have listed many patents, either pub- tinue so long as cancer remains a deadly threat. It is interest- lished or issued in the United States or internationally, that ing to note, however, that DNA patenting arouses more scru- cover cancer treatment by live microorganisms or their prod- tiny and societal concern than others, as reflected by several Bacterial Strains and Products as Anticancer Agents Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 53 recent court cases. In Monsanto Technology v. Cefetra, the [8] Leonard, A., Saltzman, D.A. Method of cancer treatment. European Court of Justice ruled on July 6, 2010, against US2010098665 (2010). [9] Bereta, M., Paulina, C. New strain of Salmonella enteric, S. typhi- Monsanto concluding that a patent on DNA (a gene encoding murium, its use and a method to obtain a therapeutic vaccine vec- an enzyme that confers insensitivity to herbicides such as tor. WO2010095966 (2010). glyphosate) does not confer protection when the DNA func- [10] Saltzman D.A, Leonard A., Mueller M.J. Method of cancer treat- tion is no longer the same for which the patent was issued. In ment. CA2701052 (2010). another case Prometheus Laboratories v. Mayo Collabora- [11] Sprott, G.D., Krishnan, L., Sad, S. Vaccine adjuvant properties of liposomes formed at elevated temperatures from the polar chloro- tive Services, where the question of patentability of natural form extractable lipids from Mycobacterium bovis. metabolic processes (such as determining the level of a drug US20080248094 (2008). metabolite through a known metabolic pathway for optimiza- [12] Garcia, U.P Process for obtaining a product containing polar lipids tion of drug delivery to the body) was raised, the Court of and mycobacterial hydrophilic substances from Mycobacterium tuberculosis used as a vaccine or immunostimulant. MX2008001575 Appeals for the Federal Circuit (CAFC) in December 2010 (2009). affirmed the patentability of such personalized medicine and [13] Wagner, T.E., Wei, Y. Combination therapy of hybrid cells with medical diagnostic claims. A more contentious patent case BCG injection for treating cancer patients. US2009087450 (2009). involves the patenting of genetic mutations that predispose [14] Marchal, G., Abolhassani, M. Recombinant Mycobacterium strain people to breast and ovarian cancers such as BRCA1 and expressing a mycobacterial FAP protein under the control of a promoter active under hypoxia and its application for cancer ther- BRCA2 gene mutations. Such patents were issued in the apy. HK1112753 (2010). United States in 1998, and were affirmed in the European [15] Jo, E.K., Kim, K.K., Shin, D.M, Lee, H.M. Pharmaceutical compo- Union in a highly restricted form in 2008. In March 2010, a sition containing Mycobacterium tuberculosis-EIS recombinant judge in the federal court in the Southern District of New protein as an active ingredient. KR20100119161 (2010). [16] Santin, A.D. Therapy with Clostridium perfringens enterotoxin to York ruled against eligibility of such patents in a case known treat ovarian and uterine cancer. US20090291047 (2009). as Association for Molecular Pathology v. United States Pat- [17] Mckerracher, L., Lasko, D. Clostridium botulinum C3 exotrans- ent & Trademark Office, ruling that isolated DNA and their ferase compositions and methods for treating tumour spreading. sequence determinations for recognizing specific mutations US20090285833 (2009). simply reflect products of nature and therefore remain unpat- [18] Nakanishi, T. Growth inhibitor of colon cancer cells. JP2009269836 (2009). entable. Although the ruling has been appealed to the CAFC, [19] Woo, S.L.C, Li, Z. Compositions and methods comprising geneti- it is becoming clear that patenting of DNA, including micro- cally enhanced obligate and facultative anaerobic bacteria for on- bial DNA for potential cancer therapy, could come under copathic therapy. WO2009111177 (2009). scrutiny in the future if the revocation of the BRCA1/2 pat- [20] Yunlong, C., Hongfu, L. Use of Clostridium butyricum in preparing medicine composition for preventing and curing intestinal cancer. ents stands. Thus patenting of live microorganisms or their CN101306020 (2008). products, whether for cancer therapy, cancer diagnostics or [21] Paterson, Y. Specific immunotherapy of cancer using live recombi- other uses, might become a controversial subject if the New nant bacterial vaccine vector. JP2007254485 (2007). York ruling is affirmed by the CAFC or ultimately by the [22] Maciag, P., Wallecha, A., Shahabi, V. Dual delivery system for U.S. Supreme Court. heterologous antigens. WO2009143167 (2009). 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