Recent Patents on Live Bacteria and their Products as Potential Anti- cancer Agents

25 Pages·2011·0.79 MB·English

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
Arsenio M. Fialho1,*, Nuno Bernardes1 and Ananda M. Chakrabarty2

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 pub-
lished/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
investigated and recognized as potential cancer treatments.
to 6.
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 hy-
poxic 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: [phone];
agents (Part A, Table 1).
Fax: [phone]; E-mails: [email protected]; [email protected]

(cid:21)(cid:21)(cid:20)(cid:21)-(cid:22)9(cid:26)(cid:19)/12 $100.00+.00 © 2012 Bentham Science Publishers
32 Recent Patents on Anti-Cancer Drug Discovery, 2012, Vol. 7, No. 1 Fialho et al.
A
Salmonella
(IA, PDMA)
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
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
treatment of cancer, comprising the administration of a live
(aroA, rfaH, and thyA). It also provides a method to treat
attenuated Salmonella typhimurium strain harbouring a
cancer including the step of administering these Salmonella
plasmid carrying a gene that encodes a truncated form of the
strains [6].
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 im-
This patent discloses the use of a bacterial strain of Sal-
mune response against cancer cells. It comprises, in a phar-
monella enterica serovar Typhimurium VNP20009 deposited
maceutically delivery system, a mixture containing two
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
of CPE. This invention also provides a method of treating
ii). Mycobacterium Strains as Anti-Cancer Agents
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
tinal disease. It comprises a Clostridium butyricum mixture
mycobacterial gene encoding a FAP protein under the tran-
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 construc-
tion 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
1. PATENTS ON BACTERIAL TOXINS AS ANTI-
with a sequence encoding a PEST-containing polypeptide.
CANCER AGENTS
This patent also discloses methods of preparing same, and
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
Claudins as Markers for Early Detection, Diagnosis, Prog-
step of administering a genetically modified Listeria strain
nosis and as Targets of Therapy for Breast and Metastatic
harbouring a foreign gene encoding a chimeric protein which
Brain or Bone Cancer
possesses a listeriolysin O fragment together with the E7
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
based on immunohistochemical detection of claudin recep-
Compositions, Methods and Uses for Stimulating Immune
tors, namely 1 and 7 (frequently underexpressed) and 3 and 4
Responses
(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
In this patent, gene expression assays together with hier-
Therapy
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
ovarian carcinoma: transforming growth factor beta receptor
The summary of this invention is described in reference
HI, platelet-derived growth factor receptor alpha, SE-
[19].
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-
I /Ep-CAM monoclonal chimeric/humanized antibody. Fi-
In this patent a treatment for multiple diseases including
nally, in this invention, cancer treatment methods using the
cancer is disclosed. As effective component, this pharmaceu-
enterotoxin CPE from Clostridium perfringens are also dis-
tical composition contains fermented fruit by lactic acid bac-
closed [28].
teria or enterobacteria [26].
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
Other products:
Plantaricin
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 bacte-
rial 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(cid:2) (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 (cid:1)-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 (cid:1)- 1.5. Exotoxin A
subunit of shiga toxin as an immunotoxin [37].
Pseudomonas Exotoxin A Mutants and Uses Thereof
Chimeric Polypeptide Comprising Fragment (cid:1) of Shiga
This invention discloses chimeric immunotoxin proteins
Toxin and Peptide of Therapeutic Interest
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 (cid:1) 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,
based on the chimeric immunotoxins constructed, discloses
Shiga Toxin (cid:1)-Subunit as a Vector for Tumor Diagnosis
compositions and methods for inhibiting the growth and pro-
and Drug Delivery to GB3 Expressing Tumors
liferation of cancers [45].
This invention describes the construction and use of a
Chimeric Toxins for Targeted Therapy
novel chimeric protein able to recognize Gb3 receptors,
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 (cid:1) 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
endometrial island cells, benign hyperplasia of prostate and
Shiga Toxin (cid:1)-Subunit/Chemotherapeutics Conjugates
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 (cid:1)-subunit moiety linked to a
Comprising an Exotoxin a Moiety having a Furin Cleavage
particular anti-cancer agent that requires intracellular uptake
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 pro-
tein [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 com-
posed 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
Stem Cell Targeting of Cancer, Methods and Compositions
transport to the endoplasmic reticulum of cells. A method for
Therefor
inhibiting or destroying mammalian cancer cells using this
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 overex-
pressed 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
This invention is related to a pharmaceutical combination
strain is disclosed. This patent also describes the method of
product for a cancer therapy containing two active ingredi-
producing arginine deiminase by fermentation of Pseudo-
ents: glutaminase and 6-diazo-5-oxo-L-norleucine (DON)
monas plecoglossicida CGMCC No.2039. Activity of argin-
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 be-
nign 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
Publication number: US2007003610;Publication date: 2007-01-04; Inventors: Chancellor Michael B; Fraser Matthew O; Chuang Yao-
neurotoxin
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 specifi-
cally 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 thera-
human cancer cells expressing DR5. The formulation is in-
peutic use in cancer treatments [60].
tended 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].